Book of Abstracts (PDF) - International Mycological Association

IMC7 

Book of Abstracts 

The 7th International Mycological Congress 

Oslo 11–17 August 2002

IMC7 Oslo, Norway 

Contents 

Contacts for Information …………… Page 2 

Organizing Committee………………… Page 3 

Main Sponsors ……………………………… Page 3 

Lectures Monday 12th ………………… Page 4 

Lectures Tuesday 13th ………………… Page 33 

Lectures Wednesday 14th …………… Page 66 

Lectures Thursday 15th ……………… Page 77 

Lectures Friday 16th …………………… Page 109 

Posters Main Theme 1 ………………… Page 139 

Contacts for information 

Technical organizer 

PLUS Convention Norway AS 

P.O.Box 1646 Vika 

N-0119 Oslo, Norway 

Tel: + 47-22 92 55 40 

Fax: + 47-22 92 55 44 

E-mail: imc7@plus-convention.no 

IMC7 Congress Secretariat 

IMC7 Congress Secretariat 

P.O. Box 24 Blindern, 

N-0314 Oslo, Norway 

Tel: + 47-22 85 46 28 

Fax: + 47-22 85 46 64 

E-mail: imc-7@bio.uio.no 





Genera Index ………………………………… Page 366 

Family Index ………………………………… Page 374 

Order Index …………………………………… Page 375 

Author Index ………………………………… Page 376 

2 Book of Abstracts

11–17 August 2002 IMC7 

The imc7 Organizing Committee: 

Organizing Committee 

Leif Ryvarden (chair) 

Trond Schumacher (vice chair) 

Gro Gulden 

Geir Hestmark 

Arne Holst-Jensen 

Klaus Høiland 

Einar Timdal 

Anna-Elise Torkelsen 

Trude Vrålstad 

Nils Hallenberg 

IMC7 Project Leader and Congress Secretariat 

Jonathan E. Colman 

Main sponsors 

The IMC7 would like to thank: 

– The International Mycological Association 

– The University of Oslo 

– The Norwegian Ministry of Foreign Affairs 

– The Norwegian Ministry of Education 

Research and Church Affairs 

– Scandinavian Airlines System (SAS) 

Exciting links to these sponsors’ home pages can be found on the IMC7 website: 

www.uio.no/conferences/imc7 


Special thank you to committee members: 

Jan Stenlid 

Søren Rosendal 

Øyvind Stensrud 

Karin-Brit Schonhowd 

Tor Erik Brandrud 

Egil Bendiksen 

Katriina Bendiksen 

Håvard Kauserud 

Isabella Børja 

– The Canadian Journal of Botany 

– TINE 

– MYCOTEAM 

3

IMC7 Monday August 12th Lectures 

1 - Character evolution in the Lasiosphaeriaceae sensu 

lato 

A.N. Miller 1* & S.M. Huhndorf 2 

1 University of Illinois at Chicago, Dept. of Biological 

Sciences, Chicago, IL 60607-7060, U.S.A. - 2 The Field 

Museum of Natural History, Dept. of Botany, Chicago, IL 

60605-2496, U.S.A. - E-mail: amiller@fmnh.org 

There are a limited number of phylogenetically informative 

morphological characters for delimiting taxa above the 

species level in many pyrenomycetes, mostly due to their 

small stature and simplistic form. Ascospore morphology 

has been used extensively throughout the filamentous 

ascomycetes for distinguishing families and genera and 

many genera in the Lasiosphaeriaceae have been 

segregated using ascospore morphology. However, other 

morphological characters, such as ascomatal wall 

characters, have been suggested, but not tested, for 

delimiting genera in this family. For example, several 

genera possess unique bombardioid walls in which the 

middle wall layer is gelatinized and several species of 

Lasiosphaeria and Cercophora possess identical threelayered 

walls in which the outer layer is composed of 

hyphal strands. To test the utility of these morphological 

characters for predicting evolutionary relationships, 

sequences of the 28S nuclear ribosomal large-subunit 

(LSU) gene were generated for taxa within the 

Lasiosphaeriaceae and selected outgroups. Sequences from 

two nuclear protein-coding genes, B-tubulin and RPB2, 

were also generated to corroborate the LSU data. 

Maximum parsimony and Bayesian analyses of separate 

and combined data sets suggest that ascospore characters 

are extremely homoplastic and not useful for delimiting 

genera in the Lasiosphaeriaceae. Ascomatal wall characters 

are, however, sometimes phylogenetically informative and 

could be used for segregating taxa. 

2 - Developing phylogenies for integrating mitotic fungi 

in the Hypocreales and Diaporthales 

A.Y. Rossman * , L.A. Castlebury & D.F. Farr 

Systematic Botany & Mycology Laboratory, USDA-ARS, 

Beltsville, MD 20705, U.S.A. - E-mail: amy@nt.arsgrin.gov 

Recent comprehensive studies of the Hypocreales and 

Diaporthales using both morphological and molecular 

characters present the opportunity to integrate the mitotic 

fungi and to evaluate character evolution of both 

teleomorphic and anamorphic states. The majority of plantassociated 

fungi are mitotic species without any known 

sexual state and limited morphology. Using molecular 

sequence data it is possible to integrate the mitotic fungi 

into the Hypocreales and Diaporthales and to demonstrate 

that a vast number of mitotic fungi are derived from within 

these sexual state lineages. In the Hypocreales, the mitotic 

fungi tend to be hyphomycetous with the exception of the 

Clavicipitaceae while in the Diaporthales they are primarily 

coelomycetous. Despite considerable effort, some mitotic 

4 


species cannot be closely linked with sexual states or even 

families and thus appear to represent divergent lineages. 

The character evolution within the Hypocreales and 

Diaporthales appear similar with fruiting bodies ranging 

from solitary perithecia to compound stromata, often 

characteristic of specific families. Ascospore morphology 

is also characteristic of specific families although 

exceptions exist in each family. Biologically the 

Diaporthales appear to be confined to plant substrata often 

as virulent plant pathogens in temperate regions while the 

Hypocreales fill much broader niches attacking insects and 

other fungi as well as plant substrata with their greatest 

diversity in tropical regions. 

3 - Remembering and dismembering Stilbella: A 

classical hyphomycete genus 

K.A. Seifert 

Eastern Cereal and Oilseed Research Centre, Agriculture 

& Agri-Food Canada, 960 Carling Ave., Ottawa, Ontario 

K1A 0C6, Canada. - E-mail: seifertk@em.agr.ca 

The 275 described taxa of Stilbum and Stilbella were 

reduced to 45 species in seven genera in my 1985 

monograph. Cladistic analysis of LSU rDNA sequences 

provides new insight into the phylogeny and taxonomy of 

these fungi. Stilbella-like anamorphs occur in four families 

of Hypocreales (Bionectriaceae, Hypocreaceae, 

Nectriaceae, Clavicipitaceae), in the Phyllachorales, 

Microascales and Leotiales. The type, S. fimetaria, is 

related to the cleistothecial Emericellopsis 

(Bionectriaceae). Previous delimitations of Tubercularia 

(teleomorphs: Nectria ss, Nectriaceae), Gliocladium 

(teleomorphs: Sphaerostilbella, Hypocreaceae), 

Rhizostilbella (teleomorph: Corallomycetella, Nectriaceae) 

and Polycephalomyces (teleomorphs: Byssostilbe, 

Clavicipitaceae) are supported by LSU data. The recently 

described Gracilistilbella (teleomorphs: Stilbocrea), is a 

monophyletic group within the Bionectriaceae. Volutella 

(teleomorphs Cosmospora, Nectriaceae), previously 

restricted to setose sporodochial species, is emended to 

include synnematous species. Stilbella annulata is related 

to the Phyllachorales. The phylogenetic distribution of 

these anamorphs suggests that synnemata are either 

plesiomorphic in the Hypocreales and sporadically 

expressed, or that they have arisen repeatedly. A strictly 

redefined Stilbella includes only two species, but implies 

recognition of many monotypic genera. This once large 

form taxon is a useful metaphor for patterns in anamorph 

taxonomy revealed by molecular data. 

4 - Ecology and evolution in the Onygenales: an 

overview based on molecular and morphological 

characters 

W.A. Untereiner 

Brandon University, 270-18th Street, Brandon Manitoba 

R7A 6A9, Canada. - E-mail: untereiner@brandonu.ca


Among the filamentous ascomycetes, few groups have 

received as much attention in molecular phylogenetic 

studies as the members of the Onygenales, an order that 

includes the most important fungal pathogens of mammals 

and the only fungi capable of degrading keratin. Parasitic 

Onygenales are thought to be derived from saprobic 

ancestors, but the origins of parasitism within the 

Arthrodermataceae and Onygenaceae, two families of 

keratinolytic Onygenales, remains obscure. Phylogenies 

based on analyses of conserved gene regions have 

demonstrated that parasitic species do not form a separate 

lineage within the Onygenales, but these sequences lack 

sufficient numbers of phylogenetically informative 

characters to resolve the branching order among recently 

diverged taxa or evaluate the evolution of ecological and 

morphological characters hypothesized to be correlated 

with the origins of parasitism. This study, which addresses 

the question of how the evolution of specific characters 

may be correlated to the evolution of parasitism in the 

Onygenales, is based on the analyses of molecular 

(mitSSU, partial nucLSU gene sequences) and 

morphological characters for nearly 50 members of this 

order. Emphasis has been placed on the inclusion of 

saprobic representatives of the Arthrodermataceae and 

Onygenaceae, and members of the Gymnoascaceae (the 

closest non-keratinolytic relatives of these families) have 

been included to determine the direction of the evolution of 

characters of interest. 

5 - Evolution of anamorph form in the Clavicipitaceae 

K.T. Hodge 

Cornell University, Dept. of Plant Pathology, 334 Plant 

Science Bldg., Ithaca, NY 14853, U.S.A. - E-mail: 

kh11@cornell.edu 

Members of the Clavicipitaceae exhibit a diverse array of 

life histories, and a diverse array of anamorph forms. Most 

members are symbiotic with plants, fungi, or arthropods. 

Among the arthropod pathogenic members, classification 

has so far been based upon the scant characters of the 

sexual stromata, and the resulting systems developed by 

Kobayasi and other authors prove to be difficult to apply. 

Increasingly, we know that they do not reflect relationships 

as revealed by molecular evidence. Asexual forms have 

seldom been considered key characters for classification, 

and indeed, anamorph connections are as yet unknown for 

about 50% of teleomorph species. Yet molecular evidence 

suggests that anamorph genera (for example, 

Harposporium, Hirsutella, and a subset of Paecilomyces 

species) may each comprise monophyletic groups 

correlated with distinctive teleomorphic characters. The 

functional significance of differing forms in the life 

histories of these organisms has barely been studied, 

although biocontrol experience suggests that anamorphs 

play important roles in pathogenicity. This study addresses 

the evolution of anamorph form, particularly among the 

arthropod pathogenic species of the Clavicipitaceae, and 

highlights the potential role of anamorph morphology and 

ecological factors in improving classification systems in 

the Clavicipitaceae. 

6 - Well supported, major groups among the 

Euascomycetes 

A. Tehler 

Naturhistoriska riksmuseet, Sektionen för 

kryptogambotanik, Box 50007, 104 05 Stockholm, Sweden. 

- E-mail: anders.tehler@nrm.se 

A data set with 1555 sequences of SSU rDNA has been 

phylogenetically analyzed. Cladistic methods using 

parsimony jackknifing for establishing group frequencies 

and the parsimony ratchet algorithm for finding most 

parsimonious trees was utilised. Until recently attempting 

to find all most parsimonious trees in large data sets, has 

been impractical, given current computational limitations. 

The parsimony ratchet method for rapid parsimony analysis 

of large data sets was found very efficient in searching and 

finding most parsimonious trees in the current large data 

set. The results show that the Glomales are sister group to 

the Dikaryomycetes. The Ascomycetes include the 

Euascomycetes and are both monophyletic. Within the 

Euascomycetes the inoperculate Euascomycetes are 

monophyletic except for the Orbiliomycetes which are 

included in an operculate, pezizalean sister group. 

Geoglossum is the sister group to the rest of the 

inoperculate euascomycetes. The Sordariomycetes, 

Dothideomycetes, Chaetothyriomycetes and 

Eurotiomycetes are each highly supported as monophyletic 

whereas the Leotiomycetes and Lecanoromycetes are both 

shown to be paraphyletic. In the consensus ratchet tree the 

Dothideomycetes, Chaetothyriomycetes, Eurotiomycetes 

and the paraphyletic assemblage Lecanoromycetes form 

the sister group to the Sordariomycetes and the 

paraphyletic assemblage Leotiomycetes, but that topology 

receives no jackknife support. 

7 - Changes in systematics of heterobasidiomycetous 

fungi during 30 years 

F. Oberwinkler 

University Tuebingen, Auf der Morgenstelle 1, 72076 

Tuebingen, Germany. - E-mail: franz.oberwinkler@unituebingen.de 

Systematics and hypotheses about their evolution have 

been revolutionized within the last few decades. Main 

impacts for new views came from (a) the detection of new 

key species, (b) the application of new techniques to study 

sofar unknown characters, (c) the reconstruction of lifehistories 

in nature and/or in culture, (d) the understanding 

of fungus-host interrelationships, and (e) the combination 

of old and new datasets for comparative interpretations. (a) 

I will present comments on species originally hidden 

outside the Basidiomycetes, but now representing key taxa 

in the heterobasidiomycetous groups, and new species with 

considerable importance for basidiomycetous phylogeny. 

(b) Clarification of monophyla referred in major parts to 

ultrastructural characters. A surprisingly high support for 

Book of Abstracts 5


new systematic arrangements came from molecular studies. 

(c) There was a renaissance for ontogenetic studies in 

culture. Thus, life-history experiments provided new 

insights in most of heterobasidiomyxcetous taxa, especially 

in smuts, tremelloid fungi and yeasts. (d) Coevolutionary 

processes play an important role in diverse 

heterobasidiomycetous taxa. Therefore, systematics of 

hosts have to be included in fungal character sets. (e) 

Fortunately, most phylogentic interpretations are now 

based on several datasets, suggesting possibilities for 

improved evolutionary hypotheses. 

8 - Ultrastructure of heterobasidiomycetous fungi 

R. Bauer 

Universität Tübingen, Lehrstuhl Spezielle Botanik und 

Mykologie, Auf der Morgenstelle 1, D-72076 Tübingen, 

Germany. - E-mail: robert.bauer@uni-tuebingen.de 

Significant ultrastructural characteristics of 

heterobasidiomycetes, such as nuclear and spindle pole 

body behaviour, septal pore apparatus, colacosomes, 

symplechosomes as well as myco- and phytoparasitic 

interactions will be discussed with special regard to their 

role as phylogenetic markers. 

9 - Heterobasidiomycetous yeasts: diversity, phylogeny 

and classification 

J.P. Sampaio 1* , R. Bauer 2 , M. Weiß 2 & M. Gadanho 1 

1 Centro de Recursos Microbiológicos, Secção Autónoma 

de Biotecnologia, Faculdade de Ciências e Tecnologia, 

Universidade Nova de Lisboa, 2829-516, Portugal. - 

2 Universität Tübingen, Institut für Biologie I, Lehrstuhl 

Spezielle Botanik und Mykologie, Auf der Morgenstelle, 1, 

D-72076, Tübingen, Germany. - E-mail: 

jss@mail.fct.unl.pt 

Basidiomycetous yeasts form a remarkably diverse group 

of fungi and are distributed throughout the three main 

lineages of the Basidiomycota: Urediniomycetes, 

Ustilaginomycetes and Hymenomycetes. Traditionally, the 

yeast stages of plant or mycoparasitic basidiomycetes and 

the unicellular forms of fruitbody-producing taxa have 

been excluded from the yeast domain. When these nonconventional 

basidiomycetous yeasts are cultivated in 

laboratory media, the usual growth form is unicellular and 

corresponds to the haploid and/or saprophytic phases. Nonconventional 

dimorphic basidiomycetes rarely develop 

their complete life cycle in culture, contrary to typical 

basidiomycetous yeasts. This artificial separation of taxa, 

mainly for historical reasons, prevents an integrated 

assessment of the diversity of these groups and the 

implementation of a natural classification system. The 

objectives of this presentation are two-fold: (i) to advocate 

a new concept of 'basidiomycetous yeasts' encompassing 

all dimorphic basidiomycetes, regardless of their life cycle 

6 


and (ii) to present and discuss new developments 

concerning the systematics of asexual (Rhodotorula) and 

sexual (Leucosporidium, Mastigobasidium, 

Rhodosporidium and Sporidiobolus) taxa of the sub-class 

Microbotryomycetidae. The polyphyletic nature of 

Rhodotorula and the relationship between Leucosporidium 

/ Mastigobasidium and the Microbotryales will be 

addressed. 

10 - Molecular phylogeny of rusts 

W. Maier 

Universität Tübingen, Spezielle Botanik/Mykologie, Auf der 

Morgenstelle 1; 72076 Tübingen, Germany. - E-mail: 

w.maier@uni-tuebingen.de 

Sequence data from nuclear large subunit ribosomal DNA 

was used to infer phylogenetic relationships of selected 

genera of the rust fungi and to test the monophyly of these 

genera. Neighbor joining analysis and a Bayesian method 

of phylogenetic inference using Monte Carlo Markov 

chains confirm that (1) Puccinia, Uromyces, Cumminsiella 

and Endophyllum have a common origin. (2) The 

autoecious Rosaceae-rusts Phragmidium, Kuehneola, 

Triphragmium, and Trachyspora are a monophyletic group, 

representing Phragmidiaceae. (3) The gasteroid genus 

Ochropsora is closely related to Tranzschelia. The 

monophyly of the following genera could be confirmed: 

Chrysomyxa, Coleosporium, Cronartium, 

Gymnosporangium, Melampsora, Phragmidium, and 

Tranzschelia, whereas the genera Puccinia, Pucciniastrum, 

Thekopsora and Uromyces are polyphyletic. The molecular 

phylogenetic hypotheses are compared to morphologybased 

systematics with special emphasis on spermogonial 

and teliospore morphology, and the usefulness of these 

characters on different systematic levels is discussed. 

11 - Evolution in smuts 

D. Begerow * & R. Bauer 

University of Tübingen, Botanisches Institut; Auf der 

Morgenstelle 1, Germany. - E-mail: 

ustilaginomycetes@uni-tuebingen.de 

Many changes in the view of smut phylogeny have been 

discussed within the last decade. An overview of the main 

lineages of that plant parasitic group is presented. Based on 

an integrated phylogeny hypotheses of the major traits of 

evolutionary mechanisms will be discussed. New 

morphological, ecological, and coevolutionary aspects of 

smut evolution are in focus of present studies and will be 

highlighted.


12 - An integrating approach to ecology, morphology, 

and systematics of smut fungi 

M. Piepenbring 

Botanical Institute, University of Frankfurt am Main, 

Senckenberganlage 31-33, 60054 Frankfurt am Main, 

Germany. - E-mail: piepenbring@em.uni-frankfurt.de 

Systematic relationships are studied in order to understand 

evolution, which is the result of the interaction of the 

organisms with environmental factors. The latter decide 

whether a combination of morphological characteristics 

contributes or not to the fitness and evolutionary success of 

the respective individual. These are aspects which 

apparently are important for the survival of certain species 

of plant parasitic smut fungi: Host species with high 

frequency of individuals growing on sites exposed to wind 

or water; Development of sori in nutrient rich parts of the 

host plant; Exposure of masses of teliospores to the 

respective vector of dispersal for a long period of time; 

Protection of diaspores against desiccation, intense light, 

and feeding when young and during dispersal; Adaptations 

to different vectors of dispersal. The integration of 

ecological, morphological, and systematic aspects is 

illustrated by Cintractia amazonica Syd. & P. Syd. in the 

relationship of Cintractia s. l. and Tilletia spp. as well as 

Erratomyces patelii (Pavgi & Thirum.) M. Piepenbr. & R. 

Bauer versus Entyloma spp. 

13 - Phylogeny of morphological and molecular studies 

in the genera Tremella and Sirobasidium 

C.-J. Chen 1* & F. Oberwinkler 2 

1 

Department of Biotechnology, Southern Taiwan 

University of Technology, 1 NanTai Street, Yung Kang 

2 

City, Tainan 71043, Taiwan. - Spezielle 

Botanik/Mykologie, Universitaet Tuebingen, Auf der 

Morgenstelle 1, D-72076 Tuebingen, Germany. - E-mail: 

c5200999@mail.stut.edu.tw 

Comparison with morphological and molecular 

phylogenies, six groups in the genera Tremella and 

Sirobasidium are concluded. They are: 1. Aurantia group: 

heterogeneously mixed with basidiomycetaceous host 

hyphae up to the subhymenium, e.g. T. aurantia, T. 

encephala; 2. Foliacea group: basidiocarps foliose and 

brown, subhymenium with numerous anastomoses, 

haustoria rarely, e.g. T. foliacea sensu lato, T. giraffa; 3. 

Fuciformis group: heterogeneously associated with 

Hypoxylon basidiocarps, e.g. T. flava, T. fuciformis; 4. 

Indecorata group: heterogeneously associated with 

pyrenomycetaceous perithecia, e.g. T. indecorata, T. 

moriformis, T. nivalis; 5. Mesenterica group: basidiocarps 

gyrose to cerebriform, yellowish, basidia oval, hyphidia 

existing, subhymenial structure loose, e.g. T. mesenterica, 

T. taiwanensis, T. tropica; 6.Sirobasidium group: chained 

basidia, basidiomata habitually associated with ascomata or 

stromata of ascomycetes, e.g. S. magnum, S. intermediae. 

The results of LSU phylogenies compared with 

morphological data for the Tremella grouping strongly 

suggest that using large subunit (LSU) rDNA sequences 

are potentially useful in taxonomic, ecological, and 

evolutionary population-level studies. Moreover, molecular 

phylogeny implies that the genus Sirobasidium developed 

from that of Tremella evolutionally. 

14 - Auriculariales and related taxa 

M. Weiß 1* , M.-A. Selosse 2 , K.-H. Rexer 3 & F. 

Oberwinkler 1 

1 Universität Tübingen, Botanisches Institut, Lehrstuhl 

Spezielle Botanik und Mykologie, Auf der Morgenstelle 1, 

D-72076 Tübingen, Germany. - 2 Institut de Systématique 

(IFR CNRS 1541), Muséum National d'Histoire Naturelle, 

43 rue Cuvier, F-75005 Paris, France. - 3 Universtität 

Marburg, FB Biologie, Spezielle Botanik/Mykologie, Karlvon-Frisch-Straße 

1, D-35032 Marburg Straße 1, D-35032 

Marburg. Straße 1, Germany. - E-mail: 

michael.weiss@uni-tuebingen.de 

We present current hypotheses of phylogenetic 

relationships within heterobasidiomycetous 

Hymenomycetes with particular reference to 

Auriculariales. Molecular phylogenetic estimations are 

discussed in correlation to morphological and ecological 

data. 

15 - Tulasnelloid and other rhizoctonia-forming fungi 

P.J. Roberts 

Royal Botanic Gardens, Kew, Surrey TW9 3AB, U.K. - Email: 

p.roberts@rbgkew.org.uk 

Tulasnella, Ceratobasidium, Oliveonia, and other genera of 

rhizoctonia-forming fungi are of phylogenetic interest since 

they lie somewhere on the boundary between the holo- and 

heterobasidiomycetes. This pivotal position appears to be 

supported by micromorphological research, ultrastructure 

research, and molecular research. However, the precise 

disposition and relationships of these fungi have yet to be 

satisfactorily resolved. Morphologically, the Tulasnellales 

appear to be a well-defined and coherent group based on 

their unique basidia. But there is an unusual plasticity of 

forms (including spores, basidia, and conidial structures) 

within the group and sequencing data has proved difficult 

to interpret. The Ceratobasidiales present equal but 

different problems, particularly in defining genera and 

species. These problems are made more critical by the 

economic importance of plant pathogenic and orchid 

endomycorrhizal rhizoctonias. For several species, any 

change in nomenclature (such as synonymization) resulting 

from systematic research may be rejected, raising 

interesting questions about the relationship between 

taxonomists and the consumers of their output. 



16 - Systemic phenomena in pine pathosystems 

P. Bonello 

Ohio State University, Dept. of Plant Pathology, 2021 

Coffey Road, Columbus, OH 43210, U.S.A. - E-mail: 

bonello.2@osu.edu 

In many herbaceous host-pathogen systems, localized 

infections by pathogens or beneficial microorganisms, or 

localized treatment with certain chemicals, induce wholeplant 

resistance to subsequent pathogenic infections. This 

phenomenon is termed Systemic Induced Resistance (SIR). 

This presentation will illustrate similar phenotypes and 

associated putative defense responses in three conifer 

pathosystems: 1) pitch canker of Monterey pine, caused by 

Fusarium circinatum; 2) Heterobasidion annosum root and 

butt rot of ponderosa pine; and 3) Sphaeropsis sapinea tip 

blight and canker of Austrian pine. An SIR phenotype was 

recently described in Monterey pine (Bonello et al. 2001. 

Systemic induced resistance in Monterey pine. Forest 

Pathology 31:99-106), while systemic changes in defensive 

secondary metabolism were observed in mature ponderosa 

pines (in preparation). Working with potted Austrian pine 

we have now observed a unique phenomenon in which 

expression of SIR, or the opposite phenotype of systemic 

induced susceptibility (SIS), depend on whether the 

challenge infections occur on the stem or on the shoot tips. 

We are now attempting to correlate this organ specific 

SIR/SIS phenomenon with the expression of specific 

putative defense responses of the host. The significance of 

these results will be discussed along with future lines of 

investigation. 

17 - Longevity and plant defence: Host-pathogen 

interactions in short- and long-lived tissues of trees 

S. Woodward 

University of Aberdeen, Department of Agriculture & 

Forestry, MacRobert Building, 581 King Street, Aberdeen 

AB24 5UA, Scotland, U.K. - E-mail: 

s.woodward@abdn.ac.uk 

Relative to many herbaceous plants, trees are subject to a 

wide range of diseases during their long life cycles. 

Different diseases may be classed as persistent, once 

established present within the host until the tree dies, or 

periodic, attacking short-lived tissues when the 

environmental conditions favour pathogen development. 

This presentation is focused on two broad categories of 

disease which affect overall tree growth and productivity: 

(1) fine root diseases and (2) secondary root diseases. The 

main host species discussed will be Pinus sylvestris and 

Picea sitchensis. In mature trees and under normal forest 

conditions, disease of fine roots are most likely to be 

periodic, killing limited sections of the fine root system, 

except where prevailing microclimatic conditions favour 

disease. Diseases of secondary roots, however, are often 

persistent, colonising woody tissues, but remaining 

8 


restricted to bark or heartwood tissues unless conditions 

favour pathogen development. This presentation considers 

the interactions between tree roots and pathogenic fungi in 

terms of host defence and the various external factors 

influencing disease development. 

18 - Pathogenicity of the Heterobasidion annosum 

complex 

Å. Olson * , M. Karlsson, M. Lind & J. Stenlid 

Dept. of Forest Mycology and Pathology, Swedish 

University of Agricultural Sciences, Box 7026, SE-750 07 

Uppsala, Sweden. - E-mail: ake.olson@mykopat.slu.se 

The outcome of an interaction between a pathogen and its 

host is dependent on the genotype of the individuals 

involved as well as environmental conditions. We have 

taking both a genetic and a genomic approach to address 

the question of what genetic factors are of importance for 

pathogenicity and host specialisation in the tree pathogen 

Heterobasidion annosum. In North America H. annosum is 

present as two intersterile groups corresponding to 

biological species with different host preferences; the Sgroup 

which has spruce, fir and hemlock as its main hosts 

and the P-group which has pines as its main hosts. In the 

genomic approach we have sequenced 1148 randomly 

picked cDNA clones from a library constructed from 

mRNA of H. annosum challenged with Scots pine roots. 

Contig analysis identified 339 unique contig sequences out 

of which 247 were assigned a putative function deduced 

from sequence similarity with proteins in public databases. 

In the genetic approach we are currently preparing a 

genetic linkage map using AFLP markers using a mapping 

population of 97 progeny isolates from a cross between a P 

and a S homokaryon. Virulence on pine has been analysed 

for the 97 progeny isolates. Further studies of SP hybrids of 

H. annosum have revealed an importance of the 

mitochondria for virulence in pine. 

19 - Phytophthora in forest trees: host-parasite 

interactions 

E.M. Hansen 1* , G.E.St.J. Hardy 2 & W. Osswald 3 

1 Botany and Plant Pathology, Oregon State University, 

Corvallis OR 97331, U.S.A. - 2 Murdoch University, Perth, 

Western Australia, Australia. - 3 Technical University of 

Munich, Freising, Germany. - E-mail: 

hansene@bcc.orst.edu 

Phytophthora is well known as a genus of agricultural 

pathogens, often destructive when soil conditions favor 

zoospore spread. Phytophthora species are also widespread 

in many forests, but little is known about their interactions 

with potential hosts. We focus this analysis on three host 

tree-pathogen systems that illustrate a range of pathogenic 

behaviors; by reviewing what is known, we hope to focus 

attention on critical gaps in our understanding of forest


Phytophthoras. P. cinnamomi is the most studied of the 

three, with a broad host range and global distribution. It is 

introduced to Australia, where it causes ecological 

destruction in some forest communities, but has negligible 

impact in others. In some hosts at least, it moves in the 

xylem well beyond the margin of phloem necrosis. P. 

lateralis, by contrast, exhibits essentially host specific 

pathogenicity in forests of the western United States. 

Native Chamaecyparis lawsoniana are aggressively 

attacked and killed as the pathogen colonizes the phloem in 

a broad advancing front that moves up the roots into the 

stem. P. quercina is associated with oak decline in Europe, 

but appears to be native, or at least of long standing in 

these forests. It is confined to the fine roots of susceptible 

oaks, but induces symptoms far beyond the colonized 

tissues. We review what is known and describe ongoing 

research into the infection biology and pathogenesis of 

these contrasting species, and host responses to them. 

20 - Induction of defense structures in conifers: Norway 

spruce as a model system and a brief phylogenetic study 

V.R. Franceschi 

School of Biological Sciences, Washington State 

University, Pullman, WA 99164, U.S.A. - E-mail: 

vfrances@mail.wsu.edu 

Norway spruce was used as a model system to study 

anatomical and chemical defense responses to Ceratocystis 

polonica and attack by its bark-beetle vector Ips 

typographus. Fungal inoculation into the phloem generated 

responses within 3 wk, including increase in polyphenolic 

parenchyma cell (PP cell) size and staining, wound 

periderm initiation, and traumatic resin duct (TD) 

formation. Fungi were not seen in samples 3 wk after 

phloem inoculation, but were in some samples 6-9 wk after 

inoculation. Inoculations into the cambium resulted in 

partial (3 wk) or complete (6, 9 wk) fungal colonization. 

This indicates that PP cells have defenses capable of 

inhibiting fungal growth. Samples taken near bark-beetle 

galleries had similar anatomical responses, validating the 

inoculation approach. In an attempt to determine signals 

involved in these reactions, we found methyl jasmonate 

(MJ) induced similar anatomical and chemical responses. 

A single MJ treatment induced swelling of PP cells, 

increase in their phenolic contents, and formation of 

additional PP cells and TDs. Treatment enhanced resin 

flow and increased resistance to C. polonica. MJ 

application to the oldest internode of 2-yr-old saplings also 

induced TD formation, and, more surprisingly, TDs were 

formed in the untreated internode. MJ was found to have 

similar effects at upregulating defense responses in other 

conifer trees, although surprising differences were found in 

some taxa. 

21 - An overview of wound response and antimicrobial 

defence in eucalypts 

C. Mohammed 1* , K.M. Barry 2 , A. Eyles 3 , K. Harrison 3 & 

M. Hall 1 

1 

CSIRO Forestry and Forest Products, GPO Box 252-12, 

2 

Hobart, Tasmania, 7001, Australia. - School of 

Agricultural Science, University of Tasmania, GPO Box 

252-12, Hobart, Tasmania, 7001, Australia. - 3 CRC for 

Sustainable Production Forestry, GPO Box 252-12, 

Hobart, Tasmania, 7001, Australia. - E-mail: 

caroline.mohammed@csiro.au 

With the expanding use of regrowth eucalypt forests and 

plantations for wood production, the profitability of 

processing operations (particularly sawmilling) will 

increasingly depend on the maximum recovery of high 

quality products from relatively small logs. The quality and 

value of eucalypt logs can be significantly affected by the 

incidence, location and extent of discoloration and defect 

resulting from kino veins, decay fungi and insect attack. 

Our group is the first internationally to carry out detailed 

studies of eucalypt wood defence. We seek to compare two 

important Australian plantation species (E. nitens and E. 

globulus) with other tree species. Our studies in the area of 

eucalypt defence initially focused on the formation of a 

reaction zone (antimicrobial defence barrier between 

healthy and decayed sapwood). Our most recent studies 

have also investigated the new tissue formed immediately 

adjacent to the wound site which hitherto has been given 

relatively little importance. Both reaction zone and 

especially wound tissue have been found to contain a 

surprising concoction of tannins, flavonoids and terpenes. 

The nature and significance of these extractives and the 

tissue changes observed in response to wounding and 

infection in eucalypts will be examined in the broader 

context of tree defence strategies. 

22 - Clonality in wood-inhabiting fungi on different 

spatial scales 

R. Vasiliauskas * & J. Stenlid 

Dept. of Forest Mycology & Pathology, Swedish University 

of Agricultural Sciences, Box 7026, SE - 750 07 Uppsala, 

Sweden. - E-mail: Rimvydas.Vasiliauskas@mykopat.slu.se 

This presentation focuses on clonal spread in woodinhabiting 

fungi and its impact on population structures 

over local and large geographic areas. The work is based 

on genetic and spatial data and reveals strikingly different 

modes and consequences of asexual propagation in a 

number of species. The following species are discussed: 1) 

Heterobasidion annosum, 2) Rhizina undulata, 3) Stereum 

sanguinolentum, 4) Amylostereum chailletii, 5) A. 

areolatum and 6) Phlebiopsis gigantea. Clonality in those 

species arises as a result of: a) mycelial growth and 

colonisation of spatially separated resource units (trees) 

over discrete territories (species 1 and 2); b) airborne 



spread of non-outcrossing spores (species 2 and 3); c) 

vectoring of vegetative mycelium by woodwasps and wood 

material (species 4 and 5); d) use of single fungal strain as 

a large-scale biocontrol agent (species 6). The results show 

that fungal clones at one extreme are small and local; at the 

other extreme, clones of the wood-inhabiting fungi are 

capable of moving between widely separated locations and 

have a pronounced impact on population structure of a 

given species on trans- or even inter-continental scales. 

Persistence of such clones in nature varies from several 

years up to several decades. They are mainly characteristic 

for forest areas under anthropogenic impact, with the 

presence of stumps, wounds on living trees, burned areas 

and exotic species. 

23 - Sexuality and genetic variation in three boreal 

Polypores possessing divergent niche requirements 

H. Kauserud * & T. Schumacher 

University of Oslo, Department of Biology, P.O.Box 1045 

Blindern, 0316 Oslo, Norway. - E-mail: 

haavarka@bio.uio.no 

The population structure and sexuality of Fennoscandian 

populations of three model species, the vulnerable 

Phellinus nigrolimitatus and Fomitopsis rosea and the 

common pioneer wood-fungus Trichaptum abietinum, was 

investigated. Molecular support for a heterothallic mating 

system in P. nigrolimitatus was found. Molecular data and 

mating studies demonstrated that T. abietinum and F. rosea 

have a heterothallic mating system. The observed genotype 

distributions in the geographic populations of the model 

species were mostly in accordance with Hardy-Weinberg 

expectations. Our results suggest that the mating 

populations are large and that panmictic conditions exist. 

Although P. nigrolimitatus and F. rosea probably have 

experienced decline in population sizes as a result of 

negative impacts from forestry, no distinct heterozygote 

deficits ascribed to inbreeding could apparently be 

detected. The molecular markers revealed little genetic 

differentiation among geographic populations of the three 

model species, indicating high gene flow and good 

dispersal ability. The geographic populations may be 

regarded as subpopulations sharing the same gene pool. A 

high number of mating types were present in one T. 

abietinum and two F. rosea populations, a feature that may 

indicate high genetic diversity. A high number of T. 

abietinum and F. rosea genets occurred on single logs. In 

P. nigrolimitatus fewer but larger genets that often 

produced more than one basidiocarp occurred. 

24 - Genetic structure of Daldinia loculata populations 

H. Johannesson 1* , A. Guidot 2 , A. Dahlberg 1 & J. Stenlid 1 

1 SLU, Forest Mycology and Pathology, Box 7026 SE-750 

07 Uppsala, Sweden. - 2 SLU, ArtDatabanken, Box 7007 

SE-750 07 Uppsala, Sweden. - E-mail: 

hanna.johannesson@mykopat.slu.se 

10 


The genetic structure of the postfire ascomycete Daldinia 

loculata was studied on both a local and a continental 

scale. We were able to show endophytic establishment of 

D. loculata by isolating mycelia from non-decayed wood 

of birches. The distribution of genets was studied at one 

Swedish forest site, and in all trees studied we found only 

one, or occassionally two, unique genets per stem. The 

relatively large genet in the stem apparently was one of the 

parents of the offspring of the stromata attached to the tree, 

while each stroma had one or several unique second 

parents. Low levels of genetic differentiation among 

Eurasian subpopulations were confirmed by analysing the 

allelic distribution nuclear gene loci of 1 Russian and 6 

Fennoscandian subpopulations. Taken together, these 

results suggests that D. loculata consists of a long-lived 

background Eurasian population of latent mycelia in nonburned 

forests, established by ascospores dispersed from 

scattered burned forest sites. When the trees die from a 

forest fire, the latently established genets starts to decay the 

wood, and the sexual reproduction is triggered. In order to 

investigate the spatial distribution and origin of the parents 

of the stromata, all vegetative genets and offspring of the 

attached stromata were sampled at one burned forest site. 

The sequence of highly variable nuclear gene loci were 

analysed from each mycelium and the high level of unique 

genets found indicates ascospore origin of all mycelia. 

25 - Population genetics and dispersal biology of 

endangered lichen species: some implications for lichen 

conservation biology 

C. Scheidegger * , J.-C. Walser, D. Yetman, S. Werth & C. 

Sperisen 

WSL Swiss Federal Research Institute, CH-8903 

Birmensdorf, Switzerland. - E-mail: scheidegger@wsl.ch 

Global populations of endangered lichen-forming fungi are 

often highly fragmented and local populations are reduced 

to critical size, e.g. in Erioderma pedicellatum. Especially 

in heterothallic species such as Lobaria pulmonaria this 

process is likely to reduce the survival probability of local 

populations if genetic diversity is reduced. For instance, in 

this species genetically depauperate populations seem to be 

unable to reproduce sexually and therefore propagation is 

limited to vegetative, symbiotic dispersal units. However, 

the low range of these relatively large dispersal units limits 

the local populations to rather low average population 

densities. The study of the dispersal and post-dispersal 

processes is therefore of prime importance to identify 

critical distances among habitats, especially in managed 

forests. PCR-based techniques were recently developed and 

allow the detection of single symbiotic propagules. Further, 

sets of markers with high levels of genetic variation will 

become available and will allow to detect rare genotypes 

which depend on conservation measures such as population 

augmentation.


26 - Genetic heterogeneity in Phialocephala fortinii 

populations along a latitudinal transect 

M.M. Piercey * , M.A. McPherson, S.W. Graham & R.S. 

Currah 

Dept. Biological Sciences, University of Alberta, 

Edmonton, Alberta, T6G 2E9, Canada. - E-mail: 

mpiercey@ualberta.ca 

Dark septate root endophytes (DSE) commonly inhabit 

roots of vascular plants in cold-stressed environments. One 

species of DSE, Phialocephala fortinii, displays very little 

host specificity and, when inoculated on some plants, a 

variety of effects can be observed including an increase in 

dry weight of the host plant, a decrease in dry weight of the 

host plant, or no observed effect. The apparent strict 

asexual mode of reproduction would indicate that there 

should be minimal variation in this fungus within a 

population, with distinct clones occupying distinct habitats. 

To assess whether genetic variation within P. fortinii is 

correlated to latitude and how this variation is structured 

within and among populations, P. fortinii was isolated from 

Salix root fragments collected from 40 x 40 m plots along a 

latitudinal transect from 49°N to 78°N. Genetic 

heterogeneity of isolated P. fortinii strains was evaluated 

by amplified fragment length polymorphism (AFLP) 

analysis and comparisons were made within populations 

and between populations. Clones were not detected within 

any population, and at all latitudes a single, distinct 

individual was found inhabiting each collected Salix root 

fragment. Genetically distinct populations were detected 

only in arctic latitudes. 

27 - Mechanisms and factors implicated in genet 

distribution and in dynamics of populations of 

ectomycorrhizal basidiomycetes 

H. Gryta 

CESAC, 29, rue Jeanne Marvig 31055 TOULOUSE Cedex 

4, France. - E-mail: gryta@ecolog.cnrs.fr 

Among ectomycorrhizal fungi -root symbionts of woody 

plants-, Basidiomycetes are the most diversified (with 

about 4500 described species) and studied ones. In natural 

environments, roots of woody plants are potentially 

connected to an assemblage of several fungal species each 

represented by more or less different individuals. To 

understand the functioning and ecological roles of 

ectomycorrhizal fungi, a frequent approach is to focus on 

local populations of these fungi to determine their genetic 

diversity and its distribution. This to subsequently infer 

how biotic and abiotic processes and determinants lead or 

not to establishment of fungal individuals, to their 

persistence as growing mycelia and, finally, to temporal 

changes in fungal populations. Contrasting with the 

strongly discussed but more often conserved sporophores 

sampling method, the constant development of molecular 

markers in the last twelve years has largely contributed to 

great advances in this approach. Accurate characterizations 

of populations have revealed large variations of 

demographic strategies (often defined as the relative parts 

of sexual reproduction and of vegetative growth in 

structuring a population) among species as well as between 

populations sampled in different environments of a done 

species. Advances and comments concerning this approach 

are illustrated with examples from works using Tricholoma 

populinum, Tricholoma scalpturatum, Hebeloma 

cylindrosporum and Suillus variegatus as models. 

28 - Global patterns of genetic variation in 

Schizophyllum commune 

T.Y. James * , J.-M. Moncalvo & R. Vilgalys 

Department of Biology, Duke University, Durham, NC 

27708, U.S.A. - E-mail: tyj2@duke.edu 

Some mushroom species can be noticeably common and 

have wide geographic distributions. One such species is the 

wood decaying split-gill mushroom Schizophyllum 

commune. Our initial study addressed the global population 

structure of S. commune and asked whether long distance 

spore dispersal could possibly explain the fungus' 

distribution and occurrence. Earlier studies by John Raper 

demonstrated that the global population is completely 

interbreeding, and more surprisingly that the many matingtypes 

are randomly distributed with regard to climate and 

geography. Using both allozyme and DNA markers, we 

demonstrated that S. commune is in fact divisible into three 

highly divergent geographic lineages: South America, 

North America, and the Eastern Hemisphere. Evidence for 

even smaller scale population substructure has been 

observed among isolated populations in the Caribbean. 

Through a selective spore-trapping method, we found that 

spores of S. commune are extremely common in the air 

flora, yet extant populations are genetically subdivided, 

perhaps arguing for a role in natural selection in 

maintenance of population differentiation. Finally, direct 

studies of the mating-type genes themselves suggest an 

explanation for Raper's observation of mating-type 

distribution is the prolonged maintenance of all possible 

mating-types within each geographic lineage through 

strong balancing selection. 

29 - Chemical organization of the Aspergillus fumigatus 

cell wall and a revisited role for GPI-anchor proteins in 

cell wall organization 

I. Mouyna, T. Fontaine, M. Bernard, S. Chabane & J.P. 

Latgé * 

Unité des Aspergillus, Institut Pasteur, Paris, France. - Email: 

jplatge@pasteur.fr 

Recent structural analysis of covalent linkages between the 

different constitutive polysaccharides of the structural 

alkali-insoluble skeleton of the cell wall of Aspergillus 



fumigatus have shown that branching of the beta, 1-3 

glucan through beta, 1-6 linkages is the first event in the 

construction of the cell wall followed by linkages of the 

branched glucan with chitin, galactomannan and beta 1- 

3,1-4 glucan. In contrast to yeast, no proteins covalently 

bound to cell wall polysaccharides that could play a 

morphogenetic role in cell wall construction, have been 

identified in A. fumigatus. Accordingly, branching enzymes 

responsible for the linkages between polysaccharides 

represent essential enzymes for the construction of the cell 

wall. During the search of such glycosyltransferases, we 

identified a beta 1-3 glucanosyltransferase playing an 

important role in cell wall biogenesis. This enzyme that is 

responsible for elongation of beta1-3 glucans, was 

glycosylphosphatidyl inositol (GPI) anchored to the plasma 

membrane. Comparative proteomic and genomic 

approaches have identified 5 GPI-protein families common 

to both A. fumigatus and yeast. None of the genes encoding 

putative polysaccharide-bound proteins in yeast have been 

found in A. fumigatus. Disruption of all the GPI-genes has 

been undertaken in A. fumigatus. Up to now, data suggest 

that the A. fumigatus GPI-proteins have enzymatic 

functions involved in the biosynthesis of fungal cell wall. 

30 - The regulation of chitin synthesis in yeast and 

pathogenic fungi: A common theme? 

C. Roncero * , M. Sanz, C. Jimenez, F. Castrejon & A. 

Duran 

CSIC/UNIVERSIDAD DE SALAMANCA, Avda. Campo 

Charro s/n. 37007-Salamanca, Spain. - E-mail: 

crm@usal.es 

Chitin synthesis is a process maintained across the fungal 

kingdom that, thanks to the power of genetic manipulation 

of yeast cells, is now beginning to be understood. Chitin 

synthesis is based on the regulation of distinct Chitin 

Synthases (CS) isoenzymes, whose number ranges from 1 

in Schizosaccharomyces pombe to 7 in some filamentous 

fungi such as Aspergillus fumigatus. This high diversity 

makes it difficult to find a unique model of regulation. 

Sequence analysis, together with the functional data 

reported, strongly support the notion that all fungi contains 

two highly divergent CS families, one of each crucial for 

cell division and the other involved in bulk chitin synthesis. 

The first family is likely to be controlled at cell cycle level, 

whereas the second one would depend on the function of 

several genes whose function has been studied in the yeast 

Saccharomyces. We review the current knowledge about 

the function of such genes and whether these genes may be 

conserved in other fungi. Finally, we discuss our recent 

results in the characterization of some of these genes in A. 

fumigatus and C. albicans. 

12 


31 - Alpha and beta (1-3) glucan synthesis in Aspergillus 

fumigatus 

A. Beauvais * , W. Morelle, M. Diaquin & J.P. Latgé 

Unité des Aspergillus, Institut Pasteur, Paris, France. - Email: 

abeauvai@pasteur.fr 

Alpha and beta (1-3)glucan are the main components of the 

cell wall of A. fumigatus. Like in other fungi, beta (1- 

3)glucan synthesis in A. fumigatus is under the control of 

FKS and RHO1 that are essential genes. Two genes AGS1 

and AGS2 encoding two putative alpha (1-3)glucan 

synthases have been now characterized in A. fumigatus. 

The predicted Ags1 and Ags2 proteins have an estimated 

molecular size of 273 kDa. Ags1p and Ags2p have three 

major hydrophobic regions separating two hydrophilic 

domains. The first hydrophilic domain is homologous to 

bacterial alpha amylases, and the second to bacterial 

glycogen synthases. AGS1 and AGS2 were disrupted in A. 

fumigatus. The ags1 and ags2 mutants have similar 

phenotype. Growth and conidiation are both reduced. 

Morphologically, abnormal sporulating structures were 

seen in liquid culture during early growth, whereas the wild 

type strain (WT) only produces mycelium. The ags1 

mutant is more sensitive to the cell wall inhibitors 

Nikkomycin and Congo Red than the WT and ags2 strains. 

No difference was seen in the carbohydrate composition of 

the alkali insoluble and soluble fractions of the cell wall of 

ags2 and WT strains. In contrast, a reduction in the alpha 

(1-3)glucan content of the alkali soluble fraction of the 

ags1 was observed. These results were in agreement with 

the immunolocalization of Ags1p and Ags2p since Ags1p 

was localized at the cell wall whereas Ags2p was found in 

the Golgi apparatus. 

32 - Beta-Glucan synthesis in S. cerevisiae: a genomewide 

synthetic lethal analysis of participating gene 

families 

G. Lesage, H. Li, A.-M. Sdicu, P. Menard & H. Bussey * 

Department of Biology, McGill University, 1205 Avenue 

Docteur Penfield, Montreal, Quebec H3A 1B1, Canada. 

We used the yeast deletion set to find synthetic lethal and 

sick interactions of genes involved in beta-glucan 

synthesis, to define networks of participating genes. We 

used deletion mutant arrays (Tong et al., 2001, Science 

294: 2364) to make double mutants in query glucan genes. 

We examined two gene families: FKS1, GSC2, FKS3 and 

KRE6 and SKN1. For FKS1, encoding a beta-1,3-glucan 

synthase, we find synthetic interactions with 82 genes, 

principally in areas of wall maintenance, cell stress, protein 

degradation, lipid and fatty acid metabolism and 

transcription. For GSC2 and FKS3, we find 20 and 83 

interactions respectively. Surprisingly, there is little 

overlap between the sets for the three FKS genes, a total of 

175 interactions involving 165 genes, reflecting the distinct 

biological contexts in which these related proteins function.


KRE6 encodes a glucanase-like protein required for beta- 

1,6-glucan synthesis, and is synthetically lethal with the 

related SKN1. KRE6 interacts synthetically with 87 genes, 

and SKN1 with 39. Again there is little overlap between 

the sets, with a total of 126 interactions among 121 genes. 

These genes are mainly in areas of cell wall maintenance, 

cell stress, protein degradation, protein modification, lipid 

and fatty acid metabolism and vesicle transport. The 

KRE6/SKN1 set show limited overlap with the FKS family 

set, consistent with their distinct functions, with a total for 

both families of 301 interactions among 286 different 

genes. 

33 - The PMTs: an evolutionarily conserved family of 

protein O-mannosyltransferases 

V. Girrbach, M. Lommel & S. Strahl * 

University of Regensburg Department of Cell Biology and 

Plant Physiology, Universitätsstrasse 31, D-93053 

Regensburg, Germany. - E-mail: sabine.strahlbolsinger@biologie.uni-regensburg.de 

Protein O-mannosylation is an essential protein 

modification in yeast. Furthermore, it is indispensable for 

cell morphology and cell wall integrity [1]. Yet, how Omannosylation 

affects cell wall architecture is still obscure. 

In yeasts and fungi O-mannosylation is initiated at the 

endoplasmic reticulum by an evolutionarily conserved 

family of protein O-mannosyltransferases, the PMTs. 

Phylogenetic analyses revealed, that this family can be 

divided into Pmt1p, Pmt2p and Pmt4p like subfamilies, 

including transferases closely related to Pmt1p, Pmt2p and 

Pmt4p, respectively. Here we present the molecular 

characterization of the PMT subfamilies of S. cerevisiae 

showing that Pmts are integral membrane proteins with 

seven membrane spanning domains which form specific 

high molecular weight complexes in vivo. Further, 

characterizing pmt mutants we found that their phenotype 

closely resembles mutants of the cell wall integrity 

signaling (PKC1-) pathway which monitors cell wall 

stability [2]. Analysis of the PKC1-pathway revealed that 

O-mannosylation is essential for induction of the this signal 

transduction cascade upon external stresses. Biochemical 

analysis of the highly O-mannosylated upstream receptors 

of the cell wall integrity pathway showed that abated Omannosylation 

affects their maturation, stability and 

function. References [1] Strahl-Bolsinger S et al. (1999) 

Biochim. Biophys. Acta 1426: 297-307 [2] Heinisch JJ et 

al. (1999) Mol Microbiol 32: 671-680. 

34 - Regulation of glycosylation and pH in the yeast 

Golgi 

N. Dean * & X.D. Gao 

Department of Biochemistry and Cell Biology, Institute for 

Cell and Developmental Biology, State University of New 

York, Stony Brook, NY 11794-5215, U.S.A. - E-mail: 

Neta.Dean@stonybrook.edu 

In yeast, glycoproteins are heavily modified by mannose in 

the Golgi. The donor for mannosylation is GDP-mannose. 

Use by lumenal mannosyltransferase requires that GDPmannose 

be transported from the cytosol to the Golgi by a 

specific transporter. Once the mannose is donated to 

protein, GDP is converted to GMP by nucleoside 

diphosphatases. GMP is an antiporter whose export from 

the Golgi is coupled to the lumenal import of GDPmannose. 

Substrate provision thus involves a cycle in 

which mannosylation acts as a sink to continuously 

generate the antiporter. As a byproduct of this cycle, 

hydrolysis of GDP to GMP generates a huge amount of 

phosphate that must be removed to prevent an overly acidic 

lumenal pH. Existence of a phosphate transporter is 

hypothesized to be critical for pH homeostasis in the Golgi, 

as mannosylation in the Golgi accounts for the vast 

majority of cellular GDP hydrolysis. We present evidence 

that ERD1 plays a key role in regulating lumenal Golgi pH. 

Erd1p is Golgi localized and is homologous to other 

phosphate transporters. erd1 mutants suffer from a number 

of Golgi defects, including glycosylation and ER protein 

retention. erd1 mutants are EGTA sensitive,show pH- and 

PO4-dependent growth defects and are suppressed by 

ERS1, a gene that functions to regulate vacuolar pH. Taken 

together our results support the model that Erd1p regulates 

the removal of lumenal phosphate that is generated through 

the consumption of nucleotide sugars during glycosylation 

in the Golgi. 

35 - Environmental stimuli suppress the Neurospora 

crassa cot-1 phenotype 

R. Gorovits * & O. Yarden 

Hebrew University of Jerusalem, Faculty of Agriculture, 

Rehovot 76100, Israel. - E-mail: gorovits@agri3.huji.ac.il 

TheNeurospora crassa colonial temperature sensitive-1 

(cot-1) gene encodes a Ser/Thr protein kinase required for 

proper hyphal elongation. The temperature-sensitive cot-1 

strain exhibits normal spreading radial growth at or below 

25 °C, but mutant colonies grow slowly with extensively 

branched hyphae at or above 32 °C. Antibodies raised 

against COT1 detect a 67-kDa polypeptide that is absent in 

extracts obtained from cot-1 grown at restrictive 

conditions. The cot-1 hyperbranching phenotype is 

accompanied by an increase in proton efflux (as 

determined by rate of medium acidification) and a 50-75% 

reduction in relative intracellular sodium content 

(determined by X-ray microanalysis). Ammending the 

growth medium with ion pump inhibitors (DES, Amiloride 

or Ouabain), NaCl (0.5-1.5M) or sorbitol (1-1.5M) 

remedies the ionic imbalance and suppresses the cot-1 

phenotype to various degrees. The COT1 67kDa 

polypeptide was detected in extragenic suppressors of cot-1 

(exhibiting partial or full suppression), but not in cot-1 

cultures suppressed by the tested environmental stimuli. 

Based on these results we suggest that impaired COT1 

function confers changes in cellular ionic homeostasis and 

that suppression of the cot-1 phenotype by genetic 

alterations versus changes in environmental conditions may 

involve different pathways. 



36 - The status of names and records of Australian 

macrofungi 

T.W. May 

Royal Botanic Gardens Melbourne, Private Bag 2000, 

South Yarra, Victoria 3141, Australia. - E-mail: 

tom.may@rbg.vic.gov.au 

In the Catalogue and Bibliography of Australian Fungi, all 

names applied to Australian fungi are arranged under 

accepted names, with a comprehensive listing of literature 

records. Treatments for basidiomycete macrofungi are 

complete, and data from the first volume is accessible online. 

Some macrofungi are recorded from Australia only as 

names on herbarium specimens. Such records will soon be 

accessible through the development of on-line specimen 

databases. Mapping schemes are generating large numbers 

of unvouchered sight records for selected macrofungi. 

Information from different types of records (literature, 

herbarium and sight) needs to be integrated and assessed. It 

is apparent that the take up of names and the quality of 

records for Australian macrofungi varies considerably. The 

take up of names refers to how widely and with what 

confidence names are applied. Quality is measured by such 

things as the amount of descriptive data supporting the 

record and whether there are vouchers. Most species based 

on Australian types have been disposed among modern 

genera, but often are known only from the type. For species 

based on extra-Australian types, numerous Australian 

records appear to be misapplied. A scheme will be outlined 

for coding the status and reliability of names and records. 

Filters can be applied according to the various purposes for 

which data might be used (such as nomenclature, 

conservation status, distribution, quarantine, identification 

or regional censuses). 

37 - Distribution of Southern Hemisphere inoperculate 

discomycetes, the influence of Gondwanaland 

P.R. Johnston 1* & I. Gamundi 2 

1 Landcare Research, Private Bag 92170, Auckland, New 

Zealand. - 2 Universidad Nacional del Comahue, Casilla de 

Correo 1336, 8400 San Carlos de Bariloche, Prov. de Rio 

Negro, Argentina. - E-mail: 

johnstonp@LandcareResearch.co.nz 

Many authors have attempted to explain present-day 

species-level distributions of Southern Hemisphere fungi 

with reference to ancient continents and geology. We 

suggest that present-day species-level distributions are 

more likely to reflect the present-day proximity of 

landmasses, or anthropogenic factors, than they are to 

reflect the ancient geology of Gondwanaland. The question 

of the influence of Gondwanaland on the distribution of 

Southern Hemisphere fungi needs to be addressed by 

considering phylogenies at higher taxonomic levels. Some 

examples are provided, including Cyttaria and 

Torrendiella. The extent to which general fungal 

14 


distribution is still influenced by the events of 20-80 

million years ago will be known only after many more 

robust phylogenies are available for groups of Southern 

Hemisphere fungi at about the level of genus or family. 

38 - Coprophilous ascomycetes of Australia 

A. Bell * & D.P. Mahoney 

Independent mycological research, 45, Gurney Road, 

Lower Hutt, New Zealand. - E-mail: AskUs@xtra.co.nz 

Coprophilous ascomycetes of Australia This short narration 

is a summary of approximately three years research into the 

coprophilous ascomycetes of Australia. The culmination of 

this project is a recently completed fully illustrated manual 

documenting these fungi. The project was funded by the 

government financed Australian Biological Resources 

Study (ABRS). A total of 180 species of ascomycete fungi 

were identified 20 of which (=11%) represent hitherto 

undescribed species. The most frequently encountered 

genera include: Podospora (31 species), Sporormiella (19 

species), Chaetomium (16 species) and Ascobolus (13 

species). This presentation will include illustrations and 

discussion of a few of the most noteworthy taxa. The 

completed publication will also include the hitherto 

unpublished research of the late Harry Dade who in his 

retirement years also worked on Australian coprophilous 

fungi. The performing of this research also highlighted the 

urgency of upgrading of keys and the great need for more 

direct observations of living fungi on their natural 

substrates. 

39 - Saprobic and mycorrhizal ammonia fungi of the 

Southern Hemisphere 

A. Suzuki 1* , T. Fukiharu 2 & C. Tanaka 3 

1 Faculty of Education, Chiba University, Chiba 263-8522, 

Japan. - 2 Natural History Museum and Institute, Chiba, 

Chiba 260-8682, Japan. - 3 Graduate School of Agriculture, 

Kyoto University, Kyoto 606-8502, Japan. - E-mail: 

asmush@e.chiba-u.ac.jp 

Saprobic ammonia fungi, such as Ascobolus denudatus, 

Peziza moravecii, Tephrocybe tesquorum and Coprinopsis 

phlyctidospora were observed both in New Zealand and 

Australia. Morphological characters of collections of each 

fungus from New Zealand and Australia broadly fit within 

the species concepts of the same ammonia fungi from the 

Northern Hemisphere. The genetic variation among the C. 

phlyctidospora collections that were obtained from the 

Netherlands, Japan, New Zealand and Australia was 

examined by analyses based on the ITS rDNA sequences 

and by inter- and intracompatibility of the isolates from 

Japan and those from New Zealand and Australia. C. 

phlyctidospora is a species complex and individuals 

currently recognized as C. phlyctidospora in the Northern 

Hemisphere and those in the Southern Hemisphere are


distinct species. Mycorrhizal ammonia fungi obtained in 

the Southern Hemisphere were Hebeloma spp. and 

Laccaria spp. Many collections of the Hebeloma spp. 

obtained from New Zealand conformed to the 

morphological species concept of H. aminophilum in 

Australia. Data from their ITS rDNA sequences supported 

the above identification. H. vinosophyllum, a species only 

recorded from Japan, is a Northern Hemisphere counterpart 

species to H. aminophilum. These results suggest that some 

ammonia fungi in the Southern Hemisphere have closely 

similar counterpart species in the Northern Hemisphere. 

40 - Southern Hemisphere truffles - friends or 

relatives? 

T. Lebel * & F. Udovicic 

Royal Botanic Gardens Melbourne, Birdwood Avenue, 

South Yarra, 3141 Victoria, Australia. - E-mail: 

teresa.lebel@rbg.vic.gov.au 

Current research utilising molecular data is focussing 

attention on the evolution of and relationships between and 

within previously stable higher taxonomic groupings. 

Preliminary data is causing many researchers to take a 

closer look at morphological characters traditionally 

thought to delimit genera, families and even orders. The 

truffles or sequestrate fungi are a polyphyletic, diverse 

group of macrofungi, well represented in Australia and 

New Zealand. Where do they fit into the changing world of 

fungal relationships? Three examples will be discussed, 

highlighting some of the challenges at different taxonomic 

levels facing researchers. A new genus of truffle based on a 

single species is described and its position within the 

Agaricales confirmed. The recently described genus 

Amarrendia is thought to be the simplified 'end-point' of a 

morphological continuum including the agaricoid Amanita 

and the secotioid Torrendia. Relationships to other 

Amanitaceae are examined using molecular and 

morphological data. In Australasia the truffle-like 

Russulales are more diverse than previously thought, with 

some 40 new species recently described. Analyses of 

morphological characters provide some support of truffle 

generic boundaries, while molecular work suggests 

multiple origins of truffles within Russula and Lactarius. 

However, it is possible that there are both lineages which 

contain both agaricoid and truffle species, as well as 

lineages with strictly truffle morphology. 

41 - Corticiaceae of Patagonia: species' richness, rarity 

and distributional patterns 

A.G. Greslebin * & M. Rajchenberg 

Centro Forestal CIEFAP, CC 14, 9200 Esquel, Chubut, 

Argentina. - E-mail: alina@ciefap.cyt.edu.ar 

The Corticiaceae (Aphyllophorales, Basidiomycota) of 

southern Argentina are reviewed, with emphasis on Tierra 

del Fuego. A total of 156 species were recorded of which 

20% are endemic, 3% austral and 77% were either 

cosmopolitan or with another distributional pattern. Within 

the endemic species we found the new genera and species 

Nothocorticium patagonicum and Rhizochaete brunnea, 

and species in Hyphodontia (8), Dendrothele (5), 

Athelopsis (2), Hymenochaete (2), Aleurodiscus (3), 

Amyloathelia (1), Ceraceomyces (1), Fibricium (1), 

Hypochniciellum (1), Leptosporomyces (1), Sistotrema (2), 

Tubulicrinis (2), Tulasnella (1) and Vararia (1). An 

assessment of rarity of these taxa is presented, based on 

their abundance, distributional area and niche specificity. 

Of the five austral taxa, three are very rare, being recorded 

only once (Dendrothele biapiculata, Epithelopsis fulva and 

Hypochniciellum oblongisporum). Some endemic taxa 

appear to be closely related to Australian/New Zealand 

taxa, sharing distinctive characters within their genera. For 

example, Aleurodiscus antarcticus, A. triviale (Argentina) 

and A. parmuliformis (New Zealand) have smooth 

basidiospores and skeletocystidia and, morphologically 

seem intermediate between Aleurodiscus and Stereum. 

Species in Hyphodontia are also discussed as well as 

noteworthy species from other genera. Our knowledge of 

these fungi is far from complete since several forest types 

with distinctive environments remain unexplored. 

42 - Rust fungi (Uredinales) and smut fungi 

(Ustilaginales) in New Zealand 

E.H.C. McKenzie 

Landcare Research, Private Bag 92170, Auckland, New 

Zealand. - E-mail: mckenziee@landcareresearch.co.nz 

This paper examines the relationship between rust fungi 

and smut fungi in New Zealand and other parts of the 

world. New Zealand is a land of immigrants, and much of 

the rust and smut mycota is also introduced. Early 

Polynesian migrants may have introduced some rust fungi 

to the northern, subtropical Kermadec Islands, but 

European introductions of host plants, together with 

contaminating fungal spores, are the principal source of 

introduced rusts and smuts. Despite strict quarantine 

enforcement new rusts and smuts are continuing to be 

introduced. Recently introduced plant material may have 

introduced gladiolus rust and a dock rust from South 

Africa. A major source of new arrivals is wind-blown rust 

spores from Australia. The native rust and smut mycota has 

close affinities to the Australian mycota. Of 125 native 

rusts, 89 (71%) are considered to be endemic while 33 

(26%) also occur in Australia; of 30 native smuts, 14 (47%) 

are endemic and 12 (40%) are also found in Australia. 

Some indigenous rusts and smuts are known only in 

outlying islands e.g., Puccinia embergeriae (Chatham Is), 

Uredo inflata (Auckland Is), Restiosporium dissimile 

(Chatham Is). Puccinia oreoboli is restricted to the New 

Zealand subantarctic islands and to the highlands of Papua 

New Guinea while the smut, Microbotryum nivale, is 

known on an isolated mountain in Central Otago, and in 

Arctic Europe and Greenland. Only two indigenous rusts 

and three smuts are known to have South American 

affinities. 



43 - Witches' broom and frosty pods: The unexpected 

relatedness of two major cacao pathogens 

G.W. Griffith 1* , J.N. Nicholson 1 , A. Nenninger 1 , R.N. 

Birch 1 & J.N. Hedger 2 

1 Institute of Biological Sciences, University of Wales 

Aberystwyth, Penglais, Aberystwyth, Ceredigion SY23 

3DA, Wales, U.K. - 2 School of Life Sciences, University of 

Westminster, 115 New Cavendish Street, London W1M 

8JS, England, U.K. - E-mail: gwg@aber.ac.uk 

The agaric Crinipellis perniciosa is a hemibiotrophic 

pathogen which causes witches' broom disease of cacao 

and has recently decimated the Brazilian cacao industry. In 

addition to the pathogenic C-biotype, other biotypes are 

found associated with unrelated plant taxa, notably lianas 

(L-biotype) and solanaceous hosts (S-biotype). The C- and 

S- biotypes are non-outcrossing and form broom symptoms 

on hosts whereas the L-biotype is outcrossing and 

asymptomatic. Genetic relatedness within and between 

biotypes can provide information about the past spread of 

the disease and the likely future direction of evolution of 

the pathogen. Phylogenetic analysis of several regions of 

the rRNA locus (ITS, partial IGS, partial 28S) revealed 

near identity between all C- and S-biotype isolates from 

diverse locations. L-biotype isolates formed a discrete but 

distinct clade. Surprisingly, analysis of sequences from 

Moniliophthora roreri, an imperfect basidiomycete 

pathogen of cacao in the New World revealed a very close 

relationship with C. perniciosa (closer than the 

morphologically very similar C. eggersii). Similarities in 

host infection between C. perniciosa and M. roreri were 

noted by Harry Evans but it is difficult to reconcile the 

gross morphological differences. Pairings between 

monokaryons of C. perniciosa and M. roreri (both lacking 

clamps) gave rise to a clamped dikaryotic mycelium 

suggestive of a hybridisation event. The nature and 

implications of these findings are discussed. 

44 - Conservation of fungi in the Southern Hemisphere 

- Challenges and progress 

P.K. Buchanan 1* & T.W. May 2 


Zealand. - 2 Royal Botanic Gardens Melbourne, Birdwood 

Avenue, South Yarra, Victoria 3141, Australia. - E-mail: 

buchananp@landcareresearch.co.nz 

Fungal conservation in all parts of the Southern 

Hemisphere lags well behind organisms of larger size, 

capable of movement, or coloured green. Other challenges 

are the inadequate inventories and knowledge of 

distribution of fungi, the few professional and amateur 

mycologists, and the simplistic world-view confined to 

'plants and animals'. The authors are not aware of any 

Southern Hemisphere government Red Data Lists for Fungi 

using IUCN criteria. But in Australasia and other southern 

regions, some progress is evident. In New Zealand, the 

16 


conservation status of fungi is being considered for the first 

time at national level with iconic threatened fungi 

including species of Claustula, Ganoderma, and Puccinia. 

In Australia, distribution of selected species of fungi is 

being mapped in the nationally coordinated programme 

'Fungimap', and two reserves have been established based 

in part on the presence of threatened fungi including 

Hypocreopsis and Hygrocybeae. Both countries have 

checklists of all recorded fungi in preparation. In southern 

South America corticioid fungi have been assessed for 

rarity. The conservation status of many South Australian 

macrofungi has also been assessed, although incomplete 

knowledge of distribution often limits definitive 

conclusions. Examples are given of threatened fungi in 

different regions and a summary presented of fungal 

conservation hemisphere-wide. 

45 - How alpine are 'alpine' fungi? 

M. Moser 

University Innsbruck, Tschnikerstr. 25, A-6020 Innsbruck, 

Austria. - E-mail: Meinhard.Moser@uibk.ac.at 

Several of the 'alpine' fungi from alpine tundra have been 

observed in lowlands but in association with Salix species; 

others can change the host genus in lower elevations. Vice 

versa 'lowland' fungi are occasionally observed in alpine 

tundra but seemingly without being able to establish 

permanent populations. Factors, which enable alpine fungi 

to survive the stress conditions of the alpine tundra, are 

either passive (growth respectively fructification in 

especially protected habitats or only during favourable 

periods or the ability of basidiomata to revive) or active 

(reduction of water content, increase of carbohydrate 

storage, special proteins and enzymes, membrane lipids 

etc.). 

46 - Cortinarius favrei: an example for phylogenetic, 

morphological, and ecological species concepts in alpine 

fungi 

U. Peintner 1* , M. Moser 1 , E. Horak 2 & R. Vilgalys 3 

1 University Innsbruck, Technikerstr. 25, 6020 Innsbruck, 

Austria. - 2 ETH Zürich, Zollikerstr. 107, 8008 Zürich, 

Switzerland. - 3 Duke University, P.O. Box 90338, Durham, 

NC 27708, U.S.A. - E-mail: Ursula.Peintner@uibk.ac.at 

Extensive analysis of closely related species from alpine, 

subalpine and montane habitats should facilitate the 

investigation of ecological, morphological and 

phylogenetic species concepts in ectomycorrhizal 

mushrooms. Cortinarius favrei is one of the most frequent 

alpine Cortinarius spp. associated with dwarf willow (e.g. 

Salix herbacea, S. polaris) and Dryas. The 

morphologically very similar C. absarokensis occurs with 

shrubby willow in subalpine habitats. In contrast, C. 

septentrionalis, C. trivialis and C. vernicosus are closely


related species occurring in boreal habitats in association 

with mostly deciduous trees. For this study, numerous 

collections of C. favrei and of C. absarokensis from 

circumpolar localities between the West-coast of the USA 

and the Russian Far-East were morphologically compared 

to each other and to closely related taxa of subgenus 

Myxacium section Myxacium. Moreover, nuclear DNA 

sequence data from the internal transcribed spacers (ITS1 

and ITS2) were generated and analyzed for the study 

group. Based on these extensive molecular and 

morphological data, phylogenetic, morphological, and 

ecological species concepts in alpine fungi are discussed. 

47 - Basidiomycetes of Greenland 

T. Borgen, S.A. Elborne & H. Knudsen * 

Botanical Museum, University of Copenhagen, 

Gothersgade 130, DK-1123 Copenhagen K., Denmark. 

Based on c. 10.000 collections of basidiomycetes from the 

Greenland Herbarium in Copenhagen (C) a checklist 

including 843 species has been established (in press). A 

quantitative analysis of the composition of the list 

regarding mode of nutrition and taxonomic structure is 

presented and compared to that from a temperate, northern 

European region. The development of special arctic 

elements in the funga as well as different ways of 

adaptation to the cold environment is shown and some 

fungal taxa acting as pioneers in young landscapes are 

pointed out. Examples of the major distributional types of 

basidiomycetes in Greenland are presented as well as 

typical representatives of an arctic(-alpine) and subarctic(subalpine) 

circumpolar distribution based on the authors 

collections from Alaska, Siberia, Svalbard, Iceland and the 

Alps as well as records from the literature. Finally, the 

biodiversity of the Greenland funga is compared to that of 

the well-investigated Greenland flora. 

48 - Mycoknowledge related to Svalbard (Spitsbergen) 

G. Gulden 

Botanical museum, P.O.Box 1172 Blindern, N-0318 Oslo, 

Norway. - E-mail: gro.gulden@nhm.uio.no 

Svalbard is today an easily accessible arctic region with 

well equipped research stations and housing facilities. 

Never the less, the mycological exploration has been casual 

and our knowledge is fragmentary. Only the lichen flora, 

comprising almost 600 recognised species, may be 

considered well studied. During the almost 170 years from 

S. C. Sommerfelt published the first account on fungi from 

Svalbard, up till today, only about 600 non-lichenised fungi 

have been recognised. They belong to all four divisions of 

the Mycota, but only a few taxonomic groups have been 

systematically sampled and studied, e.g. ascomycetes of 

the genera Lamprospora and Pleospora, gasteromycetes of 

the genera Calvatia and Bovista, agarics of the genera, 

Arrhenia and Galerina. An account on micromycetes on 

vascular plants exists (197 species) and a checklist of 

Svalbard's pyrenomycetes ss. lat. (129 species). Some 

ecological groups have been studied in detail, e.g. 

lichenicolous fungi (60 species) and dryadicolous fungi (34 

species). Furthermore, soil fungi have been isolated from 

peat soils and mycorrhiza and root-associated fungi have 

been studied in a number of vascular plants. A taxonomic 

catalogue to Svalbard plants, fungi, algae and 

cyanobacteria was published in 1996 with the intention 

also of giving a broad impression of occurrence and 

ecology of the species. For most of the fungi this is very 

tentative. 

49 - Fungal diversity in arctic Lapland and the 

Scandinavian mountains 

K. Bendiksen 1* & E. Ohenoja 2 

1 Botanical Garden and Museum, Trondheimsveien 23 B, N- 

0562 Oslo, Norway. - 2 Botanical Museum, University of 

Oulu, P.O. Box 3000, FIN-90014 University of Oulu, 

Finland. 

In Fennoscandia (North Europe) the climatic timberline is 

1200 m asl. in the central mountain massif of S. Norway, 

and decreases gradually towards the north and coastal 

areas. It reaches the sea level on the N. coast of Finnmark 

(71°N). The area above the timberline covers large parts 

(32%) of Norway. In Sweden the arctic (oroarctic) 

mountain areas are situated near the Norwegian border. In 

Finland the arctic vegetation covers only small areas in the 

northernmost Lapland. Records concerning Basidiomycetes 

from the arctic zone are sporadic. Some areas, particularly 

around the research stations, have been objects for more 

intensive studies and collecting. The following areas where 

relatively plentiful data is available are included in the 

presentation: 1) S. Norway, Hardangervidda (Finse), 

Jotunheimen, Rondane, Dovrefjell 2) N. Sweden, 

Norrbotten (Abisko) 3) NW. Finnish Lapland (Kilpisjärvi), 

the adjacent areas in Troms, Norway 4) NE. Finnish 

Lapland (Kevo, the fjells of Utsjoki and Inari), the adjacent 

areas in Finnmark, Norway About 400 identified fungus 

taxa (Basidiomycetes except wood-inhabiting 

Aphyllophorales) have been dealt with, and ecological 

aspects are discussed. There is a considerable reduction 

(60-80%) in the number of fungus species in the arctic 

zone as compared with that in the subarctic birch forests. 

The proportion of mycorrhizal species is highest (ca. 60%) 

in low arctic continental areas versus the low - middle 

arctic, slightly oceanic areas (45%). 

50 - Alpine macrofungi of North America (Rocky 

Mountains) 

C.L. Cripps 

Montana State University, Plant Sciences Dept., Bozeman, 

MT, U.S.A. - E-mail: CCripps@montana.edu 



Remarkably, macrofungi of the North American alpine 

zone have remained largely unknown until our recent 

intensive efforts in the Rocky Mountains revealed a diverse 

mycoflora. While Agaricales are well known for many 

arctic-alpine regions, only a few were reported above 

treeline in NA. To date, we have discovered over 150 

species of Agaricales from the Canadian border south into 

the Rocky Mountains, and above treeline (3300 m at 45°N, 

3600 m at 38°N). Most are typical arctic-alpine fungi at 

their furthest southern extent in NA. Predominant 

ectomycorrhizal plants are Salix reticulata, S. arctica, S. 

planifolia, S. glauca, Dryas octopetala, and Betula 

glandulosa (rare). Mycorrhizal taxa include Amanita cf 

nivalis, A. greenlandica, Russula nana, R. norvegica, R. 

delica, Lactarius nanus, L. glyciosmus, L. 

repraesentaneous, L. salicis-reticulatae, Leccinum 

rotundifolia (rare), Laccaria bicolor, L. pumila, L. 

montana, Entoloma alpicola, Cortinarius absarokensis, C. 

favrei, C. hinnuleus, Hebeloma spp., and over 28 Inocybe 

species. Bryophilous taxa include Omphalina, Rickenella, 

Galerina, Marasmius epidryas, Arrhenia auriscalpium, and 

A. (Phaeotellus) acerosus. Terrestrial saprophytes include 

Cystoderma, Lepiota, Melanoleuca, Calocybe, and 

Collybia. Most are known from other arctic-alpine habitats, 

some are alpine-subalpine, and a few appear to be new. 

Species distributions differ, with some restricted to 

northern or southern RM regions. 

51 - Basidiomycetes in arctic tundra in North Amercia 

O.K. Miller Jr. 

Virginia Tech, Dept. of Biology, Blacksburg VA 24061, 

U.S.A. - E-mail: orsonk@cs.com 

Approximately 140 species of Agaricales and 

Aphyllophorales in the Basidiomycetes have been 

described and reported from arctic tundra in North 

America. Of these about 30% are ectomycorrhizal 

associates of 14 species and two varieties of Salix, in 

addition to Dryas integrifolia, Arctostaphylos rubra, and 

Cassiope tetragona. Chief among the ectomycorrhizal 

associates are Cortinarius, Inocybe, Hebeloma, Laccaria, 

Lactarius, and Russula. Three species of basidiolichens in 

the genus Botrydina (Omphalina) are widely distributed 

and common. More than 32 genera of decomposers contain 

about 100 species. Galerina, Phaeogalera, and 

Leptoglossum have 22 species which are decomposers of 

pleurocarpous Bryophytes. Over 60 species of 

decomposers belong to the Coprinaceae, Tricholomataceae, 

Strophariaceae, and Hygrophoraceae. Distribution of 

tundra species appears to be circumpolar with restricted 

endemism. The mycoflora is discussed in relation to the 

Alaska North Slope tundra, and the arcto-alpine habitats in 

the mountainous regions of northern North America. 

18 


52 - Arctic-alpine agarics and boletes (Basidiomycota) 

past, present and future 

E. Horak 1 , O.K. Miller Jr. 2 & C.L. Cripps 3* 

1 Institute of Geobotany, Herbarium, ETH, Zollikerstrasse 

107, CH-8008 Zurich, Switzerland. - 2 Virginia Tech, Dept. 

of Biology, Blacksburg, VA 24061, U.S.A. - 3 Dept. of Plant 

Sciences, Montana State University, 215 AgBioscience 

Bldg., Bozeman, MT 59717-3150, U.S.A. - E-mail: 

ccripps@montana.edu 

By definition, arctic-alpine agarics and boletes are 

ectomycorrhizal, saprobic or parasitic mycota which 

closely interact-coexist with autochthonous associations of 

lichens, mosses and angiosperms exposed to the harsh 

ecological conditions above the upper timberline in alpine 

habitats or in tundra of Subarctica and Subantarctica. The 

first records on arctic-alpine macromycetes have been 

published by Hooker (1811, Iceland), Greville (1822, 

Scotland), Sommerfeldt (1833, Svalbard) and Heer (1936, 

Switzerland). In general, comprehensive data about 

taxonomy, ecology and distribution of arctic-alpine 

macrofungi are still scarce and limited to few localities of 

small geographic range in the Alps (Austria, Italy, France, 

Switzerland), Pyrenees (Spain), Rocky Mts. (Colorado), 

and several sites located in high circumpolar latitudes 

(Alaska, Fennoscandia, Faerøer, Greenland, Iceland, 

Russia, Svalbard). Taxonomically and ecologically the 

moss-, lichen- and peat-associated taxa of Galerina are 

today by far the best known group of arctic-alpine agarics. 

Due to lack of data, the following localities are still white 

or grey spots on the arctic-alpine map viz. the Caucasus, 

the Himalayas, northern Japan (Hokkaido), and the high 

mountain ranges in the equatorial belt of South America 

(páramos), SE-Asia and Africa. Mycological fieldwork in 

these regions is urgent. 

53 - From microscopes to molecules: a journey through 

time 

J. Campbell 

UIUC, Dept of Plant Biology, 265 Morrill Hall, 505 S. 

Goodwin Ave, Urbana, IL, U.S.A. - E-mail: 

jcampbe2@life.uiuc.edu 

The earliest references to marine fungi were made in 1846. 

In 1907 there were only 16 known species of marine fungi. 

It was not until the 1940's that any substantial interest in 

marine fungi was generated. The Halosphaeriaceae was the 

first family of pyrenomycetous marine ascomycetes to be 

proposed. The importance of morphological characters 

used to classify the family however were disputed, which 

led to inconsistencies in the listing of representative genera. 

Our understanding of the taxonomic importance of these 

characters has increased with the availability of new 

techniques. Here we look back on the journey through time 

from light microscope to electron microscopes to molecular


phylogeny using genera in the Halosphaeriaceae as 

examples of initial concepts and current perceptions. 

54 - Mangrove mycology in South East Asia: Who, 

what...and what else ? 

T.K. Tan 

Department of Biological Sciences, National University of 

Singapore, Science Drive 4, Singapore 117543, Singapore. 

- E-mail: dbstantk@nus.edu.sg 

One of the earliest records of marine fungi in south east 

Asia was that of Jones (1968), who reported Lulworthia 

floridana and Halosphaeria quadric-cornuta on drift wood 

collected in Singapore. Interest in marine mycology in the 

region was nurtured at IMC3 in Tokyo in 1983. As 

mangrove swamps constitute the extensive and 

characteristic coastal vegetation in this part of the world, 

subsequent post-IMC3 works focused primarily on the 

marine fungi in mangrove habitats. The first few papers 

were that of Gacutan and Uyenco (1983; Philippines), 

Kohlmeyer (1984, Thailand), Tan (1985, Singapore and 

Malaysia), Koch (1986, Thailand), and Jones and Tan 

(1987, Malaysia). Since then, intense interest and studies 

have yielded a wealth of information on the biodiversity of 

mangrove fungi in the region, with more than 100 new 

species and genera described. Apart from biodiversity, the 

proximity and accessibility of mangroves facilitated 

ecological studies undertaken to account for the observed 

occurrence, abundance and distribution of these fungal 

species. These included studies on fungal succession, 

vertical distribution, substrate specificity, and to a lesser 

extent, fungal interactions. Molecular taxonomy has also 

been studied in the last few years. Three IMCs later, how 

do we take stock of the progress in mangrove mycology ? 

There is certainly scope for further work on biodiversity 

and ecology. The question is: how can studies on mangrove 

fungi create a greater impact on life ?. 

55 - Diversity and phylogeny of Halophytophthora 

(Oomycetes) 

A. Nakagiri 

Institute for Fermentation, Osaka, 17-85, Juso-honmachi 

2-chome, Yodogawa-ku, Osaka 532-8686, Japan. - E-mail: 

nakagiri-akira@ifo.or.jp 

Halophytophthora species are straminopilous oomycetes 

inhabiting marine and brackish water environments as a 

first colonizer on the submerged fallen leaves. Fourteen 

species and two varieties have been described so far mainly 

from mangrove environments. They show wide variation in 

ecology and characters of asexual reproduction. For the 

purpose of investigating the phylogenetic relationship with 

other straminopiles and clarifying the species level 

taxonomy of this diverse group, 18S rDNA, D1D2 region 

of 28S rDNA and ITS1-5.8S rDNA-ITS2 were sequenced 

in 58 strains of Halophytophthora species and analyzed 

phylogenetically with other related oomycetes. The 

analyses revealed 1) the most species of Halophytophthora 

nested in the Peronosporomycetidae clade, but H. spinosa 

positioned apart from the main group but close to 

Sapromyces (Rhipidiomycetidae), which suggests at least 

removing H. spinosa to other (new) genus, 2) the 

Phytophthora-Peronospora clade nested within the main 

group of Halophytophthora as a sister group of H. vesicula 

complex, which includes H. vesicula and other 

morphologically similar species, 3) the species in the main 

group of Halophytophthora formed several clades, each of 

which corresponds to its specific character of asexual 

reproduction, 4) looking at the phylogenetic relationship 

between Phytophthora and Halophytophthora, the tree 

showed that the terrestrial or freshwater parasites might 

have evolved from the marine or brackish water saprobes. 

56 - Thai lignicolous aquatic fungi: From river to the 

sea 

S. Sivichai * , J. Sakayaroj & I. Chatamala 

BIOTEC-Mycology Laboratory, National Center for 

Genetic Engineering and Biotechnology, National Science 

and Technology Development Agency, 113 Pahonyothin 

Rd., Klong 1, Klong Luang, Pathumthani 12120, Thailand. 

Over the past 10 years our knowledge of Thai aquatic fungi 

has increased dramatically and with the description of 

many new taxa. The only previous records were those of 

Tubaki et al. (1983) on Ingoldian fungi. Although the 

major focus has been on their ecology, recent studies 

examines their molecular phylogeny, especially genera not 

assigned to an order or family for example Bathyascus, 

Torpedospora (Ascomycetes), Dendryphiella and 

Sigmoidea (anamorphic fungi). Currently some 600 higher 

aquatic fungi have been recorded for Thailand. 

57 - Marine fungi: A time to redefine? 

E.B.G. Jones 

BIOTEC, 113 Pahonyothin Road, Klong 1, Klong Luang, 

Pathumthani 12120, Thailand. - E-mail: 

bhgareth@yahoo.com 

Many workers have tried to characterize physiologically 

and ecologically what is a marine fungi? Over 1,000 fungi 

have been recorded from marine habitats and they appear 

to be well adapted for life in this environment. However, 

some species have also been recovered from freshwater 

habitats and this raises the issue of how we define this 

unique group. Recently, molecular studies have 

demonstrated that members of the Halosphaeriales 

originated from a terrestrial ancestor, rather than evolution 

from ancestral marine taxa. Similarly, a number of marine 

bitunicate ascomycetes have been shown to group with the 

terrestrial Pleosporales. It is therefore time to reconsider 



our definition of what is a marine fungus and to look 

forward as to future studies in marine mycology. 

58 - A curmudgeon's view of priorities and economics 

R.P. Korf 

Cornell University, Dept. Plant Pathology, Plant Science 

Bldg., Ithaca, NY 14853, U.S.A. - E-mail: 

RPK1@cornell.edu 

The noun curmudgeon is defined and used here in two, not 

necessarily exclusive, ways: n. 1. archaic: a crusty, illtempered, 

churlish old man. 2. modern: (i) anyone who 

hates hypocrisy and pretense and has the temerity to say so; 

(ii) anyone with the habit of pointing out unpleasant facts 

in an engaging and humorous manner. The presenter's 

theses are that we have nearly all of our priorities 

backwards, that monetary and not scientific ends drive 

many of our research efforts, and that misjudgment 

frequently drives the rest. Topics to be addressed are (1) 

Where We Stand in Inoperculate Discomycete Systematics; 

(2) Asking the Wrong Questions; (3) Money, and the 

Failure of Universities: Grant Writing; (4) Milking the 

Grant Agencies; (5) Alpha Taxonomy Remains our 

Primary Necessity; (6) Training Students: How Did We Go 

So Wrong? (7) Journals, Proliferation and Blatant Errors in 

Judgement: Choosing Between Tree-Destruction and the 

www; (8) A View of the Future: the Presenter Exposed as a 

Crypto-Optimist. 

59 - Anamorphs - teleomorphs and the disparity of 

sampling 

D.H. Pfister 

Harvard University Herbaria, 22 Divinity Ave., 

Cambridge, MA 02138, U.S.A. - E-mail: 

dpfister@oeb.harvard.edu 

Nannfeldt published his magnificent treatment of the 

Leotiales, one of the largest orders of Ascomycota, 70 

years ago and today it still provides the clearest and most 

comprehensive view that we have of the classification of 

the inopcerculate discomycetes. Prior to his study the 

literature on these fungi was scattered and family 

delimitations were unclear. Nannfeldt 1932 system has 

been nearly universally followed with minor modifications, 

most notably the recognition of a few, generally, small 

segregate families such as the Hemiphacidiaceae, 

Vibrisseaceae, Loramycetaceae, Sclerotiniaceae and 

Rutstroemiaceae. These families were segregated based on 

ascomatal morphology, anamorph differences and, in the 

case of the Rutstroemiaceae, by a combination of 

morphological features and molecular phylogenetic data. 

The intent of this paper is to review current knowledge of 

helotialean anamorphs and the life history data. The 

notable situation of the families Dermateaceae and 

Leotiaceae/Helotiaceae, which remain problematic and 

20 


largely unsampled in molecular phylogenetic data sets, will 

be discussed. The need for detailed biological studies of 

these fungi is of paramount importance in developing a 

reliable and predictive classification for this large and 

important group. 

60 - Neotropical discomycetes: Where do we stand and 

where do we want to go in systematics and biodiversity 

research? 

T. Iturriaga 

Universidad Simón Bolívar, Dpto. Biología Organismos, 

Aptdo. 89000, Sartenejas, Baruta, Edo. Miranda, 

Venezuela. - E-mail: titurri@usb.ve 

This talk will present an overview regarding the last 70 

years of knowledge about neotropical Discomycetes. Areas 

to be discussed include the principal Discomycete 

collectors in the neotropics in this period, as well as 

numbers of publications dealing with neotropical 

Discomycetes in major journals and congress presentations. 

Knowledge of the total numbers of Discomycetes for some 

neotropical areas will be compared with other groups of 

fungi from the same geographical regions. A major 

contributor towards neotropical Discomycetes knowledge 

since 1949, date of his first expedition to Venezuela, is R. 

W. G. Dennis. In his famous and widely used 'Fungus Flora 

of Venezuela and Adjacent Countries' (Dennis, 1970), he 

treated 161 species of Discomycetes. Other mentors of this 

group of fungi, as well as their dedication and relevance to 

the area, will be presented. Advantages and disadvantages 

of conducting research on neotropical Discomycetes will 

be discussed in the sense of research funding possibilities 

and the consequences that these fund restrictions have had 

on Discomycetes knowledge. A correlation between the 

number of Discomycete collections and areas where these 

have been made, in comparison with publications resulting 

from these studies, will be analyzed. Based on 

achievements over the last 70 years, a general overview 

and recommendations of the paths to be followed in the 

future will be discussed. 

61 - Molecular diversity and phylogenetic affinities of 

mycorrhizal and root-endophytic fungi of the Helotiales 

T. Vrålstad * , E. Myhre & T. Schumacher 

Div. of Botany and Plant Physiology, Dept. of Biology, 

University of Oslo, P.O. Box 1045 Blindern, 0316 Oslo, 

Norway. - E-mail: trudevr@ulrik.uio.no 

Teleomorphic species of the ascomycete order Helotiales 

are abundant in nature, and range from hardly visible to 

some more prominent representatives. However, a number 

of asexual or sterile root symbiotic (endophyic, ericoid 

mycorrhizal and ectomycorrhizal) fungi have proven to 

belong to the Helotiales, yet their taxonomy, phylogeny 

and ecological roles are largely unresolved. We have


studied the diversity and phylogenetic affinities of 

symbiotic root-associated ascomycetes of the Helotiales 

based on ITS1-5.8S-ITS2 (ITS) nrDNA sequences. 

Mycobionts were obtained from roots of ericoid plants and 

grasses and from Piceirhiza bicolorata ectomycorrhizas 

(pbECM) on conifers and hardwoods, predominantly in 

burnt and metal-polluted habitats. The mycobionts were 

sequenced through the ITS and compared to sequences of 

known helotialean taxa. We recognised 132 fungal ITSsequences 

with affinity to the Helotiales, of which 75% (54 

different ITS-genotypes) grouped within the 

Hymenoscyphus ericae aggregate including Phialophora 

finlandia. This aggregate showed stronger affinity to 

members of the Hyaloscyphaceae and Dermateaceae than 

to Hymenoscyphus fructigenus (genus-type species; 

Helotiaceae). The H. ericae aggregate may be referable to a 

generic unit, and includes a diverse group of closely related 

± darkly pigmented root-associated ascomycetes where the 

borders between intra- and interspecific ITS-sequence 

variation as well as different types of root-symbioses 

remain unclear. 

62 - How little we know about the inoperculates in 

China 

W.-y. Zhuang 

Institute of Microbiology, Chinese Academy of Sciences, 

P.O. Box 2714, Beijing 100080, China. - E-mail: 

zhuangwy@sun.im.ac.cn 

Three stages can be recognized in studies of inoperculate 

discomycetes in China. Saccardo's taxonomic treatments 

were applied by the early mycologists at the very 

beginning, the influence of Seaver's works constituted the 

second stage, and treatments by Nannfeldt and Korf were 

much involved after the 1950's. Revisionary work by Korf 

plus several other recent workers towards the natural 

classification system is accepted currently. Known species 

of the inoperculates in China increased from 67 in the 

1930's and 1940's, 81 in 1963 by Prof. S. C. Teng and 140 

in 1979 by Prof. F. L. Tai, and up to 424 in 103 genera at 

the end of 2001 in the third stage. Significant progress has 

been made through the efforts of mycologists from 

different institutions dealing with species diversity of the 

group in the past two decades and the known species 

increased dramatically. Examples revealing the species 

richness of the Hyaloscyphaceae and a number of genera in 

other families were provided. Gross morphology, detailed 

anatomic structures, and molecular data should be 

combined properly in order to discover the phylogenetic 

relationships among genera or families aiming at the 

establishment of correct species concepts and a reasonable 

taxonomic system for inoperculate discomycetes. We seem 

to be far from that goal. 

63 - Will ascus characters give meaning to future 

reclassifications of non-lichenized inoperculate 

discomycetes? 

G.J.M. Verkley 

Centraalbureau voor Schimmelcultures, P.O.Box 85167, 

NL-3508 AD, Utrecht, The Netherlands. - E-mail: 

verkley@cbs.knaw.nl 

Most families of the inoperculate discomycetes are still 

regarded as heterogeneous. Many genera are also still 

poorly delimited, although in recent years progress has 

been made by integrating molecular and phenotypic 

approaches in some monographic studies. With molecular 

tools we can rapidly trace the informative morphological 

characters. Although there is still an enormous amount of 

work to be done, we should also take action at suprageneric 

level, using a similar approach. It is clear that the 

inoperculate discomycetes are not monophyletic, and major 

reclassification is imperative, but this calls for robust multigene 

datasets, combined with congruent morphological 

character complexes that provide enough synapomorphies. 

Familiar characters such as excipular anatomy and 

gelatinous substances may fail at this point. Ultrastructural 

studies of selected taxa e.g., in the Helotiaceae and 

Hyaloscyphaceae, have shown that the ascus apical 

apparatus constitutes a promising set of microanatomical 

features and should be explored further. Although some 

fundamental differences in apical structures are only 

resolved by transmission electron microscopy, it is 

certainly feasible to screen larger groups of taxa using light 

microscopy. I will attempt to correlate morphological and 

molecular data currently available, however fragmentary 

they may be. Obviously, more data should be gathered 

before extensive conclusions can be drawn. 

64 - Progress and challenge: Helotiales in the new 

millenium 

B.M. Spooner 

Royal Botanic Gardens, Kew, Richmond, Surrey TW9 3AE, 

U.K. - E-mail: b.spooner@rbgkew.org.uk 

After 70 years of progress, radical changes to the 

classification of inoperculates proposed by Nannfeldt have 

been few. Such changes are mainly recent, based on 

powerful tools such as DNA and ultrastructural analyses. 

They include demolition of the traditional Leotiaceae and 

Sclerotiniaceae, now regrouped into 5 families and, more 

extreme, the ousting of Orbilia as representative of a whole 

new class. In contrast, molecular study of Hyaloscyphaceae 

indicates surprising coherence of seemingly disparate taxa. 

But there remain conflicts in results and uncertainty about 

the placement of many taxa. Problems include an acute 

lack of data on the species involved. Two-thirds of these 

may remain undescribed, and only a handful have yet been 

sampled for modern analyses. Small subunit RNA 

sequences, for example, are known for less than 50 genera, 



only 7% of those described so far. Few species have 

received detailed morphological study, and many are 

scarcely represented at all in the world's herbaria. 

Identification remains fraught with problems and there is 

huge disparity in knowledge between different regions. 

Less than 200 species have yet been reported from the 

tropics, for example, fewer than from some sites in 

England. Yet rate of description of new species continues 

to fall. Clearly, there is still a pressing need for description 

and morphological study, to provide the basic taxa for 

application of modern techniques and to approach 

eventually a natural system for the Helotiales. 

65 - Diversity in the genus Penicillium 

J.I. Pitt 

Food Science Australia, P.O. Box 52, North Ryde, NSW 

1670, Australia. - E-mail: John.Pitt@csiro.au 

Penicillium is a tightly circumscribed but diverse genus, 

characterised by a distinctive asexual fruiting structure, the 

penicillus. Separation from other genera is usually 

unequivocal. Diversity is reflected in occurrence in a wide 

range of habitats. Some species, especially in subgenera 

Aspergilloides and Furcatum, are primarily associated with 

soil. Some species, especially in subgenus Biverticillium, 

are common in biodegradation, i.e. decaying plants and 

plant based materials. Others, especially in subgenus 

Penicillium, are found in seeds, which includes foods such 

as cereals, or dung. Few can be considered pathogens of 

animals or plants. A few are truly ubiquitous. Penicillium 

species are most common in cooler temperate climates, a 

reflection of relatively low temperature profiles for growth. 

Penicillium is associated with two sexual genera, 

Eupenicillium and Talaromyces, with mostly soilborne 

species. Some taxonomic questions remain. The possibility 

exists that Penicillium is not truly monophyletic, and that 

the species associated with the two sexual states are distinct 

in evolutionary terms, though not obviously so in 

ecological or morphological terms. The question of 

delimitation of subgenera, based on morphology, arises 

because molecular studies suggest that the subgenera 

Aspergilloides and Furcatum are not distinct from each 

other. And finally, the perennial question of what 

constitutes a species has been fuelled by molecular studies 

in particular. 

66 - ISSR variation in Penicillium miczynskii and 

Eupenicillium shearii 

D.E. Tuthill 

University of Wyoming, PO Box 3165, Laramie, WY 82071, 

U.S.A. - E-mail: dtuthill@uwyo.edu 

There is little evidence to indicate whether species of 

Penicillium are or are not genetically cohesive. To address 

this issue, I have been performing ISSR analyses on 190 

22 


strains of P. miczynskii. These strains were isolated 

primarily from four Pinus dominated sites in southeast 

Wyoming. Additional strains were isolated from a mixed 

hardwood forest in Denmark and a Nothofagus dominated 

forest in Chile. I also collected 130 strains of E. shearii, 

primarily from Costa Rica and India. Preliminary results 

indicate that P. miczynskii strains from SE Wyoming are 

very much alike, though some variation is present. Danish 

strains of P. miczynskii show ISSR patterns very different 

from those of the Wyoming strains, and also show 

considerable variation amongst themselves. Parsimony 

analysis using ITS sequences from several of these strains 

placed most of them in a clade including the type strain of 

P. miczynskii. In contrast, the strains of E. shearii clustered 

into two groups, irrespective of origin. These two groups 

may be interpreted as distinct species. This hypothesis is 

further supported by ITS sequences, which indicate that at 

least one of these groups is not closely related to the type 

strain of E. shearii (no representative of the second group 

has been sequenced yet). These results suggest that 

variation is structured quite differently in mitotic Penicillia 

than in meiotic Eupenicillia, and that lack of a meiotic 

cycle does not impose a constraint on intraspecific 

variation. 

67 - Phenotypic diversity in Penicillium and its 

teleomorphs 

J.C. Frisvad 1* & M. Christensen 2 

1 Mycology Group, BioCentrum-DTU, Buildinng 221, 

Technical University of Denmark, DK-2800 Kgs. Lyngby, 

Denmark. - 2 Department of Botany, Aven Nelson Building, 

University of Wyoming, Laramie, U.S.A. - E-mail: 

jcf@biocentrum.dtu.dk 

The genus Penicillium contains species of an impressive 

diversity and they are consistently found all over the world. 

In a detailed analysis of species occurrence in 16 soil 

microfungal surveys, our data indicate that: 1) Penicillia 

are abundant in certain soils, accounting for nearly 50% of 

all species obtained with 10 - 71% recently described or 

undescribed; 2) the soils with highest pendiversity appear 

to be soils from tropical and temperate rain forests (upper 

Amazon, Peru; ancient conifer forest, Oregon, USA) and 

coastal annual grasslands (California, USA); and 3) a great 

many soil species are reliable indicators of habitat type. 

Eupenicillium associated Penicillia are usually soil- or 

foodborne, while the very different Talaromyces associated 

Penicillia often are with plant polymer associated. Some 

species are associated to plants and one of them even 

produce myconodules in Elder trees. One species can grow 

in saturated cupper sulphate solution. Few species are 

xerophilic, most of them are still to be described. Some 

acidophilic species can only grow below pH 3.5 and are 

associated to wood, while others are alkalinophilic. Many 

species, mostly undescribed, are psychrophilic, while 

others are thermophilic. The chemical diversity of 

Penicillium species is equally impressive, isolates in each 

species being able to produce a large number of secondary 

metabolites. Most species produce 5-10 different families 

of secondary metabolites each with 3 to 20 members.


68 - Resolving the identification of P. glabrum, P. 

purpurogenum, P. thomii and Eupenicillium lapidosum 

S. Peterson 

USDA-ARS-NCAUR, 1815 N. University Street, Peoria, IL 

61604, U.S.A. - E-mail: peterssw@mail.ncaur.usda.gov 

The identification of Penicillium glabrum and other closely 

related species presents a challenge to mycologists because 

of the phenotypic similarity of these species. To address 

this problem, DNA sequences from ITS, beta-tubulin, 

calmodulin and translation elongation factor 1 alpha were 

determined from ex type and other isolates belonging to P. 

glabrum, P. thomii, P. yezoense, P. grancanaraie, P. 

baiiolum, P. palmense, P. trzebinskii, P. terlikowskii, P. 

purpurescens, P. viridoalbus, P. oledskii and Eupenicillium 

lapidosum. Prior studies showed that in the ITS region, 

total variation among these species is two base 

substitutions, although some of these species may be 

synonyms. Ex-type isolates of E. lapidosum and P. thomii 

had identical sequences in ITS, CF, EF1 and BT; P. 

viridoalbus and P. purpurescens were identical at 3 loci 

and differed by a single base at the other suggesting 

synonymies in each case. Penicillium spinulosum also 

resembles P. glabrum and is closely related. However, it is 

a genetically and phenotypically distinct species. The 

pattern of genetic variability at multiple loci suggests a 

complex of variable and closely related species centered on 

P. glabrum. 

69 - Species delimitation of Penicillia and Aspergilli; 

dilemma's between phenotypic and molecular 

characterization 

R.A. Samson 

Centraalbureau voor Schimmelcultures, PO Box 85167, 

3508 AD Utrecht, The Netherlands. - E-mail: 

samson@cbs.knaw.nl 

The taxonomy of Penicillium and Aspergillus and their 

teleomorphs has been subject to various methods of 

investigations. With the classical morphological 

approaches using characters of conidiophores, conidia and 

sexual propagules, delimitation of many taxa has been 

unclear and debatable. Introduction of physiological and 

culture criteria has only contributed little, particularly 

because standardisation of media for cultivating 

Penicillium and Aspergillus remains problematic. On the 

other hand chemotaxonomic profiles of secondary 

metabolites are promising showing fine distinction of taxa. 

With the introduction of various molecular tools 

delimitation of taxa have been solved, although one 

particular method does not always apply for all taxa of both 

genera resulting in different taxonomies. In some cases it is 

difficult to show similarities between the phenotypic and 

molecular characterization. This is particular true for 

important groups such as Aspergillus section Nigri and 

Penicillium subgenus Penicillium. Species of these groups 

are economically important and correct identification is 

crucial. In this paper examples with taxonomic constraints 

are discussed. 

70 - A polyphasic taxonomic study of Fusarium 

avenaceum and F. tricinctum 

K.A. Seifert 1* , U. Thrane 2 , J. Bissett 1 , B. Blackwell 1 & J. 

McDonald 3 

1 Eastern Cereal and Oilseed Research Centre, Agriculture 

& Agri-Food Canada, 960 Carling Ave., Ottawa, K1A 0C6, 

Canada. - 2 Mycology Group, BioCentrum-DTU, Technical 

University of Denmark, Søltofts Plads, Building 221, DK- 

2800 Kgs. Lyngby, Denmark. - 3 Centre for Plant 

Quarantine Pests, Canadian Food Inspection Agency, 3851 

Fallowfield Road, Ottawa, Ontario, K2H 8P9, Canada. - 

E-mail: seifertk@em.agr.ca 

A common set of 50 strains of Fusarium avenaceum, F. 

tricinctum and F. stilboides were examined by colony 

characters, image analysis of sporodochial conidia and 

conidia from the aerial mycelium, substrate utilization 

profiles (BIOLOG), rep-PCR (BOX, ERIC, REP primers), 

DNA sequencing (beta-tubulin, elongation factor 1-alpha) 

and metabolite profiling (HPLC, NMR). In general, DNA 

sequencing and rep-PCR results resulted in identical 

branching patterns in dendrograms. BIOLOG and rep-PCR 

detected more strain-level variation than DNA sequencing, 

but the BIOLOG phenogram differed in several respects 

from the molecular phylograms. Most strains identified as 

F. avenaceum (or its putative synonym F. 

arthrosporioides) clustered together in the molecular 

analyses, but two small groups of strains, one from 

Australia and the other from New Zealand, may represent 

distinct phylogenetic species. A population isolated from 

Turkish wheat formed a distinct cluster. F. tricinctum and 

F. stilboides formed distinct clusters except for a single 

identified strain of each species that appeared to be more 

distantly related. Representative strains of two subspecies 

of F. avenaceum subsp. aywerte and subsp. nurragi, were 

distantly related to F. avenaceum subsp. avenaceum and 

probably represent distinct species. Analyses of combined 

data sets and correlations between molecular, 

morphological and physiological data sets will be used to 

refine species concepts for these taxa. 

71 - Towards a polyphasic and predictive taxonomy for 

small-spored Alternaria 

B. Andersen 1* & R.G. Roberts 2 

1 BioCentrum-DTU, Building 221, Technical University of 

Denmark, DK-2800, Kgs. Lyngby, Denmark. - 2 USDA, 

ARS, Tree Fruit Research Laboratory, 1104 N. western 

Avenue, Wenatchee, WA 98801, U.S.A. - E-mail: 

ba@biocentrum.dtu.dk 



Within recent years, several techniques have been used to 

study sub-generic classification in small-spored Alternaria. 

Examination of 3-dimensional sporulation patterns on 

standardized media has shown that there are more than 

seven stable patterns of sporulation. Molecular methods, 

such as RAPD-PCR, support this segregation, and 

metabolite profiling has shown that sporulation patterns are 

predictive of certain secondary metabolites. New methods 

to utilize phenotypic features unbiased have been 

developed and applied to Alternaria. Digital images of 

cultures, grown on standardized media at different 

temperatures, are captured and transformed into data 

matrices (VideometerLab). Similarly, total HPLC 

chromatograms are transformed into matrices (COW Tool). 

Each matrix is then subjected to statistical analysis. Adding 

other characters, such as pathogenicity or presence of 

specific genes, may strengthen existing species or speciesgroups 

based on morphology and reveal additional groups. 

By using multivariate statistics, it is also possible to 

combine several sets of different data derived from both 

current and new methods (e.g. RAPD and metabolites) in 

one analysis to get segregation of isolates into natural 

clusters of similar taxa. The purpose of this presentation is 

to introduce image analysis and multivariate statistics to 

analyse chemical, physiological and molecular data, 

separately and combined, in order to apply a polyphasic 

and predictive strategy to Alternaria taxonomy. 

72 - An assessment of research applications of Biolog 

MicroPlates TM 

J. Bissett * , C.A. Nolan & R. Mukagasana 

Agriculture and Agri-Food Canada, Eastern Cereal and 

Oilseed Research Centre, Ottawa, Ontario K1A 0C6, 

Canada. - E-mail: bissettj@em.agr.ca 

Biolog MicroPlates TM are employed to characterize fungal 

strains based on differential assimilation of test substrates 

and redox reactions in a 96-well test plate. The Biolog 

method is potentially advantageous in being relatively 

simple, fast and economical; and data acquisition and 

identifications can be automated using a microplate reader 

and applicable software. Several research applications of 

the Biolog system are presented: i) the reliability of the 

Biolog system as a diagnostic tool is determined for 

identification of the 'green mould' disease of the 

commercial mushroom caused by Trichoderma 

aggressivum, ii) data from Biolog MicroPlates are 

compared with ITS sequence data, and their utility assessed 

in a survey of the biodiversity of Trichoderma strains from 

Asia, and iii) more than 900 Trichoderma strains are 

characterized using Biolog microplates, and the resulting 

database used to select strains that might be exploited for 

specific bioconversions. Strain level variation is more 

evident with Biolog data than with DNA sequencing or 

RAPD techniques, and Biolog data do not consistently 

reflect phylogenies constructed from molecular data. 

However, the Biolog system is an economical alternative 

method for surveying biological diversity, and provides 

data that complements molecular data in phylogenetic 

studies. Biolog data may provide a rational for the selection 

of strains for industrial applications. 

24 


73 - Penicillium toxins in food and feed 

T. Rundberget * , I. Skaar & A. Flaaoyen 

National Veterinary Institute, PO Box 8156 Dep., 0033 

Oslo, Norway. - E-mail: thomas.rundberget@vetinst.no 

Penicillium species appear to dominate in storage damaged 

food and feed. Microbiological studies carried out at the 

National Veterinary Institute have shown that under certain 

conditions food may be severely infected by moulds, 

especially Penicillium species, and it is important to 

establish if mycotoxins are present. The Penicillium 

species identified belong to the most proficient known 

mycotoxin producing moulds. There are few analytical 

methods available for the analyses of Penicillium toxins in 

various matrices and therefore a screening method using 

liquid chromatography-mass spectrometry (LC-MS) to 

quantify the commercially available Penicillium toxins 

roquefortine C, griseofulvin, mycophenolic acid, 

verruculogen, chaetoglobosin B and penitrem A has been 

developed. Penicillium crustosum Thom., is very common 

in food spoilage worldwide. Several cases of intoxication 

caused by toxins produced by P. crustosum have been 

reported and the fungus is known to produce roqufortine C 

and the penitrems A-F. Investigations of isolates of P. 

crustosum, revealed the presence of penitrem A, but only 

small amounts of other penitrems (B-F) when the isolates 

were grown on rice. From the LC-MS traces of the rice 

extracts, two major components together with penitrem A 

could be seen. These two components have been isolated 

and identified as two new indole-alkalod isoprenoids, 

18,19-dehydrosecopenitrem A and E. 

74 - Regulations in the EU for mycotoxins in foods 

W.J. de Koe 

Food and Public Health Consultant, Hazekamp 2, 6707 

HG Wageningen, The Netherlands. - E-mail: 

wjdekoe@bird.nl 

The quality and safety of food may be threatened by a host 

of factors, including natural toxins. Of the large spectrum 

of natural toxins, mycotoxins are among the most widely 

studied and raise considerable concern, because of their 

ubiquity and potential deleterious effects on human and 

animal health. Mycotoxins are produced by certain fungi, 

which, under favorable temperature and humidity 

conditions, invade a wide range of agricultural crops in the 

field, and during post-harvest stages of production. In 1994 

the EU started the harmonization process on 

standardization of certain mycotoxins in foods after its 

Scientific Committee on Food (SCF) expressed an opinion 

on -in their view at that moment-, the most important 

mycotoxins: aflatoxins, ochratoxin A and patulin. 

Aflatoxins and ochratoxin A have been more or less 

regulated now and recently certain emerging Fusarium 

toxins such as deoxynivalenol have been added to the list 

to be regulated soon. Estimation of the actual dietary intake


of mycotoxins is essential for risk assessment and will be 

used in determining whether there may be a relationship 

between observed adverse effects in humans and exposure 

to a particular mycotoxin. In order to apply the control on 

mycotoxins and to set standards for trading purposes, it is 

necessary to have collaborative tested and validated 

methods of anaylsis and sampling. The coherent approach 

of how EU-wide standards will be established will be 

addressed. 

75 - Ink cap sex and mushroom development 

U. Kües 1* , R.C. Bertossa 2 , A.P.F. Bottoli 2 , D. Ciardo 2 , P.- 

H. Clergeot 2 , Yi Liu 2 , S. Loos 2 , G. Rupricht-Robert 2 , P.J. 

Walser 2 , M. Künzler 2 & M. Aebi 2 

1 

Molecular Wood Biotechnology, Institute for Forest 

Botany, Georg-August University Goettingen, Buesgenweg 

2 

2, D-37077 Goettingen, Germany. - Institute for 

Microbiology, ETH Zurich, Schmelzbergstr. 7, CH-8092, 

Switzerland. - E-mail: ukuees@gwdg.de 

Sexual development in C. cinereus occurs on the dikaryon 

under control of the mating type loci. A mating type 

proteins, homeodomain transcription factors, induce 

fruiting body initiation under appropriate light, temperature 

and nutritional conditions. Development starts with intense 

localized branching leading to a small loose structure 

(primary hyphal knot). Light and A genes are needed to 

transform it into a compact round aggregate (secondary 

hyphal knot) in which cap and stipe tissues differentiate. 

Once a primordium is fully established, light and the B 

mating type proteins (pheromone and their receptors) 

induce karyogamy and fruiting body maturation. Next to 

studying nutritional signalling, we identified some genes 

acting at the first steps in fruiting. One uncharacterised 

gene acts in primary hyphal knot formation, gene cfs1 for a 

potential cyclopropane fatty acid synthase in the transition 

to secondary hyphal knots. Work in B. Lu's lab presented 

us two galectin genes, whose first expression correlates 

with formation of primary (cgl2) and secondary hyphal 

knots (cgl1). Structural analysis within the fruiting body 

suggests these protein to function in hyphal aggregation. 

Due to control by A mating type proteins, light, C and N 

sources, their promoters provide us with an ideal system to 

identify cis- and trans-acting elements in transcriptional 

regulation. Supported by the ETH Zurich, the Swiss 

National Science Foundation and the Deutsche 

Bundesstiftung Umwelt. 

76 - How mushrooms work 

N.P. Money 

Department of Botany, Miami University, Oxford, Ohio 

45056, U.S.A. - E-mail: moneynp@muohio.edu 

With characteristic eccentricity, A. H. R. Buller (1931) 

determined the weight-lifting capacity of Coprinus fruiting 

bodies. From the mass raised by developing basidiocarps, 

he estimated that hyphae elongating within the stipe 

generated a pressure of 0.07 MPa (or two-thirds of an 

atmosphere). When all of the fruiting bodies emerging in a 

cluster exert pressure of this magnitude, their combined 

force is sufficient to crack compacted soil or decaying 

wood (and to dislodge paving slabs positioned over their 

mycelia in urban areas). For example, a pressure of 0.07 

MPa applied over an area of 0.01 square meters (= 10 × 10 

cm), produces a total force of 700 N. This is sufficient to 

lift the combined mass of the author of this presentation 

plus all seven volumes of Buller's Researches on Fungi (= 

71 kg). Interest in these classical experiments has been 

revived by biomechanical research involving precise 

measurement of the forces exerted by single hyphae. The 

instrumentation used in these investigations is also 

effective for studying the mechanical behavior of 

multicellular fungal organs, and valuable information on 

the origin and control of force during fruiting-body 

emergence has been obtained during preliminary 

experimentation. Once the mushroom has surfaced, a 

variety of developmental adaptations optimize the 

discharge of spores from the gills and dispersal in air 

currents flowing beneath the cap. Recent work on 

basidiocarp form and function is also featured in this 

presentation. 

77 - Forceful invasion of corn leaves 

H.B. Deising * , S. Werner, J.A. Sugui & M. Wernitz 

Martin-Luther-Universität Halle-Wittenberg, 

Landwirtschaftliche Fakultät, Phytopathologie und 

Pflanzenschutz, Ludwig-Wucherer-Str. 2, D-06099 Halle 

(Saale), Germany. - E-mail: deising@landw.uni-halle.de 

The corn pathogen Colletotrichum graminicola causes the 

anthracnose disease of several cereals and grasses. To 

infect the leaf, conidia germinate and differentiate a 

specialized infection cell called an appressorium (Deising 

et al. 2000). When an appressorium matures it accumulates 

osmotically active compounds to yield high concentrations. 

The resulting turgor pressure of more than 5 MPa is 

translated into forces of up to 17 µN by single appressoria 

(Bechinger et al. 1999; Bastmeyer et al. 2002). These 

figures indicate the importance of the rigidity of the fungal 

cell wall. We performed targeted inactivation of three 

chitin synthase genes of the corn anthracnose fungus and 

found that only CgCHSC is essential for vegetative growth 

and pathogenic development. To analyze the role of cell 

wall-degrading enzymes in the infection process we 

generated mutants defective in CgSNF1, a gene involved in 

activation of catabolite-repressed genes (Tonukari et al. 

2000). Reduced growth rates on complex corn cell walls 

and reduced rates of infection suggest that cell walldegrading 

enzymes assist forceful invasion by C. 

graminicola. Bastmeyer, M., Deising, H.B., Bechinger, C. 

2002. Annu. Rev. Biophys. Biomol. Struc. 31:167-175 

Bechinger, C., Giebel, K.-F., Schnell, M., Leiderer, P., 

Deising, H.B., Bastmeyer, M. 1999. Science 285:1896- 

1899 Deising, H.B., Werner, S., Wernitz, M. 2000. 

Microbes Infect. 2:1631-1641 Tonukari, N.J., Scott-Craig, 

J.S., Walton, J.D. 2000. Plant Cell 12:237-248. 



78 - Sexless superguns - the enigmatic life history and 

infection processes of the zoosporic nematode pathogen 

Haptoglossa 

G.W. Beakes 1* & S.L. Glockling 2 

1 University of Newcastle upon Tyne, Newcastle upon Tyne 

NE1 7RU, U.K. - 2 Northern Illinois University, DeKalb, IL 

60115, U.S.A. - E-mail: G.W.Beakes@ncl.ac.uk 

The biflagellate zoosporic nematode pathogen Haptoglossa 

is placed in the Oomycete fungi, although its precise 

phylogenetic position has still to be resolved. All 

Haptoglossa species are characterized by having sausagelike 

thalli, which often completely fill the body cavity of 

their host. Most species produce discharge tubes which 

breech the nematode cuticle and through which the spores 

are released at maturity. Haptoglossa species fit into two 

major groups: zoosporic and aplanosporic. The latter group 

always seem to produce two or more different types of 

infection cell of varying karyology. No clear evidence of a 

sexual stage has been found, although internal nuclei-filled 

compartments have been observed within the thallus of one 

aplanosporic species, which is suggestive of a sexual stage. 

Synaptonemal complex-like structures have been seen in 

the nuclei of differentiated cysts of a zoosporic species. 

Therefore the exact nature of the life cycle remains 

enigmatic. The other characteristic structural feature of this 

genus is the production from cysts or aplanospores of 

complex infection gun cells. Barron likened them to 

ballistic missiles, although they actually function like 

hypodermic syringes with retracted needles. They are some 

of the most complex structures observed in any zoosporic 

fungi. Whilst the fundamental structural components have 

been conserved they show enormous phenotypic plasticity 

from one species to another. 

79 - Bug murder in the rainforest: A fatal attraction of 

sex versus celibacy with a potential for discovering new 

biocontrol agents 

N.L. Hywel-Jones 

BIOTEC-Mycology, National Center for Genetic 

Engineering and Biotechnology, National Science and 

Technology Development Agency, 113 Pahonyothin RD., 

Khlong 1, Khlong Luang, Pathum Thani 12120, Thailand. - 

E-mail: nigelhj@biotec.or.th 

In nature insect fungi are faithful in respect to those they 

kill. In agriculture: anything goes, promiscuity the order. In 

nature: promiscuity the exception. Work aimed at 

developing biocontrol agents is mostly with isolates from 

artificial ecosystems. And much work is with a few species 

of Beauveria, Metarhizium, Nomuraea and Paecilomyces. 

Sex rarely rears its ugly head in agro-ecosystems: I posit 

that asexual populations from agro-ecosystems become 

clonal and adopt a promiscuous attitude to their hosts 

murder using general-purpose pathogenicity genes. In 

nature, sexual forms are faithful to those they kill - 

26 

pathogenicity genes more focused. The ancestral pathogen 

that moved from forest to field 15000 yrs ago had to adapt 

or die. A cicadellid pathogen from forest may have infected 

a leafhopper in a nearby rice field. That hopper may have 

died in a field where hoppers were rare but caterpillars 

common. Survival of the fittest favours a pathogenicity 

gene that could infect Homoptera and Lepidoptera. The 

cosy world of nature favours sex with fidelity where 

murder is the order. Agro-ecosystems favour asexual 

promiscuity when it comes to bug murder. 15000 years of 

agriculture has given the few biocontrol agents now being 

tested: the random consequence of sex and murder moving 

from forest to field. Isolations from insects in forest or 

from characterised soil isolates may provide a more 

targeted approach to discovering new biocontrol agents in 

the future. 

80 - Ectomycorrhizal development in wet alder carr 

forests: above- and below-ground diversity 

J. Baar 


Applied Plant Research, Wageningen University and 

Research Center, P.O. Box 6042, 5960 AA Horst, The 

Netherlands. - E-mail: j.baar@ppo.dlo.nl 

The aim of the study was to investigate ectomycorrhizal 

development in wet Alder carr forests in The Netherlands. 

In North-Western Europe, wet Alder carr forests are mostly 

found in peatlands and along streams forming an important 

component of wetland ecosystems. The soils are 

predominantly fed by groundwater and permanently 

waterlogged. The dominant tree species in wet Alder carr 

forests is Alnus glutinosa (Black alder). The present study 

showed that the majority of A. glutinosa roots in various 

wet Alder carr forests in The Netherlands were associated 

with ectomycorrhizal fungi, even in anoxic waterlogged 

soils. Ectomycorrhizal root tips and hyphae were observed 

up to two meters from the A. glutinosa trees in waterlogged 

soil. Ectomycorrhizal development under wet and oxygen 

limited conditions was related to radial oxygen loss from A. 

glutinosa roots. Field observations showed that the 

diversity of ectomycorrhizal sporocarps was higher in wet 

Alder carr forests in a peatland than along a stream. 

Moreover, PCR-based techniques showed that the 

abundance and diversity of ectomycorrhizal fungi below 

ground was higher in the peatland than along the stream. In 

the wet Alder carr forests in the peatland, a relatively high 

redox potential of the waterlogged soil was measured 

together with radial oxygen loss from roots of the dense 

understory vegetation indicating that ectomycorrhizal 

development in waterlogged soils is stimulated by oxygen 

input of the surrounding vegetation.


81 - Mycorrhizal fungi in wetlands: the role of the 

extramatrical mycelium 

A.V. Rice 1* , M.N. Thormann 1 , B.J. Wilson 1 , H.D. Addy 2 & 

R.S. Currah 1 

1 Department of Biological Sciences, University of Alberta, 

CW-405, Edmonton, AB T6G 2E9, Canada. - 2 Department 

of Biological Sciences, University of Calgary, 2500 

University Drive, N. W., Calgary, AB T2N 1N4, Canada. - 

E-mail: arice@ualberta.ca 

Wetland plants are generally considered non-mycorrhizal 

but recent surveys have confirmed that mycorrhizal fungi 

are common in wetland plants and that all of the usual 

types (VAM, orchid, ericoid, and ectomycorrhizas along 

with dark septate endophytes) can be found in appropriate 

host species. Among these fungi, we are interested 

primarily in the ascomycetous associates if peatland plants 

that have dematiaceous hyphae, a group that includes 

Phialocephala fortinii (DSE) and Oidiodendron maius 

(ericoid), among others. Phialocephala fortinii is a 

dematiaceous hyphomycete common in the roots of a 

variety of wetland plants. The ecological role of P. fortinii 

is poorly understood but it is an enzymatically diverse 

species, capable of degrading various organic nitrogen (N) 

and carbon (C) sources, including complex phenolic 

polymers, such as lignin, that are important constituents of 

peat. In many acidic bogs, P. fortinii is absent from the 

roots of ericaceous shrubs where Oidiodendron maius is 

the most frequent isolate. Our research demonstrates that 

O. maius is also an enzymatically diverse species, capable 

of utilizing organic C and N, and a proficient decomposer 

of Sphagnum fuscum, the primary component of bog peat. 

Our research suggests that the extramatrical mycelium of 

selected mycorrhizal fungi and other root endophytes may 

be important in nutrient cycling within wetland habitats. 

82 - Diversity of mycorrhizas in wet ecosystems, what 

we should consider: from a case study on alder EM to a 

new approach to screen AM 

F. Buscot 1* , C. Renker 1 , K. Pritsch 2 & J.-C. Munch 2 

1 University of Jena, Institute of Ecology, Dornburger 

Strasse 159, D-07743 Jena, Germany. - 2 GSF-Research 

Center for Environment and Health GmbH, Insitute of Soil 

Ecology, Neuherberg, D-85758 Oberschleissheim, 

Germany. - E-mail: francois.buscot@uni-jena.de 

The mycorrhizal status in wet ecosystems is complex, 

including non truly mycorrhizal associations with dark 

septate fungi to plants like poplars or alders forming both 

AM and EM. In the latter case, the scarce available results 

indicate that AM presence decreases and EM diversity 

increases from wet pioneer to dryer advanced succession 

phases. The aim of the paper is to present partial results 

and new technical developments on the basis of which 

exhaustive studies of the mycorrhizal status in wet 

ecosystems could be performed. A first point concerns a 

comparison between above and below ground diversity of 

EM at two sites of an alder forest near a lake in Northern 

Germany. By combining identification of fruiting bodies, 

morphotyping of the EM and analysis of the polymorphism 

in the ITS region of both kinds of samples, it was possible 

to demonstrate an increase of the EM diversity from the 

lake to the higher site of the stand. Direct investigations of 

AM diversity in the field are difficult for technical reasons. 

Identification on the basis of the spore morphology is 

possible, but spores only partially reflect the diversity of 

the AM themselves. AM anatomy is not convenient to 

determine the fungal partners at the 'species' level. Several 

methods based on molecular biology have been proposed, 

but the analyses are often hampered by contaminating 

fungi. We have developed an approach combining a Nested 

PCR with a restriction, which partially resolves the 

difficulty. 

83 - Contributions of arbuscular mycorrhizal fungi to 

plant performance in wet ecosystems: studies with 

Lythrum salicaria 

K.J. Stevens * , S.W. Spender, R.L. Peterson & R.J. Reader 

Department of Botany, University of Guelph, Guelph, 

Ontario, N1G 2W1, Canada. - E-mail: 

kjsteven@uoguelph.ca 

The impact of arbuscular mycorrhizal (AM) fungi on plant 

performance in wetland areas was investigated through 

studies on Lythrum salicaria (L.). Under field conditions 

plant performance and AM colonization levels varied with 

water availability and there was a negative correlation 

between plant performance and AM colonization levels. 

Partially submerged plants produce numerous, free-floating 

adventitious roots along submerged stem lengths. The 

adventitious roots, the primary root system, and AM fungal 

hyphae represent three possible pathways for resource 

acquisition. We tested the hypothesis that, to sustain shoot 

growth, a reduction in the ability of one or more pathways 

to acquire resources will require an increase in resource 

uptake in the remaining pathway(s). While greenhouse 

studies revealed that individually AM fungi did not have a 

significant effect on plant performance, the results did 

indicate a relationship among the three pathways 

supporting our hypothesis. Greenhouse studies were 

conducted to determine if, in inundated soils, AM fungi 

contribute to plant performance across a range of 

phosphorus availabilities. AM fungi did not enhance 

growth in inundated soils; in some cases performance was 

limited and it is unlikely that AM fungi enhance 

phosphorus nutrition in inundated soils. We suggest that 

because of the large differences between terrestrial and 

aquatic habitats, AM fungi may be performing different 

roles in each habitat. 



84 - Mycorrhization of endangered wetland plant 

species 

B. Fuchs 1 & K. Haselwandter 2* 

1 University of Salzburg, Department of Botany, 

Hellbrunnerstrasse 34, A-5020 Salzburg, Austria. - 

2 University of Innsbruck, Department of Microbiology, 

Technikersrasse 25, A-6020 Innsbruck, Austria. - E-mail: 

Kurt.Haselwandter@uibk.ac.at 

Arbuscular mycorrhizal (AM) fungal diversity in soil 

seems to influence plant biodiversity. As information 

concerning the mycorrhization of endangered plants is 

lacking, we determined the mycorrhizal status of Serratula 

tinctoria (Asteraceae), Betonica officinalis (Lamiaceae), 

Drosera intermedia (Droseraceae) and Lycopodiella 

inundata (Lycopodiaceae), occurring at two different sites 

(bog and fen meadow) in the county of Salzburg, Austria. 

In addition to AM fungi, infection by dark septate 

endophytes (DSE) was quantified. The infection intensity 

of AM fungi and DSE appeared to be higher in the fen 

meadow than in the peat bog. The roots of S. tinctoria and 

B. officinalis were heavily infected by AM fungi and both, 

vesicles as well as arbuscules were observed over the 

vegetation period. L. inundata showed AM fungal 

infection, too; in spring vesicles were observed frequently, 

in autumn they were less numerous. In D. intermedia roots 

AM fungal infection intensity was lower than in the species 

mentioned before; however, fungal structures typical for 

AM fungal infection were observed, especially in spring. S. 

tinctoria and B. officinalis were heavily infected by DSE, 

while L. inundata was just slightly infected and D. 

intermedia did not seem to be infected by DSE at all. 

85 - Molecular genetics of gibberellin biosynthesis in 

Gibberella fujikuroi 

B. Tudzynski * & M. Mihlan 

Universität Münster, Institut für Botanik, Schlossgarten 3, 

48149 Münster, Germany. - E-mail: 

bettina.tudzynski@uni-muenster.de 

As well as being phytohormones, gibberellins (GAs) are 

present in some fungi and bacteria. GAs were first 

discovered in the fungus Gibberella fujikuroi, from which 

gibberellic acid (GA3) and other GAs are produced 

commercially. Although higher plants and the fungus 

produce identical GAs, important differences in the 

pathways and enzymes used have become apparent with 

the identification of the genes for GA-biosynthesis in 

Arabidopsis thaliana and G. fujikuroi. These profound 

differences indicate that higher plants and fungi have 

evolved the complex biosynthetic pathways to GAs 

separately and not by horizontal gene transfer. In G. 

fujikuroi, the 7 genes of the gibberellin (GA)-biosynthetic 

path-way including four cytochrome P450 monooxygenases, 

are located in a gene cluster. To study the function 

of several genes of this cluster, we used a gene replacement 

28 


approach followed by GC-MS and HPLC analysis. Most of 

the genes encode multifunctional enzymes. The availability 

of the genes allows the study of molecular mechanisms of 

gene regulation. Because the GA biosynthesis is regulated 

by nitrogen metabolite repression, we cloned several genes 

which are involved in nitrogen regulation, such as the 

general nitrogen regulators, areA and nmr. Gene 

replacement of areA led to a significant reduction of 

gibberellin formation by repressing the expression of the 

GA-pathway genes. In contrast, NMR does not play the 

role as a general counterpart of AREA as expected. 

86 - Involvement of ethylene in a plant - Botrytis cinerea 

interaction 

Y. Elad * , Z. Lapsker, I. Kolesnik, N. Korolev & B. 

Kirshner 

Dept. of Plant Pathology, The Volcani Center, P. O. B. 6, 

Bet Dagan 50250, Israel. - E-mail: 

elady@volcani.agri.gov.il 

The role of ethylene was studied in the interaction of B. 

cinerea with the host plants tomato, French bean and 

Arabidopsis thaliana. Infected resistant Arabidopsis 

produce less ethylene as compared with infected sensitive 

plants. Ethylene promotes the disease development 

whereas inhibition of plant ethylene production resulted in 

disease suppression. Increased plant tissue reactive oxygen 

species (ROS) levels resulted in elevated ethylene and 

severe disease. The actual contact with pathogen cells (e.g. 

dead conidia) induced ethylene and ROS in plants. This 

effect was also obtained by leaf injury, H2O 2, pH reduction, 

oxalic acid and botrydial; the compounds and effects are 

produced by the pathogen. Antioxidants reduced ethylene 

production and disease severity. Interestingly, ethylene 

promoted conidia germination and subsequent penetration 

to the host. Ethylene signaling mutants of Arabidopsis 

showed extremely high susceptibility to B. cinerea; it is 

possible that the pathway that leads to susceptibility is 

independent of other ethylene signal transduction pathway. 

B. cinerea has the potential to produce ethylene, thus it is 

possible that the ethylene originates not only from the 

plant. In conclusion, in B. cinerea - plants interaction 

ethylene induces auto-catalytic production of the same 

hormone in exposed tissues thus promoting the 

deterioration of the infected tissue and the ROS 

development and vice versa. 

87 - Abscisic acid biosynthesis genes in Botrytis cinerea 

V. Siewers * & P. Tudzynski 

Institut fuer Botanik, Westf.Wilhelms Universitaet, D- 

48149 Muenster, Germany. 

Botrytis cinerea causes the grey mould disease in more 

than 200 plant species. Like several other phytopathogenic 

fungi, B. cinerea has been shown to produce different kinds


of phytohormones in axenic culture. Although the impact 

of this biosynthetic capacity on host-parasite interaction is 

unclear, it has been suggested that production of the 

phytohormone abscisic acid (ABA) is supporting the 

infection process. A molecular approach has been initiated 

to study the role of phytohormones in the pathogenicity of 

B. cinerea. Our aim is to clone genes encoding enzymes of 

the biosynthetic pathways, especially the ABA pathway, to 

study their expression and to analyse the pathogenicity of 

deletion mutants. Therefore, a cDNA library of an ABA 

overproducing strain was established and differentially 

screened. Several cDNA clones derived from genes 

expressed during ABA production were characterised. 

88 - Ethylene production and ethylene-induced genes in 

Botrytis cinerea 

V. Chague & A. Sharon * 

Department of Plant Sciences Tel Aviv University, Tel Aviv 

69978, Israel. - E-mail: amirsh@tauex.tau.ac.il 

We defined the biosynthetic pathway of ethylene in the 

pathogenic fungus Botrytis cinerea, and characterized the 

conditions that affect ethylene production in vitro. During 

the first 48 h of culture the fungus uses methionine to 

produce a-keto g-methylthiobutyric acid (KMBA). In 

darkness, KMBA accumulates in the medium and no or 

very low ethylene levels are produced. In light KMBA is 

photo-oxidized and ethylene is released. Ethylene 

production rates in the light were the highest after 24 h (7 

ml/g/h) and declined thereafter, whereas in the dark 

ethylene was detected only after 48 to 96 h of culture. A 

cDNA library was constructed from fungal mycelium that 

was grown for 24 h in the light in methionine-enriched 

medium. The library was differentially screened with 

mRNA probes from light (ethylene +) and dark (ethylene -) 

grown cultures. Over 20 differential clones were identified. 

These clones represent transcription factors, glycolysis 

enzymes, ribosomal proteins and unknown ESTs. Northern 

hybridization was conducted to confirm the differential 

expression pattern of the clones and to further define the 

expression pattern of clones that were highly induced by 

ethylene. This work provides the first molecular evidence 

for ethylene response in fungi. 

89 - Alteration of plant-pathogen interaction by 

phytohormones 

M. Al-Masri 1 , A. Sharon 2 & R. Barakat 1* 

1 

Hebron University, College of Agriculture, PO Box 40, 

2 

Hebron, Palestinian Authority, Israel. - Tel Aviv 

University, Dept. of Plant Sciences, Tel Aviv 69978, Israel. 

- E-mail: rbarakat@netvision.net.il 

The effect of plant growth regulators on in vitro growth of 

the pathogenic fungus Sclerotinia sclerotiorum and on 

white mold disease that is caused by the pathogen was 

investigated. Naphthalene acetic acid at concentrations of 

200-400 µg/ml inhibited the fungal growth in culture and 

reduced white mold severity on bean and cucumber plants. 

Gibberellic acid at concentrations of 50-250 µg/ml 

promoted both mycelium growth and white mold disease 

severity on plants. Methyl jasmonate at concentrations of 

75-250 µg/ml inhibited mycelium growth in culture and 

suppressed bean and cucumber white mold. Absiscic acid 

at concentrations of 100-300 µg/ml decreased mycelium 

growth but promoted disease development on bean and 

cucumber plants. Ethylene released from ethephon (200- 

600 µg/ml) increased bean and cucumber white mold 

severity. S. sclerotiorum has the capacity to produce 

ethylene in culture. Ethylene production by S. sclerotiorum 

reached peak (400 µl/g/h) after 6 days of incubation 

followed by a decline to 155 µl/g/h after 10 days. The 

ethylene biosynthesis inhibitor aminoethoxvinyleglycine 

(AVG) suppressed white mold severity on bean and 

cucumber plants at concentrations up to 300 µg/ml. The 

results demonstrate a variable effect of plant hormones on 

the development of white mold. The changes in disease 

development may be due to both the effect of the plant 

hormones on the susceptibility of the plant to infection as 

well as due to a direct effect on the fungus. 

90 - Differential effect of auxin-compounds produced by 

two Pythium species with different pathogenicity 

G. Le Floch 1* , P. Rey 1 , M.I. Salerno 2 , N. Benhamou 3 & Y. 

Tirilly 1 

1 Laboratoire de Microbiologie et Sécurité Alimentaire, 

ESMISAB technopôle Brest Iroise 29 280 Plouzane, 

France. - 2 Laboratorio de Proteccion Forestral, CISAUA- 

Facultad de Ciencias Agrarias y Forestales UNLP 60 y 

119 CC31 (1900) La Plata, Argentina. - 3 Département 

Recherche en Sciences de la Vie et de la Santé, Pav. Ch.E. 

Marchand, Université Laval Sainte-Foy Québec GIK7P4, 

Canada. - E-mail: gaetan.lefloch@univ-brest.fr 

Pythium ability to produce phytohormones-like compounds 

has been frequently assumed to be involved in plant 

Pythium relationship. This study shows that within Pythium 

genus, two fungi: P. oligandrum and Pythium group F, 

exert different effects on plant growth; both of them 

produce auxin compounds through the tryptamine pathway 

under the same cultural conditions. Indeed, the ability of 

Pythium group F to produce indole-3-acetic acid (IAA) in 

the immediate vicinity of roots had no positive effect on 

plant development. On the contrary, P. oligandrum 

production of tryptamine (TNH 2) was associated with 

increased plant growth. Although analysis revealed marked 

differences in auxin-compounds produced by both Pythium 

in the plant nutrient solution, their differential influence on 

tomato growth likely results from different fungal-plant 

relationships. Pythium group F is a minor pathogen: it 

causes symptomless infections, and then yield losses in 

tomato soilless cultures; its hydrolytic enzymes, have a 

destructive, though limited, impact on the root cell-walls of 

the outer cortical area. So, one may hypothesise that the 

effect of auxins on already damaged tissues dramatically 

disturbs the host physiology. As a consequence, abnormal 

root swellings and irregular plant development was 



observed. On the contrary, P. oligandrum, increased root 

development by inducing neither damages nor disturbance 

of TNH 2 absorption by roots, tryptamine has a positive 

impact on plant development. 

91 - Use of molecular biological techniques to directly 

monitor fungi with laccase activities in different soil 

compartments 

P. Luis 1* , F. Martin 2 & F. Buscot 1 

1 Institute of Ecology, Department of Environmental 

Science, University of Jena, Dornburger Str.159 D-07743, 

Jena, Germany. - 2 UMR INRA/UHP 1136, Centre INRA de 

Nancy, F-54280 Champenoux, France. - E-mail: 

patricia_luis@yahoo.fr 

Fungal oxidative exo-enzymes without substrate specificity 

play a central role in the cycling of organic matter in soils. 

Due to their broad ecological impact and their favorable 

gene structure, i.e. four highly conserved regions encoding 

copper binding sites, laccases appeared to be the best 

appropriate enzymes to develop a technique to monitor 

fungi with an oxidative potential in soils without taking 

them into culture. Several degenerated primers, specific for 

basidiomycetes, were designed to perform single and 

nested PCR and assessed on DNA extracts from pure 

cultures. A single PCR allowing amplification of laccase 

gene fragments of about 200 bp gave highest success rates 

and was chosen for an application on two forest and one 

agricultural soils. Diverse fragments of around 140 and 200 

bp were amplified and the polymorphism revealed by 

analyzing their sequence indicated that the method allows 

to monitor laccase genes from a broad spectrum of 

basidiomycetes. The analyses also showed a certain 

specificity of the distribution of fungal laccase genes in the 

soils and within the horizons, which probably reflects a 

specific distribution of corresponding fungal species. For 

the forest soil horizons, the higher diversity of laccase 

genes appeared in the litter which contained mainly 140 bp 

fragments and decreased with the depth. This work 

illustrates the potential of molecular biological tools to 

screen fungal genes with a precise function in complex 

environment compartments. 

92 - Phosphate uptake by communities of arbuscular 

mycorrhizal fungi 

I. Jakobsen 

Risø National Laboratory, P.O. Box 49, DK-4000 Roskilde, 

Denmark. - E-mail: iver.jakobsen@risoe.dk 

Arbuscular mycorrhizal fungi (AMF) constitute an 

important component of ecosystems by their ability to 

influence nutrient capture and structure of plant 

communities. The crucial role of the root-external 

mycelium in the soil-plant P transport by AMF has been 

unambiguously demonstrated by means of growth systems 

30 


encompassing one or more mesh-enclosed soil 

compartments. The mesh excludes root growth, but allows 

for in-growth of hyphae and thereby for the direct 

measurement of hyphal uptake of P isotopes added to the 

root free soil. Examples will be given on the successful use 

of such experimental system to demonstrate the P uptake 

by communities of AMF in various field studies. There is 

increasing evidence for marked inter- and intraspecific 

differences between AMF in their rate and pattern of 

mycelial growth and in their effectiveness of P acquisition, 

but the relationship between the P uptake and the diversity 

of an AMF community is only poorly understood. Future 

research challenges within this area will be discussed, 

including the identification of the dominating components 

of AMF communities in roots and the subsequent test of 

the hypothesis that it is these dominating AMF, which are 

responsible for the major proportion of P uptake by the 

communities. 

93 - Determining mycorrhizal versus saprotrophic 

status of fungi by isotopic methods 

E.A. Hobbie 

University of New Hampshire, Morse Hall, Durham NH 

03824-3525, U.S.A. - E-mail: erik.hobbie@unh.edu 

Mycorrhizal status of fungi is often difficult to determine 

because of difficulties in culturing functional symbioses. In 

addition, the possibility of facultative mycotrophy means 

that mycorrhizal status may vary in different field 

situations. Recent studies indicate that isotopic patterns of 

nitrogen (N-15), carbon (C-13), and C-14 (radiocarbon) 

often distinguish between mycorrhizal and saprotrophic 

fungi, and may therefore be a potent new tool to examine 

fungal functioning in situ. N-15 patterns in mycorrhizal 

fungi are correlated with proteolytic capabilities and N 

dynamics between fungi and plant hosts. C-13 patterns in 

mycorrhizal fungi appear to indicate source C and have 

therefore been used to determine plant hosts in mycorrhizal 

fungi of broad host specificity. Saprotrophic fungi are 

consistently enriched in C-13 relative to source C in litter 

and wood, suggesting that saprotrophic fungi primarily 

incorporate carbohydrate-derived C (C-13 enriched), with 

little incorporation of lignin-derived C (C-13 depleted). 

Radiocarbon indicates the age of fungally-assimilated C 

with a precision of about 1 year, and can therefore readily 

distinguish between mycorrhizal fungi (relying almost 

exclusively on current photosynthate) and saprotrophic 

fungi (relying on litter and wood of a range of ages). 

Continued improvement in our ability to interpret isotopic 

patterns in fungi will require culture studies to better 

understand C and N isotope fractionation during 

metabolism.


94 - Weathering, a new role of ectomycorrhizal fungi in 

ecosystems 

R. Landeweert 1* , E. Hoffland 1 , M. Smits 2 , L. van Schöll 1 , 

T.W. Kuyper 1 & N. van Breemen 2 

1 Wageningen University, Sub-dept. of Soil Quality., 

P.O.Box 8005, NL-6700 EC Wageningen., The 

Netherlands. - 2 Wageningen University, Laboratory of Soil 

Science and Geology., P.O.Box 37, NL-6700 AA 

Wageningen., The Netherlands. - E-mail: 

renske.landeweert@bb.benp.wag-ur.nl 

Ectomycorrhizal fungi play a major role in the nutrient 

acquisition of trees. Mycelial uptake and transport of 

dissolved nutrients as well as fungal access to organic N 

and P sources have received much attention. Recent 

research suggests a third role of ectomycorrhizal fungi: 

mobilisation of P and other essential plant nutrients directly 

from minerals through excretion of organic acids. The 

mobilised cations are subsequently translocated to the 

ectomycorrhizal trees and thus by-pass the soil exchange 

complex. The primary source of all essential plant nutrients 

except nitrogen is the weathering of minerals. Low 

molecular weight organic acids excreted by plant roots and 

microorganisms are considered to be the most important 

biological weathering agents in soils. Commonly released 

into their environment by different fungal species are 

oxalic and citric acid, both strong chelators of trivalent 

metals such as Al3+. As the concentration of Al3+ in the 

soil solution diminishes through chelation, weathering rates 

of Al silicates such as feldspars increase. The awareness 

that ectomycorrhizal fungi contribute to this process has 

increased since the discovery of hypha-shaped tunnels 

inside mineral grains. The contribution of ectomycorrhizas 

to mineral weathering may change traditional ideas on 

nutrient cycling and soil formation in forest systems. Latest 

results on measurements of fungal weathering activities 

will be presented and ecological implications will be 

discussed. 

95 - Decomposition of humic substances, lignin and 

organopollutants by soil-litter decomposing 

basidiomycetes 

M. Hofrichter 1* , K.T. Steffen 2 & A. Hatakka 2 

1 International Graduate School Zittau, Unit Environmental 

Biotechnology, Markt 23, D-02763 Zittau, Germany. - 

2 University of Helsinki, Dept. Applied Chemistry & 

Microbiology, Viikki Biocenter 1, PO box 56, FIN-00014 

Helsinki, Finland. - E-mail: Martic@gmx.net 

Humic substances (HS) are ubiquitous nonliving organic 

materials in all terrestrial and aquatic environments, and 

represent the major fraction of organic matter in soils and 

sediments. Consequently, microbial degradation of HS is 

essential for maintaining the carbon cycle. Despite this 

fact, only less is known about the particular 

microorganisms which decompose humic matter. Our main 

objective has been to examine the potential of litterdecomposing 

basidiomycetes colonizing forest soils to 

disintegrate HS and other recalcitrant compounds (lignin, 

organopollutants). Within the scope of several screening 

test, we have selected Nematoloma frowardii, Collybia 

dryophila, Stropharia coronilla and Agrocybe praecox as 

particularly active species degrading different HS, lignin 

and/or several organopollutants in liquid as well as in 

sterile and nonsterile solid-state cultures. These agaric 

fungi produce manganese peroxidase (MnP) as the 

predominant oxidative enzyme which is secreted into their 

microenvironment in response to high Mn 2+ titers. In 

combination with certain low-molecular mass effectors, 

isolated MnP was found to attack recalcitrant aromatic 

substances unspecifically via one-electron abstractions 

giving rise to instable radicals, which tend to disintegrate 

spontaneously into polar fragments and CO2. 

96 - Arbuscular mycorrhizal communites in rare and 

common Pulsatilla spp.: towards restoration of 

endangered plant species 

R. Sen 1* , M. Öpik 2 , M. Moora 2 , U. Kõljalg 2 & M. Zobel 2 

1 Department of Biosciences, Division of General 

Microbiology, P.O. Box 56, FIN-00014 University of 

Helsinki, Finland. - 2 Institute of Botany and Ecology, Tartu 

University, 40 Lai Str., Tartu-51005, Estonia. - E-mail: 

robin.sen@helsinki.fi 

In the context of plant conservation it can be hypothesized 

that the presence of compatible arbuscular mycorrhizal 

fungi (AMF) directly contribute to the growth and survival 

of endangered plants. Here, we investigated a congeneric 

rare vs. common species pair, Pulsatilla patens and P. 

pratensis, in Estonian soils. Seeds were sown in soil 

inocula from four localities and germination, seedling 

establishment and root colonizing AMF species 

communities were assessed. A lack of soil treatmentrelated 

differences in P. pratensis germination and seedling 

establishment rates strongly contrasted with the 

significantly higher establishment rates of the rare P. 

patens in soil inoculum from a Scots pine forest where both 

target species co-exist in the understorey. Distinct 

communities of AMF species, identified following rDNA 

(SSU) sequence and phylogenetic analyses, were detected 

in roots of established plants and test seedlings grown in 

soil inocula from an agricultural landscape and a Scots pine 

forest. In seedling roots of both species exposed to forest 

soil inoculum, SSU sequences exhibited high homology to 

woodland Glomus sequences, earlier detected on 

Hyacinthoides non-scripta in a deciduous UK woodland 

site. The data provides support for the involvement of 

specific AMF in early seedling establishment of Pulsatilla 

species. Greater host species-linked dependency towards 

particular AMF may explain the loss of the rare P. patens 

in mycobiont limited disturbed or managed landscapes. 



97 - Net transfer of water between plants connected by 

common mycorrhizal networks 

J.I. Querejeta 1* , L. Egerton-Warburton 2 & M.F. Allen 1 

1 University of California Riverside, CCB University 

Laboratory BLDG RM 203, Riverside CA 92521, U.S.A. - 

2 Chicago Botanic Gardens, 1000 Lake Cook Rd, Glencoe 

IL 60022, U.S.A. - E-mail: nachoq@ucrac1.ucr.edu 

Common mycorrhizal networks (CMNs) may play a major 

role in mediating water parasitism associated with 

hydraulic lift. Neighboring plants that share a CMN with 

the lifting individual may be able to derive the greatest 

benefit from this process. The ability of CMN to directly 

transfer significant amounts of water between plants was 

evaluated in the obligate mycorrhizal species coast live oak 

(Quercus agrifolia). Mycorrhizal (ecto- + arbuscular) oak 

seedlings were grown in microcosms comprising two 

adjacent compartments separated by air gaps and mesh 

barriers that could be crossed by hyphae but not by roots. 

Oak seedlings in one of the compartments had access to a 

taproot chamber with unlimited supply of water and 

conducted hydraulic lift. Following an 11 day drought 

period, during which watering was withdrawn from the 

upper two compartments, we detected a net transfer of 

water from the lifting oaks to neighboring plants through 

CMNs. Quercus, Salvia and Eriogonum seedlings in the 

adjacent compartment showed up to 20% higher leaf water 

content compared with their experimental counterparts 

with severed hyphal connections at the onset of the drought 

period. Isotopic data obtained after injecting deuteriumdepleted 

tracers into the taproot chamber also indicated that 

neighboring droughted seedlings received up to 19% of 

their water from the lifter oaks through CMNs. 

32 

Book of Abstracts

IMC7 Tuesday August 13th Lectures 

98 - Fungal biodiversity: What do we know? What can 

we predict? 

G.M. Mueller 1* , J.P. Schmit 2 & R. Courtecuisse 3 

1 Dept. of Botany, Field Museum of Natural History, 1400 

S. Lake Shore Dr. Chicago, IL 60605, U.S.A. - 2 Dept. of 

Plant Biology, U. Illinois, 265 Morrill Hall, Urbana, IL 

61801, U.S.A. - 3 Dept. de Botanique, Univ. de Lille, B.P. 

83, FR-59006 Lille Cedex, France. - E-mail: 

gmueller@fmnh.org 

This years IMC comes roughly 10 years after published 

reassessments that greatly increased the estimated numbers 

of fungal species. We feel it timely to review these 

estimates based on a decade of data gathering and thinking 

on the topic. Each talk in the symposium will present the 

current status of known diversity for a major group of fungi 

with a geographical focus. Contributions will cover macro-, 

micro-, lichenized, aquatic, soil-inhabiting, and arthropodassociated 

fungi, plus a summary that integrates each of 

these presentations. Each talk will address the following 

questions for its fungal group: How many species are 

known worldwide? How accurate is this number? How is 

this group geographically distributed? Which geographic 

areas are well versus poorly known? What are the species 

overlap between different geographic regions? And what 

are the estimated centers of diversity? Thus, each 

presentation will function both as a meta-analysis of 

current data and a gap assessment to indicate where future 

research efforts should be concentrated. In addition to 

providing a summary of the state of our knowledge on 

fungal diversity, a goal of this symposium is to help inspire 

future work on fungal diversity and biogeography so that 

significant progress toward our understanding of these 

issues will be made by the next IMC. 

99 - Global diversity and distribution of macrofungi 

G.M. Mueller 1* , J.P. Schmit 2 , R. Courtecuisse 3 , L. 

Ryvarden 4 , K.-H. Larsson 5 , J.M. Trappe 6 , R.E. Halling 7 , 

M. Rajchenberg 8 , T.W. May 9 , J. Cifuentes 10 , D.E. 

Desjardin 11 , K. Hjortstam 12 , Q.X. Wu 13 , P. Leacock 13 , D.J. 

Lodge 14 , T. Iturriaga 15 , R. Watling 16 , S.A. Redhead 17 & B. 

Buyck 18 

1 Dept. of Botany, Field Museum of Natural History, 1400 

S. Lake Shore Dr. Chicago, IL 60605, U.S.A. - 2 Dept. of 

Plant Biology, U. Illinois, 265 Morrill Hall, Urbana, IL 

61801, U.S.A. - 3 Dept. de Botanique, Univ. de Lille, B.P. 

83, FR-59006 Lille Cedex, France. - 4 Botany Div., Biology 

Inst., Univ. of Oslo, P. O. Box 1045, Blindern, NO-0316 

Oslo 3, Norway. - 5 University of Goteborg, P.O. Box 460 

SE 405 30 Goteborg, Sweden. - 6 Dept. of Forest Science, 

Oregon State Univ., Corvallis, OR 97331, U.S.A. - 7 Inst. of 

Systematic Botany, New York Botanical Garden, Soujtern 

Blvd at 200th St Bronx, NY 10458-5126, U.S.A. - 8 Centro 

Forestal CIEFAP, C.C. 14, 9200 Esquel, Chubut, 

Argentina. - 9 Royal Botanic Gardens Melbourne, Privat 

Bag 2000, South Yarra, Victoria, 3141, Australia. - 

10 Sección de Micología, Herbario FCME, Facultad de 

Ciencias, UNAM, Apto. postal 70-399, C.P. 04510, Ciudad 

Universitaria, Mexico. - 11 Dept. Biology, San Fransisco 

State Univ., San Fransisco, CA 94132, U.S.A. - 12 

Malaregatan 12, SE-441 35 Alingsås, Sweden. - 13 Dept. of 

Botany, Field Museum of Natural History, Chicago, IL 

60605, U.S.A. - 14 Cent. for Forest Mycology Res. USDA- 

Forest Service, FPL, P.O. Box 1377, Luquillo, PR 00773- 

1377, U.S.A. - 15 Depto. de Biologia de Organismos, Univ. 

Simon Bolivar, Apartado 89000, Sartenejas, Baruta, Edo. 

Miranda, Venezuela. - 

16 Caledonian Mycological 

Enterprises, Crelah, 26 Blinkbonny Ave, Edinburgh, EH4 

3HU, Scotland, U.K. - 17 Agriculture Canada, Eastern 

Cereals & Oilseeds Res. Centre, William Saunders Bldg., 

Central Experimental Farm, Ottawa, Ont. K1A 0C6, 

Canada. - 

18 Laboratoire de Cryptogamie, Museum 

National d'Histoire Naturelle, 12, rue Bufon, FR-750005, 

France. - E-mail: gmueller@fmnh.org 

We have compiled data on macrofungal diversity and 

distribution patterns for major geographical regions of the 

world. For this presentation, we have defined macrofungi 

to include ascomycetes and basidiomycetes with large, 

easily observed sporocarps that fruit epi- or hypogeously. 

Co-authors provided data on a particular taxonomic group 

or geographic area. We then employed a meta-data analysis 

to investigate species overlaps between areas, levels of 

endemism, centers of diversity, and estimated percent of 

species known for each geographic area for each taxonomic 

group and for the combined macrofungal data set. Thus, we 

provide both a meta-analysis of current data and a gap 

assessment to help identify research needs. 

100 - Diversity of saprobic microfungi 

K.D. Hyde 1* , P.W. Crous 2 , S. Lee 2 , S. Lumyong 3 , E.H.C. 

McKenzie 4 & B. Paulus 5 

1 Centre for Research in Fungal Diversity, Department of 

Ecology & Biodiversity, The University of Hong Kong, 

Pokfulam Road, Hong Kong, China. - 2 Department of Plant 

Pathology, University of Stellenbosch, Private Bag X1, 

Matie Land 7602, South Africa. - 3 Department of Biology, 

Chiang Mai University, Chiang Mai, Thailand. - 4 Landcare 

Research, Private Bag 92170, Auckland, New Zealand. - 

5 James Cooke University, Cairns, North Queensland, 

Australia. - E-mail: kdhyde@hkucc.hku.hk 

Recent studies in Australasia, Hong Kong, South Africa 

and Thailand have provided new data towards quantifying 

the magnitude of fungal diversity. High global fungal 

species numbers rely heavily on there being a relatively 

high ratio of unique fungi to host plant species. Data on 

host specificity for saprobic fungi is sparse, but a study of 

four New Zealand native plant genera; Agathis (1 species), 

Metrosideros (x taxa), Nothofagus (5 taxa) and 

Rhopalostylis (2 species) has found that 7 out of 10 

saprobic microfungi described on Agathis are unique to 

Agathis in New Zealand, 0 of 2 are unique to Metrosideros, 

16 of 20 are unique to Nothofagus, and 6 of 9 are unique to 

Rhopalostylis. Figures for fungi on Proteaceae and 

Restoniaceae in Cape Fynbos, S. Africa are also presented. 

Is this enough diversity to account for the estimated 

numbers of fungal species in the world? This paper will 



explore techniques available to assess microfungal 

numbers in a habitat or region and present results from 

some of the studies. It will concentrate on efforts to access 

whether saprobic microfungi are host or tissue specific and 

comment on reasons for these relationships. Databases can 

be used to establish the numbers of fungi on different plant 

hosts. As databases become more comprehensive it will be 

possible to examine the true extent of specificity amongst 

saprobic microfungi. 

101 - Biodiversity of soil-inhabiting fungi 

W. Gams 

Centraalbureau voor Schimmelcultures, P.O. Box 85163, 


Gams@CBS.KNAW.NL 

We count filamentous fungi living in soil layers but 

exclude those from the often more diverse litter. The 

Centraalbureau voor Schimmelcultures (CBS) preserves a 

great diversity of these fungi. The database contains 5768 

isolates from soil or roots, distributed over appr. 2430 spp., 

including unpublished names and a few double counts for 

anamorph and teleomorph names. 2430 - (131 zoosporic + 

89 macromycete basidiomycetes) = 2210 spp. of soil fungi 

(? 70% of those known). Basidiomycetes often account for 

the majority of fungal activity in a soil but are better 

assessed in mycorrhizal and wood/litter-decomposing 

inventories. Recently, the general increment in the numbers 

of soil fungi is only slightly accelerated, after CBS has 

broadened its accession policy and since the advent of 

molecular taxonomic tools. Particularly some poorly 

defined genera and species yield new taxa. Among 

thousands of isolates from any soil novelties will be found. 

Soil fungi spread easily and are regarded as cosmopolitans. 

Even in tropical forests many taxa are similar to those 

found in temperate latitudes. Molecular studies tell us that 

continental barriers determine a phylogeographical 

differentiation for slimy-spored species of Fusarium (now 

138 in CBS) and Trichoderma (now 49 in CBS). These are 

predicted to increase in number of known taxa by not more 

than 100%. In the more ubiquitous genera Penicillium and 

Aspergillus (now ca. 300 and 200 spp.) an increment of 

about 100% is anticipated. 

102 - Global diversity and distribution of fungi in 

aquatic habitats 

C.A. Shearer 1* , E. Descals 2 , J.W. Fell 3 , J. Kohlmeyer 4 , L. 

Marvanová 5 , D.E. Padgett 6 , D. Porter 7 , H. Raja 1 , J.P. 

Schmit 1 , H. Voglmayr 8 & B. Volkmann-Kohlmeyer 4 

1 Dept. of Plant Biology, University of Illinois, Rm. 265 

Morrill Hall, 505 S Goodwin Ave, Urbana, IL 61801, 

U.S.A. - 2 IMEDEA (CISC-Univ. Illes Velears), calle Mique, 

Marques, 21, 07190 Esporles, Mallorca, Spain. - 3 RMAS- 

BLR Dept., S209 Grosvenor Bldg. 4600 Rickenbacker 

CSWY, Miami, FL 33149, U.S.A. - 4 Institute of Marine 

Sciences, University of North Carolina, Morehead City, 

34 


NC 28557, U.S.A. - 5 Czech Collection of Microorgansms, 

Masaryk Unive\rsity Brno, Tvrdého 14, 602 00 Brno, 

Czech Republic. - 6 Biology Dept., Univ. of North Carolina- 

Wilmington, Wilmington, NC 28403, U.S.A. - 7 Dept. of 

Botany, Univ. of Georgia, Athens, GA 30602, U.S.A. - 

8 University of Vienna, Institute of Botany, Rennweg 14, A- 

1030 Wien, Austria. - E-mail: carolshe@life.uiuc.edu 

We have compiled data on the diversity and distribution 

patterns of fungi and fungal-like straminipiles that occur in 

aquatic habitats ranging from freshwater to marine. Major 

groups included in this assessment are: Chytridiomycetes, 

Ascomycetes, Basidiomycetes, Hyphochytriomycetes and 

Oomycetes. Co-authors provided data on a particular 

taxonomic and/or ecological group. We then analyzed data 

to determine the total number of species, the number of 

species per habitat and/or substrate type, species overlap 

among habitats and substrates, and latitudinal and 

continental distribution patterns in diversity. We also 

estimated levels of endemism and centers of diversity for 

the various taxonomic/ecological groups. 

103 - Biodiversity of lichens 

T. Feuerer 

Institut für Allgemeine Botanik, Ohnhorststr. 18 22609 

Hamburg, Germany. - E-mail: feuerer@botanik.unihamburg.de 

Estimates for lichen diversity on the planet range between 

13500 to 19000 species. A more precise estimate is hardly 

possible, for several reasons: First, the species concept of 

several lichen groups is still unsettled. Second, there are 

numerous unresolved taxonomical problems, especially in 

crustose lichens, and third, the lichen flora in several areas 

of the world - especially in the tropics - is still underexplored. 

However, there is already an enormous amount 

of data from all over the world, which needs to be 

compiled and validated to reach a better estimate of species 

diversity. Lichen information is already accessible on-line 

from several, heterogeneous sources (see e.g. e.g. ITALIC, 

http://dbiodbs.univ.trieste.it/, 

http://biobase.kfunigraz.ac.at/lichen/medlich2.html). To 

interconnect the different sources and to provide 

corresponding software tools is part of the ongoing project 

'Global biodiversity of lichens' by T. Feuerer and G. 

Rambold (http://www.checklist.de/). Until the end of 2003, 

data from all 193 states and 300 adjacent geographical 

units will be available (For an experimental version see 

http://141.84.65.132/ChecklistsDe/Lichens/index.html). In 

support of this project, the International Association for 

Lichenology has established a Commitee, which 

coordinates and gathers lichen diversity information for the 

global checklist project.


104 - Biodiversity of arthropod-associated fungi: What 

do we know? What can we predict? 

A. Weir 1* , M. Hughes 1 , M.M. White 2 , M.J. Cafaro 2 , S.-O. 

Suh 3 & M. Blackwell 3 

1 Faculty of Environmental & Forest Biology, SUNY 

College of Environmental Science & Forestry, 350 Illick 

Hall, 1 Forestry Drive, Syracuse NY 13210, U.S.A. - 

2 Department of Ecology and Evolutionary Biology, 

University of Kansas, Room 8002 Haworth Hall, 

Lawrence, KS 66045, U.S.A. - 3 Department of Biological 

Sciences, Louisiana State University, Baton Rouge, LA 

70803, U.S.A. - E-mail: aweir@syr.edu 

One of the greatest uncertainties in attempting to estimate 

global species richness of fungi is being able to draw 

inferences from the known number of taxa associated with 

hyperdiverse groups such as arthropods. Knowledge of the 

diversity and biogeography of these interactions is 

improving, but for many groups remains an imprecise 

foundation for extrapolation. Arthropods and fungi are 

closely associated in nearly all types of terrestrial, 

freshwater, and marine environments. Here we discuss only 

those fungi that use arthropods as a resource. Associations 

along the mutualist-pathogen continuum vary, and these as 

well as life history traits are discussed for each major group 

of fungi in relation to their known and potential diversity. 

We review the current state of knowledge regarding 

diversity, host relations, and biogeography of 

Laboulbeniales, Trichomycetes, Entomophthorales, 

Septobasidium, Cordyceps, and endosymbiotic yeasts. For 

the most diverse and one of the best biogeographicallystudied 

groups, Laboulbeniales, patterns of diversity and 

host utilization at different geographical scales are 

analyzed. On the bases of these datasets, species discovery 

patterns and host associates, and the latest estimates of 

global insect species richness, we extrapolate as to the 

potential magnitude of the biodiversity of insect associated 

fungi and discuss ways in which the estimates may be 

refined. We also address the major lacunae that require 

coordinated effort from systematists. 

105 - Fungal diversity: Estimates, predictions and 

future challenges 

J.P. Schmit 1* , G.M. Mueller 2 & R. Courtecuisse 3 

1 Dept. of Plant Biology, Unvirsity of Illinois, 265 Morrill 

Hall, 605 S. Goodwin, Urbana, IL 61801, U.S.A. - 2 Dept. of 

Botany, Field Museum of Natural History, 1400 S 

Lakeshore Dr. Chicago, IL 60605, U.S.A. - 3 Dept. de 

Botanique, Univ. de Lille, B.P. 83, FR-59006, Lille Cedex, 

France. - E-mail: jpschmit@life.uiuc.edu 

This presentation summarizes the data from the other 

presentations in this symposium. Based on that 

information, we present an estimate of the total number of 

fungal species currently known to science. We also make a 

conservative estimate of the number of fungi yet to be 

discovered. This is done by using data from geographic 

regions which are relatively well explored for fungi to 

estimate the number of new taxa yet to be found in poorly 

known areas. We also identify where there are gaps in our 

knowledge, both geographically and taxonomically. 

106 - Comparative methods for managing phylogenetic 

uncertainty 

M. Pagel 

Reading University, AMS, Whiteknights, Reading RG6 6AJ, 

England, U.K. - E-mail: m.pagel@rdg.ac.uk 

Comparative analyses across species must be based upon 

good phylogenetic information to distinguish similarity that 

arises from common ancestry from that which arises from 

parallel or convergent evolution. This means that 

phylogenies are necessary for studying ancestral states, for 

calculating rates of evolution, and for investigating 

correlations among two or more traits. I will briefly 

describe these sorts of analyses, giving examples, then go 

on to describe a difficulty that arises in comparative 

studies. The difficulty is that phylogenies are seldom 

known with certainty and different phylogenies can give 

different answers to the comparative question. A relatively 

new technique in comparative studies make it possible to 

take phylogenetic uncertainty into account. Bayesian 

methods for inferring phylogenies, based upon Markov 

Chain Monte Carlo methods, allow the investigator to draw 

a random sample of phylogenetic trees, from the universe 

of possible trees. The comparative hypothesis can then be 

tested in each tree, in effect, removing phylogenetic 

uncertainty from the hypothesis. These methods are 

becoming increasingly easy and practical to implement on 

desktop computers. I give examples from several areas of 

research. 

107 - Major fungal lineages are derived from lichen 

symbiotic ancestors 

F. Lutzoni 1* , M. Pagel 2 & V. Reeb 1 

1 Duke University, Department of Biology, Box 90338, 

Durham, North Carolina, U.S.A. - 2 University of Reading, 

School of Animal and Microbial Sciences, Whiteknights, 

Reading, U.K. - E-mail: flutzoni@duke.edu 

About one-fifth of all known extant fungal species form 

lichen symbioses. Lichens are widely believed to have 

arisen independently on multiple occasions, accounting for 

the high diversity and mixed occurrence of lichenized and 

non-lichenized (42% and 58%, respectively) fungal species 

within the Ascomycota. Depending on the taxonomic 

classification chosen, 15-18 orders of the Ascomycota 

include lichen-forming taxa, and 8-11 of these orders 

(representing about 60% of the Ascomycota species) 



contain both lichenized and non-lichenized species. Here 

we report a phylogenetic-comparative analysis of RPB2, 

and the small and large subunit of the nuclear rRNA genes 

from about 60 species representing about 75% of the 

species diversity of the Ascomycota, a phylum that 

includes > 98% of known lichenized fungal species. Using 

a Bayesian phylogenetic tree sampling methodology, 

combined with a statistical model of trait evolution, we 

take into account uncertainty about the phylogenetic tree 

and ancestral state reconstructions. Our results, based upon 

a sample of 19,900 phylogenetic trees, show that lichens 

evolved earlier than believed, and that gains of 

lichenization have been infrequent during Ascomycota 

evolution, but have been followed by multiple independent 

losses of the lichen symbiosis. As a consequence, major 

Ascomycota lineages (Ascosphaerales, Eurotiales, 

Onygenales and Chaetothyriales) of exclusively nonlichen-forming 

species are derived from lichen-forming 

ancestors. 

108 - Evolution of interkingdom host-jumping of 

Cordyceps and the Clavicipitaceae 

J.W. Spatafora * & G.-H. Sung 

Oregon State University, Dept. of Botany and Plant 

Pathology, 2082 Cordley Hall, Corvallis, OR 97331-2902, 

U.S.A. - E-mail: spatafoj@bcc.orst.edu 

The genus Cordyceps includes over 300 described species 

of fungi that are pathogens of arthropods and fungi. It is a 

member of the Clavicipitaceae (Ascomycota, Hypocreales) 

which also includes symbionts of plants, mainly of the 

Poaceae. Current subfamilial classification of the 

Clavicipitaceae comprises the Clavicipitoideae, which 

includes all of the grass symbionts, and the 

Cordycipitoideae, which includes all of the pathogens of 

arthropods and fungi. This represents an evolutionary 

hypothesis that implicitly assumes inter-Kingdom host 

jumps are rare and irreversible. To test this hypothesis we 

initiated a molecular phylogenetic study of the 

Clavicipitaceae with a particular emphasis on Cordyceps. 

We have assembled a four-gene region dataset that 

includes data from the nuclear SSU and LSU rDNA, betatubulin, 

and elongation factor 1-alpha for 75 taxa. 

Phylogenetic analyses reject the monophyly of Cordyceps 

and the Cordycipitoideae and do not support host jumps as 

being rare and irreversible. Three clades of clavicipitaceous 

fungi, which all contained pathogens of animals and fungi, 

were resolved in these analyses. Pathogen of animals is 

resolved as the primitive symbiosis for the Clavicipitaceae 

with a single jump to the Poaceae that was most likely 

followed by a reversal to animals. Multiple jumps have 

occurred from animals to fungi with at least one reversal to 

animals. Character coding of host affiliation and testing 

character state reconstructions will be discussed. 

36 


109 - Molecular phylogenetics and breeding systems in 

the ascomycete fungi 

M.L. Berbee 

Department of Botany, University of British Columbia, 

Vancouver BC V6T 1Z4, Canada. - E-mail: 

berbee@interchange.ubc.ca 

By a combination of phylogenetic approaches, we are 

studying the evolution of breeding systems among fungi in 

the Ascomycota. Within the family Pleosporaceae, 

combined data from the glyceraldehyde-3-phosphate 

dehydrogenase gene and ribosomal ITS regions provide 

good resolution. Our phylogenetic analysis shows that the 

'asexual' species and the selfing species are nested among 

outcrossing sexual species, indicating that outcrossing 

sexuality was the ancestral condition for the family. In 

sexual filamentous ascomycetes, opposite mating type 

information at the MAT1 locus regulates mating and the 

opposite mating type genes each have a clonal, nonrecombining 

phylogenetic history. Many species in the 

Pleosporaceae have no known sexual states, but where 

tested, these supposedly asexual fungi have intact, 

functional mating type genes. We used PCR to amplify and 

sequence fragments of the opposite mating type genes from 

supposedly asexual species in the genus Alternaria, in the 

Pleosporaceae. Each haploid fungal isolate had just one 

mating type, but both mating types were present in each 

species. We sequenced the ribosomal ITS regions for 

isolates of opposite mating types, for three asexual species 

and four known related sexual species. Analysis of the 

probability of substitution patterns indicated that, if 

sexuality had been lost, it was lost after the most recent ITS 

substitutions had evolved in each species. 

110 - Trends in morphological evolution in 

homobasidiomycetes 

D.S. Hibbett * & M. Binder 

Biology Department, Clark University, Worcester MA 

01610, U.S.A. - E-mail: dhibbett@black.clarku.edu 

Homobasidiomycetes display an incredible diversity of 

fruiting body forms, ranging from simple corticioid forms, 

which lie flat on their substrates, to elaborate, 

developmentally integrated forms, such as stinkhorns. 

More than half of all described homobasidiomycetes have 

pileate-stipitate fruiting bodies, with gills or other 

configurations of the hymenophore. A functional (if not 

phylogenetic) distinction can be made between 

gasteromycetes, which lack ballistospory and produce 

spores internally, and hymenomycetes, which retain 

ballistospory and produce spores externally. Much of the 

research in higher-level homobasidiomycete systematics in 

the last century has focused on tracing patterns of evolution 

in fruiting body forms, and has revealed extensive 

convergence. Our recent analyses concern trends in the 

evolution of fruiting body morphology. Examples of the


questions that concern us are: 1) Are simple resupinate 

forms evolutionarily more labile than complex forms? 2) Is 

there a general trend toward the evolution of pileatestipitate 

forms? 3) Can we reject the hypothesis that the 

loss of ballistospory is irreversible? To address these 

questions we are employing binary and multistate 

maximum likelihood methods to develop and test models 

of fruiting body evolution, using a large (ca. 500 species) 

phylogenetic tree derived from rDNA sequences. 

111 - Concepts and approaches during 50 years of 

biological control of fungal plant pathogens 

J.M. Whipps 

Horticulture Research International, Wellesbourne, 

Warwick, CV35 9EF, U.K. - E-mail: 

john.whipps@hri.ac.uk 

There are now more than 80 products near to or on the 

market that have biological control activity against plant 

pathogens in soil, root, aerial and post-harvest 

environments. Most of these been developed relatively 

recently in response to environmental concerns which have 

forced reductions of fungicide and fumigant use and 

availability. Bacterial products are dominated by 

Pseudomonas, Burholderia and Bacillus species and fungal 

products by Gliocladium and Trichoderma species. With 

the exception of products based on Agrobacterium 

radiobacter for control of crown gall, Phlebiopsis gigantea 

for control of stem and root rot of pine and some 

Trichoderma-based products, few have been on the market 

for 10-20 years or more. Indeed, several have come and 

gone. Key features for achieving a successful biocontrol 

product must be cost-effective and reproducible disease 

control. In the last 15 years, the importance of 

understanding the ecological interactions between a 

biocontrol agent, its target pathogen and host plant before 

this can be achieved has gradually become appreciated. 

Against this background, several successful biocontrol 

agents have been selected, screened for activity, 

characterised and then undergone registration and 

marketing. Reassessment of the concepts and processes 

involved in natural disease suppression has also provided 

novel biological approaches to disease control. 

112 - Biological control of fungal plant pathogensrecent 

advances and future perspectives 

C. Alabouvette 

INRA-CMSE UMR BBCE-IPM, BP 86510 21065 DIJON 

Cedex, France. - E-mail: ala@dijon.inra.fr 

Two different approaches have been described to control 

plant pathogens: either introduce a selected biological 

control agent or enhance naturally occurring biological 

control. The first approach has been the most commonly 

followed during the 30 last years. Most emphasis was 

devoted to the study of the modes of actions of selected 

antagonists and less to the study of the conditions required 

for successful application. More recently, the development 

of molecular tools led to the creation of transformed 

biological agents possessing several modes of action, or 

having other beneficial traits such as a greater ability to 

survive or colonize the target. These 'improved strains' are 

supposed to possess an enhanced biological control 

activity, but demonstrations in nature of these advantages 

are still missing. Biological control of fungal pathogens 

remains very limited due to the narrow specificity of the 

biological control agents and the inconsistency of their 

efficacy. Today there is a renewed interest for studying the 

effects of organic matter and compost amendment on the 

soil inoculum potential, with the objective of enhancing the 

natural potential of suppressiveness that exist in any soil. 

Whatever the method used, biological control should be 

part of an integrated pest management strategy. Research in 

this field requires an holistic approach taking into account 

not only the microbial interactions and the plant defense 

reactions but also the whole agronomical system. 

113 - Premier bacteria and fungal plant disease 

suppression 

D.M. Weller 

USDA-ARS, Washington State University, 367 Johnson 

Hall, Pullman, Washington 99164, U.S.A. - E-mail: 

wellerd@mail.wsu.edu 

Take-all, caused by Gaeumannomyces graminis var. tritici, 

is an important root disease of wheat worldwide. Take-all 

decline (TAD) is the spontaneous decrease in the incidence 

and severity of take-all that occurs with monoculture of 

wheat or other susceptible host crops after one or more 

severe outbreaks of the disease. In Washington State, USA, 

TAD develops in wheat fields because of the build up of 

strains of Pseudomonas fluorescens, which produce the 

antifungal metabolite 2,4-diacetylphloroglucinol (DAPG). 

DAPG producers are abundant in other suppressive soils. 

Whole-cell repetitive sequence-based (rep)-PCR with the 

BOXA1R primer distinguished 17 different genotypes (A 

through N) within worldwide collections of DAPG 

producers. Genotype D, which is primarily responsible for 

TAD in Washington soils, aggressively colonizes wheat 

and barley and suppresses take-all when applied at very 

low doses (as few as 10 2 per seed or gram of soil). Field 

studies have verified that introduced populations of 

genotype D strains are sustained throughout the growing 

season, survive between crops, and reestablish on the roots 

in successive years above the threshold required for 

suppression of take-all. 



114 - Risk assessment of fungal biocontrol agents - How 

can RAFBCA help? 

T.M. Butt 1* , H. Strasser 2 & G. Tomimatsu 3 

1 1. School of Biological Sciences, University of Wales 

Swansea, Singleton Park, SA2 8PP, U.K. - 2 2. Institute of 

Microbiology, Leopold Franzens University Innsbruck, 

Technikerstrasse 25, A-6020 Innsbruck, Austria. - 3 3. U.S. 

Environmental Protection Agency, Office of Pesticide 

Programs/Biopesticides and Pollution Prevention, 

Arlington, VA 22202, U.S.A. - E-mail: 

t.butt@swansea.ac.uk 

Fungal biocontrol agents (BCAs) secrete a wide range of 

metabolites. Many have been shown to play an important 

role in pathogenesis-antagonism and the successful control 

of the target organism. Current registration procedures for 

products that purport biocontrol of plant pests and diseases 

require risk characterization and evaluation of known, and 

potential exposures to human health and nontarget 

organisms in the environment. Evaluation of all the 

metabolites produced by BCAs would be expensive and 

time-consuming. One of the objectives of the EU-funded 

RAFBCA project is to develop the methods and tools to 

assess the risks of fungal BCA metabolites. This paper will 

describe: (1) some of the methods and tools being 

developed for high throughput analysis such as biosensors, 

and microtitre plate assays and (2) studies that show that 

the quantities of secondary metabolites produced by fungi 

in-vivo are usually much less than those secreted in 

nutrient rich liquid media. Additionally, information on 

intra-species and inter-species variability in the production 

of selected metabolites will be provided, as well as data on 

the spatial-temporal distribution of fungal metabolites. 

Furthermore, strategies will be presented which could 

standardise the risk assessment of fungal BCAs. 

115 - Molecular approaches to studying fungal 

interactions in relation to biocontrol 

M. Lorito 1* , M. Ruocco 1 , D. Piacenti 1 , R. Ciliento 1 , Z.X. 

Lu 2 , F. Scala 1 , A. Zoina 1 , J.K. Jansson 2 & S. Woo 1 

1 Department of Plant Pathology, University of Naples, Via 

Università, 100, 80050 Portici, Italy. - 2 Department of 

Plant Pathology, Plant Pathology and Biocontrol Unit, 

Swedish University of Agricultural Sciences, Uppsala, 

Sweden. - E-mail: lorito@unina.it 

Trichoderma-based biofungicides are a reality in 

commercial agriculture, although they represent only a 

niche market in comparison to that of chemicals. However, 

more than a dozen formulations are available today as 

registered products in the USA and Europe, and the 

application of these biocontrol agents, both as biopesticides 

or biofertilizers, is steadily increasing also in organic 

farming. The modes of action of these beneficial fungi are 

many and very complex. Therefore several research 

strategies, also at a molecular and gene level, have been 

38 

applied to identify the main genes and compounds involved 

in this complex, three-way interaction among the 

antagonist, plant and pathogen. We have used different 

reporter systems and genetic manipulation to identify 

molecules that are involved in the cross-talk between 

Trichoderma and its host, and have found opportunities for 

developing new disease control methods. In addition, we 

have been able to monitor the interaction in vivo and in situ 

between Trichoderma and its host and correlate the 

occurrence of mycoparasitism to the biocontrol effect. 

116 - Weed biocontrol by fungal plant pathogens - 

research approaches and practical application 

M. Vurro 

Institute of Sciences of Food Productions, Viale Einaudi, 

51 - 70125, Bari, Italy. - E-mail: ma.vurro@area.ba.cnr.it 

Weeds are among the major pest constraints for many 

human activities, and their management represents the 

major cost of agriculture. Pathogens that infect weeds have 

been considered to partially replace traditional control 

methods, using two main approaches, the classic and the 

inundative method, but their success has been quite limited. 

The best results are mainly against alien weeds of pasture 

or forest, but also some mycoherbicides are available as 

commercial products. In despite of the partial success, in 

the last years some argumentations have further risen the 

interest in weed biocontrol, e.g.: increased consumption of 

organic products, ban of dangerous herbicides, lack of 

registered herbicides for niche crops, forbidden use of 

chemicals in anthropical environments (parks, 

archeological sites), resistance. To not disappoint these 

expectations, more efficacious agents are needed. Different 

strategies could be used to enhance effectiveness of fungal 

pathogens, including formulations, distribution systems 

(precision agriculture), selection of better characters and 

genetic enhancement, or integrated approaches, as mixing 

fungal pathogens, combining bacteria or bioactive 

metabolites with fungal pathogens, or managing traditional 

and biological methods. The presentation will focus mainly 

on the different strategies of weed biocontrol using fungal 

plant pathogens and will discuss the possibilities to 

improve the herbicidal properties of weed pathogens. 

117 - Xerophilic fungi, a physiological wonder 

J.I. Pitt 


Food Science Australia, P.O. Box 52, North Ryde, NSW 

1670, Australia. - E-mail: John.Pitt@csiro.au 

Xerophilic fungi are defined as those that can grow below a 

water activity of 0.85, which means the ability to grow in 

60% glucose, 45% glycerol or 20% NaCl - all (w/w). Few 

organisms on earth, other than halophilic bacteria, are able 

to grow under these conditions. To grow at reduced water 

activities, microorganisms must balance the external


environment by an equivalent water activity inside, as 

growth requires turgor. Bacteria concentrate K+ ions and 

certain amino acids to achieve this, while fungi have 

evolved the ability to use polyols - glucose, mannitol and in 

the limit glycerol - as the internal solute. At its limit, 

Xeromyces bisporus, the most xerophilic known organism, 

is able to grow at a water activity of 0.65, equivalent to a 

solution containing 65% (w/w) glycerol. As X. bisporus is 

known to accumulate glycerol to balance the outside 

environment, that means that life processes, including all 

enzyme functions, must be carried out in a thick glycerol 

syrup. X. bisporus and the yeast Zygosaccharomyces rouxii 

are capable of sexual reproduction at 0.7 water activity, i.e. 

in 60% (w/w) glycerol. A few of these fungi are able to 

grow in very concentrated NaCl solutions, but use the same 

internal solute system, so are correctly classified as 

halophilic xerophiles. These remarkable fungi, a number of 

genera and species, are all Ascomycetes, and may only 

have evolved once. Some further details of these 

fascinating fungi will be provided in this paper. 

118 - Molecular ecology of anaerobic fungi (Anaerobic 

fungi-do we know them all?) 

G.W. Griffith 1* , E. Ozkose 2 , D.R. Davies 3 & M.K. 

Theodorou 3 



3DA, Wales, U.K. - 2 KSU, Agriculture Faculty, 46100 

Kahramanmaras, Turkey. - 3 Institute of Grassland and 

Environmental Research, Plas Gogerddan, Aberystwyth, 

Ceredigion SY23 3EB, Wales, U.K. - E-mail: 

gwg@aber.ac.uk 

Anaerobic fungi are important components of the rumen 

ecosystem but their physiology and enzymology have 

attracted more interest than their ecology. As such, 

information relating to the mechanisms and dynamics of 

spread between hosts and niche differentiation between 

species remain unanswered. It is not clear how many 

species may coexist in a single host animal, nor whether 

the various species differ in their patterns of substrate 

colonisation. We have conducted detailed analysis of 

fungal populations from rumen digesta and faeces of 

silage-fed cows, and have investigated the effect of 

environmental conditions on survival of these fungi in 

faeces. Using the MPN technique with various substrates as 

an enrichment source, quite different fungal taxa were 

isolated. For instance, Caecomyces spp. were more 

abundant on cellobiose. In the course of this work we also 

isolated and characterised a new genus of anaerobic fungus 

Cyllamyces (type species C. aberensis). Difficulties in 

reliable morphological identification led us to use PCR- 

RFLP of the ITS region of the rRNA locus as a tool for 

identifying fungi isolated from MPN tubes. Sequence 

analysis of these regions was used to validate restriction 

enzyme patterns. Ambiguous RFLP patterns were found to 

be attributable to between-repeat polymorphisms within the 

ITS region of single isolates. ITS2 sequences from a single 

isolate diverged by 5% or more and there was more 

variability between repeats than between quite unrelated 

isolates. 

119 - Psychrophilic fungi - do they really exist? 

J.C. Frisvad 

Mycology Group, BioCentrum-DTU, Building 221, 


Denmark. - E-mail: jcf@biocentrum.dtu.dk 

Many food-borne Penicillia and some soil-borne species 

grow well at low temperatures (0-5 °C) but most of these 

species also grow quite well at 30 °C and could thus be 

regarded as psychrotrophic. Isolates from Svalbard and 

Antarctis have also been known species that were 

psychrotrophic and well known. The only new species of 

Penicillium described from Antarctis, P. antarcticum, is 

psychrotrophic to mesophilic. Some psychrophilic lichens 

and anamorphic ascomycetes like Humicola marvinii with 

optimum temperatures at 15 °C have been reported. A 

rather large number of psychrotrophic Penicillia have been 

isolated from Alpine or polar regions and some of them can 

be regarded as real psychrofiles. These fungi hardly grows 

(or do not grow at all) at 25 °C, and have an optimum 

temperature for growth near 15 °C. In few samples from 

Zachenberg in Greenland all Penicillia appeared to be new 

species, except one known taxon P. nordicum, known from 

refrigerated meat in Europe. Taxonomically the new 

species belonged to subgenus Furcatum and have some 

resemblance to common soil-borne species in series 

Canescentia. All species produced a surprisingly large 

number of bioactive secondary metabolites, including 

different combinations of kojic acid, penicillic acid, 

griseofulvin, chaetoglobosins and tryptoquivalins. Nearly 

all species produced some closely related cyclic peptides, 

and we hypothesise that these peptides are involved in 

adaptation to cold temperatures. 

120 - Halophilic fungi - are they widespread in the 

fungal kingdom? 

P. Zalar 1 , G.S. de Hoog 2 , A. Plemenitas 3 & N. Gunde- 

Cimerman 1* 

1 University of Ljubljana, Biotechnical Faculty, Biology 

Dept., Veèna pot 111, SI-1000 Ljubljana, Slovenia. - 

2 Centralbureau voor Schimmelcultures, P.O.Box 85167, 

3508 AD Utrecht, The Netherlands. - 3 University of 

Ljubljana, Medical Faculty, Institute of Biochemistry, 

Vrazov trg 2, SI-1000 Ljubljana, Slovenia. - E-mail: 

nina.gunde-cimerman@uni-lj.si 

Recently the novel observation was made that fungi, 

representing the only kingdom not so far known to sustain 

extremely saline natural conditions, populate manmade 

eutrophic and oligotrophic salterns, saturated or nearly 

saturated with NaCl. The majority of isolates from all 

sampled eutrophic salterns were black meristematic yeast 

like fungi: Hortaea werneckii, Phaeotheca triangularis, 

Aureobasidium pullulans and Trimmatostroma salinum. 

They were lacking in the oligotrophic ones, where 

saprophytic members of the genus Cladosporium prevail. 



These species are all closely related, belonging to the 

single order Dothideales. A large number of g. 

Cladosporium strains, along with reference strains of 

frequently isolated species were sequenced and their 

morphology and physiology are currently studied. The 

resulting clusters contain ubiquitous isolates originating 

from different extreme environments and at considerable 

phylogenetic distance away a separate branch, consisting of 

saltern isolates. From both types of hypersaline 

environments, xerophilic nonmelanized filamentous fungi, 

known primarily as food contaminants were also frequently 

isolated: Wallemia sebi, Penicillium and Aspergillus with 

teleomorphic stage Eurotium. Further analyses of these 

isolates show in some cases distinct differences in 

xerophily, contrary to the general belief, that with fungi it 

is determined primarily by the water activities of the 

culture media and not by the chemical nature of the solute. 

121 - Resolving differences between thermophilic and 

mesophilic Paecilomyces 

J.J. Luangsa-ard 1* , L. Manoch 2 , N.L. Hywel-Jones 1 & R.A. 

Samson 3 

1 BIOTEC, 73/1 Rama VI Rd, Bangkok 10400, Thailand. - 

2 Kasetsart University, Bangkhen, Bangkok 10900, 

Thailand. - 3 CBS, Uppsalalaan 8, 3508 AD Utrecht, The 

Netherlands. - E-mail: jajen@biotec.or.th 

Paecilomyces Bainier was erected for the thermotolerant 

Paecilomyces varioti. Brown & Smith (1957) broadened 

the concept of the genus to include mesophilic species off 

insects. Samson (1974) provided a modern morphologybased 

treatment of the genus. With few, plastic 

morphological features available for study confident 

identification of many Paecilomyces is problematic. The 

availability of molecular phylogenetics provides the 

opportunity to combine morphology and sequences to more 

fully resolve differences at the genus/species level. Isolates 

from the Samson Monograph were sequenced (18S, ITS 

and â-tubulin gene) to compare mesophilic and 

thermophilic Paecilomyces. Thermophilic section 

Paecilomyces form a separate clade to the mesophilic 

section Isarioidea. This work accepts the placement of 

section Paecilomyces in the Eurotiales and the placement 

of section Isarioidea in the Hypocreales. Paecilomyces 

varioti forms four clades and is apparently a cryptic 

species. Two of these clades have a Byssochlamys 

teleomorph. Evidence suggests that the thermophilic 

Talaromyces/Penicillium may be derived from a 

thermophilic Byssochlamys/Paecilomyces ancestor. 

122 - Enzyme discovery from extremophilic fungi 

L. Lange 

Novozymes, 1AMs.04, Smørmosevej 25, 2880 Bagsværd, 

Denmark. - E-mail: lla@novozymes.com 

40 


Discovery of enzymes from fungi living in extreme 

environments has primarily focused on thermophiles. 

Among the thermophilic fungi very rich enzyme producers 

have been found and also industrially exploited, e.g. from 

Humicola and Scytalidium. However, recent discovery 

efforts in our group have shown that much more is still to 

be found! The presentation will include data on new 

enzymes found from thermophiles (e.g. from Trichophaea 

saccata) but also new data of enzyme discoveries from 

other extreme environments as the gut flora of the termite 

larvae and the Iberian snail, from psychrophilic fungi, from 

xerophiles (as Talaromyces) and from alakali tolerant 

Stilbella. The presentation will further cover suggestions to 

new molecular methods of discovery from fungi from 

extreme environments, building on the rapidly rising 

amount of fungal sequence information available. 

123 - Lichen 'oases' in the Namib Desert 

A.F. Hunt 1 , P.D. Crittenden 1* , C.M. Scrimgeour 2 & M.A. 

Sutton 3 

1 University of Nottingham, School of Life & Environmental 

Sciences, University of Nottingham, Nottingham NG7 2RD, 

U.K. - 2 Scottish Crops Research Institute, Invergowrie, 

Dundee, DD2 5DA, U.K. - 3 Centre for Ecology and 

Hydrology, Bush Estate, Penicuik, Midlothian, EH26 0QB, 

U.K. - E-mail: pdc@nottingham.ac.uk 

Lichenized filamentous fungi dominate large areas of the 

coastal Namib Desert, the most arid region in Southern 

Africa. The existence of these lichen 'fields' is due to 

coastal fog depositing sufficient water for lichen growth. 

The most well-developed lichen communities are 

dominated by the fruticose lichen Teloschistes capensis 

which in this region is at the northernmost part of its range. 

While fog events occur along much of the Namibian coast, 

Teloschistes fields are very localized. The most extensive 

occurrence of T. capensis is inland from Cape Cross, the 

location of one the world's largest seal (Cape fur seal) 

colonies. We show that NH3 emitted from the seal colony 

can be detected over the desert from 32 µg m -3 at 250m 

from the colony to 0.2 and 0.02 µg m -3 at 3.6 and 26 km, 

respectively. The NH 3 is enriched in 15 N, and δ 15 N in T. 

capensis declines from +3.6‰ at 2.1 km from the colony to 

-5.5 and -7.2‰ at 11 and 26 km, respectively. Uptake of 

NH4 + from simulated fogwater can be demonstrated under 

laboratory conditions and this is greatly enhanced by the 

addition of PO 4 3- . Chemical analysis of fogwater revealed 

that the seal colony is also a source of PO 4 3- aerosol in 

addition to NH3. Anecdotal evidence suggests that many 

members of the Teloschistales have an ecological 

preference for eutrophicated habitats and we hypothesise 

that NH 3 and/or PO 4 3- output from the seal colony has 

promoted the development of Teloschistes fields in this 

area.


124 - Fungi and lichen: microbial ecology in Antarctic 

rock desert 

C. Ascaso 1* , A. de los Rios 1 & J. Wierzchos 2 

1 Centro de Ciencias Medioambientales, CSIC, Madrid, 

Spain. - 2 Servei de Microscòpia Electrònica, Universitat de 

Lleida, Spain. 

Fungi and prolichens account for a substantial proportion 

of the endolithic biomass of rock. To explore the 

development and survival of epilithic and endolithic fungi, 

visualisation technologies have to be stretched to their full 

potential. These techniques have enabled the observation of 

biologically transformed minerals in rocks from Antarctica, 

including diagenetic iron hydroxide nanocrystals and clays 

around chasmoendolithic fungi in sandstone, and calcium 

oxalate and silica deposits close to fungal cells in granite. 

As for any ecosystem, the study of the rock microhabitat, 

requires previous knowledge of the components and the 

processes that take place within it. The structure and 

function of each component of the lithic microecosystem 

needs to be established by quantifying and identifying the 

fungi present in each lithobiontic niche and defining the 

mineralogical features of these hidden microhabitats. Once 

we have selected the techniques to observe these 

components in situ and locate the presence of water, the 

questions that need to be addressed are: how are the fungi 

organised in the fissures and cavities, which fungi are 

present and how many are there, what water relationships 

are there and what effects do fungi have on the substrate's 

minerals? This last question is crucial, since mechanical 

and chemical changes in minerals and mineralisation of 

fungal cells can lead to physical and/or chemical traces 

(biomarkers), even after the death of the microorganism. 

125 - Exploring the transcriptome of the 

ectomycorrhizal symbiosis 

F. Martin * , S. Duplessis, A. Kohler & D. Tagu 

UMR IaM, Centre INRA de Nancy, 54280 Champenoux, 

France. - E-mail: fmartin@nancy.inra.fr 

Studies of the ectomycorrhizal symbiosis have highlighted 

the role of transcriptional regulation in controlling the 

morphological stages of symbiosis differentiation. To 

examine gene-activity changes associated with the 

development of the ectomycorrhizal symbiosis, we have 

performed expression profiling using poplar, eucalypt and 

Pisolithus cDNA arrays. A marked change in the gene 

expression in the mycobiont and the host-plants was 

observed at multiple levels: (a) a general activation of the 

protein synthesis machinery probably supporting an intense 

cell division/proliferation, (b) an increased accumulation of 

transcripts coding for cell wall proteins in hyphae and roots 

probably involved in the symbiotic interface formation, and 

(c) the upregulation of energy metabolism in colonised 

roots. This data suggests a highly dynamic environment in 

which symbionts are sending and receiving signals, are 

exposed to high levels of stress conditions and are 

remodeling their tissues. With multiple EST/cDNA array 

programmes dealing with ectomycorrhizal associations, we 

will have in a near future an unparalleled opportunity to 

ask which genetic features are responsible for 

common/divergent traits involved in this symbiosis. A few 

of the many possible breakthroughts will be in 

characterisation of common transcriptional and 

transduction networks and new insights into unique 

metabolic routes critical for mycorrhiza functioning. 

126 - Responses of mycorrhizal fungi to heavy metals: a 

cellular and molecular investigation 

L. Lanfranco 1* & P. Bonfante 2 

1 Dipartimento di Biologia Vegetale Università di Torino, 

Viale Mattioli 25, 10125 Torino, Italy. - 2 Istituto per la 

Protezione delle Piante, Sezione di Torino, CNR, Viale 

Mattioli 25, 10125 Torino, Italy. - E-mail: 

luisa.lanfranco@unito.it 

Within the fungal kingdom mycorrhizal fungi represent 

direct links between plants and soil and are often needed to 

ensure plant survival in heavy metals polluted areas. A 

sterile mycelium PSIV, an ascomycete establishing ericoid 

mycorrhiza, was used to understand how zinc ions affect 

the cellular mechanisms of fungal growth. Chitin, the most 

characteristic shape-determinant of fungal wall, was 

quantified and located by specific probes. Conspicuous 

changes in hyphal morphology were observed in zinctreated 

mycelium, where hyphal walls were thicker and 

chitin labelling was more intense. Chitin synthase genes 

belonging to two classes were differentially expressed. 

Morever, molecular mechanisms leading to protection 

against heavy metals were investigated in an arbuscular 

mycorrhizal fungus (Gigaspora margarita BEG34) and led 

to evidence of a structurally novel metallothionein. The 

gene, designated GmarMT1, encodes a functional 

polypeptide capable of conferring increased tolerance 

against cadmium and copper as revealed by 

complementation assays in yeast. The GmarMT1 RNA is 

expressed in both presymbiotic spores and symbiotic 

mycelia even in the absence of metal exposure, but it is 

significantly less abundant in the latter stage. An opposite 

pattern was observed upon copper exposure, which 

upregulated GmarMT1 expression in symbiotic mycelia, 

but not in germinated spores. Part of this work is supported 

by the European Project GENOMYCA (QLK5-CT-2000- 

01319). 



127 - Dissecting carbon metabolism in AM fungi: 

Implications for symbiosis and biotrophy 

B. Bago 1* , P.E. Pfeffer 2 , P. Lammers 3 & Y. Shachar-Hill 3 

1 CSIC, CIDE, Cami de la Marjal s/n, Albal (Valencia), 

Spain. - 2 USDA, ERRC, 600 E. Mermaid Ln. 19038 

Wyndmoor, PA, U.S.A. - 3 NMSU, Dept. Chemistry and 

Biochemistry, NMSU, 88033 Las Cruces NM, U.S.A. - Email: 

berta.bago@uv.es 

The obligate biotrophic nature of arbuscular mycorrhizal 

fungi (AMF) has long been known but never overcome, 

making all efforts of culturing these organisms 

continuously under axenic conditions unsuccessful. This 

has considerably limited progress in basic research and 

biotechnological strategies for the large scale production of 

AM fungal inoculum. All cytochemical, biochemical, 

metabolic and genome investigations carried out the last 25 

years indicated that AMF resemble saprophytes (e.g. 

genome size, metabolic capabilities, capacities for DNA, 

RNA and protein synthesis, etc.), however we are still 

unable to make these fungi complete their life cycle in the 

absence of a suitable host plant. One of the reasons 

proposed for this failure is the existence of a metabolic lack 

in their C metabolism. However, neither assays of involved 

enzymatic activities revealed the origin of such a metabolic 

blockage, nor did any of the numerous C sources assayed 

in synthetic media induce the fungus to fulfil its life cycle 

axenically. In recent years the application of a range of 

experimental techniques including AM monoxenic 

cultures, molecular biological methods, NMR spectroscopy 

and in vivo microscopy has greatly contributed to our 

understanding of C metabolism in AMF, both under 

asymbiotic and symbiotic conditions. The results have shed 

some light in the difficult, but fascinating question of the 

obligate biotrophic nature of AM fungi. 

128 - Carbohydrate and nitrogen dependent generegulation 

in the ectomycorrhizal fungus Amanita 

muscaria 

U. Nehls * , A. Bock, R. Kleber & R. Hampp 

Universitaet Tuebingen/Physiologische Ökologie der 

Pflanzen, Auf der Morgenstelle 1/72076 Tuebingen, 

Germany. - E-mail: uwe.nehls@uni-tuebingen.de 

Carbohydrate and nitrogen support has a profound impact 

on gene regulation and fungal physiology in saprophytic 

ascomycetes. As saprophytic fungi, ectomycorrhizal fungi 

live in environments with a different carbohydrate and 

nitrogen support (e.g. extramatrical hyphae versus 

mycorrhizal hyphae). To understand ectomycorrhizal 

fungal physiology in soil and the symbiotic structure, the 

effect of carbohydrate and nitrogen nutrition on fungal 

gene expression is of special interrest. Northern blot 

analysis was carried out for selected genes of the 

ectomycorrhizal fungus A. muscaria. The fungus was 

grown in liquid culture at different carbohydrate and 

42 


nitrogen conditions as well as in ectomycorrhizas. Genes 

investigated so far encode a monosaccharide transporter, a 

phenylalanine ammonium lyase, an excreted protease and 

an amino acid transporter. These studies revealed an 

interconnection between carbohydrate and nitrogen 

dependent regulation of gene expression in A. muscaria 

sharing homologies but also some differences to 

ascomycetes. Ectomycorrhizal hyphae are not uniform but 

consists of two 'fungal networks', the Hartig net that is 

attached to root cortical cells (forming together the 

plant/fungus interface), and hyphae that are ensheating the 

infected root. Since both structures have different functions 

with regard to carbohydrate and nitrogen uptake, storage 

and partitioning, we also compared the expression of 

fungal genes in both hyphal networks. 

129 - Functional genomics of fungal-plant 

communication in the arbuscular mycorrhizal 

symbiosis 

V. Gianinazzi-Pearson 1* , L. Brechenmacher 1 , M. 

Tamasloukht 2 , D. van Tuinen 1 , G. Bécard 3 , S. LaCamera 1 , 

S. Gianinazzi 1 & P. Franken 2 

1 UMR 1088 INRA/Université Bourgogne BBCE-IPM, 

INRA-CMSE, BP 86510, 21065 Dijon Cedex, France. - 

2 Max-Planck-Institut for Terrestrial Microbiology, Karlvon-Frisch-Strasse, 

35043 Marburg, Germany. - 3 Equipe 

de Mycologie Végétale, UMR 5546 CNRS/Université Paul 

Sabatier, BP17, 31326 Castanet-Tolosan, France. 

Fossil data indicate that arbuscular mycorrhizal (AM) fungi 

already colonized early land plants so that AM formation 

probably belongs to one of the earliest developmental 

programs plants evolved. At the same time, rRNA 

sequence analysis has shown that AM fungi form a 

monophyletic group (Glomeromycota) of which little is 

known about the biology of the possible progenitor. 

Analyses of the symbiotic programme in AM interactions 

have targeted fungal genes involved in cell wall synthesis, 

nutrient metabolism or membrane transport, and plant 

genes associated mainly with interactions with pathogenic 

organisms or symbiotic rhizobia. Several non-targeted 

approaches have also been used to further identify genes 

involved in AM formation and/or function but very limited 

information has been gained so far about fungal gene 

expression. This has prompted us to adopt alternative 

strategies of transcriptome analysis in a more extensive 

search for genes involved in different stages of AM 

development, based on suppressive subtractive 

hybridisation and large scale cDNA sequencing. 

Expression profiling has identified ESTs which correspond 

to fungal genes that are induced by host root exudates in 

pre-symbiotic phases or in mycelium during symbiotic root 

interactions. Such global approaches of functional 

genomics of AM will contribute to a better molecular 

understanding of fungal-plant communication in these 

widespread symbiotic associations and of how they 

promote plant vigour.


130 - Mycorrhiza-bacteria interactions and functioning 

in boreal forest soils 

R. Sen 

Division of General Microbiology, Department of 

Biosciences, P.O. Box 56, FIN-00014 University of 

Helsinki, Finland. - E-mail: robin.sen@helsinki.fi 

Boreal forest trees are highly dependent on root symbiotic 

ectomycorrhizal (ECM) fungi for growth in low pH, ligninrich 

podzolic soils. In Scots pine seedling microcosm 

studies, extensive mycorrhizal fungal colonization of the 

constituent upper humus (O) and underlying mineral (E 

and B) horizons was visualised and horizon-specific ECM 

identified following ITS-RFLP/sequence phylogenetics. 

Soil N and P mobilisation and uptake was confirmed in 

analyses of mycorrhizosphere compartment-specific gene 

and/or enzyme expression and linked to organic acid 

production. Bacterial biofilms in O horizon 

mycorrhizospheres were shown to be mainly comprised of 

forest soil specific gram -ve bacteria, Bacillus spp. and 

Crenarchaea based on carbon profiling and 16S sequence 

phylogenetics. Preferential organic acid utilisation by 

bacteria in the external mycorrhizosphere highlights 

mycorrhizal fungal involvement in weathering of E and B 

horizon minerals. Mycorrhizospheres developed in the O 

horizon did not host Pseudomonas fluorescens which was 

further confirmed in the rapid loss of marker gene tagged 

P. fluorescens introduced into mycorrhizal silver birch root 

systems. However, Scots pine mycorrhizospheres 

developed on petroleum contaminated soils do support 

catabolic plasmid harbouring fluorescent pseudomonads 

and increased hydrocarbon oxidation activity. Based on 

these data it is hypothesised that, in functional terms, the 

mycorrhizosphere behaves as an external rumen/gut of the 

host tree. 

131 - The Paxillus involutus / Betula pendula symbiosis: 

Gene expression in ectomycorrhizal root tissue 

T. Johansson * , A. Le Quéré, D.G. Ahrén, B. Söderström & 

A. Tunlid 

Microbial Ecology, Ecology Building, Lund University, S- 

223 62 Lund, Sweden. - E-mail: 

tomas.johansson@mbioekol.lu.se 

Ectomycorrhizas (ECM) are symbiotic associations formed 

between plants and soil fungi. To identify genes and 

metabolic pathways specifically expressed in the 

mycorrhizal root tissue, 3,555 Expressed Sequence Tags 

(ESTs) were analyzed in a cDNA library constructed from 

ECM formed between the basidiomycete Paxillus involutus 

and Betula pendula (birch). In parallel, cDNA libraries 

from saprophytically growing fungus (3,964 ESTs) and 

from axenic plants (2,532 ESTs) were analyzed. By 

assembly of all ESTs (10,051), 2,284 contigs were 

identified, each representing a unique transcripts of either 

fungal or plant origin. Of those, 650 contigs (28%) were 

uniquely expressed in the mycorrhizal tissue. Based on 

homology to sequence information in the GenBank (nr) 

protein database a majority of contigs could be assigned 

putative functional and metabolic roles. By comparing 

redundancies between libraries, transcripts related to 

protein synthesis were found down-regulated, whereas 

transcripts related to cell rescue, defense, cell death and 

ageing were up-regulated in the mycorrhizal root tissues as 

compared to the free-living fungus. Furthermore, the 

mycorrhizal root tissue displayed an up-regulation in 

transcripts related to nucleotide metabolism and carbon 

utilization as compared to the saprophytically growing 

fungus, whereas transcripts related to amino-acid 

metabolism and lipid, fatty-acid and isoprenoid metabolism 

were down-regulated. 

132 - Nutrient-regulated expression of functionally 

diverse surface proteins in truffles 

S. Ottonello * , B. Montanini & A. Bolchi 

Dipartimento di Biochimica e Biologia Molecolare- 

Università di Parma, Parco Area delle Scienze 23/A-43100 

Parma, Italy. - E-mail: s.ottonello@unipr.it 

Inorganic nitrogen is often the most limiting nutrient in the 

rhizosphere and N-availability is one of the environmental 

cues that influence ectomycorrhizae formation. This 

presentation will focus on four distinct, N-status-regulated 

surface protein genes from Tuber borchii. Two of them 

code for transmembrane proteins involved in inorganic-N 

internalization that resemble related transporters from other 

(symbiotic and non-symbiotic) fungi. They specifically 

respond to N-shortage, but do so at a surprisingly slow rate. 

Much more intense responses (to both N and C starvation) 

were measured for the genes encoding two other Surface 

Proteins, which harbor secretion signal peptides at their Ntermini, 

are both loosely associated to the cell wall, and 

bear only a very restricted (TbSP1) or no (TbSP2) 

resemblance to polypeptide sequences found in databases. 

The TbSP2 gene, which contains starvation stress response 

elements in its promoter, codes for a cysteine-rich, 11 kDa 

structural protein of as yet unknown function. The product 

of the TbSP1 gene, instead, is a calcium-activated 

phospholipase A2 that is both secreted and cell wallassociated 

in pre-symbiotic mycelia, but also accumulates 

in symbiosis-engaged hyphae as well as in fruibodies. 

Generalized surface remodeling and lipid-mediated events 

thus appear to predominate in the Tuber response to 

nutrient shortage. The results of ongoing experiments 

aimed to understand the physiological significance of such 

events will be discussed. 



133 - Rarity of mushroom species, a numerical 

approach 

G. Straatsma 1* , I. Krisai-Greilhuber 2 & S. Egli 3 

1 

Applied Plant Research, Mushroom Research Unit, PO 

Box 6042, 5960 AA Horst, The Netherlands. - 2 Institute of 

Botany, Department of Mycology and Cryptogams, Vienna, 

Austria. - 

3 Swiss Federal Research Institute WSL, 

Birmensdorf, Switzerland. - E-mail: 

g.straatsma@ppo.dlo.nl 

Abundances and yearly frequencies of mushroom species 

are tightly correlated in the data sets on a forest plot in 

Switzerland and on several forest and grasland plots in 

Austria. The geographic range of species seems correlated 

with abundance and yearly frequency. Species that score 

low on the three parameters can be called rare. None of the 

plots studied for 21 and 7 years, respectively, seems to 

have shown its full species richness. Many (?) rare species 

may thus have gone unnoticed. Rare species run the risk of 

extinction. Therefore rare species need to be considered for 

a 'red list' (according to IUCN criteria). The red lists of the 

Netherlands and Germany were evaluated for the presence 

of rare species. The fungal biota of both countries are well 

studied. The evaluation was done by comparing the red 

lists with distribution data. Distribution data of the 

Netherlands have been published (Arnolds, Dam & Dam- 

Elings, 1995). German distribution data were estimated on 

a basic estimate that the whole country holds 6000 species 

and on the basis of distribution data of higher plants, 

mosses, birds, mammals, beetles and butterflies. The 

impression is that very rare mushroom species are 

underrepresented on the red lists of both countries. 

134 - Diversity of soil fungi in different Namibian 

biomes 

C. Görke 

Spezielle Botanik & Mykologie, University of Tübingen, 

Auf der Morgenstelle 1, D-72076 Tübingen, Germany. - Email: 

claudia.goerke@uni-tuebingen.de 

Within the framework of the BIOTA project (Biodiversity 

Monitoring Transect Analysis in Africa) the biodiversity of 

soil fungi is the aim of this survey. The scientists of the 

frame project (Southern Africa) study the influence of 

climatic changes and management systems on different 

organisms along the rainfall gradient spanning from the 

Cape of Good Hope to the Angolan border. This 

presentation concentrates on results of Namibia, where 

sampling started in 2001. Soil samples were taken in 

different biomes of Namibia: dry forest, thorn bush 

savannah, Nama Karoo and the Namib. The soil fungi were 

isolated by the soil washing method. Most cultures belong 

to the Fungi Imperfecti (Hypho- and Coelomycetes), not so 

many are Asco- or Zygomycetes. Only a few Oomycetes 

and one Basidiomycete have been observed up to now. 

Members of the genus Aspergillus are frequent. In nearly 

44 


half of the soil samples of the dry forest of northern 

Namibia a member of the A. niger-group (A. niger 

respectively A. japonicus) was isolated, but nowhere else. 

Fewer cultures were gained from the soil of the Nama 

Karoo. In the first sampling the dominating isolate was a 

Coelomycete, most likely a member of the genus Phoma. 

However, in this area the samples of 2001 were taken 

before the onset of the raining season, while the raining 

season in the other areas was well under way. In 2002 the 

sampling was after the annual rain was fallen. 

135 - Arbuscular mycorrhizae in Namibian grasses: a 

comparison between North and South 

E. Uhlmann * , C. Görke & F. Oberwinkler 

University of Tuebingen, Spezielle Botanik & Mykologie, 

Auf der Morgenstelle 1, 72076 Tuebingen, Germany. - Email: 

elisabeth.uhlmann@uni-tuebingen.de 

Arbuscular mycorrhizae (AM) are studied within the 

framework of the BIOTA Southern Africa project. One aim 

is an inventory of species along the rainfall gradient from 

the Cape of Good Hope to the Angolan border. In addition 

to this, different land management systems are studied with 

respect to their influence on biodiversity. This survey is 

focussed on Namibia, comparing the South (with an annual 

rainfall of approx. 150 mm) to the North (with 450 mm 

rain/year). Grasses were chosen as host plants of AM as 

they occur along the whole transect and are associated with 

AM fungi under normal conditions. Morphological studies 

revealed different rates of mycorrhization in the North and 

South. Molecular studies (sequencing of the ITS region of 

nuclear ribosomal genes) of grass roots infected with 

arbuscular mycorrhizal fungi showed that several AM 

species (eg Glomus intraradices) occur along the whole 

transect while others occur only in the North or the South. 

These results are backed by morphological identification of 

spores isolated from surrounding soils. Comparing 

different management systems in the South, differences in 

rate of mycorrhization can be detected; although no 

differences were noted in species composition of AM 

fungi. 

136 - Biodiversity of endophytic fungi from Pinus 

tabulaeformis in China 

L.-D. Guo 

Institute of Microbiology, CAS, P.O. Box 2714, Beijing, 

China. - E-mail: guold@sun.im.ac.cn 

In a survey of biodiversity of endophytic fungi from Pinus 

tabulaeformis in China, A total of 25 taxa and 6 

morphotypes of mycelia sterilia were found. In the 6 

morphotypes, 18 isolates were arbitrarily selected from 

white morphotype and identified into various taxonomic 

levels based on rDNA sequence analysis. The 5.8S gene 

and ITS regions of rDNA from the 18 white morphotypes


were amplified and sequenced. Phylogenetic analysis based 

on the 5.8S gene sequences showed that these 18 white 

morphotypes belonged to the Ascomycota. Further 

identification of the white morphotypes to lower taxonomic 

levels was conducted by means of sequence similarity 

comparison and phylogenetic analysis of both the 5.8S 

gene and ITS regions. The origins of WMS9 and WMS10 

were inferred to be the genus Lophodermium of the 

Rhytismataceae, and the origins of WMS11, WMS13, and 

WMS18 were the Rhytismataceae. WMS2, WMS3, 

WMS4, WMS5, and WMS6 were identified to the genus 

Rosellinia, WMS1 to the genus Entoleuca, and WMS14 to 

the genus Nemania of the Xylariaceae, and the origins of 

WMS7, WMS8, WMS12, WMS15, WMS16, and WMS17 

were the Xylariaceae. The value of using DNA sequence 

analysis in the identification of endophytic fungi is 

discussed. 

137 - Assessing diversity of leaf-inhabiting pathogenic 

coelomycetes in south-east Europe 

T.V. Andrianova 

M.G. Kholodny Institute of Botany, NAS Ukraine, 

Tereshchenkivska, 2, Kiev 01601, Ukraine. - E-mail: 

tand@darwin.relc.com 

Leaf-inhabiting pathogenic coelomycetes are difficult to 

monitor in nature because of their microscopic size, large 

number of inadequately characterized and classified 

morphs. About 1100 species of these anamorphic fungi 

have been recorded from south-east Europe. Fungal 

diversity assessment in these temperate and submeridional 

regions allow one to predict an eventual total of around 

15300 microfungi on the 5100 known vascular plants. 

Some names need critical revision at species rank, other 

anamorphic names need reassessment in the light of 

teleomorph names and the application of one name for the 

holomorph. An eventual 1500-2000 leaf-inhabiting 

pathogenic coelomycetes can thus be expected for southeast 

Europe. Main centres of biodiversity are the 

Carpathian mountains and Crimean peninsula. About 500 

leaf-inhabiting coelomycete pathogens were observed in 

mountain forests, subalpine meadows, forest-steppes and 

steppes of Crimea. The eastern Carpathian virgin broadleaf 

and conifer forests, and alpine plant communities are 

characterized by over 260 of these pathogens, but this 

number is not final. The species richness of leaf-inhabiting 

coelomycetes is based on a combination of abiotic factors 

and host-plants diversity. Decline of the total population of 

these fungi, at 43% in the Carpathians, and accumulation of 

species with agressive pathogenic characters has been 

observed in unstable ecosystems. Study of these fungi can 

provide a way to monitor stability of natural ecosystems. 

138 - Functional biodiversity of grassland saprotrophic 

fungi 

L.J. Deacon 1* , C.H. Robinson 1 , B.W. Bainbridge 1 & J.C. 

Frankland 2 

1 King's College London, Division of Life Sciences, 

Franklin-Wilkins Building, 150 Stamford Street, London 

SE1 9NN, U.K. - 2 Centre for Ecology and Hydrology 

Merlewood, Windermere Road, Grange Over Sands, 

Cumbria LA11 6JU, U.K. - E-mail: 

lewis.deacon@kcl.ac.uk 

The aim of this project is to characterise the functional 

biodiversity of saprotrophic decomposer fungi in a 

grassland community. The presence of all saprotrophic 

fungal species in a grassland community may not be 

necessary to maintain the function of the ecosystem, i.e. 

some species may be functionally redundant in the process 

of decomposition because several species may break down 

the same carbon and nitrogen substrates. Fungal isolates 

obtained from an upland grassland soil in Scotland, were 

selected to cover not only the most abundant species 

(common isolations), but also a cross section of the fungal 

community (occasional isolations) from different treatment 

plots (control, lime, nitrogen and nitrogen plus lime) at the 

field site. Isolates were tested for their ability to grow on 

specific substrates, in defined and semi-defined media. 

Primarily the activity of cellulolytic enzymes, ligninolytic 

enzymes, pectinase, amylase and chitinase were tested on 

solid media. Twelve isolates were chosen for further 

screening for the utilisation of smaller molecular weight 

carbon and nitrogen sources using the BIOLOG system. 

Further work is in development to test the paired isolates 

ability to utilise and compete for natural substrates 

available in the grassland. 

139 - Guatemalan Macrofungi: diversity and uses 

R.E. Flores 1* , M.C. Bran 1 , O. Morales 1 & M. Honrubia 2 

1 Universidad de San Carlos, DIGI/Facultad CCQQ y 

Farmacia. Edif T-12 Ciudad Universitaria 01012, 

Guatemala. - 2 Universidad de Murcia, Depto. Biología 

Vegetal (Botánica). Campus de Espinardo 30100, Spain. - 

E-mail: rfloresa@yahoo.com 

Many fungal species from North to South America and 

vice versa found place in Guatemala due to its geographical 

position and orographic relief. Many of them are 

mycorrhizal with pines, oaks, alders and fir. In addition, 

there is also a large diversity of saprophitic species, 

especially in the tropical areas. Mayan people in Guatemala 

have used mushrooms as part of their rituals and as a 

source of food since many centuries ago. Currently 

mushrooms are used as food and as medicine but 

exceptional uses as insecticide or decorative elements are 

reported. The main researches on macrofungi in Guatemala 

have been directed for 10 years ago by the University of 

San Carlos. Currently its Mycological Herbarium contains 



a large collection of saprophitic and mycorrhizal dried 

samples from almost all the regions of the country. The 

most important collections of mycorrhizal mushrooms 

come from the highlands because of their diversity and 

knowledge of the Mayan people. Species described only 

for North America as Boletus edulis, Catathelasma 

ventricosa, Gomphus floccosus, Lactarius salmonicolor, 

Rhizopogon evadens and others were found. New and 

possible endemic species are present in the genera 

Amanita, Boletus, Cortinarius, Lactarius, Russula, 

Tylopilus, but more studies are necessary. Interesting 

findings on distribution of species in the Caribbean area 

and south east of the country have also been reported. 

140 - Effect of host tree species on fungal community 

composition on a tropical rain forest in Panama 

A. Ferrer 

State University of New York-ESF, 706 W. Michigan Ave 

Urbana, Illinois 61801, U.S.A. - E-mail: 

astridferrer@hotmail.com 

Wood-inhabiting ascomycetes and basidiomycetes 

represent a diverse group of taxa which play an important 

role in decomposition and nutrient cycling. Little is known 

about how communities of these organisms are organized 

or how tree host influences community structure. To 

determine whether host tree species influences the 

composition of fungal communities, the ascomycetes and 

basidiomycetes present on three host species: Prioria 

copaifera, Quararibea asterolepis, and Trichilia 

tuberculata were sampled on a plot on Barro Colorado 

Island, Panama. Fungal diversity was high. Sampling of 

181 trees and branches found dead over three years yielded 

75 ascomycetes and 112 basidiomycetes species. The 

highest diversity of both fungal groups was found on 

Prioria, but the asymptote of species accumulation curves 

was not reached for any host tree. Ordination of fungal 

communities revealed distinct differences between hosts, 

but strong similarity within hosts across years. The most 

abundant fungal species of both ascomycetes and 

basidiomycetes were generalists found on all three hosts. 

However, Quararibea harbored more species unique on 

this host. Randomization tests revealed that there were 

significantly fewer host generalist fungi than expected for 

ascomycetes but not for basidiomycetes. These results 

indicate that host composition plays a role in structuring 

both ascomycete and basidiomycete communities, but that 

the most successful species are capable of colonizing 

multiple host species. 

141 - Conservation of biodiversity: Effects of varying 

levels of green-tree retention on ectomycorrhizal fungus 

diversity 

D.L. Luoma * & J.L. Eberhart 

Department of Forest Science, Oregon State University, 

Corvallis, OR 97331, U.S.A. - E-mail: luomad@fsl.orst.edu 

46 


The Demonstration of Ecosystem Management Options 

(DEMO) experiment examines the effects of green-tree 

retention on mycorrhizal fungi, vegetation, wildlife, 

insects, soils, hydrology, and social perceptions of forested 

landscapes. Ectomycorrhizal (EM) fungi were sampled 

from three replicated treatment blocks. Sporocarp 

production and ectomycorrhizae were assessed before and 

after trees were cut. 197 EM species were identified from 

sporocarps (57 hypogeous, 140 epigeous) and 188 EM 

morphotypes were described from soil cores. Hypogeous 

sporocarp production of the genera Gautieria and 

Rhizopogon was greatly reduced by heavy thinning as was 

mushroom production in the genera Cortinarius, Inocybe, 

and Russula. Moderate thinning retained higher levels of 

sporocarp production. The cumulative number of EM types 

was estimated for each treatment via the Abundance-based 

Coverage Estimator (ACE). The ACE attempts to account 

for species that are present in a population but not observed 

in the sample. Fewer EM types per soil core in the most 

heavily cut areas translated into a severe reduction in the 

rate at which species accumulated. The rarer species were 

disproportionately affected in the heavily cut areas and EM 

diversity was greatly lowered, especially as compared to 

the moderately thinned areas. Both the sporocarp data and 

the EM data suggested that diversity indices responded in a 

non-linear manner to varying levels of green-tree retention. 

142 - Hygrocybe and Cuphophyllus as ecological 

indicators 

J.B. Jordal 

Ressurssenteret i Tingvoll, N-6610 Øksendal, Norway. 

A total of 46 species and 7 varieties of Hygrocybe sensu 

lato are known from Norway. Most species are been found 

during investigations of seminatural grasslands. Hygrocybe 

species are also found in rich forests, fens, and in alpine 

areas. Data on Hygrocybe species from 650 grassland 

localities have been collected in the years 1992-2001, with 

a total of 3650 records excl. duplicates. Some species show 

a southern distribution pattern, e. g. H. intermedia. Studies 

of vertical distribution show that there are lowland species, 

e. g. H. irrigata, indifferent species, e.g. H. conica, H. 

pratensis, and 'summer farm species' with the highest 

frequency in north boreal (subalpine) region: e.g. H. 

nitrata, H. turunda. There are species occuring mainly in 

calcareous areas, e.g. H. colemanniana. H. vitellina seems 

to have an oceanic distribution pattern, some other species 

may be suboceanic. The frequencies of the grassland 

species are presented. Most rare grassland species seem to 

be good indicators of species rich localities with a long 

grazing continuity. The fragmentation of Hygrocybe 

localities in the cultural landscape is a continuing process. 

The county of Møre og Romsdal has a much higher density 

of species rich localities than Denmark and the 

Netherlands. It is a challenge to save the Hygrocybe 

localities for future generations.


143 - Autecology of Hygrocybe spp. in temperate 

grasslands 

G.W. Griffith 1* , G.L. Easton 1 , A.W. Jones 1 , A. Stott 2 , N. 

Ostle 2 & R. Bol 3 



3DA, Wales, U.K. - 2 Centre for Ecology and Hydrology, 

Merlewood Research Station, Grange-Over-Sands, 

Cumbria, LA11 6JU Cumbria, LA11 6JU, England, U.K. - 

3 Institute of Grassland and Environmental Research, North 

Wyke Research Station, Okehampton, Devon EX20 2SB, 

England, U.K. - E-mail: gwg@aber.ac.uk 

Members of the genus Hygrocybe are ubiquitous and 

colourful components of many undisturbed and nutrientpoor 

grasslands in Northern Europe. Through surveys of 

the distribution of Hygrocybe spp. and of other 

macrofungal genera showing similar patterns of 

occurrence, a picture is gradually emerging of the more 

important waxcap grassland sites, and of those species in 

greatest need of protection. As part of a UK-based Soil 

Biodiversity Programme 

(http://mwnta.nmw.ac.uk/soilbio/sourhope.htm), we have 

monitored the effect of various management regimes on 

fruiting of Hygrocybe spp. Fine-scale mapping combined 

with genetic analysis (AFLP/ISSR) is being used to 

measure the extent of individual genets, with speciesspecific 

PCR probes being used to establish the vertical 

location of mycelia. Analysis of the natural abundance of 

the stable isotopes in fruitbodies showed that grassland 

Hygrocybe spp. show significant depletion for 13 C(-28 to - 

30‰) and enrichment for 15 N(+12 to +18‰), a pattern that 

sets them apart from most macrofungi previously 

examined. Furthermore, the other macrofungi associated 

with these undisturbed grasslands (eg Clavariaceae, 

Geoglossaceae) have very similar isotope signatures 

despite being taxonomically unrelated. Experiments using 

plant litter enriched in 15 N are currently underway and will 

further clarify our understanding of the role of these fungi 

in nutrient cycling and explain why they are so adversely 

affected by many agricultural practices. 

144 - What is Agaricus cossus? - Phylogeny of the white 

Hygrophorus species in Northern Europe 

E. Larsson * & S. Jacobsson 

Göteborg University, Dep of Biology, Box 461, SE 405 30 

Göteborg, Sweden. - E-mail: ellen.larsson@systbot.gu.se 

The genus Hygrophorus Fr. comprises about 40 species in 

the Scandinavian countries. They are characterized by 

medium to large, fleshy basidiomes that are whitish or 

colored in gray, brown, yellowish orange to red. All 

species grow in woodlands and are obligate mycorrhiza 

formers. There are at least ten white to whitish species in 

the Nordic countries. In traditional classification color or 

color- changes, viscidity, odor etc. of the basidiome have 

been the most important characters. However, these 

characters are often vague and also dependent on 

environmental conditions. For these reasons great 

taxonomical and nomenclatural problems among the white 

Hygrophorus species exists. The internal transcribed spacer 

regions 1 and 2 and the first 900 basepairs of the 5' region 

of the nuclear large subunit ribosomal DNA was sequenced 

for 35 ingroup and outgroup taxa. Phylogenetic analysis 

shows that Hygrophorus is a well-defined genus but the 

white species do not form a monophyletic group. Host 

specificity seems to be high among the analyzed taxa. 

145 - Tropical Hygrophoraceae: convergent evolution 

in Hygrocybe, and support for retention of Hygroaster 

based on molecular evidence 

S.A. Cantrell 1* , D.J. Lodge 2 , J.-M. Moncalvo 3 , R. 

Vilgalys 3 , K.K. Nakasone 4 & G.W. Griffith 5 

1 Science & Technology, Universidad del Turabo, PO Box 

3030, Gurabo, PR 00778, U.S.A. - 2 Center for Forest 

Mycology Research, USDA-FS, FPL, PO Box 1377, 

Luquillo PR 00773-1377, U.S.A. - 3 Dept. Botany, Duke 

Univ., Box 90339, Durham, NC 27708-0339, U.S.A. - 

4 Center for Forest Mycology Research, USDA-FS, FPL, 

One Gifford Pinchot Dr., Madison, WI 53705-2398, U.S.A. 

- 5 Institute of Biological Sciences, Univ. of Wales 

Aberystwyth Penglais, Aberystwyth, Deredigion SY23 3DA, 

Wales, U.K. - E-mail: sharonac@coqui.net 

The family Hygrophoraceae is highly diverse in tropical 

regions where all the genera are represented 

(Camarophyllopsis, Cuphophyllus, Hygroaster, Hygrocybe, 

Hygrophorus and Humiditis). Hygroaster and Hygrocybe 

Section Firmae are restricted to the tropics and subtropics. 

Using ribosomal DNA sequences, we have obtained a 

preliminary phylogeny. Despite having ornamented rather 

than smooth spores, the genus Hygroaster, is a good 

member of the Hygrophoraceae. With the exception of 

Sections Firmae and Glutinosae, the molecular phylogeny 

is concordant with the current division of Hygrocybe into 

two subgenera: Hygrocybe and Pseudohygrocybe. Section 

Glutinosae forms a separate clade from the other genera, 

supporting recognition of the genus Gliophorus, but more 

taxon sampling is needed. Within Hygrocybe ss., Section 

Firmae is included in the subgenus Hygrocybe clade and is 

basal to sections Hygrocybe and Chlorophanae. The most 

basal members of section Firmae (H. hypohaemacta) and 

Chlorophanae (H. glutinipes) that we have sampled have 

similar macro- and micromorphology, except for the 

presence of dimorphic basidia and spores in Section 

Firmae. Section Firmae should be reclassified in subgenus 

Hygrocybe. Although section Firmae may be derived from 

section Coccineae in subgenus Pseudohygrocybe as 

suggested by Heinemann, our results suggest that 

convergent evolution is responsible for the similarity of 

micromorphology in terminal species in the Sections 

Firmae and Coccineae. 



146 - Recent advances in molecular phylogeny of 

temperate Hygrophoraceae and concordance with 

morphology and ecology 

A.E. Kovalenko 1* , J.-M. Moncalvo 2 , R. Vilgalys 2 , R.H. 

Petersen 3 , K.W. Hughes 3 & D.J. Lodge 4 

1 V. L. Komarov Botanical Institute, Russian Academy of 

Sciences, 2 Prof. Popov Street, 197376, St. Petersburg, 

Russia. - 2 Department of Botany, Duke University, 

Durham, North Carolina, 27708, U.S.A. - 3 Department of 

Botany, University of Tennessee, Knoxville, Tennessee, 

37996-1100, U.S.A. - 4 Centr. for Forest Mycology 

Research, USDA-FS, FPL PO Box 1377, Luquillo PR 

00773-1377, U.S.A. - E-mail: alkov@AK3010.spb.edu 

Phylogenetic relationships within Hygrophoraceae 

(Basidiomycota) were studied using parsimony analysis of 

nuclear LSU, ITS1-2 and SSU rDNA. The analysis 

included 114 sequences (30 LSU; 23 SSU; and 61 ITS of 

41 species) from Russia, the USA and Europe. For 

comparisons, a phylogenetic reconstruction was also made 

using morphological and ecological data (30 characters in 

total). Results from the molecular analyses are largely 

concordant with each other as well as with the analysis of 

morphological end ecological data. However, the ITS 

sequences varied too much between representatives of 

different genera, so this region is most appropriate for 

phylogenetic studies within genera of the Hygrophoraceae. 

The results suggested that the Hygrophoraceae appears to 

be monophyletic group. The family contains 9 separate 

clades that support the following genera: Cuphophyllus, 

Neohygrocybe, Camarophyllopsis, Humidicutis, 

Gliophorus, Pseudohygrocybe, Hygrocybe, 

Neohygrophorus and Hygrophorus. The taxonomic 

position of some species should be reevaluated. Including 

several representatives of the Tricholomataceae s.l. (i.e. 

Omphalina) in the analysis demonstrated that some of them 

may be related to the Hygrophoraceae and probably should 

be transferred to this family. (This study was partly 

supported by grants from Hesler Endowment Fund and 

Russian Foundation for Basic Research). 

147 - Biogeographical implications of Trichomycete 

distributions 

R. Lichtwardt 

University of Kansas, Department of Ecology & 

Evolutionary Biology, Lawrence, Kansas 66045, U.S.A. - 

E-mail: licht@ku.edu 

Known distributions of Trichomycetes worldwide fall into 

several patterns depending upon their host types, habitats, 

and host specificity. It is hypothesized that in all cases 

autonomous dissemination of the gut fungi does not occur 

except within boundaries of their immediate host 

populations, and that dispersals over greater distances are 

the result of particular active or passive mechanisms. 

Examples of vicariant distributions of the symbionts in 

48 


both the Northern and Southern Hemispheres are 

presented. The hypothesis that this fungus-arthropod 

association is very ancient is supported by historical 

biogeography. 

148 - Progress toward a rDNA based phylogeny of the 

Harpellales 

M.M. White 

University of Kansas, Dept. Ecology and Evolutionary 

Biology, Lawrence, Kansas 66045, U.S.A. - E-mail: 

trichos@ku.edu 

The Harpellales (Trichomycetes), gut fungi, are unique 

endosymbionts associated with larval aquatic insects 

worldwide. Their evolutionary relationships have been 

difficult to infer because of the paucity of morphological 

characters and unculturability of most of the 35 known 

genera. Previous DNA studies have included at most four 

genera of culturable Harpellales. This study used thalli 

taken from guts (mixed genomic template) to generate 18S 

and 28S rDNA sequences to infer the phylogeny of the 

Harpellales using cladistic analyses. Sixteen genera of 

Harpellales were included, with 64 of 72 sequences from 

unculturable samples. The genus Orphella fell outside an 

otherwise monophyletic group of Harpellales, more closely 

allied to the Kickxellales. The two largest genera, Smittium 

and Stachylina, are polyphyletic, perhaps masked by 

convergent and limited morphological characters. The 

cladograms did not reveal any pattern that corresponds 

with the two most important morphological criteria, 

trichospore shape and zygospore type, as well as 

appendage number for both. The current two family system 

of classification is not supported, suggesting that the 

Legeriomycetaceae may need to be dropped as a rank. 

However, the need to add more sequence data, pursuit of a 

protein gene, and continued collections for subsequent 

cladistic analyses are discussed. 

149 - Systematics of non-culturable orders of 

Trichomycetes based on molecular markers 

M.J. Cafaro 

University of Kansas, Dept. Ecology and Evolutionary 

Biology, Lawrence, Kansas 66045, U.S.A. - E-mail: 

matcaf@ku.edu 

The Eccrinales and Asellariales are morphologically 

diverse orders of the class Trichomycetes (Zygomycota). 

These gut fungi inhabit a wide range of hosts: Crustacea, 

Insecta and Diplopoda in varied habitats (marine, 

freshwater and terrestrial). The order Eccrinales is 

characterized by unbranched, nonseptate, multinucleate 

thalli and sporangiospores that are formed basipetally from 

the thallus apex; they live attached to the cuticle lining of 

the digestive tract of the host by a secreted, basal holdfast. 

The order Asellariales, on the contrary, has branched,


septate thalli that produce arthrospores in their fertile 

branches. They attach through holfast cells that have 

unique morphology commonly used for species 

identification. Sexual reproduction is not known for either 

group. No species of either order has been axenically 

cultured so far. Most of their taxonomy, if not all, is based 

on a few micromorphological characters. Molecular 

markers have been developed to study the relationship of 

these orders to other groups of fungi. Ribosomal gene (18S 

and 28S) sequence analyses do not support a close 

association of these orders to the Harpellales, their putative 

sister taxon. 

150 - Recent advances in trichomycete ecology and 

physiology 

C.E. Beard 1* , P.H. Adler 1 & J.W. McCreadie 2 

1 Clemson University, Department of Entomology, Clemson, 

SC, 29634-0365, U.S.A. - 2 University of South Alabama, 

Department of Biological Sciences, Mobile, AL, 36688, 

U.S.A. - E-mail: cbrd@clemson.edu 

Although the systematics of trichomycetes represents a 

fairly mature level of knowledge, studies of trichomycete 

ecology and physiology have lagged behind. To address 

this imbalance, we have investigated the ecology of 

trichomycetes and aspects of their physiology. We asked 

whether the distributions of trichomycetes in time and 

space were predictable. A two-year study revealed that the 

prevalence of trichomycetes inhabiting black flies is related 

to season. In some streams, Harpella melusinae was most 

prevalent in spring and fall. Efforts to relate prevalence to 

environmental characteristics showed that the prevalence 

of H. melusinae is greatest in mountain streams. Prevalence 

of this species in the black fly Simulium tuberosum is 

related to stream pH and conductivity. We also investigated 

physiological aspects of establishment of trichomycetes in 

hosts. Zygospores of H. melusinae and Stachylina spp. are 

rare, but formation can be stimulated by elevated levels of 

potassium and pH (as KOH). Smittium megazygosporum 

has a disjunct distribution and large zygospores (150 µm 

long). Large zygospores might enable trichomycetes to 

persist in the environment. The role of zygospores in the 

life history of trichomycetes remains unclear. Smittium 

culisetae inhabits specific regions of the host hindgut, 

which might be related to the timing of spore germination 

as spores transit the gut. Nutritive characters of the hindgut 

regions might determine which areas are habitable. 

151 - Dispersal mechanisms in species of Harpellales 

A.M. Rizzo * & S.T. Moss 

University of Portsmouth school of Biological Sciences, 

King Henry St. Portsmouth PO5 3EN, U.K. - E-mail: 

alan.rizzo@port.ac.uk 

Fungi of the Harpellales (Trichomycetes) live attached to 

the mid-gut (Harpellaceae) and hind-gut 

(Legeriomycetaceae) of their dipteran hosts. The hosts 

include species of both simuliid and culiciid vectors for 

river blindness and malaria respectively. Transmission of 

these fungi within host populations is by means of 

asexually produced sporangia termed trichospores. These 

spores are produced by the thallus within the host gut. 

Light microscopic images of these spores from 

representative genera will be shown and an explanation 

made of the mode of transmission and infection. Some 

members of the Harpellales have a chlamydosporic stage 

which is associated with the eggs of host Diptera. These 

spores are thought to provide a means of dispersal to new 

or ephemeral habitats. Data relating to the occurrence of 

these spores at various collection sites will be presented. 

Light and scanning electron micrographs showing the 

structure of this stage will also be provided for three 

species of the Harpellales: Genistellospora homothallica, 

Harpella melusinae, Smittium sp. Results of infection trials 

in order to establish positively a process for reinfestation 

within the host by these spores will be given. The 

mechanism(s) by which species of the Harpellales invade 

the tissue of the adult fly is unknown. Transmission 

electron micrographs of pupae and gravid adults will be 

provided to show a possible pathway of infection. 

152 - Selected habitats of Xylaria species of Mexico 

F. San Martín 1* , J.D. Rogers 2 & P. Lavín 1 

1 BIOTA, Sierra Hermosa 617 Fr. Villa Real. Cd. Victoria. 

Tam. 87027, Mexico. - 2 Washington State University, 

Pullman, WA 99164-6430, U.S.A. 

Due to its diverse floristic composition, México is, perhaps, 

the area of the New World with more species of Xylaria. In 

this contribution, 50 species of Xylaria are reported 

growing on different habitats, i.e., on wood and fruits of 

Quercus spp., on Guazuma ulmifolia, Liquidambar 

styraciflua and Magnolia spp. fruits, on pods and pod-like 

structures, on fallen leaves, on wood of several dicots and 

monocots, on different fruit remains and soil and dung. 

Morphologic and host relationships are presented for the 

following species of Xylaria: X. adscendens, X. alata, X. 

albisquamula, X. amphithele, X. apiculata, X. aristata, X. 

castilloi, X. coccophora, X. delicatula, X. dichotoma, X. 

diminuta, X. duranii, X. enteroleuca, X. equina, X. 

eugeniae, X. feejeensis, X. cf. filiformis, X. gracillima, X. 

guazumae, X. inaequalis, X. ianthino-velutina, X. ianthinovelutina 

large-spored variety, X. jaliscoensis, X. juniperus 

var. asperula, X. lancea, X. liquidambar, X. longiana, X. 

longipes, X. magniannulata, X. magnoliae, X. cf. 

magnoliae, X. mexicana, X. michoacana, X. microceras, X. 

multiplex, X. oxyacanthae, X. palmicola small-spored 

variety, X. perezsilvae, X. phyllocharis, X. poitei, X. 

rhizomorpha, X. quercinophila, X. sp. aff. hypoxylon, X. sp. 

aff. longiana, X. sp. cubensis complex, X. scabriclavula, X. 

cf. scruposa, X. striata, X. subcoccophora, and X. 

tumulosa. 



153 - The Xylariaceae of Ecuador - an update 

T. Læssøe 

Botanical Institute, Copenhagen University, Denmark. - Email: 

thomasl@bot.ku.dk 

Besides the early effort by Lagerheim and Patouillard 

(1890s) not much was known about Ecuadorean 

Xylariaceae until the current effort initiated in 1987. Even 

today many areas of high potential remain unexplored but 

at least a rough picture of the xylariaceous biodiversity can 

now be presented. Ca. 17 genera have been recorded, and 

Xylaria rules with more than 98 species, many of which 

lack formal names. Camillea has the highest country record 

known with 23 spp. Hypoxylon is also represented with ca. 

23 spp., but a high proportion are only known from single 

records, so this number should increase. The recent survey 

has documented the presence of Entonaema pallida in the 

country. Anthostomella and Rosellinia remain poorly 

known, but R. evansii has been found as new to South 

America. It is clear that the Amazon basin has the highest 

diversity with more than 110 species, but many of the 

novelties discovered occur at higher altitudes. Also the 

Choco region of the northwest and the southern provinces 

have produced promising results. Alnus acuminata was 

investigated in order to establish whether it shares 

xylariaceous fungi with its temperate relatives, and both 

Hypoxylon multiforme and H. fuscum were recorded 

together with the more subtropical/tropical Daldinia 

childiae. Mycogeographically it is interesting to note that 

members of the genera Phylacia, Thamnomyces and 

Leprieuria so far only have been recorded within the 

Amazon basin. 

154 - Where are the Xylariaceae? The Southeast Asia 

connection 

M.A. Whalley 1* & S. Thienhirun 2 

1 School of Biomolecular Sciences, Liverpool John Moores 

University, Byrom Street, Liverpool, L3 3AF, U.K. - 

2 Forest Products research Division, Forest Research 

Office, Royal Forest Department, Bangkok 10900, 

Thailand. - E-mail: mawhalley@hotmail.com 

The family Xylariaceae is truly cosmopolitan but the 

greatest concentration of species occurs in the tropics. In 

the past it has always been acknowledged that tropical 

South and Central America house the greatest diversity. 

However, recent surveys in Thailand, Malaysia and Papua 

New Guinea suggest that the 'species gap' between the old 

and the new tropics is not as exaggerated as previously 

suggested. The lack of collectors, and indeed specialists in 

Southeast Asia, has contributed significantly to underrecording 

in the region. For example Camillea has always 

been considered to be a South American and Central 

American genus but two new species have recently been 

described from Malaysia and C. tinctor has been found to 

be regularly collected in Malaysia, Thailand and Papua 

50 


New Guinea. A total of 12 genera of Xylariaceae are 

reported from Malaysia and at least 15 taxa are considered 

to be previously undescribed. The Xylariaceae is now 

known to be well represented in Thailand with 17 genera 

recognised. Studies in Papua New Guinea also reveal a 

strong presence there. It is of some significance that in each 

of the countries surveyed a considerable number of the 

collections of Hypoxylon and Xylaria are probably 

undescribed whilst others are infrequently reported. The 

range of species reported from the region will be discussed, 

together with variation associated with habitat and host, 

and the findings compared with currently available records 

from Africa and tropical America. 

155 - Can lifestyle be used to find the rare or the new 

among the Xylariaceae? 

A.J.S. Whalley 1* & P. Sihanonth 2 

1 School of Bimolecular Science, Liverpool John Moores 

Universitys, Byrom Street, Liverpool, L3 3AF, U.K. - 

2 Department of Microbiology, Chulalongkorn University, 

Bangkok 10330, Thailand. - E-mail: 

bmsawhal@livjm.ac.uk 

Although the Xylariaceae are seen primarily as wood 

inhabitants genera such as Hypocopra, Poronia, 

Podosordaria, and Wawelia are specifically dung 

inhabitants. Wawelia is also xerophilic. Other members of 

the family are litter, fruit or seed inhabitants and others are 

associated with soil or insects. The majority are 

saprotrophic but an increasing number of taxa are being 

recognized as phytopathogens through latent invasion in 

stressed hosts. The Xylariaceae also exhibit an outstanding 

presence as endophytes, especially in the tropics. Many 

taxa also have a strong host preference and some seem to 

be host specific. Within the wood inhabiting species not 

only is host important but also the nature of the host. Some 

species grow only on wood which retains its bark others 

only on decorticated wood and some only on well-rotted 

and water soaked wood. Thus the nature of the substratum 

and the habitat strongly influence the species which will be 

found. Studies on endophytic Xylariaceae show that some 

of the species isolated appear not to have been recorded 

from that vicinity and in some cases they appear to be 

previously undescribed. There must be teleomorphs 

somewhere? A number of xylariaceous taxa have been 

found to be associated with mangrove ecosystems and 

apparently even marine situations. It is proposed that host, 

habitat and climate all interact to dictate which taxa are 

present and that a knowledge of these conditions should 

prove useful in detecting rare and new species. 

156 - Rosellinia morphology: the first step towards 

molecular studies 

L. Petrini 

Scientific Advisor, Tèra d'sott 5, CH-6949 Comano, 

Switzerland. - E-mail: lpetrini@swissonline.ch


The genus Rosellinia is cosmopolitan in its distribution and 

is reported from hardwoods, dicotyledonous plants, 

coniferous litter and occasionally from monocotyledonous 

hosts. The stromata are usually uniperitheciate and 

embedded in a persistent or evanescent subiculum. Species 

of Rosellinia can be characterised by ascospore shape, 

combined with stromatal, subicular and anamorphic 

characters. About 70 taxa are validly described, belonging 

roughly to six spore shape groups. (1) R. necatrix-type 

spores are 30-120 µm long, slender, with short germ slits 

and tapering ends surrounded by a slimy sheath. (2) Five 

taxa possess ellipsoidal spores. Taxa belonging to groups 

1-2 are associated with a Dematophora anamorph. (3) The 

third group is characterized by 60-120 µm long, broadly 

rounded ascospores with a long germ slit, slimy sheath or 

caps. (4) In the fourth group ascospores range from 25-50 

µm, are ellipsoidal, with a long germ slit and one or two 

cellular, semiglobose appendages and slimy sheath. (5) 

About five taxa possess ascospores with two large, conical, 

cellular appendages. (6) The largest and most 

heterogeneous group includes approximately 40 taxa and 

has 10-60 µm long, ellipsoidal ascospores with long, short 

or without germ slit, with slimy caps or sheath, 

occasionally with a cellular appendage. The subiculum is 

evanescent. Groups 3-6 are characterised by a 

Geniculosporium anamorph. 

157 - Interesting xylariaceous fungi from Taiwan 

Y.-M. Ju 1* , J.D. Rogers 2 & H.-M. Hsieh 1 

1 Institute of Botany, Academia Sinica, Nankang, Taipei, 

Taiwan 115, Taiwan. - 2 Department of Plant Pathology, 

Washington State University, Pullman, WA 99164-6430, 

U.S.A. - E-mail: yumingju@gate.sinica.edu.tw 

Results from cultural and anamorphic studies on some 

recently collected xylariaceous fungi from Taiwan are 

highly interesting and are valuable in settling taxonomic 

problems related to these particular taxa. Penzigia 

cranioides is the type species of the genus Penzigia. The 

Xylocoremium anamorph produced in culture indicates that 

P. cranioides is in fact a Xylaria species and Penzigia is 

considered in synonymy with Xylaria. Poronia pileiformis 

had been reported to have an anamorph typical of Xylaria 

and could be accommodated in the genus Podosordaria. 

Our Taiwan collection, however, produced a Lindquistia 

anamorph; we thus feel justified in accepting the species in 

Poronia. A new Theissenia species produced elongated 

conidia which were sympodially borne on conidiogenous 

cells. This fungus has bipartite stromata and evanescent 

asci lacking an apical ring. It may represent an interface 

between Xylariaceae and Diatrypaceae. We accept two 

sections in the genus Hypoxylon: sections Hypoxylon and 

Annulata. Hypoxylon ravidoroseum belongs to section 

Hypoxylon of the genus Hypoxylon. One collection is 

peculiar in having a thickening on the ascospore perispore 

and in having both papillate and umbilicate ostiolar 

openings. A closer examination revealed that the 'papillae' 

of the papillate ostiolar openings are indeed homologous to 

the overlying stromatal tissue of ostiolar discs in 

Hypoxylon section Annulata which are pushed away during 

perithecial development. 

158 - New evidence on the taxonomy and the secondary 

metabolism of Daldinia and Hypoxylon 

M. Stadler 

Bayer AG Pharma Research PH-R EU LSC NP, P.O.B. 

101709-42096 Wuppertal, Germany. - E-mail: 

marc.stadler@t-online.de 

During the past years, we evaluated the taxonomy of 

Daldinia Ces & De Not. was by a polyphasic taxonomic 

approach, using classical morphological studies of 

teleomorphic and anamorphic characters in combination 

with scanning electron microscopy, HPLC-based 

secondary metabolite profiling and different methods of 

PCR fingerprinting. Several new species were discovered, 

and some new chemotaxonomically relevant secondary 

metabolites were identified. Once the evaluation of most 

European Daldinia species had been accomplished, our 

studies were meanwhile extended to tropical Daldinia 

species and to those members of the allied genus 

Hypoxylon Bull. that occur in temperate climates. Besides a 

short review of previous work, recent results on the 

taxonomy of Daldinia and novel metabolites from 

Hypoxylon will be presented. References: [1] Stadler, M. et 

al. 2001. Mycotaxon 77, 379-429. [2] Stadler, M. et al. 

2001. Phytochemistry 56, 787-793. [3] Wollweber, H. & 

Stadler, M. 2001. Z. Mykol. 67, 3-53. [4] Stadler, M., et al. 

2001. Mycotaxon 80, 167-177. [5] Stadler, M., 

Baumgartner, M. & Wollweber, H. 2001. Mycotaxon 80, 

179-196. [6] Stadler, M. et al. 2001. Mycological Research, 

105, 1191-1205. [7] Stadler, M. et al. 2002. Mycological 

Progress 1, 32-41. [8] Mühlbauer, A. et al. 2002. Submitted 

to Mycological Progress. 

159 - Transposable elements in F. oxysporum: impact 

on gene expression, genome organization and evolution 

M.J. Daboussi 

Université Paris-Sud, IGM, Bat400, 91405 Orsay cedex, 

France. - E-mail: daboussi@igmors.u-psud.fr 

The F. oxysporum genome harbors diverse representatives 

of most known families of transposable elements (TEs). 

Different active transposons have been identified 

determining a broad spectrum of mutations. The TEmediated 

changes range from changes in the size and the 

arrangement of the whole genome to changes in single 

nucleotides. In addition, some TE-induced mutations, such 

as read-out transcription, or local rearrangements due to 

aberrant transposition process may be of evolutionary 

significance to their host by bringing new regulatory 

elements in proximity to exonic sequences. Analysis of the 

distribution of some elements in natural populations 

revealed that their dynamics within and between species 

appeared to be the result of different evolutionary 

processes: vertical transmission, stochastic loss, 

inactivation via a RIP-like mechanism, burst of 

amplification, and horizontal transmission. These 



investigations showed the capacity of TEs to restructure the 

F. oxysporum genome in interesting ways and their 

potential as valuable tools for tracing populations. 

160 - Fusarium species big and small, and the interface 

between systematics and population genetics 

D.M. Geiser 

Penn State University, Department of Plant Pathology, 

University Park, PA 16802, U.S.A. 

Multilocus phylogenetic approaches have revolutionized 

our view of species in fungi, particularly in the genus 

Fusarium. Using these approaches, phylogenetic units can 

be identified that correspond to coherently defined species, 

within which varying degrees of phylogenetic structure are 

observed. Deciding where to draw the line between species 

and subspecific lineages requires consideration of a variety 

of phenotypic, biogeographic and ecological factors. In this 

talk, I will compare and contrast several fusaria that have 

been well-characterized phylogenetically, discussing how 

species are usefully delimited and what we know about 

genetic structure within those species. Case studies will 

include Fusarium avenaceum (little phylogenetic 

structure), F. hostae (some phylogenetic structure), F. 

proliferatum (much phylogenetic structure) and species 

lineages of the F. solani complex (varying levels of 

phylogenetic structure). As has been demonstrated in other 

groups of fungi, phylogenetic partitions within species may 

be useful as landmarks for defining the boundaries within 

which population genetic processes occur in Fusarium. The 

DNA sequence databases of Fusarium species that have 

emerged from these studies are also extremely useful as 

tools for identification. 

161 - Phylogenetic analysis of Fusarium based on 

partial TEF-1α gene sequences 

R. Kristensen * , M. Torp, A.K. Knutsen, B. Kosiak & A. 

Holst-Jensen 

National Veterinary Institute, Ullevålsveien 68, P.O.Box 

8156 Dep., 0033 Oslo, Norway. - E-mail: 

ralf.kristensen@vetinst.no 

In an attempt to develop rapid molecular detection methods 

for economically important mycotoxin producing Fusarium 

species, we have sequenced a part of the translation 

elongation factor 1α (TEF-1α) gene of more than 13 

species of the sections Sporotrichiella, 

Roseum/Arthrosporiella, Gibbosum and Discolor. This 

gene was chosen because it was expected to provide a high 

degree of phylogenetic signal and because the gene 

phylogeny and the taxon phylogeny in related taxa has 

been reported to be highly concordant. Our sequence data 

were combined with additional data retrieved from 

sequence databanks, aligned and subjected to phylogenetic 

analyses (PAUP*). The resulting phylogeny largely 

52 


supports the subdivision of the taxa into the four sections, 

but a few exceptions were observed indicating that the 

sections Sporotrichiella and Discolor are polyphyletic. The 

taxa representing the section Roseum/Arthrosporiella were 

found to form a monophyletic clade, but our results suggest 

that it may be more appropriate to include some of the taxa 

currently classified in the sections Sporotrichiella and 

Discolor in section Roseum/Arthrosporiella. We found a 

high degree of concordance between the clades and taxa 

known to produce DON/nivalenol and T-2/HT-2 

mycotoxins. The delimitation of taxa is discussed in light 

of the observed phylogeny. 

162 - Evolution of the Fusarium graminearum species 

complex 

K. O'Donnell * & T. Ward 

USDA-ARS-NCAUR, 1815 N. University, Peoria, IL 61604, 

U.S.A. - E-mail: kodonnell@mail.ncaur.usda.gov 

Fusarium head blight or scab of cereals is one of the most 

devastating plant diseases worldwide. These pathogens 

cause significant reduction in seed quality and yields and 

often contaminate seeds with trichothecene and estrogenic 

mycotoxins. Genealogical concordance phylogenetic 

species recognition was used to investigate species limits 

within this complex in order to understand the host range, 

geographic distribution and mycotoxin potential of these 

fusaria. Because trichothecene chemotypes are not well 

correlated with the species phylogeny, we investigated the 

evolutionary history of the trichothecene mycotoxin gene 

cluster. Results of these phylogenetic analyses 

demonstrated that polymorphism within these virulenceassociated 

genes is trans-specific and appears to have been 

maintained by balancing selection acting on chemotype 

differences that originated in the ancestor of this important 

group of plant pathogens. 

163 - Medicinal mushrooms: Past, present and future 

S.P. Wasser 

Institute of Evolution, University of Haifa, Mt. Carmel, 

Haifa 31905, Israel. - E-mail: 

spwasser@research.haifa.ac.il 

Medicinal mushrooms have been known in Oriental 

medicine for hundreds of years as beneficial for health. In 

2001, the value of world mushroom production and 

medicinal mushroom products was estimated to be worth 

approximately 18 billion US dollars. Mushrooms make up 

a vast and yet largely untapped source of powerful new 

pharmaceutical products. Many if not all Basidiomycetes 

mushrooms contain biologically active polysaccharides in 

fruit bodies, cultured mycelium, and culture broth. The data 

about mushroom polysaccharides are summarized for 651. 

These polysaccharides are of different chemical 

composition; the main ones comprise the group of β-


glucans. β-(1->3) linkages in the main chain of the glucan 

and further β-(1->6) branch points are needed for their 

antitumor action. Most of the antitumor clinical evidence is 

from commercial polysaccharides lentinan, PSK (krestin), 

and schizophyllan. Mushroom polysaccharides prevent 

oncogenesis, show direct antitumor activity against various 

allogeneic and syngeneic tumors, and prevent tumor 

metastasis. Cultivation and development of edible and 

medicinal mushrooms can positively generate equitable 

economic growth that had already an impact at national and 

regional levels. This impact is expected to continue 

increasing and expanding in the 21st century Therefore, 

sustainable research and development of mushroom 

production and mushroom product can become a "nongreen 

revolution" (Chang, 1999). 

164 - Beneficial effects of edible mushrooms in Japan 

T. Ikekawa 

Japanese Association for Integrative Medicine(JAIM), 

Sanshin Bldg 2-15-14, Uchikanda, Chiyoda-ku, Tokyo 101- 

0047, Japan. - E-mail: info@asahimedical.net 

The aqueous extracts of edible mushrooms showed so high 

growth-inhibitory activity aganist solid sarcoma 180. 

Among many antitumor polysaccharides, a glucan from 

Lentinus edodes was used clinically as an injection of a 

narrow adaptation in Japan. However, substances having 

the antitumor activity by p.o. were a low molecular weight 

protein- bound polysaccharide from one of the most 

popular edible mushrooms, Flammulina velutipes. An 

epidemiological study in Nagano Prefecture, Japan 

indicated that the cancer death-rate of farmers producing F. 

velutipes as a main occupation was remarkably lower than 

that of common people in the Prefecture. Another edible 

mushroom, Hypsizygus marmoreus had also high antitumor 

activity and preventive effect for tumor metastasis. Cancer 

preventive effect of the mushroom was proved by 

comparison of carcinogenesis between mice bred with an 

feed containing the dried fruitbodies and mice bred with an 

ordinary feed. The mechanism of cancer inhibitory and 

preventive activities of edible mushrooms was due to 

immunopotentiation and antioxidant activity. Thus, the 

intake of mushrooms proved to be effective in cancer 

prevention and growth-inhibition. Based on these results, 

EEM (Extracts of Edible Mushrooms), a suppliment was 

proved to have a clinical effectiveness. It has been said in 

traditional Oriental medicine from the ancient times that 

Medicine and Food have the same origin. This is identical 

with our conclusion from mushroom-studies. 

165 - Identification of species and strains of medicinal 

mushrooms using repetitive elements 

A.S.M. Sonnenberg * , J.J.P. Baars & P.M. Hendrickx 

Applied Plant Research, P.O. Box 6042 5960 AA, Horst, 

The Netherlands. – E-mail: 

A.S.M.Sonnenberg@PPO.DLO.NL 

A comparison of a previously isolated retrotransposon of 

the gypsy-family from the white button mushroom 

Agaricus bisporus with published retrotransposons of the 

matsutake mushroom (Tricholoma) showed a high 

similarity in translated sequence of the reverse transcriptase 

gene. Based on conserved sequences degenerate primers 

were designed and used to amplify similar sequences in a 

number of gilled mushrooms that are known for their 

medicinal potentials. From strains of species examined 

(Pleurotus ostreatus, Agaricus subrufescens, A. blazei, 

Stropharia rugoso-anulata, Ganoderma lucidum), 

fragments were amplified with the expected length. 

Sequence analyses showed that all fragments indeed 

represented the relevant part of the reverse transcriptase 

gene of retrotransposons. Southern analyses showed that 

most clones hybridized only to the DNA of the species 

from which the clone was isolated, thus provide speciesspecific 

probes. Transposons were subsequently used to 

study relationships between strains within a species. Using 

transposon-tagging to map individual transposon copies, 

phenograms were constructed of a collection of wild strains 

of A. bisporus. The relationships found within this 

collection agree with previously constructed phenograms 

using mitochondrial DNA and RFLP markers. This study 

shows that retrotransposons are, in principle, useful to 

identify mushroom species and individual strains within 

species. 

166 - Standard methods for the determination of 

immunomodulatory properties of medicinal 

mushrooms 

L.J.L.D. Van Griensven 1* , H.F.J. Savelkoul 2 & D.E.A. 

Florack 1 

1 Plant Research International, POB 16, 6700 AA 

Wageningen, The Netherlands. - 2 Immunology & Cell 

Biology, Dept. Animal Sciences, Wageningen UR, The 

Netherlands. - E-mail: l.j.l.d.vangriensven@plant.wagur.nl 

A variety of edible fungi have been used in the past 

hundreds of years to prevent or cure a variety of diseases, 

e.g. various malignancies, infectious diseases, 

cardiovasculair diseases, hypertension and even stomach 

ache after serious drinking. Many serious studies were 

carried out mostly in China and Japan into the effects of 

mushroom derived polysaccharides, i.e. beta-glucans. on 

solid tumor growth. Ganoderma lucidum, Lentinus edodes, 

Pleurotus spp. and many others have been described as 

potentially anti-tumorigenic based on changes of in vivo or 

in vitro transplantability of laboratory tumor cell lines in 

mice. In recent years studies have been carried out into 

immunomodulation as the basic mechanism behind those 

anti-tumor effects. However, comparison of the outcome of 

the different experiments is almost impossible 1. due to 

inadequate identification procedures of the fungal species 

and strains used and 2. due to different methods of 

measuring immune activity. The present contribution aims 

to internationally standardize methods employed 1. in 

isolation and characterization of fungal beta-glucans, 2. 

measuring modulation of both innate and adaptive mouse 



resp. human immunity in vitro and ex vivo. Reliable 

methods will be proposed. 

167 - Genoprotective activities of mushrooms 

Y.L Shi 1 , I.F.F. Benzie 2 & J.A. Buswell 1* 

1 Department of Biology, The Chinese University of Hong 

Kong, Shatin, New Territories, Hong Kong SAR, China. - 

2 Department of Nursing & Health Sciences, The 

Polytechnic University of Hong Kong, Kowloon, Hong 

Kong SAR, China. - E-mail: jabuswell@cuhk.edu.hk 

Selected mushroom species have been examined as a 

source of natural antioxidants for offsetting the adverse 

effects of 'reactive oxygen species' linked to numerous 

pathological conditions including coronary heart disease, 

various types of cancer and neurodegenerative conditions. 

Aqueous extracts of the sporophores of Agaricus bisporus, 

Auricularia auricula, Flammulina velutipes, Hypsizygus 

marmoreus, Ganoderma lucidum, Lentinula edodes, 

Pleurotus sajor-caju and Volvariella volvacea were 

assessed for ability to prevent H 2O 2-induced oxidative 

damage to cellular DNA using the Comet assay. 

Genoprotective effects were observed with cold (20 °C) 

and hot (100 °C) water extracts of A. bisporus and G. 

lucidum fruit bodies but not with the other mushroom 

species tested. The genoprotective effect of A. bisporus is 

associated with a heat-labile protein present in the 

mushroom fruit body. This protein has been purified and 

identified as tyrosinase on the basis of its catalytic and 

electrophoretic properties, and inhibition studies. 

Genoprotection is dependent upon the enzymic 

hydroxylation of tyrosine to L-DOPA and subsequent 

conversion of this metabolite to dopaquinone and other 

oxidation products that enhance the oxidation status of the 

cell. These findings indicate that some edible mushrooms 

represent a valuable source of biologically active 

compounds with potential for protecting cellular DNA 

from oxidative damage. 

168 - Lentinan as a possible "oral vaccine" against 

tumor development 

A.-T. Yap & M.-L. Ng * 

National University of Singapore, Department of 

Microbiology, 5 Science Drive 2, Faculty of Medicine, 

Singapore 117597, Singapore. - E-mail: 

micngml@nus.edu.sg 

Lentinan, a purified b-glucan isolated from Lentinus 

edodes has remarkable anti-tumor and anti-viral activities 

in animal models when given through intravenous or intraperitoneal 

injection. We investigated the efficacy of oral 

administration of lentinan using the AKR mice model. 

Results obtained showed that pre-feeding the mice for 7 

days at an optimum dose of 3mg/mouse was most effective 

against tumor induction. A tumor inhibition rate of 94.44% 

54 


was observed. The mice remained tumor-free several 

months after this treatment. In addition to feeding, passive 

transfer of lentinan-primed lymphocytes from AKR mice 

was able to restrict tumor development in immunodeficient 

mice. Lymphocytes removed from the spleens of 

either lentinan-fed or buffer-fed AKR mice were inoculated 

simultaneous with human colon carcinoma cells into both 

the nude mice (athymic mice) and SCID (severe combined 

immuno-deficient) mice. Large tumors developed in mice 

that received lymphocytes from the buffer-fed AKR mice 

whereas very small or no tumors were found in mice 

receiving lymphocytes from the lentinan-fed AKR mice. 

Electron microscopy studies and TUNEL assays indicated 

that the number of tumor cells was reduced by the host 

immune system through apoptosis. DNA fragmentation, 

increment in proteases' activities and cytokine production 

were also observed. Tumor cells isolated from lentinan-fed 

mice displayed higher levels of caspase 3 and caspase 8 as 

compared to that of the buffered-fed control mice. 

169 - Application of inhibition ELISA using antibodies 

to antitumor polysaccharides in screening of mushroom 

possessing immunomodulating activities 

K. Minato 1* , S. Kawakami 2 , T. Hashimoto 2 , H. Ashida 2 & 

M. Mizuno 2 

1 Miyagi Agricultural College, Sendai 982-0215, Japan. - 

2 Graduate School of Science and Technology, Kobe 

University, Kobe 657-8501, Japan. - E-mail: 

minato@miyanou.ac.jp 

Mushrooms have recently become attractive as health 

beneficent foods, and as a source material for the 

development of drugs. Many investigators have isolated 

and identified antitumor polysaccharides from edible 

mushrooms. We have attempted to quantify exactly and 

easily the antitumor polysaccharide in mushrooms by 

enzyme linked immunosorbent assay (ELISA). And, the 

inhibition assay of ELISA using antibodies to certain 

polysaccharides has been established to determine the 

contents of antitumor polysaccharides. It has been 

recognized that lentinan from Lentinula edodes and βglucan 

from Grifola frondosa (GGF) were the antitumor 

polysaccharides possessing strong immunomodulating 

activities. In this study, we applied the inhibition assay of 

ELISA using antibodies against lentinan and GGF in order 

to screen the mushrooms containing a high level of these 

antitumor polysaccharides. And, the relationship between 

their contents and immunomodulating effect of each 

mushroom on cytokine production, such as tumor necrosis 

factor-α (TNF-α), and nitric oxide (NO), was investigated. 

It was ascertained that Lentinula edodes, Flammulina 

velutipes, Grifola frondosa, and Meripilus giganteus 

containing the positive polysaccharides to anti-lentinan and 

anti-GGF antibodies at high level augmented the 

production of TNF-α and NO from mouse peritoneal 

macrophages. These results showed that an inhibition assay 

of ELISA was a powerful approach to determine of 

antitumor polysaccharides in mushrooms.


170 - TNF-α and NO production from macrophages is 

enhanced through up-regulation of NF-κB by 

polysaccharides purified from Agaricus blazei Murrill 

S. Kawakami 1* , K. Minato 2 , T. Hashimoto 1 , H. Ashida 3 & 

M. Mizuno 1 

1 

Graduate School of Science and Technology, Kobe 

University, Kobe, 657-8501, Japan. - 2 Miyagi Agricultural 

3 

College, Sendai, 982-0215, Japan. - Faculty of 

Agriculture, Kobe University, Kobe, 657-8501, Japan. - Email: 

ksachiko@ans.kobe-u.ac.jp 

The Basidiomycete fungus Agaricus blazei Murrill has 

medically been used as a health food for the prevention of 

cancer. We chromatographically purified polysaccharide 

from alkaline-soluble fraction of A. blazei, which showed 

the highest activity of TNF-α and NO production from 

macrophage cell line, RAW264.7. The result of time course 

experiments on TNF-α and NO production from 

RAW264.7 stimulated with the purified polysaccharide 

was indicated that TNF-α was produced 5 h earlier than 

NO. To investigate the effect of the purified polysaccharide 

on mRNA expression of TNF-α and iNOS, the level of 

mRNA was evaluated by RT-PCR. The levels of TNF-α 

mRNA was induced at 0.5 h after stimulation, peaked at 3 

h. On the other hand, those of iNOS mRNA was induced at 

3 h and thereafter maintained the same levels. Furthermore, 

DNA binding activity of NF-κB was rapidly increased even 

at 0.5h after treatment, and peaked at 2h. When RAW264.7 

was stimulated with the purified polysaccharide 

simultaneously in the presence of anti-TNF-α antibody, 

TNF-α mRNA expression did not affect and that of iNOS 

was down-regulated. Taking consideration into these 

results, it was ascertained that TNF-α production from 

RAW264.7 stimulated with the purified polysaccharide 

was enhanced through up-regulating NF-κB, and then the 

newly produced TNF-α enhanced NO production with 

autocrine pathway. Thus, the purified polysaccharide 

possessed the immunomodulating activity and might have 

anti-tumor activity. 

171 - Synergistic molecular interactions in the 

Trichoderma biocontrol system 

M. Lorito * , M. Ruocco, D. Piacenti, R. Ciliento, S. 

Lanzuise, V. Scala, A. Zoina, F. Scala & S. Woo 

Dept. ARBOPAVE- Plant Pathology, University of Naples, 

Via Università, 100, 80055 Portici (NA), Italy. - E-mail: 

lorito@unina.it 

Recent studies have shown that the antagonist mechanism 

of Trichoderma spp. is much more complex than expected 

because of the numerous factors involved and the variety of 

mechanisms activated during the interaction with the host 

and the plant. Most of the research at a molecular level has 

been focused on the role of fungal cell wall degrading 

enzymes and antibiotics in the mycoparasitic relationship 

with phytopathogenic fungi. Pathogenicity factors have 

been identified among chitinases and glucanases, and from 

this, several opportunities have arisen for developing new 

biotechnologies for disease control. We have investigated 

the interaction between enzymes and antibiotics and found 

that Trichoderma applies these factors in synergistic 

combinations during attack. Synergism has also been found 

among different types of enzymes, and when the encoding 

genes are transgenically expressed in plants or other 

biocontrol microbes. Also synergisms appears to be the 

mode that some chitinases and glucanases interact in 

releasing mycoparasitism inducers from the host cell wall 

during the early recognition phase. Finally, a high level of 

synergism has also been found when Trichoderma or its 

antifungal compounds (enzymes or antibiotics) are applied 

in combination with Pseudomonas spp. and other 

biocontrol agents. We show that functional synergism 

between differently acting compounds is critical in 

biocontrol and is a key to understand mycoparasitic 

mechanisms. 

172 - Chitinases and plant induced systemic resistance: 

new insights in T. asperellum (T. harzianum 203) 

A. Viterbo 1* , M. Shoresh 2 , I. Yedidia 2 , O. Ramot 2 & I. 

Chet 1 

1 Weizmann Institute, Dept. of Biological Chemistry, 

Rehovot, 76100, Israel. - 2 Hebrew University, Otto 

Warburg Center, Rehovot, 76100, Israel. 

Two main modes of action can be discerned in the 

biocontrol mechanism of some Trichoderma species: 

mycoparasitism and plant systemic induced resistance. In 

order to better understand the role of chitinases in 

mycoparasitism of T. asperellum, we have studied 

regulation of the expression of the endochitinase chit36 and 

the N-acetylglucosaminidases coding for Chit73 and 

Chit102 using Northern blots and a promoter-GFP reporter 

system. Despite their close sequence similarity, chit73 and 

chit102 are differentially regulated under stress conditions 

and by chitin degradation products. chit36 expression is upregulated 

in vivo during pathogen confrontation. Induced 

plant resistance by Trichoderma was studied using a 

spatially separated challenge system and real-time PCR. 

We could show that the ethylene and jasmonic acid 

signaling pathway are triggered in cucumber roots 

inoculated with T. asperellum. As the result of this 

different PR proteins and antibacterial phenolic compounds 

are systemically induced in the leaves of the plant. These 

preliminary data provide initial insights at the molecular 

level on the Trichoderma-plant interaction. 



173 - Role of β-1,3-glucanase from Trichoderma 

harzianum cect 2413 in antagonism against 

phytopathogenic fungi 

A.M. Rincón & T. Benítez * 

Departamento de Genética, Facultad de Biología, 

Universidad de SevillaUniversidad de Sevilla, Avenida 

Reina Mercedes s/n, Spain. - E-mail: tahia@us.es 

T. harzianum CECT 2413 grown under conditions of 

simulated mycoparasitism secreted several β-1,3glucanases; 

the most abundant, BGN13.1, was purified and 

the corresponding gene, bgn13.1 was isolated. bgn13.1 is 

strongly induced by chitin, autoclaved mycelia or plant 

extracts and is only expressed at very narrow pH ranges 

(around 5.5). There is a weak expression of the gene under 

carbon starvation, and under some stress conditions. The 

promoter sequence has putative boxes which respond to 

stress, carbon (but not nitrogen) starvation, regulation by 

pH and regulation under mycoparasitism (MYC). The 

presence of MYC boxes in the promoter and the gene 

induction by chitin strongly suggests that BGN13.1 is 

involved in antagonism. BGN13.1 was overexpressed in T. 

harzianum and tobacco plants. T. harzianum transformants 

were more aggressive than the wild type in in vitro assays 

against Botrytis, Phytophthora and Rhizoctonia but 

maximal growth inhibition did not correlate with maximal 

levels of β-1,3-glucanase activity. Combination of 

BGN13.1 transformants with transformants which 

overexpress other enzymes showed synergistic effects 

against fungal strains, but the efficiency of both single 

transformants or their mixture vary according to the 

antagonized strain. Plant protection experiments with 

different Rhizoctonia strains, T. harzianum and tobacco 

plants, both wild types and BGN13.1 transformants are 

currently in progress. 

174 - Signaling cascades involved in the mycoparasitic 

process by Trichoderma 

A. Herrera-Estrella 1* , A. Mendoza-Mendoza 1 , V. Olmedo- 

Monfil 1 , I. Chet 2 & B. Horwitz 3 

1 Cinvestav, Km 9.6 libramiento Norte Carretera 

Irapuato/León. AP 629. Irapuato, Gto., Mexico. - 2 The 

Hebrew University of Jerusalem, P.O. Box 12, Rehovot 

76100, Israel. - 3 Technion - Israel Institute of Technology, 

Haifa 32000, Israel. - E-mail: aherrera@ira.cinvestav.mx 

In the presence of a fungal host T. atroviride produces 

hydrolytic enzymes and coils around the host hyphae. In a 

biomimetic assay, different lectins induce coiling around 

nylon fibers; coiling in the absence of lectins can be 

induced by applying cAMP or the heterotrimeric G protein 

activator mastoparan. We isolated a T. atroviride G alpha 

subunit gene (tga1). Transgenic lines generated that 

overexpress the Ga protein show very delayed sporulation 

and coil at higher frequency. Furthermore, transgenic lines 

that express an activated mutant protein with no GTPase 

56 


activity do not sporulate and coil at a higher frequency. 

Lines that express an antisense version of the gene are 

hypersporulating and coil at much lower frequency in the 

biomimetic assay. Loss of Tga1 in these mutants correlates 

with the loss of GTPase activity stimulated by the peptide 

toxin mas-7. In contrast, cAMP levels and cAMPdependent 

protein kinase activity drop when diffusible host 

signals are encountered and the mycoparasitism related 

genes ech42 and prb1 are highly expressed. Herewith we 

show that the prb1 is subjected to Nitrogen Catabolic 

Repression. Accordingly, induction of prb1 transcription 

by Rhizoctonia solani cell walls requires the release from a 

repressed condition, which is determined by nitrogen 

availability. prb1 transcription in response to nitrogen 

limitation, may be mediated by a MAPK signaling cascade. 

175 - Components involved in regulation and signaling 

of the mycoparasitic response of Trichoderma atroviride 

S. Zeilinger 1 , C.K. Peterbauer 2 , K. Brunner 1 , V. Seidl 1 , 

R.L. Mach 1 & C.P. Kubicek 1* 

1 Microbial Biochemistry and gene Technology Group, 

Institute of Chemical Engineering, TU Wien, 

Getreidemarkt 9/166 A-1060 Wien, Austria. - 2 Institute of 

Biochemical Technology and Food Technology, University 

of Agriculture, Muthgasse 11, A-1190 Wien, Austria. - Email: 

ckubicek@mail.zserv.tuwien.ac.at 

T. atroviride (former T. harzianum) strain P1 acts as a 

mycoparasite and is applied as biological control agent 

against a number of plant pathogenic fungi. The 

mycoparasitic interaction is host-specific and not merely a 

contact response. It is thus likely that signals from the host 

are recognised and provoke antifungal activities which are 

accompanied by morphological changes and the secretion 

of hydrolytic enzymes. To understand the signalling 

mechanisms involved in this process, we have begun to 

investigate the cis- and trans-acting factors controlling 

expression of the chitinases, using nag1 as a model because 

its disruption leads to a complete loss of expression of all 

chitinases. Three motifs were detected and characterized in 

vivo and in vitro. A gene (seb1) encoding a protein binding 

to one of these motifs (a CCCCT-box) was cloned from T. 

atroviride and its biochemical and functional 

characterization will be reported. Further, towards an 

elucidation of the link between chitinase gene expression 

and the mycoparasitic response, we have cloned key 

components of the cAMP and MAP kinase signaling 

pathways in T. atroviride and started their functional 

characterization by gene disruption. First results from this 

will be presented.


176 - Taxonomic perspectives on some ascomycetous 

endophytes of mycorrhizas 

R.S. Currah 

Department of Biological Sciences, University of Alberta, 

Edmonton, Alberta, T6G 2E9, Canada. - E-mail: 

r.currah@ualberta.ca 

Roots are habitat for a wide variety of fungi including a 

large number of ascomycetes. Several groups of 

ascomycetous anamorphs consistently appear during 

isolation protocols using traditional culturing methods or 

where diversity estimates are made using DNA analyses. 

Among the taxa recovered, Phialocephala fortinii, 

Leptodontidium orchidicola, Scytalidium vaccinii and 

similar species (teleomorphs apothecial or suspected as 

such), Oidiodendron spp., and Geomyces pannorus 

(teleomorphs cleistothecial) are frequent Leotiomycetes. 

Other taxa include Heteroconium chaetospira and species 

of Phialophora, Cladosporium, Phoma, and large numbers 

of unidentified or partially characterized sterile taxa. In 

most cases, the nature of the association between these 

fungi and the host plant roots is unknown but their well 

developed saprobic abilities suggest their role as mutualists 

with plants is probably not mycorrhizal in the usual sense. 

Continuing isolation work along with morphological, 

enzymatic and molecular characterization, coupled with 

resynthesis studies with host plants, will lead to a clearer 

understanding of the niches occupied by these fungi in 

roots and mycorrhizas. 

177 - Non-mycorrhizal root endophytes - aspects on 

their ecology 

A. Jumpponen 

Kansas State University, Division of Biology, Manhattan, 

KS66506, U.S.A. - E-mail: ari@ksu.edu 

Non-mycorrhizal root endophytes are a heterogeneous 

group of fungi with possible beneficial associations with 

their host plants. Recent data from a primary successional 

sub-alpine ecosystem and a secondary successional 

temperate grassland ecosystem suggest greater abundance 

of non-mycorrhizal endophytes than mycorrhizal fungi. 

The abundance of mycorrhizal and non-mycorrhizal, rootinhabiting 

fungi may fluctuate according to physiology or 

phenology of the host. Two competing hypotheses are 

presented: 1) these fungi are parasitic and tap into the host 

photosynthate translocation during seasonal changes in 

host physiological activity, 2) these fungi are actively 

involved in controlling host photosynthate reallocation. To 

identify the dominant fungi in rhizosphere, a shotgun 

cloning experiment was conducted using environmental 

DNA from soil and rhizosphere samples. Fungi in the 

shotgun cloned rDNA libraries represented ascomycetes, 

basidiomycetes, chytridiomycetes and zygomycetes. 

Detected fungal communities were concluded to be very 

similar in soil and rhizosphere. This is interpreted to 

indicate that root-inhabiting fungi frequently possess 

extramatrical hyphae extending to the soil. Alternatively, 

large proportion of the root and rhizosphere inhabiting 

fungi are common soil fungi and facultatively colonize host 

tissues. The potential of soil-inhabiting fungi having 

biotrophic and/or endophytic phases will be discussed. 

178 - Ectendomycorrhizas - specialized associations 

between members of the Pinaceae and ascomycete fungi 

R.L. Peterson 

University of Guelph, Department of Botany, Guelph, 

Ontario, Canada. - E-mail: lpeterso@uoguelph.ca 

Ectendomycorrhizas (ECM) are formed in two genera, 

Pinus and Larix, when roots are colonized by a few 

ascomycetous fungal species, primarily Wilcoxina mikolae 

and Wilcoxina rehmii. Two dematiaceous species and 

Sphaerosporella brunnea have also been reported to form 

ECM with these genera. Some of these fungi are capable of 

forming typical ectomycorrhizas with other conifer genera 

and with angiosperm species. Reports of other genera of 

conifers and angiosperms possessing ECM are based on 

field collections of roots of unknown age and thus may 

represent senescing ectomycorrhizas. ECM are common in 

Pinus and Larix seedlings in nurseries, regenerated 

seedlings on degraded sites, and in some natural forest 

sites. The identifying structural characteristics of ECM are 

a limited mantle, an Hartig net, and intracellular hyphae 

forming hyphal complexes. The latter form soon after 

Hartig net development. Intracellular hyphal complexes 

occur in epidermal and cortical cells and are surrounded by 

host-derived plasma membrane and an interfacial matrix of 

unknown composition. Although ultrastructural features of 

the fungus-root cell interface suggest that these may be 

sites of nutrient exchange, there is no evidence for this. It is 

known that some of the fungi involved are able to 

breakdown complex carbohydrates but carbon transfer to 

young seedlings has not been demonstrated. Suggestions 

have been made that ECM may be important in the 

establishment of seedlings in harsh environments. 

179 - Community structure and phylogeny of ericoid 

and epacrid mycorrhizal fungi 

S.M. Berch 1* , T.R. Allen 2 , M.L. Berbee 2 & C.B. McLean 3 

1 Ministry of Forests Research Branch Laboratory, PO 

9536, Victoria BC V8W 9C4, Canada. - 2 University of 

British Columbia, Department of Botany, Vancouver BC 

V6T 1Z4, Canada. - 3 University of Melbourne, Burnley 

College, 500 Yarra Boulevard, Richmond, Victoria 3121, 

Australia. - E-mail: shannon.berch@gems7.gov.bc.ca 

To understand the community structure of ericoid 

mycorrhizal fungi in the roots of ericaceous and 

epacridaceous plants, researchers have cultured hundreds 

of fungal isolates from thousands of mycorrhizal root 



pieces. Because many isolates do not sporulate and 

therefore cannot be identified, researchers have turned to 

molecular tools: RFLP and sequence analysis. We have 

mapped the fine-scale distribution of the fungi cultured 

from roots and found that in most cases the same fungus 

did not grow out of adjacent 2mm root segments. However, 

not all ericoid or epacrid root endophytic fungi are 

culturable. We have found two different communities of 

fungi when isolating and cultivating versus cloning PCRamplified 

DNA from the same roots. We have also 

accessed the 200+ sequences in GenBank of ericoid and 

epacrid mycorrhizal fungi and root endophytes from a 

number of research groups and carried out phylogenetic 

analyses. These analyses indicate there are only 7 ericoid 

mycorrhizal fungi or fungal groups in the parts of 3 

continents that have been studied thus far. These same 7 

fungi can also be found in the roots of a single plant on a 

site on northern Vancouver Island, British Columbia. 

180 - Ascomycetous mycorrhizae from prescribed burn 

in ponderosa pine forest from Northwest, USA 

K.E. Fujimura 1* , J.E. Smith 2 , T.R. Horton 3 , N.S. Weber 4 & 

J.W. Spatafora 5 

1 University of Northern British Columbia, 3333 University 

Way, Prince George, BC V2N 4Z9, Canada. - 2 2USDA 

Forest Service, Pacific Northwest Research Station, 

Forestry Sciences Laboratory, 3200 Jefferson Way, 

Corvallis, OR 97331, U.S.A. - 3 SUNY, 1 Forestry Dr., 

Syracuse, NY 13210, U.S.A. - 4 Oregon State University, 

Richardson Hall, Corvallis, OR 97331, U.S.A. - 5 Oregon 

State University, Dept. of Botany, Corvallis, OR 97331, 

U.S.A. - E-mail: fujimurk@unbc.ca 

Post-fire Pezizales include members that fruit as early as 

two weeks and others as late as two years after a fire. 

Despite frequent occurrence, little is known about their 

ecological function. Ponderosa pine forests have a natural 

fire regime that includes low intensity fires at a 15 to 30 

year return interval. Our objective is to determine if 

members of the post-fire Pezizales associated with 

ponderosa pine forests are mycorrhizal, which may aid in 

re-generation of these forests. Because fungi are patchily 

distributed over the landscape, and the location of 

associated mycorrhizal root tips difficult to determine, we 

collected soil cores directly beneath Pezizalean sporocarps 

from three sites that had been burned to mimic the natural 

fire regime. Root tips from the soil cores were sorted into 

morphological groups. Restriction fragment length 

polymorphism (RFLP) patterns were obtained for each 

morphological group to compare to RFLP patterns of 

collected sporocarps. Twenty three patterns were found but 

did not match any of the five genera and ten species of 

Pezizalean sporocarps collected. Sequence analysis using 

the small ITS region of the nrDNA was done for 

comparison with Ascomycetous sequences, representing 

both mycorrhizal and reputed non mycorrhizal species. 

Matches were found with Geopora spp. and Wilcoxina spp. 

58 


181 - Mycorrhiza-associated sterile fungi in a 

Mediterranean environment: systematic heterogeneity 

versus ecological convergence? 

M. Girlanda * , S. Ghignone & A.M. Luppi 

Dipartimento Biologia Vegetale dell'Università and IPP- 

CNR, V.le Mattioli 25, 10125 Torino, Italy. - E-mail: 

mariangela.girlanda@unito.it 

Mediterranean ecosystems, featuring high plant diversity 

and co-dominance, offer an interesting scenario for the 

study of root-endophytic dark sterile mycelia (DSM) in 

different hosts. Investigation of ectomycorrhizal Pinus 

halepensis and endomycorrhizal Rosmarinus officinalis 

plants in Italy indicated that neighboring healthy 

individuals of these two hosts harbor several DSM 

morphotypes. Sequence analyses of nuclear ITS and 18S 

rDNA regions of the dominant morphotypes allowed their 

identification as a unique spectrum of DSM, being quite 

diverse and distinct from those so far described in other 

host plants investigated. Recognition of systematic 

affinities was possible with varying degrees of resolution 

for the different morphotypes, ranging from species-level 

identification to placement within Ascomycete subclasses. 

Association of these fungi with both hosts spanned at least 

eleven years; they thus appear to be true root colonizers 

coexisting with the accredited mycorrhizal symbionts. In 

spite of their taxonomic diversity, such DSM displayed 

some morphophysiological convergence justifying their 

recognition as an ecological group. One morphotype was 

identified as Rhizopycnis vagum D.F. Farr, a recently 

described fungus so far known as a root pathogen of crop 

plants. Studies of population genetics of this fungus and 

pathogenicity assays are unraveling the actual range of its 

ecological plasticity. 

182 - Mutualistic colonization of roots by endophytic 

fungi 

B. Schulz 1* , A.-K. Römmert 1 , U. Dammann-Tugend 1 , M. 

Götz 2 , T. Lange 3 , M. Oros-Sichler 2 & C. Boyle 1 

1 

Institut für Mikrobiologie, Technische Universität 

Braunschweig, Spielmannstr. 7, D-38106 Braunschweig, 

Germany. - 2 Biologische Bundesanstalt, Messeweg, D- 

38104 Braunschweig, Germany. - 

3 Institut für 

Pflanzenbiologie, Technische Universität Braunschweig, 

Mendolssohnstrasse, 38106 Braunschweig, Germany. - Email: 

b.schulz@tu-bs.de 

Fungal colonization of plant roots is often mutualistic, a 

well known example being that of mycorrhiza, but also 

interactions of dark septate endophytes (DSE) and the 

mycorrhizal-like basidiomycete Piriformaspora indica 

with roots of their hosts may counteract the negative effects 

of stress or improve growth of the host. We compared 

endophytic colonizations of the roots with those of aboveground 

plant organs. Endophytic infections of Fusarium 

spp. in the shoots of bean and barley were asymptomatic,


intercellular, remained localized and had no significant 

effects on yield, carbohydrate metabolism, stress tolerance 

or induced resistance of the host. In contrast, endophytic 

colonization of the roots of bean with Fusarium sp. and 

larch with Cryptosporiopsis sp. and Phialophora sp. (a 

DSE), was extended and both inter- and intracellular and 

improved growth of the host. A culture extract of 

Phialophora sp. not only improved growth of the host, but 

also led to increased branching of the roots of the treated 

seedlings. The mutualistic effects may in part be due to the 

capability of the endophytes to synthesize IAA. We 

hypothesize that fungal colonization of the roots is more 

likely to be mutualistic than that of the above-ground 

organs: a) endophytic root colonization in contrast to that 

of the shoots is frequently systemic, b) the shoot 

synthesizes its own metabolites and c) microorganisms in 

roots are in close contact with an environment harboring 

many degradative products. 

183 - Is Asia the centre of origin for the megagenus 

Cordyceps? 

N.L. Hywel-Jones 






Cordyceps is panglobal with many anamorphs: most poorly 

known. Of its anamorphs, Beauveria, Hirsutella, 

Metarhizium, Nomuraea, Paecilomyces and Verticillium 

have been assessed as potential biocontrol agents. 

Beauveria, Metarhizium and Paecilomyces have species 

that seem to have lost the Cordyceps form, becoming 

panglobal and with wide host ranges. Significantly, all 3 

genera are linked to Cordyceps in Asia (China, Hong 

Kong, Japan, Korea and Thailand). Cordyceps of 

Beauveria and Metarhizium are from Coleoptera while that 

of Paecilomyces is from Lepidoptera. These insect orders 

have the most Cordyceps: 88 spp. from Thailand with 43 

off Coleoptera (15) and Lepidoptera (28). This is ca. 2x the 

number for N. America. Although major surveys of 

Cordyceps are known from several countries (Brazil, 

Congo, Ecuador, Ghana, Japan, Korea, N. America, Sri 

Lanka, Taiwan) only 13 of 88 species from Thailand are 

reliably named to species. Significantly, comparative 

surveys in Thailand and Japan indicate that both countries 

have a rich but disparate biodiversity. A survey of Thai 

Cordyceps has no slowing in discovery of new 

records/species: few countries enjoy such scrutiny. 

However, I posit that significant surveys from other 

countries suggest that if Cordyceps producing Beauveria, 

Metarhizium or Paecilomyces were present there should, 

by now, be records. That there are not implies an important 

role for East Asia as a possible Centre of Origin for 

Cordyceps and its anamorphs. 

184 - Changes in nutritional mode within lineages of 

insect-associated fungi 

M. Blackwell 

Biological Sciences, Louisiana State University, Baton 

Rouge, LA 70803, U.S.A. - E-mail: mblackwell@lsu.edu 

Diverse groups of insects support the growth of fungal 

parasites. They range from deadly pathogens to obligate 

biotrophs that derive nutrition without apparent harm to 

their insect hosts. Some of the biotrophs have reduced 

thallus morphology resulting in a loss of characters that 

would be useful in phylogenetics. The absence of 

morphological characters puzzled eminent mycologists 

such as Roland Thaxter, who studied the fungi. More 

recent phylogenetic studies of the fungal parasites of 

insects, including termites and beetles, support several 

independent origins of fungal associates of insects. 

Furthermore, the studies suggest that one group of 

endosymbionts arose from within a lineage of lethal 

pathogens. Other examples provide evidence that certain 

fungal biotrophs probably arose from within lineages that 

included mycoparasites with complex life histories. The 

reliance on insects to disperse mycoparasitic fungi to 

fungal hosts may have lead to the fungal host being 

bypassed with a host shift to the insect as the sole host. 

Although some hypotheses (Cope's Law), propose that 

obligate parasites are evolutionary dead ends, this may not 

always be so. 

185 - Field studies employing a recombinant 

mycoinsecticide (Metarhizium anisopliae) reveal that it 

is rhizosphere competent 

R.J. St. Leger * , G. Hu & F.M. Freimoser 

University of Maryland, Maryland, 20742, U.S.A. - E-mail: 

rl106@umail.umd.edu 

In the summer of 2000, we released genetically altered 

insect pathogenic fungi onto a plot of cabbages infested 

with Trichoplusia ni (cabbage looper caterpillars) at a field 

site in Maryland. The transformed derivatives of 

Metarhizium anisopliae ARSEF 1080, designated GPMa 

and GMa, carried the Aequorea victoria gfp gene alone 

(GMa) or with additional Pr1 protease genes (GPMa). The 

study: a) confirmed the utility of gfp for monitoring M. 

anisopliae in field populations over time; b) demonstrated 

little dissemination of transgenic strains, and produced no 

evidence of transmission by non-target insects; c) found 

that recombinant fungi were genetically stable over one 

year under field conditions, and d) determined that 

deployment of the transgenic strains did not depress the 

culturable indigenous fungi. Monitoring the fate 

(survivorship) of transformants under field conditions 

revealed that they decreased over several months, except in 

the inner rhizosphere demonstrating that rhizospheric soils 

are a potential reservoir for M. anisopliae. Overall, our 

results place a sharp focus on the biology of the soil/root 



interphase as a site where plants, insects and pathogens will 

interact to determine fungal efficacy, cycling and survival. 

However, the rhizospheric effect was less marked for 

GPMa, and overall it showed reduced persistence in soils 

compared to GMa. 

186 - Entomophthorales host-pathogen relationships: 

ecology, pathobiology and molecular characterization 

J. Eilenberg * , A.B. Jensen, C. Nielsen & L. Thomsen 

Department of Ecology, The Royal Veterinary and 

Agricultural University, Thorvaldsensvej 40, DK 1871 Frb. 

C., Denmark. - E-mail: Jei@kvl.dk 

Fungi from the order Entomophthorales are pathogenic to 

arthropods, especially insects. They are highly specialized 

and provide us with valuable insight in the development of 

host-pathogen relationships and are also potential 

biological control agents. Attention will be given to three 

host-pathogen systems, which recently were studied with 

respect to ecology, pathobiology and molecular 

characterization. 1) Entomophthora muscae complex and 

hosts from Diptera. Morphological and pathobiological 

characters allow us to discriminate between the limited 

number of recognized species, but the inclusion of 

molecular data strongly suggest that each host-pathogen 

system is independent. Each fly species thus harbors its 

own selection of genotypes 2) Strongwellsea spp. and hosts 

from Diptera. This genus disperses conidia from abdominal 

holes in the still living hosts. We have found infections in 

many dipteran families, including in the arctic. 

Morphological and pathobiological characters indicate a 

high diversity and specialization. 3) Pandora neoaphidis, 

Neozygites fresenii and Entomophthora planchoniana and 

hosts from Aphididae. These fungi are adapted to the 

holocyclic life-cycle of many host aphids. Further, a recent 

study has revealed that two clonal populations of an 

anholocyclic aphid host each harbored only one of these 

pathogens. 

187 - The occurrence and stability of the entomogenous 

hyphomycete fungi Metarhizium anisopliae and 

Beauveria bassiana 

T.M. Butt * , C.S. Wang & N. Patel 

School of Biological Sciences, University of Wales 

Swansea, Singleton Park, SA2 8PP, U.K. - E-mail: 

t.butt@swansea.ac.uk 

There is considerable interest in the development of 

entomogenous, hyphomycete fungi, such as Metarhizium 

anisopliae and Beauveria bassiana, for the control of 

arthropod pests. The successful deployment of these 

biocontrol agents will depend on a thorough understanding 

of their ecology and population dynamics. Biochemical and 

molecular markers have been developed to study: (1) the 

occurrence of indigenous strains, (2) the efficacy and fate 

60 


of released (exotic) strains, (3) impact of fungi on nontarget 

organisms, and (4) fungal stability. The latter has 

received considerable attention because it influences 

sporulation, virulence, host specificity and ecological 

fitness which, in turn, influences production costs and 

efficacy. In vitro studies show that some strains of 

entomogenous fungi are more stable than others. Some of 

the changes observed may be due to parasexual 

recombination, gene silencing or gene deletion. 

Recombinants were recovered from insects co-infected 

with two strains of either M. anisopliae or B. bassiana. 

188 - Biodiversity of thermophilic fungi 

J. Mouchacca 

Laboratoire de Cryptogamie, M.N.H.N., 12, rue Buffon, 

75005 PARIS, France. - E-mail: mouch@mnhn.fr 

Critical knowledge on the biodiversity of thermophilic 

fungi has been hindered by the proposal of several 

unwarranted nomenclatural and taxonomic decisions which 

have led to a misunderstanding of the taxonomic concepts 

of some thermophiles. The situation has been further 

aggravated by the current chaotic state in binomial citations 

of common members of the group. The systematics of 

thermophilic fungi has been recently re-appraised in a 

series of contributions which have appeared almost 30 

years after the last major monograph on their taxonomy, 

biology and economic importance (Cooney and Emerson, 

1964). To date, about 50 new names have been introduced 

at both species and varietal levels; a total almost equal to 

earlier names reported for the group. However, some of 

these introductions proved not to be justified. Indeed, some 

name changes do not appear to be supported by sound 

taxonomic arguments. Thermophilic fungi now acceptable 

as valid from a nomenclatural point of view comprise about 

40 taxa. Thus some Mucorales, several Ascomycetes and 

hyphomycetes and a single agonomycete could appear to 

be true thermophiles. However, from a taxonomic point of 

view, the status of some species still requires further study. 

Once these basic studies have been completed, they will 

guarantee that a sound database exists for thermophilic 

fungi. Such taxonomic stability will also ensure that any 

new discoveries of interest have the potential for eventual 

industrial application. 

189 - Heat and the physiology of thermophilic fungi 

B. Jensen 

Department of General MicrobiologyUniversity of 

Copenhagen, Sølvgade 83H DK-1307 Copenhagen, 

Denmark. - E-mail: boj@mermaid.molbio.ku.dk 

The thermophilic fungi compose a small ecological group 

defined solely on temperature requirement for growth, 

ranging from 20 °C to at or above 50 °C. There are only 

about 40 known species of thermophilic fungi and for only


a small number of these fungi the parameters for growth 

have been investigated. Thermophilic fungi are found in 

the environment eg compost pilesand bird nests where they 

will be exposed to different and shifting stress factors like 

high temperatures and nutritional limitations. In higher 

organisms, thermal adaption requires a concerted action at 

different levels whereas non-motile microorganisms 

exclusively have to cope with this type of changes by 

cellular mechanisms. The permissive temperature for a 

thermophilic fungus is above the heat-shock temperature 

for mesophilic fungi. This means that even the 

housekeeping proteins of the thermophiles may need some 

kind of heat-shock adaption at its optimal growth 

termperature. This implies a general heat-adaptive 

mechanism already constitutively at work in thermophiles 

compared to mesophiles. Different aspects of growth at 

elevated temperatures will be evaluated and a verification 

of maximum growth upto 62 °C of a thermophilic fungal 

strain will be presented. The stress response of different 

thermophilic fungal species/isolates will be compared in 

order to determine, whether the thermophilic fungi can be 

characterized by other criteria than the temperature range 

for growth. 

190 - Functional diversity of thermophilic fungi in 

mushroom compost 

G. Straatsma * & L.J.L.D. Van Griensven 

Applied Plant Research, Mushroom Research Unit, PO 

Box 6042, 5960 AA Horst, The Netherlands. - E-mail: 

g.straatsma@ppo.dlo.nl 

The species richness of thermophilic fungi in mushroom 

compost (and its ingredients) was 22 species. Mixed 

ingredients, starting to self heat, were species rich as was 

the alternatively surveyed substrate of pigpen litter 

(sawdust based). We assume that more species were 

present and ways of finding them will be discussed. The 

final product, mushroom compost, is an artificial, strongly 

manipulated, ecosystem. Scytalidium thermophilum 

dominates and Talaromyces thermophilus and 

Thermomyces lanuginosus can only be isolated after the 

selective killing of Scytalidium. Scytalidium's high growth 

rate and its cellulolytic ability are probably important for 

its success. Talaromyces and Thermomyces tolerate higher 

temperatures but grow relatively slow. Under mesophilic 

conditions, after inoculation with mushroom mycelium, 

Scytalidium stimulates the extension rate of growth of the 

mushroom mycelium. Also other organisms, some pure 

cultured thermophilic fungi, like Myriococcum 

thermophilum, and enrichment cultures of prokaryotes in 

compost, are stimulatory to mushroom mycelium. The 

interactions of these species with the substrate and with the 

other species present will be discussed. We will try to 

identify the key characteristics of species related to these 

interactions. Our research contributes to the general 

understanding of composting (turnover, emissions, 

stabilization) and the rational use of agricultural wastes for 

new products. 

191 - An industrialists view on thermophilic fungi as 

sources for industrial applications 

S. Landvik 

Novozymes A/S, Smoermosevej 25, 1B1.01, 2880, 

Denmark. - E-mail: salk@novozymes.com 

Many industries need enzymes that are active at, or can 

resist, high temperatures in compliance with the nature of 

the industrial processes and substrates. If the enzyme can 

be added in an existing industrial step without justification 

of temperature or pH, savings of costs and efforts can be 

done. Many bacteria and Archaea from thermal areas have 

been explored for the industry, and have provided it with 

enzymes which are thermoactive and stable at above 

boiling temperatures. Fungi can not live in as high 

temperatures, and generally do not display as thermoactive 

enzymes as some prokaryotes do. On the other hand, fungal 

enzymes are generally more acid stable than bacterial 

enzymes, which is another important factor for many 

industries. Also, fungi can produce enzymes with different 

functions than some bacterial enzymes, and are, despite 

their less thermophilic nature, of large interest for the 

industry. However, can we expect that thermophilic fungi, 

with optimum growth temperatures up to only ca 20-30 °C 

above that of non-thermophilic fungi, make enzymes that 

are more thermostable than the enzymes from these fungi? 

The general question 'Do fungal growth profiles correlate 

with enzyme characteristics' is both of academic and 

industrial interest and can be enlighted with thermorelated 

studies, as this physiological niche probably is more 

explored than any other physiological growth niche. 

192 - Settling differences: Evolution of recognition 

genes 

G. May * , S. Sherrer, A. Munkasci, A. Baumgarten & R. 

Spangler 

Dept. EEB, U. Minnesota, USA, 1987 Upper Buford Circle, 

U.S.A. - E-mail: gmay@gmay.email.umn.edu 

Mating compatibility genes, vegetative incompatibility 

genes and in plants, genes conferring resistance to 

pathogens have evolved under similar evolutionary 

pressures and consequently often display common 

genealogical patterns of diversification. Fungi sense their 

biotic environment as a series of encounters with other 

organisms. The recognition genes they carry help to 

classify such biotic interactions as same or different - same 

or genotype, species, or a compatible mate. Similarly, in 

host plant-pathogen interactions, plant fitness can be 

dependent upon recognition and response to fungal 

invaders. Because the biotic environment directly affects 

organismal fitness, genetic systems allowing recognition 

and response to other genotypes within a species and to 

other species have evolved. We will discuss our research 

results in mating genes as well as plant resistance genes in 



order to illustrate of common and different evolutionary 

themes. 

193 - Pheromone receptors of the basidiomycete 

Schizophyllum commune distinguish single amino acid 

differences among active mating pheromone ligands 

T.J. Fowler * , M.F. Mitton, E.I. Rees & C.A. Raper 

University of Vermont, Microbiology and Molecular 

Genetics Dept., 208 Stafford Hall, 95 Carrigan Drive, 

Burlington, VT 05405, U.S.A. - E-mail: 

tfowler@zoo.uvm.edu 

The linked, multigenic loci B-alpha and B-beta of the 

homobasidiomycete Schizophyllum commune contribute to 

the specification of more than 20,000 mating types. Nine 

versions each of B-alpha and B-beta have been described 

by genetic methods; only portions of a few of these 

versions have been described by molecular techniques. 

These versions carry different but related sets of genes 

encoding lipopeptide pheromones and 7-transmembranedomain 

receptors. We sequenced the linked pair B-alpha3- 

B-beta2 and we can account for all known mating activities 

of a B-alpha-B-beta complex for the first time. There are 

eleven pheromone genes and two receptor genes within the 

B-alpha3-B-beta2 complex. The amino acid sequences of 

predicted pheromones and receptors from B-alpha3-Bbeta2 

were compared with other S. commune mating 

pheromones and receptors to decipher how specificity is 

achieved in their interactions. These pheromones and 

receptors can be functionally expressed in Saccharomyces 

cerevisiae and show the same specificity as seen in S. 

commune. Using a combination of heterologous and 

homologous expression systems, we can show that single 

amino acids in some pheromones control which receptors 

(i.e. mates) are activated for pheromone response. The 

receptors likewise can be manipulated to have altered 

pheromone recognition or to be active in the absence of 

pheromone. Thus, a finely tuned set of molecules maintains 

outbreeding while prohibiting self-activation of the mating 

process. 

194 - Mating recognition in ascomycetes 

R. Debuchy 

Institut de Génétique et Microbiologie, Bâtiment 400, 

Université Paris-Sud, F-91405 Orsay, France. - E-mail: 

debuchy@igmors.u-psud.fr 

In heterothallic Euascomycetes, mating is controled by the 

mating-type alleles (idiomorphs) which allow compatible 

strains to recognize each other and to proceed to 

fertilization. Fertilization is not immediately followed by 

karyogamy. Instead, the parental nuclei divide mitotically 

several times inside the female organ and are intermingled 

in plurinucleate cells. Then nuclei from male and female 

origin, i. e. nuclei with opposite idiomorphs, recognize 

62 


each other and form dikaryotic hyphae. This step involves 

at least two key events: internuclear recognition (IR) and 

cellularization. The dikaryotic hyphae divide mitotically, 

maintaining a strict ratio of 1:1 of each parental nucleus. 

Eventually pairs of nuclei fuse and meiosis and ascospore 

formation ensue immediately. A major challenge is to 

understand the relations between the idiomorphs and the 

developmental events of the sexual cycle. Podospora 

anserina is used as a model system for investigating the 

functions of its mat+ and mat- idiomorphs and the 

molecular mechanism of IR. The mat+ sequence contains 

one gene, FPR1 and the mat- sequence contains three 

genes: FMR1, SMR1 and SMR2. FPR1, FMR1 and SMR2 

encode transcriptional factors which control fertilization 

and IR. A model, based on genetics, has been proposed for 

the control of IR by the mat genes and will be presented. IR 

is followed by a developmental arrest which is overcome 

by SMR1. The current hypotheses about the molecular 

mechanism of IR will be discussed. 

195 - Gene deletion and unidirectional mating type 

switching in Ceratocystis 

T.C. Harrington 1* , J. Steimel 1 , D.L. McNew 1 & S. 

Kanematsu 2 

1 Iowa State University, Department of Plant Pathology, 

Ames, Iowa 50011, U.S.A. - 2 National Institute of Fruit 

Tree Science, Department of Apple Research, Morioka 

020-0123, Japan. - E-mail: tcharrin@iastate.edu 

Four of the five major clades within Ceratocystis have 

homothallic species. Within the C. coerulescens clade there 

are two closely related species with contrasting mating 

systems: C. eucalypti is heterothallic, while MAT-2 strains 

of C. virescens are self-fertile due to unidirectional mating 

type switching. Most strains of C. virescens recovered from 

nature are self-fertile and behave as MAT-2 in crosses, 

progeny from selfings or from crosses segregate in a 1:1 

ratio for mating type, and MAT-1 progeny are self-sterile. 

Mating type idiomorphs and flanking regions were 

sequenced for both mating types of these two species. In C. 

eucalypti, the idiomorph has either the MAT1-1-2 and the 

MAT1-1-1 genes or only the MAT1-2-1 gene, depending on 

mating type, typical of a heterothallic ascomycete. All 

strains of C. virescens have the MAT1-1-2 and MAT1-1-1 

genes (as in MAT-1 strains of C. eucalypti), but self-fertile 

strains also have the MAT1-2-1 gene and another copy of 

the MAT1-1-1 gene. Half of the progeny recovered from 

selfings retain this arrangement of four mating type genes 

in the idiomorph and are self-fertile, but the other half 

(MAT-1, self-sterile) are missing a 4.7 kb fragment that 

includes the MAT1-2-1 gene and one copy of the MAT1-1-1 

gene. The inherited deletion irreversibly changes the 

mating type to MAT-1 and apparently allows mating with 

MAT-2 nuclei. This is the first sequencing of the DNA 

fragment deleted in unidirectional mating type switching.


196 - Examining the relationship between nonself 

recognition, heterokaryon incompatibility and hyphal 

fusion in Neurospora crassa 

N.L. Glass 1* , Q. Xiang 1 , S. Sarkar 1 , D.J. Jacobson 1 & N.D. 

Read 2 

1 Plant and Microbial Biology Department, University of 

California, Berkeley, CA, 94720-3102, U.S.A. - 2 Institute of 

Cell and Molecular Biology, University of Edinburg, 

Edinburgh, EH9 EJH, U.K. - E-mail: 

Lglass@uclink.berkeley.edu 

Filamentous fungi grow by tip extension, branching and 

hyphal fusion to form a hyphal network. Filamentous fungi 

can also undergo hyphal fusion between individuals to 

make vegetative heterokaryons. Recognition of nonself in 

such heterokaryons is mediated by genetic differences at 

het loci. Heterokaryons or transformants that contain 

alternative het alleles show severe growth inhibition, 

conidiation suppression and death. We use Neurospora 

crassa to study the molecular mechanism of hyphal fusion 

and the consequences of fusion between het incompatible 

individuals. Live cell imaging of hyphal anastomoses 

showed a self-signaling phenomenon which initiates the 

multi-stage process of fusion. We identified four mutants 

that fail to undergo hyphal fusion. Two mutations are in 

genes encoding signal transduction components; these 

mutants also suppress heterokaryon incompatibility 

mediated by the mat locus. Nonself recognition during 

heterokaryon incompatibility has been shown to be 

mediated by the formation of a HET heterocomplex. 

Suppressor analyses identified two additional loci that are 

required for mediating heterokaryon incompatibility. One 

of these mutants suppresses heterokaryon incompatibility 

at both the mat and het-c locus. By these analyses, we hope 

to shed light on nonself recognition mechanisms in 

filamentous fungi that underlie heterokaryon 

incompatibility and explore its relationship to the hyphal 

fusion process. 

197 - Somatic incompatibility in basidiomycetes. 

Genetic analysis in Heterobasidion 

J. Stenlid * , Å. Olson, N. Högberg & M. Lind 

Dept Forest Mycology and Pathology, Swedish University 

of Agricultural Sciences, Box 7026, S-750 07 Uppsala, 

Sweden. - E-mail: Jan.stenlid@mykopat.slu.se 

In basidiomycetes, intraspecific recognition of self and 

non-self is mediated through the somatic incompatibility 

(SI) system (similar to the vegetative compatibility system 

in ascomycetes). It is expressed in the vegetative phase of 

growth both in homokaryons and in heterokaryons when 

dissimilar genotypes meet. In the homokaryotic stage, the 

system can be overridden by mating. SI involves postfusion 

cell death, and depending on the species involved, 

activation of phenol oxidases, melanin production, 

formation of clear zones of collapsed aerial hyphae, hyphal 

knot cells or dense so called barrages. The genetic control 

of SI has been studied in less than ten species. In members 

of the genus Phellinus, SI was under control of one major 

gene, while 3-4 genetic loci have been implied in 

Heterobasidion, Collybia and Pleurotus. In this 

presentation we describe a QTL approach to characterise 

the genetic and molecular background to SI in 

Heterobasidion annosum. A genetic map was constructed 

using AFLP markers for a cross between a S and a P 

homokaryon of H. annosum. Siblings were mated with the 

parent mycelia and resulting heterokaryons, differing in 

one sib-related nucleus, were paired in all combinations. 

Putative loci for SI were mapped based on the 

incompatibility reactions. Implications for future work will 

be discussed. 

198 - Overview of fungal lignin degradation 

C.A. Reddy 

Dept. of Microbiology & Molecular Genetics, Michigan 

State University, East Lansing, MI 48824-4320, U.S.A. - Email: 

reddy@msu.edu 

Lignocellulose, a major component of woody biomass, is 

the most abundant renewable organic material in the 

biosphere and has important applications as a resource for 

producing pulp and paper, animal feeds, and industrial 

chemicals. Lignin, which limits access of microbial 

enzymes to cellulose and hemicellulose in woody 

materials, is a key limiting factor not only in the efficient 

conversion of the latter to useful materials but also in 

global carbon cycling. White-rot basidiomycetes are 

efficient lignin degraders in natures and produce one or 

more of three major classes of lignin-modifying enzymes 

(LMEs) designated: laccases, lignin peroxidases (LiPs), 

and manganese peroxidases (MnPs). LMEs are 

extracellular, non-specific, metalloenzymes and catalyze 

radical-mediated oxidative degradation reactions. 

Cumulative evidence indicates that a large number of 

isoenzymes of LiPs, MnPs, and laccases are produced by 

various ligninolytic fungi and that different isoenzymes 

appear to be encoded by individual genes which are 

distributed on multiple chromosomes. Regulation of 

production of LMEs appears to be subject to complex 

interplay of nutritional, environmental, temporal, and 

genetic factors. An overview of the studies on the 

physiology and molecular biology of LMEs as well as 

applications of ligninolytic fungi for bioremediation of 

priority environmental pollutants will be presented. 



199 - Biochemical and molecular aspects of lignin 

degradation by Pleurotus species 

A.T. Martínez 1* , F.J. Ruiz-Dueñas 1 , M. Perez-Boada 1 , P. 

Ferreira 1 , S. Camarero 1 , F. Guillen 1 , M.J. Martínez 1 , T. 

Choinowski 2 & K. Piontek 2 

1 CIB, CSIC, Velazquez 144, E-28006 Madrid, Spain. - 

2 ETHZ, Universitätstr. 16, Zurich, Switzerland. - E-mail: 

ATMartinez@cib.csic.es 

Pleurotus species are investigated due to their ability to 

degrade lignin selectively. Their ligninolytic system is 

different from that of the model white-rot basidiomycete 

Phanerochaete chrysosporium. Extracellular 

oxidoreductases have been characterized from Pleurotus 

eryngii including versatile peroxidase (VP), aryl-alcohol 

oxidase (AAO) and laccases. These enzymes are of 

biotechnical interest for degradation of lignin, aromatic 

compounds and dyes. P. eryngii laccases oxidize lignin via 

natural redox mediators and contribute to oxygen activation 

by redox cycling of lignin-derived quinones. AAO and VP, 

which have been recently cloned, crystallized and 

expressed in Escherichia coli and Emericella nidulans, are 

characteristic of the ligninolytic system of Pleurotus (and 

Bjerkandera) species. AAO provides hydrogen peroxide 

for peroxidase activity and generation of active oxygen 

species. VP represents a third type of ligninolytic 

peroxidase combining the catalytic properties of lignin 

peroxidase and manganese peroxidase (first described in P. 

chrysosporium) due to an hybrid molecular architecture 

including sites for oxidation of both Mn(II) and aromatic 

substrates. AAO and VP molecular models were obtained 

by homology modeling (using crystal structures as 

templates), and the VP crystal structure has been recently 

solved. Future studies include confirmation of active sites 

by site-directed mutagenesis, and modulation of catalytic 

properties by protein engineering techniques. 

200 - Role of reactive oxygen species in wood decay by 

fungi 

K.E. Hammel 

USDA Forest Products Lab, One Gifford Pinchot Drive, 

Madison, WI 53705, U.S.A. - E-mail: 

kehammel@facstaff.wisc.edu 

Extracellular reactive oxygen species (ROS) have long 

been thought to have a biodegradative function in wood 

decay by fungi, but the nature of these species and the 

mechanisms for their production have not been elucidated. 

We monitored the hydroxylation of a synthetic aromatic 

polymer, phenethyl polyacrylate, to estimate the magnitude 

of extracellular ROS production by two brown rot 

basidiomycetes, Gloeophyllum trabeum and Postia 

placenta, on cellulose. The results showed that ROS 

production was sufficient to account for much of the 

cellulose depolymerization in the cultures. Both fungi 

produced extracellular 2,5-dimethoxyhydroquinone, a 

64 


metabolite that rapidly reduces ferric iron and dioxygen, 

thus yielding hydroxyl radicals via the Fenton reaction. We 

purified and characterized a G. trabeum flavoprotein 

NADH:quinone reductase that probably drives this 

chemistry by regenerating the hydroquinone. The gene that 

encodes the reductase shares substantial similarity with 

quinone reductase genes of other fungi, including some 

nonlignicolous species. 

201 - Biotechnological applications of wood decay fungi 

E. Srebotnik * , M. Weisgram & K. Messner 

Institute of Chemical Engineering, Getreidemarkt 9, A- 

1060 Vienna, Austria. - E-mail: 

esrebot@mail.zserv.tuwien.ac.at 

The ligninolytic systems of white-rot fungi have great 

potential for applications in wood-processing such as 

pulping, pulp bleaching and wood composite manufacture. 

For example, the use of white-rot fungi to treat wood chips 

prior to mechanical pulping (biopulping) is already 

approaching industrial scale. Furthermore, the low 

specificity of these systems allows for the conversion of 

various aromatic pollutants and industrial wastes such as 

contaminated soil (bioremediation). These processes 

involve the use of white-rot fungi or their ligninolytic 

enzymes to treat polycyclic aromatic hydrocarbons (PAH), 

polychlorinated biphenyls and other hazardous xenobiotics. 

Whether to apply living fungi or isolated enzymes will 

depend on the specific properties of the material to be 

treated. For example, biopulping takes advantage of the 

fact that white-rot fungi not only produce a complete set of 

enzymes but can also transport these enzymes into wood 

chips and create the appropriate physiological conditions 

for enzymatic reactions. However, processes involving 

living organisms are relatively difficult to control. Thus, 

the use of isolated enzyme systems would be preferable in 

those cases, where the compounds to be treated are freely 

accessible such as in soil extracts. This presentation will 

give a brief overview of biotechnological applications of 

white-rot fungi and their ligninolytic enzymes and discuss 

in more detail the use of laccase for the bioremediation of 

PAH. 

202 - Some approaches to the evaluation of the white 

rot basidiomycetes ligninolytic activity 

V.I. Elisashvili * , E. Kachlishvili, N. Tsiklauri, T. 

Khardziani & M. Bakradze 

Institute of Biochemistry and Biotechnology, 10 km 

Agmashenebeli kheivani,380059 Tbilisi, Georgia. - E-mail: 

velisashvili@hotmail.com 

Ligninolytic activity of basidiomycetes from different 

taxonomic groups has been extensively. The production 

patterns and levels of laccase and Mn-dependent 

peroxidase (MnP) differed among species and strains of the


same species studied. Production of MnP was generally 

optimal in SSF of grapevine cuttings sawdust (GCS), but 

was repressed by agitation of fungi grown in submerged 

liquid culture. Conversely, basidiomycetes laccase activity 

was generally enhanced by agitation. The highest laccase 

activity (up to 400-800 nkat/ml) was revealed in a 

submerged fermentation of mandarin juice production 

wastes by the strains of genera Cerrena, whereas extremely 

high Mn-dependent peroxidase activity (50-180 nkat/ml) 

was found in a SSF of the GCS by Pleurotus ostreatus and 

Lentinus edodes. Among the factors tested, only carbon 

sources and low-molecular-weight aromatic compounds in 

the media have been shown to significantly influence 

ligninolytic activity of basidiomycetes. However, the 

response of different fungi strains is greatly depending on 

chemical nature of the carbon source. Concerning nitrogen 

source only increased basidiomycetes growth at presence 

of additional nitrogen accounts for the higher levels of 

enzymes activity. This work shows that different 

approaches might be used to regulate basidiomycetes 

ligninolytic activity in general or activity of certain 

enzyme, however, caution is necessary when evaluating 

ligninolytic capabilities of the white rot fungi. 


IMC7 Wednesday August 14th Lectures 

203 - Role of reticulation events in the co-evolution of 

Epichloë grass endophytes 

C.L. Schardl * , C.D. Moon & K.D. Craven 

University of Kentucky, S-305 ASCN, Lexington, Kentucky 

40546, U.S.A. - E-mail: schardl@uky.edu 

The Epichloë (anamorph = Neotyphodium) species are 

fungal endophytes of grasses spanning an evolutionary 

continuum from antagonistic sexual species to mutualistic 

asexual species. The asexual endophytes are strictly seedborne 

(vertically transmitted), whereas the sexual 

endophytes can transmit contagiously (horizontally). Most 

of the sexual species are highly host specific, but their 

occasional jumps between host tribes can generate strictly 

seed-borne lineages. The fact that the resulting endophytes 

express no male or female reproductive structures, and no 

contagious spores on their new hosts probably reflects their 

relatively short history of coevolution with those grasses. 

Although such endophytes are asexual, they remain 

capable of parasexual interactions with horizontallytransmitted 

Epichloë spp., giving rise to aneuploid or 

polyploid hybrids. Out of 32 phylogenetically 

distinguishable asexual lineages, 21 were hybrids of two or 

three biologically and phylogenetically distinct sexual 

species. Among endophytes in certain host genera the 

likely host jump and subsequent hybridization events can 

be discerned. For example, Epichloë bromicola apparently 

jumped from Bromus into Hordeum, then underwent 

several hybridizations with Epichloë typhina; both the nonhybrid 

and hybrid descendants occur in extant Hordeum 

populations. Likewise it appears that an asexual endophyte 

in a Lolium ancestor underwent at least five hybridizations 

during the emergence of three host species. 

204 - Fitness costs in hybridization events between 

allopatric Tilletia species 

L.M. Carris 

Washington State University, Department of Plant 

Pathology, Pullman, WA 99164-6430, U.S.A. - E-mail: 

carris@wsu.edu 

A closely related group of Tilletia species infects wild and 

cultivated hosts in the grass subfamily Pooideae. This 

group of smut fungi includes the wheat common and dwarf 

bunts, T. tritici, T. laevis, and T. controversa. A study of 

genetic relationships among isolates of common and dwarf 

bunts based on RAPD analysis supported natural 

hybridization as an explanation for the existence of isolates 

with reciprocal characteristics of spore morphology and 

germination. Nuclear rDNA sequence and RAPD analyses 

showed T. bromi, a bunt that infects brome grasses growing 

as weeds in wheat fields in the Pacific Northwestern USA, 

to be the species most closely related to the wheat bunts. 

The ability of T. controversa to form interspecific hybrids 

with T. bromi was examined. Tilletia controversa was 

hybridized with T. bromi and T. laevis in a susceptible 

66 


cultivar of wheat under greenhouse conditions. Tilletia 

controversa x T. bromi progeny exhibited reduced fitness 

relative to T. controversa x T. laevis progeny as measured 

by basidiospore mortality, basidiospore fusion, and 

pathogenicity in wheat. Additionally, no evidence of 

natural hybridization or introgression based on RAPD 

analysis was detected between allopatric populations of T. 

controversa and T. bromi. Our data suggest reproductive 

compatibility is retained among species of Tilletia infecting 

pooid hosts, but progeny produced between species with 

distinct host ranges are unlikely to survive under natural 

conditions. 

205 - Stumps as a new niche for interspecific 

hybridization in the Heterobasidion spp. complex. 

Ecological instability vs. the genetic stability of hybrids 

M.M. Garbelotto 1* , W. Otrosina 2 , J. Stenlid 3 , P. Gonthier 4 

& G. Nicolotti 4 

1 University of California, 151 Hilgard Hall, Berkeley, CA 

94720, U.S.A. - 2 USDA Forest Service, Forestry Sciences 

Laboratory, Athens, GA 30602, U.S.A. - 3 University of 

Uppsala, Box 7026, 750 07 Uppsala, Sweden. - 4 University 

of Turin, via L. da Vinci 44, 10095 TO, Italy. - E-mail: 

matteo@nature.berkeley.edu 

The five known taxa in the Heterobasidion annosum Coll. 

complex display a marked host preference. This niche 

partitioning has strengthened the genetic isolation among 

these species, in spite of their potential interfertility. 

Through field and greenhouse experiments, we show that 

host preference is lost on tree stumps as well as in pine 

trees exposed to high levels of inoculum. An analysis of 

Heterobasidion species composition in stumps shows 

significant sympatry between species. A comparative 

analysis in trees shows no shift in species composition. 

Logging has thus greatly enhanced the potential for contact 

between species. An analysis of Heterobasidion species 

composition from the airspora, shows ratios resembling 

species composition from stumps rather than from trees. 

This finding underlines the impact of stumps on 

Heterobasidion species composition. Effects of stump 

creation is variable. Where mating barriers between species 

are strong (e.g. the Alps), little evidence of hybridization 

and gene introgression have been found. Where partial 

interfertility is present (e.g. California), hybrids and 

evidence of past and present gene introgression can be 

found. Although hybrids do not appear to have enhanced 

pathogenicity we show through inoculation experiments 

that they can be as fit as one of the parental species, 

especially on tree stumps. We finally show that hybrids 

form diploid hyphae, thus ensuring long-term genetic 

stability.


206 - Influence of selection pressure on the outcome of 

current interspecific gene flow between Ophiostoma 

species 

C.M. Brasier 1* , M. Paoletti 2 & K.W. Buck 2 

1 Forest Research Agency, Farnham, Surrey GU10 4LH, 

U.K. - 2 Biology Department, Imperial College, London 

SW7 2AZ, U.K. - E-mail: clive.brasier@forestry.gsi.gov.uk 

The Dutch elm disease fungus Ophiostoma novo-ulmi has 

recently invaded areas of the northern hemisphere 

previously occupied by another DED pathogen, O. ulmi. 

This has enabled interactions between the two species, 

resulting in a series of unusual evolutionary events. O. 

novo-ulmi has physically replaced O. ulmi, forcing the 

latter into extinction. While doing so, O. novo-ulmi has 

spread as a series of single mating type single vegetative 

compatibility type (vic) clones. Locally, these clones have 

then become rapidly genetically diverse (c. 5-10 years), 

with many new vic and other phenotypes appearing. The 

mechanism for this rapid change from clonality to 

heterogeneity was unknown. Evidence based on cosegregation 

of AFLP markers and cloning and sequencing 

of mating type loci indicates that O. novo-ulmi has 

acquired both the new vic genes and the A-mating type 

locus from O. ulmi via horizontal gene flow. The O. ulmi 

vic and mat A genes appear to have been acquired by O. 

novo-ulmi under selection pressure imposed by deleterious 

viruses. Other 'less useful' O. ulmi genes acquired by O. 

novo-ulmi, such as genes for pathogenicity or for ceratoulmin 

(toxin) production, appear not to be selected, 

probably because they render O. novo-ulmi less fit. 

Furthermore, only c2% of 'foreign' DNA appears to be 

acceptable if an introgressed O. novo-ulmi genotype is to 

survive. 

207 - Evolutionary processes and genome organisation 

among emerging interspecific hybrids in Phytophthora 

species 

D.E.L. Cooke 1* , J.M. Duncan 1 , W.A. Man in't Veld 2 & 

C.M. Brasier 3 

1 Host Parasite Co-evolution, Scottish Crop Research 

Institute, Invergowrie, Dundee, DD2 5DA, U.K. - 2 Plant 

Protection Service, Department of Mycology, PO Box 

9102, 6700 HC Wageningen, The Netherlands. - 

3 Pathology Branch, Forest Research, Alice Holt Lodge, 

Farnham, Surrey, GU10 4LH, U.K. - E-mail: 

dcooke@scri.sari.ac.uk 

Whilst common in plants, reports of interspecific 

hybridisation amongst fungi are scarce. The risks, however, 

should not be underestimated. Hybridisation may 

accelerate the evolution of plant pathogenic fungi resulting 

in rapid adaptation to new plant species or environments. 

Increasing global trade in plants and, inadvertently, their 

pathogens, creates opportunities for such events. The genus 

Phytophthora comprises over sixty plant pathogenic 

species, many with broad host ranges and some responsible 

for severe crop epidemics or for destabilising terrestrial 

ecosystems. This, alongside their global distribution, 

cryptic nature and flexible diploid genetic system serves to 

increase the risks and threats of hybridisation within the 

genus. Cytological behaviour, additive nucleotide bases in 

the ITS region and AFLP and isozyme profiles of a new 

aggressive Phytophthora pathogen of alder trees spreading 

in Europe have shown that it comprises a swarm of 

heteroploid interspecific hybrids between a P. cambivoralike 

species and an unknown P. fragariae-like taxon. 

Developmental and cytological instability and marked 

phenotypic and genotypic variation all indicate a recent 

hybridisation. Moreover, mtDNA typing suggests that this 

occurred more than once. Evidence for the continuing 

evolution (or stabilisation) of the hybrid genomes will be 

presented and the challenge of delimiting such hybrid taxa 

in a way that is practicable for quarantine legislation and 

diagnosis will be considered. 

208 - Evolution of interspecific Melampsora hybrids 

exhibiting new host specificities 

G. Newcombe 1* & P. Frey 2 

1 University of Idaho, Dept. Forest Resources, Moscow ID 

83844-1133, U.S.A. - 2 INRA, INRA Forest Pathology, 

54280 Champenoux, France. - E-mail: 

georgen@uidaho.edu 

Interspecific hybridization has been an important tool in 

plant domestication. Domestication in Populus, and other 

forest trees, is just in its infancy, and hybridization has 

been the primary means by which poplar breeders have 

produced desirable plantation phenotypes. Are the 

pathogens of Populus undergoing parallel domestication? 

The answer appears to be yes for Melampsora, and no for 

Venturia. For Melampsora, interspecific hybridization of 

M. medusae and M. occidentalis now matches that of the 

respective hosts (i.e., Populus deltoides and P. 

trichocarpa). Multiple, exapted genes for rust resistance 

from both host species have been suddenly overcome in P. 

trichocarpa x P. deltoides hybrids yielding new and 

complex pathogenic variation. Venturia, and other 

ascomycetous pathogens of Populus are similar to a point. 

Multiple, exapted genes for resistance to Venturia (and 

Taphrina, and Linospora as well) are also known (from a 

common-garden study of P. trichocarpa x P. deltoides and 

P. trichocarpa x P. maximowiczii pedigrees). However, 

although European and native species of Venturia do cooccur 

in the Pacific Northwest of the U.S., they have not 

hybridized. Instead, they remain confined to their 

respective hosts (i.e., the European Populus nigra and the 

native P. trichocarpa, respectively). 



209 - Phylogenetic relationships in Homobasidiomycetes 

R.G. Thorn 1* , M. Binder 2 & D.S. Hibbett 2 

1 Department of Biology, University of Western Ontario, 

London, Ontario, N6A 5B7, Canada. - 2 Department of 

Biology, Clark University, 950 Main Street, Worcester, 

Massachusetts 01610-1477, U.S.A. - E-mail: 

rgthorn@uwo.ca 

The time since IMC6 has seen a dramatic increase in 

molecular phylogenetic studies of homobasidiomycetes. 

This talk offers the opportunity for a general overview of 

recent results from our labs as well as published and 

unpublished results from other research groups. The eight 

major clades of homobasidiomycetes identified by Hibbett 

et al. (1997, PNAS 94:12002) have generally held up to 

more recent analyses but, despite the increased 

phylogenetic information of additional sequence data, 

support for basal divisions, particularly the polyporoid 

clade, remains weak and placement of some terminal taxa 

remains equivocal. A picture is emerging of rapid 

radiations among the basal lineages, echoed by rapid 

radiations among more recently evolved groups such as the 

secotioid and gasteroid members of agaric and bolete 

clades. The consequences of the former are the difficulty of 

firmly establishing the relationships among major clades 

and the evolution of fundamental ecological characters 

such as ectomycorrhizal habit and rot type. The 

consequences of the more recent radiations in fruiting body 

morphology include the potential for havoc in our 

classification system and great fun in the mycology 

classroom. In any case, molecular phylogenetic studies of 

homobasidiomycetes are both answering questions of 

systematics and ecology, and raising many new ones. 

210 - Phylogenetic relationships in the euagarics 

(Agaricales): insights into morphological and ecological 

evolution in mushrooms 

J.-M. Moncalvo 1* , R. Vilgalys 1 & S.A. Redhead 2 

1 Department of Biology, Duke University, Durham, NC 

27708, U.S.A. - 2 Systematic Mycology and Botany Section, 

Eastern Cereal and Oilseed Research, Agriculture and 

Agri-Food Canada, Ottawa, Ontario K1A 0C6, Canada. - 

E-mail: jeanmarc@duke.edu 

Molecular phylogenetics support the view that gilled 

mushrooms have evolved multiple times from 

morphologically diverse ancestors, making the Agaricales 

polyphyletic. Molecular data also show that poroid, 

sequestrate (e.g., puffballs and secotioids) and reduced 

(e.g., cyphelloids) forms have evolved several times from 

gilled basidiocarps. Here we present results from molecular 

phylogenetic analyses that sampled about one tenth of the 

total number of species known in the largest natural group 

of homobasidiomycetes, the euagarics clade (ca. 8400 

known species). In many cases, it was possible to resolve 

natural relationships of several gilled fungi for which 

68 


taxonomic position has been controversial in the past, and 

also to unambiguously resolve among the euagarics the 

systematic placement of many gasteromycetes and reduced 

forms. The mapping of characters onto phylogenetic trees 

indicates that ecology, biochemistry and/or physiology 

rather than morphology often support natural groups of 

euagarics. Newly discovered phylogenetic affinities 

include for instance relationships of the true puffballs 

(Lycoperdales) with the Agaricaceae, of Panellus and the 

poroid fungi Dictyopanus and Favolaschia with Mycena, 

and of the reduced fungus Caripia with Gymnopus. 

Examples of newly discovered monophyletic groups of 

euagarics include the clades resupinatus, lentinuloideae, 

Tricholomataceae, Mycenaceae, Agaricaceae, psychedelia, 

and Pleurotaceae. 

211 - Phylogeny and ecology - what can primitive 

homobasidioid fungi tell us? 

E. Langer 

Universitaet Kassel, FB 19, FG Oekologie, Heinrich-Plett- 

Str. 40, 34132 Kassel, Germany. - E-mail: 

ewald.langer@uni-kassel.de 

With special emphasis on Aphyllophorales, this study was 

carried out to verify hitherto published phylogenies of 

Hymenomycetes by nuclear large subunit rDNA sequences 

and comparative morphology. Preferably Aphyllophorales 

with dolipores having the imperforate parenthesome-type 

were used to remedy the imbalance in taxa composition of 

former analyses. The results support the hypothesis of a 

monophyletic origin of homobasidiomycetes having 

dolipore parenthesomes perforated by many small holes, 

including gilled, non-gilled mushrooms and 

gasteromycetes. Only a minor part of the formally settled 

orders or families could be verified by this analysis in full 

respect. Especially the Agaricales ss Singer contain many 

different related Aphyllophorales. A monophylum 

comprising non-gilled mushrooms from certain 

Corticiaceae and Hymenochaetales, having solely dolipores 

with imperforate parenthesomes is the sistergroup of the 

homobasidiomycetes having dolipores with perforated 

parenthesomes. The borderline between 

heterobasidiomycetes and homobasidiomycetes was 

detected to be unsharp, because of repeatedly loss of 

secondary ballistospores in a monophylum nesting the 

Tulasnellales, Botryobasidiales and cantharelloid taxa. 

212 - Phylogenetic relationships among corticioid fungi 

K.-H. Larsson 1* , E. Larsson 1 & U. Kõljalg 2 

1 Goteborg University, Botanical Institute, P.O. Box 461, 

SE 405 30 Göteborg, Sweden. - 2 University of Tartu, 

Institute of Botany and Ecology, 40 Lai str., 51005 Tartu, 

Estonia. - E-mail: karl-henrik.larsson@systbot.gu.se


Basidiomycetes with effused, skin-like basidiomata are 

often classified together as corticioid fungi. Recent 

molecular phylogenetic studies have shown that 

homobasidiomycetes have evolved into at least eight 

lineages. In these earlier studies only a few corticioid 

representatives were included but already from a restricted 

sampling it is obvious that corticioid fungi are 

phylogenetically highly diverse. We performed a 

phylogenetic analysis of the homobasidiomycetes with a 

strong emphasis on corticioid forms. As characters we used 

nucleotide sites from 5.8S and 28S in the nuclear ribosomal 

DNA repeat. We recovered the eight clades earlier 

identified and in addition four new clades composed 

primarily of corticioid species. We hypothesise that 

increased sampling of corticioid fungi will reveal still other 

unique lineages and potential examples are given. The 

implications for the interpretation of homobasidiomycete 

evolution and for the design of future studies are discussed. 

213 - Phylogeny of corticioid fungi with russuloid 

characteristics (the genus Gloeocystidiellum in a 

phylogenetic perspective) 

E. Larsson * & K.-H. Larsson 

Göteborg University, Dep. of biology, Box 461, SE 405 30 

Göteborg, Sweden. - E-mail: ellen.larsson@systbot.gu.se 

The classification of fungi has traditionally relied on 

macro- and microanatomical features of the basidiome. The 

genus Gloeocystidiellum is characterized by the corticioid 

basidiomata, the presence of SA+ gloeocystidia and 

amyloid often ornamented spores. Phylogenetic analyses, 

based on nuclear rDNA, of 127 species within the russuloid 

lineage recovered 13 supported clades and six single 

branches. Species that have been combined to 

Gloeocystidiellum occur in 7 of the 13 clades. Other results 

support monophyly of the russuloid clade, and our data 

suggest that the presence of a gloeoplerous system, 

primarily observed as gloeocystidia, is a synapomorphy of 

russuloid taxa. In addition, compatibility studies were 

performed to discern biological species within the 

Gloeocystidiellum porosum / clavuligerum complex and to 

characterize the genus type G. porosum. Four compatibility 

groups were detected. Phylogenetic analysis of the 

complex revealed a great sequence divergence between G. 

porosum and G. clavuligerum. Based on morphological 

data, mating tests and phylogenetic analyses, three species 

in the complex could be characterised; G. kenyense, G. 

clavuligerum and G. porosum. Gloeocystidiellum bisporum 

was shown to be a parthenogenetic haploid species, 

evolved from G. clavuligerum. The results indicate that 

Gloeocystidiellum includes only G. porosum and a few 

closely related species. 

214 - Phylogenetic study of Thelephorales - the story of 

explanatory power of systematics 

U. Kõljalg * & K. Abarenkov 

University of Tartu, 40 Lai Str., 51005 Tartu, Estonia. - Email: 

ukoljalg@ut.ee 

Molecular phylogenies of Thelephorales and its genera will 

be presented. Based on these phylogenies the explanatory 

power of systematics to answer specific questions on 

autecology of Thelephorales will be demonstrated. In order 

to make systematics more powerful and attractive, 

database-driven Web sites of different taxa should be 

compiled which include all types of biological information. 

Such databases will provide facility to study taxon ecology, 

phylogeny, etc. in silico by other fields. The information 

facility for thelephoroid fungi is built using the PHP 

scripting language and the MySQL relational database. 

Access to database and a database administration is done 

over the Web. Currently the database-driven Web site 

includes molecular identification of resupinate thelephoroid 

fungi using ITS1, 5.8S and ITS2 regions of nuclear rDNA 

and BLAST similarity search. An example how it may be 

used for the identification of root mycobionts will be 

shown. In future the database will provide additional 

facilities for interactive identification of taxa based on 

morphology, provide information on nomenclature, 

phylogeny, ecology, etc. 

215 - Phylogenetic taxonomy of Hymenochaete and 

related genera (Hymenomycetes) 

E. Parmasto 1* , U. Kõljalg 2 , E. Larsson 3 & S. Rummo 2 

1 Institute of Zoology and Botany, Estonian Agricultural 

University, 181 Riia St., 51014 Tartu, Estonia. - 2 Institute 

of Botany and Ecology, Tartu University, 40 Lai St., 51005 

Tartu, Estonia. - 3 Botanical Institute, University of 

Göteborg, Box 461, SE-405 30 Göteborg, Sweden. - Email: 

e.parmasto@zbi.ee 

Studies by Wagner and Fischer (2001, 2002) have 

demonstrated that 19 species of Hymenochaete s.l. studied 

(except H. tabacina) form, together with 2 Hydnochaete 

and 2 Cyclomyces species, a monophyletic genus in the 

order Hymenochaetales. We examined the taxonomy of 

this group adding 16 more species of Hymenochaete, 

Coltriciella tasmanica and Hydnochaete olivacea. 

Parsimony analysis of molecular (LSU nucrDNA 

sequences), morphological and combined datasets was 

carried out, including representatives of other genera of 

Hymenochaetales s.str. As an outgroup, 6 corticioid and 

polyporoid (non-hymenochaetoid) species, characterized 

by dolipore septa with continuous parenthesomes, were 

used. About 35% of the Hymenochaete species have been 

included in this study. Most hymenochaetes comprise a 

clade of closely related species; a separate clade (a new 

genus?) is formed of 4 species with dendrohyphidia or 



denticulate setae. Two species of Hymenochaete and 

Hydnochaete olivacea can be added to the monotypic 

genus Pseudochaete. H. villosa is different from the other 

Hymenochaete species and belongs to a clade together with 

Coltricia and Coltriciella. As a rule, a clade ought to be 

named a genus only when there are morphological 

characters making the species group recognizable without 

molecular analysis. Within Hymenochaete s.str., such 

groups are lacking. Avoidance of schizotaxia (splitting) is 

advisable irrespective of the use of morphological or 

mainly molecular characters. 

216 - Building ecology: Influence of distribution, 

detection and control of biodeterioration 

J. Mattsson 

Mycoteam AS, P.O. Box 5 Blindern, 0313 Oslo, Norway. - 

E-mail: johan@mycoteam.no 

Under normal circumstances do mould and wood-decaying 

fungi not have favourably condition for growth in 

buildings, mainly due to restricted access of water. 

However, it might occur local areas in constructions where 

the physical conditions are favourable enough for 

microbiological activity. Understanding the building 

ecology with respect to influence of distribution and 

activity of different fungi, gives possibilities for a 

successful detection, evaluation and control of the 

damages. By laboratory and field experiments, it has been 

proven what physical conditions that are necessary for 

fungal activity in buildings. The main factors for growth 

are humidity, temperature and the time of exposure. Other 

aspects might also have impact, but often to less extent. 

Due to the great variations of microclimate in a 

construction, it can be a significant change of the building 

ecology in few millimetres. This variation often leads to 

difficulties in the work of building survey. Since the 

damage development might be caused by small variations 

in building ecology, the detection and understanding of the 

physical conditions are important in order to control if 

remediation has desired effect. Subsequently, the risk of 

failure in handling damages caused by mould fungi is high 

- often with a significant impact on the indoor air quality. 

Guidelines for accomplishing building survey are needed in 

order to give a better understanding of the building ecology 

and thereby ensure an optimal result. 

217 - Studies on the dry rot fungus, Serpula lacrymans 

J.W. Palfreyman 1* , J.S. Gartland 1 , N.A. White 1 , J. Bech- 

Andersen 2 & D.E.L. Cooke 3 

1 School of Contemporary Science, University of Abertay 

Dundee, Bell Street, Dundee, Scotland, U.K. - 2 Hussvamp 

Laboratories AB, Copenhagen, Denmark. - 3 Mycology, 

Bacteriology and Nematology Unit, SCRI, Invergowrie, 

Dundee, Scotland, U.K. - E-mail: mltjwp@tay.ac.uk 

70 


Molecular and morphological parameters of Serpula 

lacrymans isolates from various sites in the built 

environment in Europe and Australia were compared to 

similar parameters of 'wild' isolates from India, the Sumava 

Mountains (Czech Republic) and Mount Shasta (U.S.A.). 

The Indian, Czech Republic and all of the building isolates 

bar one showed identity in both molecular and 

morphological features. The Australian and the U.S. 

isolates (BF-050 and USA'94 respectively) showed specific 

morphological differences and could be separated on the 

basis of RAPD-PCR with the U.S. isolate being least 

closely related to the S. lacrymans type strain of FPRL12C. 

ITS sequence data revealed 2 base differences between 

FPRL12C and BF-050 in the 673 sequenced, 9 differences 

between FPRL12C and USA'94 and 16 differences 

between USA'94 and the closely related organism Serpula 

himantioides. The possible evolutionary relationships 

between the various isolates are discussed along with 

suggestions for the origin of S. lacrymans as a scourge of 

the built environment in many temperate areas of the 

world. We will also report on the use of Indian, US and 

Czech Republic Isolates of Trichoderma in the biocontrol 

of decay by S. lacrymans. 

218 - Moulds in Buildings 1998-2002 

S. Gravesen 

Danish Urban & Research Institute, box119, 2970 

Hørsholm, Denmark. - E-mail: sug@sbi.dk 

Results of a multidisciplinary research programme: 

"Moulds in buildings, 1998 - 2002" are presented. The 

purpose of the programme was to gain more knowledge 

concerning conditions for mould growth in buildings and 

the ensuing health effects on people. Furthermore the aim 

was to create basic knowledge about safe and cost efficient 

solutions for solving problems and for preventing mould 

growth in buildings during planning, operation, 

maintenance and renovation. - A clear correlation was 

demonstrated to exist between the level of moulds in floor 

dust and a number of irritative symptoms from eyes and 

upper airways including the general symptoms such as 

headaches, dizziness and difficulties in concentration. - The 

correlation between exposure to moulds from floor dust nd 

symptoms was typically stronger among pupils with 

asthma and hay fever. This condition was especially 

pronounced for irritation of the mucous membranes of the 

eyes. - Experiments have shown that wood and wood-based 

materials; wallpaper and materials with glue (starch) have 

the lowest resistance to mould infestation. - Plaster and 

concrete, which have a very modest content of organic 

material, can be infested with mould growth, provided that 

the materials have a high humidity (water activity) 

preferably 95% RH. *Danish Moulds in Buildings- 

Consortium.


219 - Mycotoxins 

K.F. Nielsen 

The Mycology Group, BioCentrum-DTU, Technical 

University of Denmark, Building 221, DK-2800 Kgs. 

Lyngby, Denmark. - E-mail: kfn@biocentrum.dtu.dk 

Mycotoxins produced by the building associated funga 

have been proposed as one of the agents, which are 

responsible for the adverse health effects, observed among 

people living or working in damp and water damaged 

buildings. The vehicle for exposure to mycotoxins in the 

indoor environ-ment is presumable the fungal spores and 

fragments of these. The quantities of thee mycotoxins 

present on building materials are quite low (ng-µg/cm 2 

range), and air concentrations generally so low that that 

they can not be measured. Penicillia do not produce many 

metabolites when growing on building materials compared 

with agar-media and foods. Aspergillus versicolor is a 

complex species, with at least 3 sub-groups and produces 

sterigmatocystins up to 30 µg/cm 2 . However generally the 

concentrations are in the ng/cm 2 range. Interestingly 4-7 

alkaloids previously seen from A. ochraceus are often 

produced. For Stachybotrys chartarum some evidence of 

building associated mycotoxicosis exists, as it in buildings 

produces high quantities of a number of metabolic families. 

These includes spiriocyclic drimanes 

(spirodihydrobenzofuranlactam) which are complement 

inhibitors (stachybotrys lactones and lactams) and have, 

fibrionolytical activity, neurotoxic effects etc. Additionally 

30% of the iso-lates produces macrocyclic trichothecenes 

which are highly cytotoxic to mammalian cells and are 

known to give severe haematoxic effects. 

220 - Biodiversity of allergenic indoor fungi colonising 

dust particles and surfaces in New Zealand households 

N.W. Waipara 1* , M. di Menna 2 , C. Marshall 3 , P. Howden- 

Chapman 3 & J. Crane 3 

1 HortResearch, Canterbury Research Centre, P.O.Box 51, 

Lincoln, New Zealand. - 2 AgResearch, Ruakura Research 

Centre, Private Bag 3123, Hamilton, New Zealand. - 

3 Wellington School of Medicine & Health Sciences, 

University of Otago, PO Box 7343, Wellington, New 

Zealand. - E-mail: nwaipara@hortresearch.co.nz 

Fungal species isolated from indoor environments have 

been shown to produce toxic volatile organic compounds 

(VOC) as well as produce allergenic spores, which can 

trigger respiratory and related illnesses. A series of 

mycological surveys of households were undertaken in four 

climatic regions of New Zealand to determine the 

biodiversity of fungi present in indoor environments, in 

particular, the presence of mycotoxic and allergenic fungal 

species. Fungi were isolated from vacuumed floor dust 

samples and from wall surfaces using 3M Petrifilms. 

Isolation results showed ubiquitous taxa, such as 

Aspergillus spp., Cladosporium spp., Epicoccum nigrum, 

Alternaria spp., and Penicillium spp., were frequently 

obtained in both dust and surface samples. An overall 

higher species diversity was obtained from dust samples 

than was observed from wall surfaces, however, the 

dominance of Cladosporium spp. on surfaces across all 

households may have adversely reduced the sensitivity of 

the Petrifilm method to detect many slow growing genera. 

Significant biodiversity differences were observed between 

the fungal species isolated from dust in different climatic 

regions. For example, Aspergillus spp. were only dominant 

in houses situated in regions with a dry warm climate. 

Many households were found to be contaminated with 

mycotoxic and VOC producing fungi such as Aspergillus 

fumigatus, A. ochraceus, Penicillium verrucosum, and 

Stachybotrys chartarum. 

221 - Chiral MVOC's - a new tool for studying moulds 

in building constructions? 

O.-A. Braathen 1* , C. Lunder 1 , N. Schmidbauer 1 , J. 

Mattsson 2 & C. Ahlén 3 

1 NILU, P.O.Box 100, 2027 Kjeller, Norway. - 2 Mycoteam, 

P.O.Box 5 Blindern, 0313 Oslo, Norway. - 3 SINTEF 

Unimed, 7465 Trondheim, Norway. - E-mail: oab@nilu.no 

MVOC-measurements can be used as a non-destructive 

way to detect moulds in closed spaces in buildings. Most of 

the VOCs produced by moulds can also be found in 

emissions from building materials and various household 

products. It is therefore hard to find MVOCs that have 

microbiological activities as the only emission source. In 

order to find a good tracer for moulds in sick buildings, 

chiral volatile organic compounds emitted by 

microbiological activity (c-MVOC) have been studied in 

indoor air. For a chiral VOC-compound, there will be at 

last two enantiomers ('mirror images' that have equal 

chemical properties, but different physical properties). 

Man-made VOCs most often occur in racemic ratio 

(50:50). When there is microbiological activity involving 

enzymic reactions, one of the enantiomers will dominate, 

giving a non-racemic ratio (for instance 80:20 or 100:0). 

Air samples were adsorbed on Tenax TA, followed by 

thermal desorption and combined gas 

chromatography/mass spectrometry (GC/MS) and 

separation of enantiomers on suitable chromatographic 

columns. In this study six of the most common moulds in 

buildings with water damages were studied both at 

controlled conditions in the laboratory (grown on gypsum 

board and plywood) and in mould-infected buildings. This 

study has shown that some chiral-MVOC can be used as a 

tracer for microbiological activity (mould) in buildings. 



222 - Ectomycorrhizal fungal community structure in 

soil as identified from soil DNA extracts 

R. Landeweert 1* , C. Veenman 2 , T.W. Kuyper 1 & E. Smit 2 

1 

Wageningen University, Sub-dept. of Soil Quality., 

P.O.Box 8005, NL-6700 EC Wageningen, The Netherlands. 

2 

- National Institute for Public Health and the 

Environment, P.O.Box 1, NL-3720 BA Bilthoven, The 

Netherlands. - E-mail: renske.landeweert@bb.benp.wagur.nl 

The various functional roles of ectomycorrhizal fungi in 

ecosystems can only be recognised when the mycelial 

distribution of individual species in soil is revealed. 

Collectively, the structure, function and activity of the 

ectomycorrhizal mycelium determine the nutrient 

mobilisation and translocation capacities of different fungal 

species. Though its ecological significance was recognized 

by many mycologists, identification of mycelium in soil 

remained impossible due to technological limitations. At 

present however, molecular identification techniques based 

on total DNA extracts can be used to detect and identify 

fungal hyphae in various substrates. In order to detect and 

identify ectomycorrhizal fungal mycelium in soil, 

molecular identification techniques were applied to sieved 

soil samples from which all root tips were removed. A 

basidiomycete-specific primer pair was used to amplify 

fungal ITS sequences from total soil DNA extracts. 

Amplified basidiomycete DNA was cloned and sequenced 

and a selection of the obtained clones was analysed 

phylogenetically. Through molecular identification of the 

soil mycelium, the ectomycorrhizal fungal diversity was 

determined in three vertical soil columns taken from a 

podzol profile. Latest results will be presented on the 

application of molecular identification as well as 

quantification techniques to determine fungal presence in 

soils. 

223 - Spatial structure in ectomycorrhizal fungal 

communities: a look belowground 

E.A. Lilleskov 1* , T.R. Horton 2 , D.L. Taylor 3 , P. Grogan 4 & 

T.D. Bruns 1 

1 University of California, Berkeley, 111 Koshland Hall, 

Berkeley, CA 94720, U.S.A. - 2 SUNY-ESF, 350 Illick Hall, 

1 Forestry Drive, Syracuse, NY 13210, U.S.A. - 3 University 

of California, Berkeley, Plant Conservation Research 

Center, UC Botanical Garden, 200 Centennial Drive 

#5045, Berkeley, CA 94720, U.S.A. - 4 Cranfield University, 

Silsoe, Bedfordshire MK45 4DT, U.K. - E-mail: 

elilleskov@fs.fed.us 

Ectomycorrhizal fungal (EMF) communities are highly 

diverse at the stand level. Although many communities 

have been characterized, little work has been done to 

examine their spatial structure. Analysis of spatial patterns 

might point toward processes structuring communities. We 

examined EMF community data from studies carried out in 

72 


mature and post-fire conifer forests. To characterize the 

distribution of communities and populations, we used 

various measures of spatial pattern: indices of dispersion, 

mantel tests, and standardized variograms. Mantel tests 

indicated higher community similarity with decreasing 

distance, but this pattern was only evident at small spatial 

scales (usually < 2 m). Indices of dispersion and 

variograms indicated that most species showed significant 

clumping, but the degree and scale of clumping varied 

widely. There was evidence of clumping in both mature 

and post-fire communities. The former could be the result 

of genet size, contagious establishment effects that are 

either endogenous (e.g., local spore rain) or exogenous 

(e.g., patchy resources or disturbance), or patchiness in 

initial inoculum. The latter might be the result of 

patchiness in the inoculum source, given the presence of 

resistant propagules in the species exhibiting patchiness. 

To distinguish among alternative mechanisms for patch 

formation, further investigation is needed of genetic 

structure of EMF populations at the mycelial level. 

224 - Community composition of arbuscular 

mycorrhizal fungi in a tropical forest 

R. Husband 1* , E.A. Herre 2 & J.P.Y. Young 1 

1 Department of Biology, University of York, PO Box 373, 

YO10 5YW, U.K. - 2 Smithsonian Tropical Research 

Institute, Apartado 2072, Balboa, Panama. - E-mail: 

rh19@york.ac.uk 

It has been proposed that arbuscular mycorrhizal (AM) 

diversity may be linked to, and even influence, plant 

community structure. In intact tropical ecosystems little is 

known of the AM fungal population, so it has not been 

possible to assess the role of these fungi. Therefore we 

have used molecular techniques to investigate the diversity 

and distribution of the AM fungi colonising tree seedling 

roots in the tropical forest on Barro Colorado Island (BCI), 

Republic of Panama. We used differences in small subunit 

ribosomal RNA genes to identify groups of AM fungi 

colonising different host species of various ages at different 

sites. The overall AM fungal diversity was found to be very 

high although the majority of types belong to a single 

family. The mycorrhizal population showed significantly 

high spatial heterogeneity and non-random associations 

with the different hosts. Moreover there was a strong shift 

in the mycorrhizal communities over time. The high 

diversity and huge variation detected across time points, 

sites and hosts, implies the AM fungal types are 

ecologically distinct and thus may have the potential to 

influence recruitment and host composition in tropical 

forests.


225 - Organization of genetic variation within 

glomalean individuals 

T.E. Pawlowska * & J.W. Taylor 

University of California, Berkeley, CA 94720-3102, U.S.A. 

- E-mail: tpawlows@nature.berkeley.edu 

Understanding the organization of genetic variation within 

individuals of arbuscular mycorrhizal fungi (Glomales) is a 

prerequisite for the study of population genetics in this 

group. Polymorphism of rDNA arrays within individual 

glomalean spores inspired generally believed yet unproven 

speculation that these fungi are largely heterokaryotic. We 

tested this hypothesis using monoxenically cultured 

Glomus etunicatum representing a natural population from 

a maize field in California. In addition to the rDNA ITS1- 

5.8S-ITS2 region, a putative gene encoding catalytic 

subunit of DNA polymerase alpha (POL) was identified as 

a variable genetic marker. To test for homo- vs. 

heterokaryosis, we examined variant sorting of these two 

markers in spores formed clonally in cultures initiated from 

single spores. The markers were PCR-amplified from 

individual spores, cloned and sequenced. In the initial 

screen of the G. etunicatum experimental population, we 

detected four variants of the rDNA ITS region and 16 POL 

variants. We analyzed 16 to 200 clones per spore in 42 

spores representing five single-spore cultures. The patterns 

of the rDNA ITS and POL variant sorting in these cultures 

were consistent with the homokaryotic model of nuclear 

organization. The presence of distinct rDNA variants 

within individual nuclei implies that concerted evolution, 

which is responsible for homogenization of dispersed 

rDNA arrays in genomes of other organisms, does not 

operate efficiently in Glomales. 

226 - Diversity and distribution of ericoid mycorrhizal 

fungi in a Mediterranean forest 

S. Perotto * , R. Bergero & M. Girlanda 

Dipartimento Biologia vegetale dell'Univesità and IPP- 

CNR, V.le Mattioli 25 -10125 Torino, Italy. - E-mail: 

silvia.perotto@unito.it 

Ericaeous plants are widespread on the globe and colonize 

substrates ranging from humid mor-humus substrates to 

arid sandy soils. This variety of habitats opens intriguing 

questions on the biodiversity of their mycorrhizal 

associates. Ericoid mycorrhiza (EM) has been regarded for 

long time as a highly specific plant-fungus interaction, 

featuring a very restricted number of fungal species. More 

recent studies by research groups worldwide have 

challenged this view as molecular analyses of EM fungi 

suggest a greater genetic diversity and a larger number of 

fungal species than previously thought. An interesting 

feature of several EM fungi is the occurrence of Group I 

introns in the small rDNA subunit, which further increases 

their genetic diversity but which may also represent a 

potential problem for RFLP analyses. The molecular 

analysis of ericoid fungi has also lead to deeper 

understanding of their ecology and relationships with 

plants, and has revealed that ericaceous plants can be very 

promiscuous, with multiple occupancy of their thin roots. 

In addition, some EM fungi seem also able to colonise 

plants from very distant taxa. We have studied EM fungusplant 

relationships in Mediterranean forests, which are 

complex environments where high biodiversity in plant 

species and mycorrhizal types (arbuscular, orchid, ericoid, 

ecto- and ectendo-mycorrhiza) occur. In these 

environments, ectomycorrhizal and EM plants were found 

to share similar root endophytes. 

227 - Evolution of secondary metabolite pathways: nonribosomal 

peptide synthetases and polyketide 

synthetases 

S. Kroken 1* , N.L. Glass 1 , J.W. Taylor 1 , B.G. Turgeon 2 & 

O. Yoder 2 

1 University of California at Berkeley, 111 Koshland Hall, 

Berkeley, CA 94720-3102, U.S.A. - 2 Torrey Mesa Research 

Institute, 3115 Merryfield Row, Suite 100, San Diego CA 

92121-1125, U.S.A. - E-mail: kroken@nature.berkeley.edu 

Our aim is to identify signaling factors (in pathways 

required for hyphal network formation, sporulation, 

vegetative growth and environmental sensing) present in 

filamentous ascomycetes, and virulence factors necessary 

for pathogenicity in plant pathogens. We hypothesize that 

independent lineages recruited similar genes from their 

non-pathogenic ancestors for novel roles in plant 

pathogenesis. To test this hypothesis, we are performing 

comparative genomics of saprobes (Neurospora crassa and 

Aspergillus fumigatus) and plant pathogens (Cochliobolus 

heterostrophus, Botrytis cinerea, Fusarium verticillioides 

and F. graminearum). The first analyses identified genes 

that encode polyketides (PKs) and non-ribosomal peptides 

(NRPs). N. crassa has 7 polyketide synthases (PKSs) and 3 

non-ribosomal peptide synthetase (NRPSs), whereas the 

other filamentous ascomycete genomes have many more of 

these genes. Phylogenies of PKSs and NRPSs each feature 

a large clade that includes genes previously described as 

virulence factors. However, N. crassa has six PKSs and 

two NPRSs that group with the virulence clade, indicating 

that these genes have other roles. Based on these 

comparisons, we will select genes in N. crassa for 

mutational analysis. In parallel with these mutational 

analyses, metabolite profiling between wild-type N. crassa 

and mutants will be performed to match PKS- and NRPSencoding 

genes with secreted metabolites, which will be 

characterized and tested for biological activities. 



228 - Evolution of mating type genes in filamentous 

ascomycetes 

S. Poeggeler 

Ruhr-Uni-Bochum, 44780 Bochum, Germany. - E-mail: 

stefanie.poeggeler@ruhr-uni-bochum.de 

To gain a deeper insight into the evolution of reproductive 

life-cycles from filamentous ascomycetes, a comprehensive 

sequence analysis of PCR-amplified sequences 

corresponding to A- and a- specific mating-type sequences 

was undertaken. The study included nine homothallic 

(compatible) and eight heterothallic (incompatible) 

members of the genera Neurospora and Sordaria. Distance 

and parsimony trees based on gene fragments from the mat 

a-1 and mat A-1 genes were compared with trees derived 

from partial DNA sequences of the gpd glyceraldehyde-3phosphate 

dehydrogenase gene. In contrast to the 

sequences from the gpd gene, mating-type genes show 

striking sequence differences suggesting that these genes 

evolve very rapidly. Strong inter-relationships were found 

among homothallic, as well as among heterothallic 

members of both genera, indicating that in each genus, a 

change from one reproductive strategy to another might 

result from one single event. Pair-wise comparisons 

between pheromone genes and pheromone receptor genes 

of the heterothallic species Neurospora crassa and the 

homothallic Sordaria macrospora revealed an extremely 

low degree of nucleotide conservation in these genes, 

indicating that these, like mating-type genes, evolved also 

very rapidly. 

229 - Phylogeny of Zygomycetes: multi-gene 

approaches and the putative role of horizontal gene 

transfer 

K. Voigt 

Friedrich Schiller Universitaet, Institute of Microbiology, 

Department of General Microbiology and Microbial 

Genetics, Fungal Reference Centre, Neugasse 24, D-07743 

Jena, Germany. - E-mail: b5kevo@rz.uni-jena.de 

The Zygomycetes encompass microscopic fungi which 

form zygospores in sexual interactions. The most 

prominent and largest order are the Mucorales comprising 

saprotrophic and facultatively parasitic species, among 

those, the soil fungi Chaetocladium brefeldii and 

Parasitella parasitica which parasitize on other 

mucoralean fungi during the establishment of plasma 

bridges and the unidirectional transfer of genes to the host. 

This parasexual interaction utilizes the pheromone trisporic 

acid for the identification of compatible hosts, similarily to 

sexual interactions. One of the last steps of trisporic acid 

biosynthesis is catalysed by the 4-dihydromethyltrisporate 

dehydrogenase. The gene encoding this aldo-keto reductase 

were found in all families of the Mucorales, Mortierellales, 

Entomophthorales and Kickxellales. The sequences were 

analysed and used for the reconstruction of phylogenetic 

74 


trees. In order to study the evolution of Zygomycetes in 

multi-gene approaches sequences for the nuclear-encoded 

genes actin and beta-tubulin were also determined and 

applied in concatenated analyses of tree constructions. 

Phylogenetic analysis in the context of available sequence 

data (approx. 6300 nucleotide positions per species) 

revealed that current classification schemes for the 

mucoralean fungi are highly unnatural at the family and, to 

a large extent, at the genus level. 

230 - RAPD and microsatellite analysis of the local and 

global population structure of aflatoxigenic Aspergillus 

species 

N. Tran-Dinh 1 , J.I. Pitt 1 & D.A. Carter 2* 

1 Food Science Australia, P.O. Box 52, North Ryde NSW 

1670, Australia. - 2 School of Molecular and Microbial 

Biosciences, University of Sydney, Maze Crescent, 

Darlington, NSW 2006, Australia. - E-mail: 

dee.carter@staff.usyd.edu.au 

The mycotoxigenic species Aspergillus flavus and A. 

parasiticus have been well characterised with regard 

aflatoxin biosynthesis. Comparatively little is known, 

however, on their natural population structure in the 

environment. We have developed new molecular markers 

based on RAPDs and microsatellites to analyse large 

populations of these fungi on crops and in soils. 

Investigation of an Australian field population revealed 1) 

a new species of aflatoxigenic fungus, related but 

genetically distinct from A. flavus was common in this 

region. This is the same as the "Group 2" A. flavus, 

previously reported by Geiser et al. (PNAS 94: 388- 

393,1998); 2) recombination has occurred in the A. flavus 

population, but the A. parasiticus population and the 

population of the Group 2 A. flavus species are clonal; 3) 

no clear distinction between toxigenic and nontoxigenic 

strains in any of the species. Seven microsatellite markers 

were then used to analyse a large, worldwide collection of 

isolates. Globally, A. flavus was found to be cosmopolitan 

and there was no geographic substructuring within this 

species. In contrast, A. parasiticus was geographically 

restricted, being very rare throughout all of Asia. Group 2 

A. flavus isolates were likewise restricted, and all but two 

came from the southern hemisphere. The microsatellite 

data also indicated that genetic diversity was higher in A. 

flavus than in either the Group 2 or the A. parasiticus 

populations.


231 - Phylogeny of arbuscular mycorrhizal fungi 

(Glomales) 

D. Redecker 

Institute of Botany, University of Basel, Hebelstr.1, 4056 

Basel, Switzerland. - E-mail: dirk.redecker@unibas.ch 

Approximately 150 species of arbuscular mycorrhizal fungi 

(AMF) have traditionally been placed in the order 

Glomales in the Zygomycota. Recently the phylum 

Glomeromycota was established for them. Although this 

ubiquitous group of symbiotic fungi was shown to be at 

least 460 million years old, its diversification apparently is 

relatively low. According to rDNA phylogenies, the 

Glomeromycota are a sister group to Asco- and 

Basidiomycota. Taxonomy and identification of AMF has 

traditionally relied on spore morphology, but it has become 

increasingly clear that spore morphology is a relatively 

poor predictor of AMF species diversity. DNA sequences 

and sequence signatures have become important characters 

to define new taxa. Several lineages of AMF were shown 

to be deeply divergent within the Glomeromycota. Two 

new genera Archaeospora and Paraglomus were 

established for them, but the polyphyly of the remaining 

genus Glomus necessitated further taxonomic changes. The 

genetics of arbuscular mycorrhizal fungi is remaining 

mysterious, even more after they were shown to be 

heterokaryotic and predominantly non-recombining. 

Therefore it is problematic to apply evolutionary species 

concepts that have been successfully used in other fungi. 

232 - Fungi and the Northwest Forest Plan 

M.A. Castellano, T.A. Dreisbach * , T. O'Dell & R. Molina 

USDA Forest Service, 3200 Jefferson Way, Corvallis, 

Oregon 97331, U.S.A. - E-mail: Tdreisbach@fs.fed.us 

During the late 1980s and early 1990s a series of lawsuits 

severely decreased the harvest level of old-growth forests 

in the Pacific Northwest. At the center of the controversy 

was the old-growth dependent northern spotted owl (Strix 

occidentalis), a rare species protected by the Endangered 

Species Act. A regional forest conference to address the 

problem led to the development pf the Northwest Forest 

Plan. The plans goal was to provide for a sustainable 

timber harvest as well as the persistence of old-growth 

forest dependent species. Many rare species requiring oldgrowth 

forest habitat were analyses for protection under the 

guidelines and recommendations of the plan. Four hundred 

presumptive old-growth dependent species, including 234 

fungal species, were identified that needed additional 

protection under a legally binding document called the 

Record of Decision that defines how the Northwest Forest 

Plan was to be implemented. We will briefly review the 

process that led to creation of the Northwest Forest Plan, 

then explore the challenges of implementing it and 

examine the early impact it has had on research of forest 

fungi in the Pacific Northwest United States. 

233 - Putting Australian fungi on the map 

T.W. May 

Royal Botanic Gardens Melbourne, Private Bag 2000, 

South Yarra, Victoria 3141, Australia. - E-mail: 

tom.may@rbg.vic.gov.au 

Relatively few of Australia's numerous fungi have been 

thoroughly documented. Many species are yet to be 

described, yet there are very few Australian mycologists 

and comparatively few specimens of fungi in Australian 

herbaria. However, three current government initiatives are 

stimulating progress in cataloguing and understanding the 

diversity of the Australian mycota. Fungi have been 

included in the program of the Australian Biological 

Resources Study, which publishes the Fungi of Australia 

series. A second key initiative which embraces fungi is the 

current effort to comprehensively database specimens in 

state botanical herbaria (the Australian Virtual Herbarium). 

Thirdly, Fungimap, a partnership between government and 

community groups, is drawing on recorders across 

Australia to rapidly advance knowledge of the distribution 

and conservation status of selected macrofungi. Research 

on fungi can also result from their myriad interactions with 

other biota. A prominent example is the need to effectively 

manage endangered mammals that subsist largely on native 

truffles. This has stimulated taxonomic and ecological 

research on Australian truffles, which have been revealed 

as among the most diverse in the world. Continued 

progress in documenting Australia's mycota depends on 

government support for the maintenance and creation of 

mycological positions within herbaria, and the further 

integration of fungi into national biodiversity and 

conservation programs. 

234 - Fungi within the UK Biodiversity Action Plan 

J.C. Duckworth 1* , S.E. Evans 2 & L.V. Fleming 3 

1 Plantlife, 21 Elizabeth Street, London SW1W 9RP, U.K. - 

2 BMS Conservation Officer, Joseph Banks Building, Royal 

Botanic Gardens, Kew, Surrey TW9 3AE, U.K. - 3 Joint 

Nature Conservation Committee, Monkstone House, City 

Road, Peterborough PE1 1JY, U.K. - E-mail: 

jenny.duckworth@plantlife.org.uk 

The development and inclusion of fungi in the UK 

Biodiversity Action Plan (BAP) is briefly reviewed. The 

extent to which the conservation of fungi has been 

advanced by this process is considered. The inclusion of 

some fungi (27 taxa) represents a significant achievement 

for a taxonomic group which is traditionally underrepresented 

in the setting of conservation priorities. 

Nevertheless, fungi remain the least well represented in 

comparison with other taxonomic groups. The action taken 

since the publication of the plans is summarised. 

Implementation of the plans includes a review and 

expansion of survey work on these target species which has 

resulted in an increase in our knowledge about their UK 



distribution. It has also facilitated scientific studies into the 

taxonomy and ecology of targeted species. The progress to 

date enables some reflections to be made on the 

appropriateness of the taxa chosen for action plans and the 

criteria used to select them. Observations are made on how 

the priorities and the process might be improved and 

adjusted to changing circumstances and continually 

improving knowledge. The influence of the process on the 

support given to the conservation of fungi by the 

government and non-government sectors is considered. 

However, the benefits arising from the inclusion of fungi in 

the UK BAP have not been confined solely to the target 

species. A much higher priority for, and a greater 

partnership approach to, the conservation of fungi is now 

evident. 

235 - The importance of fungi in IUCN Programmes 

R. Courtecuisse 1 & C. Perini 2* 

1 Département de Botanique, Faculté de Pharmacie, B.P. 

83, F-59006 Lille Cedex, France. - 2 Dipartimento di 

Scienze Ambientali, via P.A. Mattioli 4, I-53100 Siena, 

Italy. - E-mail: perini@unisi.it 

IUCN (World Conservation Union) represents the world's 

oldest and largest global conservation body ; it is a 

powerful and well-structured organisation, devoted to 

nature conservation all over the world, seeking to 

influence, encourage and assist societies throughout the 

world to conserve the integrity and diversity of nature and 

to ensure that any use of natural resources is equitable and 

ecologically sustainable. Activities involve several 

thousands of people, distributed in about 150 countries. 

Fungi have been rather recently considered in IUCN scope, 

especially in establishing a Specialist Group for Fungi 

(within the Species Survival Commission), which aims to 

list the threats to Fungi, to consider the conservation 

strategies and to promote actual conservation actions for 

threatened Fungi. Testing and improving the red list criteria 

and categories have been another recent priority. The 

current investigations will be browsed and some example 

of current actions, some of them in collaboration with other 

organisms or councils (for example ECCF, European 

Council for the Conservation of Fungi) will be given. The 

Bern Convention case (for Europe), involving relations 

between several European states and demonstrating some 

difficulties in such international actions will illustrate this 

purpose. Finally global perspectives for future conservation 

of Fungi will be considered. 

236 - Conservation based on genetic diversity 

J.-M. Moncalvo 1* , J.E. Johnson 2 & R. Vilgalys 1 

1 Department of Biology, Duke University, Durham, NC 

27708, U.S.A. - 2 Department of Biological Sciences, 

Cameron University, Lawton, OK 73505, U.S.A. - E-mail: 

jeanmarc@duke.edu 

76 


Conservation biology requires knowledge about biological 

diversity. Classic estimates of biological diversity use 

species (or genera) as proxies. The development of largescale 

molecular phylogenies provides a unique opportunity 

to estimate biological diversity at the genetic level by using 

the branching order and branch lengths from phylogenetic 

trees. Both among-taxa and among-areas phylogenetic 

diversity can be estimated and compared with the use of 

the 'phylogenetic index of diversity' (PD). We will contrast 

PD with classic estimates of biological diversity in various 

groups of mushrooms for which extensive geographic 

sampling and molecular phylogenies are available, 

including genera Amanita, Pleurotus, Xeromphalina, and 

Ganoderma. There are at least two advantages of PD over 

traditional measures of estimating biological diversity: 1) 

PD does not require a priori knowledge of taxonomic 

circumscription (which is generally subjective in fungi), 

and 2) PD takes into account genetic distances, therefore 

emphasizes genetic breath when estimating diversity. 

237 - The Convention on Biodiversity triggers 

biodiversity research programs? 

S.S. Dhillion * & M.A. Støen 

Department of Biology and Nature Conservation, 

Agricultural University of Norway & SUM, University of 

Oslo, IBN, pb5014, NLH, N- 1432 Ås, Norway. - E-mail: 

shivcharn.dhillion@ibn.nlh.no 

Both member and non-member states have responded in 

varied ways to the Convention on Biodiversity. This paper 

presents the influence of the CBD in triggering biodiversity 

programs, and explores if regulatory conditions and 

expertise exist for their implementation. To date the Thai 

government has been exemplary in setting up biodiversity 

research and competence building programs, and drafting 

regulations on biodiversity even though it is not a signatory 

to the CBD. Now it plans to form a Committee on National 

Biological Diversity Conservation and Utilization to 

oversee and harmonize all biodiversity activities in the 

country. The cross-sectorial and participatory approach 

employed by Thailand is exemplary, but not devoid of 

conflicts. Malaysia and the Phillippines, a signatory to the 

CBD, have had specific research grant programs for 

biodiversity launched after ratifying the CBD. The 

Philippines has now enacted one of the most stringent 

biodiversity laws in the world. The enforcement of 

regulations in all three countries is weak and not all 

government, academic, and private institutions are well 

informed of protocols required for biodiversity studies. To 

a large extend both Thailand and the Philippines rely on 

foreign collaboration skills to catalog species. Nevertheless 

all three countries have adopted distinct modes of 

addressing CBD objectives and are much ahead of most 

nations in doing so.

IMC7 Thursday August 15th Lectures 

238 - Capturing mycological data, the Caribbean 

experience 

M. Rodríguez Hernández 

Jardín Botánico Nacional, Carretera del Rocio km 3.5, 

Calabazar, Boyeros, La Habana, CP 19230, Cuba. - Email: 

hajb@ceniai.inf.cu 

Since 1997 a team of mycologists, mainly from Cuba and 

the UK (but with additional contributors from Belgium, 

Puerto Rico, Trinidad & Tobago and Venezuela) 

participated in the UK Darwin Initiative project 'Fungi of 

the Caribbean'. This project aimed to: establish 

infrastructure necessary for a regional fungal identification 

service in the Caribbean; carry out field work in Cuba and 

Trinidad & Tobago, augmenting mycological reference 

collections in both countries; produce a computarized 

checklist of Caribbean fungi based on all this information. 

To achieve the last of these aims, it was necessary to 

capture information from living and preserved reference 

collections, and from bibliographic sources, and to enter 

that information into a suite of previously established 

relational databases (Biological Records, Nomenclature & 

Taxonomy, Bibliographic). The main software used for 

data entry, data storage and output was the 

SMARTWAREII database system. The resulting 

publication, Fungi of the Caribbean, of almost 1000 pages, 

was based on almost one hundred and fifty thousand 

(150,000) different biological records of fungi and 

associated organisms. Many practical problems can be 

encountered during capture of mycological data. To ensure 

high quality keyboarding, curation and output, suitable 

training is therefore necessary for all the team, and a clear 

set of data entry protocols must be supplied before work 

starts. 

239 - PLANTCOD - to the database formation in the 

Komarov Botanical Institute on biodiversity of plants 

and fungi in Russia 

A.L. Lobanov 1 , Yu.K. Novozhilov 2 , A.E. Kovalenko 2* & 

D.W. Minter 3 

1 Zoological Institute of the Russian Academy of Sciences, 3 

Universitetskaya embankment, St. Petersburg, Russia. - 

2 V.L. Komarov Botanical Institute of the Russian Academy 

of Sciences, 2 Prof. Popov Street, 197376, St. Petersburg, 

Russia. - 3 CABI Bioscience, Egham, Surrey, U.K. - E-mail: 

alkov@AK3010.spb.edu 

Practically all the databases created in the Komarov 

Botanical Institute RAS have the taxonomical blocks. In 

the majority of these blocks the modification of 

ZOOCOD3 (Lobanov et al., 1999) concept is used to 

reflect the hierarchical classifications of plants and fungi in 

the flat relational tables. The field used for detailed 

representation of classification are the following: 

LATNAM (Latin name of a taxon), RANCOD (taxonomic 

code of a rank), SYSCOD (digital systematic code), SYN 

(code of synonymy), ABBR (unique mnemonic code of a 

taxon). The fields KEY (unique digital code of a taxon) and 

PARENT (value of KEY for the nearest parent) work with 

the Tree View Windows control to represent a hierarchy of 

taxa in a form of tree. Though these two fields adequately 

reflect multilevel hierarchy they are inconvenient for fast 

search of all descendants belonging to the same taxon. To 

decide the problem, the filling rules are changed for the 

SYSCOD field. The field SYSCOD is filled by the special 

utility, but the search of descendants of the given taxon is 

executed (with lightning speed and extremely simple) by 

superposition of the filter with a completely optimised 

condition. (This study was supported by grant 02-07- 

90439, Russian Foundation for Basic Research; scientific 

program 'Information system on biodiversity' 

43.073.1.1.2510, Ministry of Industry and Science, Russia; 

Darwin Initiative awards, UK Government). 

240 - Information systems can improve the efficiency of 

biodiversity research, promoting collaboration and 

sharing of information 

G. Hagedorn 

Federal Biological Research Center for Agriculture and 

Forestry, Institute for Plant Virology, Microbiology and 

Biological Safety, Königin-Luise-Straße 19, D-14195 

Berlin, Germany. - E-mail: g.hagedorn@bba.de 

Given the extent to which basic biodiversity information is 

missing - organisms being undescribed or described so 

poorly that identification is impractical - we urgently need 

to improve the efficiency of biodiversity research. To some 

degree biodiversity informatics can contribute, providing 

more efficient workbench applications and efficient sharing 

of information through web services and (xml-based- 

)interoperability data standards. - Information sharing 

covers sharing basic infrastructure resources like 

nomenclatural or reference data (which can be combined 

with project-oriented work on taxonomic assessment, 

descriptions, or reference indexing tasks). Significantly, it 

also covers sharing assessment and analysis data between 

generations of researchers, leading mycology into an age 

where updates of previous monographic works concentrate 

on new results rather than laboriously repeating work 

already done. Thought has to be given to intellectual 

property rights and copyright issues. The newly introduced 

database copyright could endanger such efficiency 

improvements and stifle future research. - The 

development of information systems capable of fulfilling 

this promise is a huge task itself. Information models need 

to be modular to simplify analysis. Applications should be 

developed as independent components that cooperate in a 

framework. Such a framework, the DiversityWorkbench 

suite of applications, is currently being conceptualized and 

developed (see http://www.diversitycampus.net/). 



241 - Use of nomenclatural and bibliographic 

mycological data 

P.M. Kirk 

CABI Bioscience, Bakeham Lane, Egham, Surrey TW20 

9TY, U.K. - E-mail: p.kirk@cabi.org 

Nomenclatural and bibliographic databases are important 

tools in mobilizing mycological information, especially 

within the subject area of biodiversity information. The 

nomenclator is fundamental to biosystematics research, 

listing names which have been published in the literature. 

The service it provides to biosystematists, as both 

producers and users of names, is obvious but most users of 

names have a somewhat different requirement. The two 

types of databases can be conveniently and favourably 

linked through the 'potential taxon' model of Berendsohn. 

This can deliver the service that users of names require; not 

the association of a name with a specimen (the type of the 

name) but the association of a name with an accepted 

taxon, usually a species. The global database of fungal 

names (http://www.indexfungorum.org/) originated as a 

nomenclator but a partial implementation of the potential 

taxon model allowed for the inclusion of taxonomic 

opinion, with heterotypic synonyms retrieved through a 

single-cycle recursive join. This simple and pragmatic 

approach has been very successful. The services it can and 

does provide will be demonstrated and possible 

enhancements and linkages will be explored. Any database 

today should seamlessly deliver to ink-on-paper and the 

Internet; the latter is the future. 

242 - Use of mycological data for producing a checklist, 

US experience 

D.F. Farr 

USDA, Systematic Botany and Mycology Laboratory, 

BARC-West, Rm. 304, Bldg. 011A, Beltsville MD 20705, 

U.S.A. - E-mail: davef@nt.ars-grin.gov 

The Systematic Botany and Mycology Laboratory has been 

a pioneer in the use of computer databases to manage 

mycological information. We found that success depends 

on several factors: clear goals, efficient user interfaces for 

data entry and continuous management of personal 

resources. Once the big push to complete a computer 

database project is over, the organization is faced with the 

need to maintain the database. To be successful over the 

long term database projects need to form an important 

component of the organization's reference materials and 

collection's management tools. Now that many mycological 

databases are available on the web we need to begin to look 

at ways in which a single user interface can be used to 

retrieve and integrate information from various Web sites. 

One way that this could be done is to extract data from a 

remote database on an as needed basis. The term 'Web 

Services' is being applied to a standard set of procedures 

that are being developed to handle this type of interaction. 

78 


These procedures can greatly simplify the exchange of 

information between remote databases and it is important 

that mycologists make use of these procedures to further 

enhance the utilization of the databases that we have 

developed. 

243 - NZFUNGI - a biodiversity information system for 

fungi in New Zealand 

J.A. Cooper 

Landcare Research, PO Box 40, Lincoln 8152, New 

Zealand. - E-mail: Cooperj@landcareresearch.co.nz 

Landcare Research is New Zealand's foremost institute in 

systematics and environmental research in terrestrial 

ecosystems, and is the custodian of the national collections 

and associated databases of plants, insects, fungi, bacteria 

and nematodes. NZFUNGI is part of an ongoing project to 

unlock our extensive data on biodiversity. It comprises a 

cross-indexed information facility which integrates a 

number of existing and new resources on taxonomy, 

pathology, literature, specimens, cultures, descriptions, 

images and keys. Integration of these data was facilitated 

using a taxonomic framework provided by the extensive 

authority files of CABI Bioscience, together with a number 

of standard authority files and database structures (e.g. 

TDWG data standards, dictionaries, and the IOPI 

taxonomic information model). Software systems for these 

data are being developed to maximize the opportunities 

provided by the recent and rapidly development enabling 

technologies of XML/SOAP and web services. Adoption of 

this technology is allowing us to address some important 

issues. For example, it allows us to provide secure and 

sophisticated application interfaces that will operate 

anywhere on the net, and to consider the virtual integration 

of distributed data resources and functionality. The 

presentation will provide a demonstration and discussion of 

work to-date. 

244 - MycoKey - a database and a mycological expert 

system 

T. Læssøe 1* & J.H. Petersen 2 

1 Botanical Institute, Copenhagen University, Oester 

Farimagsgade 2D, DK-1353 Copenhagen K, Denmark. - 

2 Department of Systematic Botany, Institute of Biological 

Sciences, University of Aarhus, Nordlandsvej 68, DK-8240 

Risskov, Denmark. - E-mail: ThomasL@bot.ku.dk 

MycoKey is a user friendly genus identification system for 

fungal genera. It is based on the relational database 

software 'Forth Dimension' and will be distributed in both 

mac and PC versions. A beta version will be available at 

the congress. At present MycoKey includes 530 genera of 

Basidiomycota (excluding rusts and smuts) known from 

Northern Europe. MycoKey is based on a character matrix 

of more than 1.000 characters, 17 interrelated files and


thousands of lines of custom programming. The result is an 

application with floating menues and a fully illustrated 

interface hiding the underlying complexity for the user. It 

contains information on authors, synonyms, type species 

and references. It also includes generic descriptions and not 

least numerous photographs of key taxa. With the '4D 

engine' MycoKey is compiled into a multi-platform, stand 

alone application for sale. MycoKey is also served by 4D 

and Webstar for use on the internet, see 

http://www.mycokey.com/. 

245 - A phylogenetic overview of the groups of fungi 

and fungus-like organisms that attack nematodes 

R.G. Thorn 

Department of Biology, University of Western Ontario, 

London, N6A 5B7, Canada. - E-mail: rgthorn@uwo.ca 

This talk will provide an introduction to the groups of 

nematode-destroying 'fungi', a guild of fungi and funguslike 

organisms that attack living nematodes and consume 

them as food. These organisms may be important in 

regulating nematode populations in nature and, in certain 

circumstances, may act as biological control agents of 

nematode pathogens and parasites of crop plants and 

livestock. Over 400 species of predatory and parasitoid 

nematode-destroying fungi are known, including 

approximately 150 species of Basidiomycota 

(Hyphodermataceae and Pleurotaceae), 130 species of 

Ascomycota (Clavicipitaceae, Hypocreaceae and 

Orbiliaceae), 100 Zygomycota (Cochlonemataceae, 

Helicocephalidaceae, Meristacraceae and Zoopagaceae), 10 

Chytridiomycota (Catenariaceae), and 40 Oomycota 

(Stramenopila: Myzocytiopsidaceae). In predatory species, 

one spore germinates to form a mycelium capable of 

attacking many nematodes; in parasitoids, each propagule 

attacks a single nematode. In the nematode-destroying 

Ascomycota and Basidiomycota, possession of a sexual 

state is apparently correlated with the predatory habit. 

Although the ability to attack and kill nematodes has arisen 

multiply (as has a marvellous diversity of means to achieve 

this end), this has not been a random process. Where well 

known, groups of nematode-destroying fungi are 

apparently monophyletic. 

246 - Teleomorph-anamorph connections in Orbiliaceae 

D.H. Pfister * & K.F. LoBuglio 




With the establishment of the teleomorph anamorph 

relation between certain of the hyphomycetous nematode 

trappers and the Orbiliaceae several phylogenetic and 

biological questions have come to light. Using molecular 

phylogenetic methods it has been suggested previously that 

the Orbiliaceae occupies a phylogenetic postion near the 

base of the Euascomycota tree along with the Pezizales. 

While the nematode trapping life style exists in several 

fungal groups there is no evidence of nematode trapping in 

the groups most closely related to the Orbiliaceae. We will 

present further molecular sequence data, particularly from 

the LSU rDNA and the RPB2 genes supporting the 

placement of the Orbiliaceae in this basal position. The 

implications of this specialized type of life strategy as 

represented in a basal member of the Euascomycota will be 

discussed. 

247 - Beyond Saccardo: a new generic system for 

nematode-trapping orbiliaceous hyphomycetes 

M.S. Scholler 1* & G. Hagedorn 2 

1 Purdue University, Department of Botany & Plant 

Pathology, U.S.A. - 2 Biologische Bundesanstalt fuer Land- 

und Forstwirtschaft, Koenigin-Luise-Strasse. 19, 14195 

Berlin, Germany. - E-mail: scholler@purdue.edu 

Perfect states are unknown for the majority of orbiliaceous 

nematode-trapping fungi. Therefore, generic concepts for 

anamorphs have been developed in which the morphology 

of conidia and conidiophores were preferentially used for 

generic delimitation. The mode and morphology of the 

trapping device, however, was only considered for species 

delimitation. A new concept is proposed with the mode of 

trapping device as the main morphological feature for 

generic delimitation. This new concept includes four 

genera, viz. Arthrobotrys Corda em. forming adhesive 

networks, Drechslerella Subram. em. forming constricting 

rings, Dactylellina M. Morelet em. forming stalked 

adhesive knobs, and Gamsylella gen. nov. producing 

adhesive columns and unstalked knobs. The concept 

corresponds well with molecular (rDNA sequences:18S, 

ITS I, II) and some ecological, physiological, and 

biological features. Also, chlamydospore formation, 

distribution, and host range are, to some degree, genusspecific. 

248 - Nematode endoparasites with verticillium-like 

anamorphs 

W. Gams 1* & R. Zare 2 

1 Centraalbureau voor Schimmelcultures, P.O. Box 85167, 

3508 AD Utrecht, The Netherlands. - 2 Dep. of Botany, 

Plant Pests and Diseases Res. Inst., P.O. Box 1454, Tehran 

19395, Iran. - E-mail: gams@cbs.knaw.nl 

Verticillium comprises many unrelated anamorphs. The 

type and the best-known plant-pathogens are 

Phyllachorales. Verticillium-like anamorphs of 

Clavicipitaceae, previously in sect. Prostrata, are 

distributed over several genera. Many are entomogenous or 

fungicolous, some discrete groups are nematode 

endoparasites. A phylogeny-based distinction separates 



species that attack free-living nematodes with adhesive 

conidia and parasites of nematode eggs and cysts. The two 

most important genera, Lecanicillium and Pochonia (syn. 

Diheterospora) differ by formation of 

dictyochlamydospores, stronger cyanophily of the mostly 

short conidia, and absence of crystal formation in the latter. 

In Lecanicillium (teleom. Torrubiella, Cordyceps), only 

few taxa are mildly nematode-parasitic. P. chlamydosporia 

(teleom. Cordyceps) and related species are efficient egg 

parasites of Heterodera and Meloidogyne species, 

promising in biological control. Species of Simplicillium 

(teleom. Torrubiella), lacking verticillate phialides, fall 

phylogenetically outside Lecanicillium; they also parasitize 

nematode eggs. Species with adhesive conidia form a 

discrete genus, Haptocillium. Its species were so far known 

as V. balanoides, but at least 7 species are distinct, the most 

frequent being H. sphaerosporum. Contrasting with 

Drechmeria coniospora, Haptocillium species attack a 

wider array of free-living nematodes and are promising 

biocontrol candidates. 

249 - Novel biologically active metabolites from 

Pochonia chlamydosporia and preliminary studies on 

their distribution in Verticillium-like anamorphs 

M. Stadler 1* , V. Hellwig 1 & H.V. Tichy 2 

1 Bayer AG, Pharma Research, P.O.Box 10 17 09, D-42096 

Wuppertal, Germany, Germany. - 2 TÜV ISB, Engesserstr. 

4b, D-79108 Freiburg, Germany. - E-mail: 

marc.stadler@t-online.de 

Extracts from the fungus Pochonia chlamydosporia var. 

catenulata strain P0297 showed interesting antiviral and 

antiparasitic activities. As active principles, we identified 

the known fungal metabolites, Monorden and Pseurotin A, 

besides several novel natural products, for which we 

propose the trivial names Pochonins A-E. The isolation and 

characterisation of these compounds is summarised in a 

concurrent poster. The current paper deals with studies on 

the distribution of secondary metabolites in strain P0297 

and further Verticillium-like anamorphs from public 

collections by HPLC-based secondary metabolite 

fingerprinting, aided by morphological comparisons and 

Minisatellite PCR, using the methodology recently 

developed in our studies of Xylariaceae [Stadler et al, 

Mycological Research 105, 1191-1205, 2001]. According 

to preliminary results, the production of resorcylic acid 

lactones is restricted to particular species of the genus 

Pochonia sensu Gams & Zare [Nova Hedwigia 72 (3-4), 

329-337, 2001], while these compounds were not detected 

in several species now accomodated in Lecanicillium and 

Haptocillium. Therefore, the recent segregation of the 

genus Pochonia from other Verticillium-like anamorphs 

based on morphological and molecular methods by Gams, 

Zare and co-workers also appears justified from a 

chemotaxonomical point of view. 

80 


250 - Rhizosphere biology of nematophagous fungi 

H.-B. Jansson * , J.J. Bordallo, L.V. Lopez-Llorca & J. 

Salinas 

Depto. de Ciencias Ambientales y Recursos Naturales, 

Universidad de Alicante, Apdo 99, E-03080 Alicante, 

Spain. - E-mail: hb.jansson@ua.es 

Nematophagous fungi can infect, kill and digest living 

nematodes. Most of these fungi can also live 

saprophytically and some even have mycoparasitic 

abilities. Since most plant-parasitic nematodes attack plant 

roots, the rhizosphere biology of nematophagous fungi is 

important from a biological control point of view. We 

previously knew that nematophagous fungi were more 

abundant in the rhizosphere than in bulk soil. In this study, 

barley and tomato axenic plants were inoculated with the 

nematode-trapping Arthrobotrys oligospora or the eggparasitic 

fungus Pochonia chlamydosporia (=Verticillium 

chlamydosporium). Roots were sequentially sampled, cryosectioned, 

and observed under light- or cryo-scanning 

electron microscopes. Both fungi grew inter- and 

intracellularly, formed appressoria when penetrating plant 

cell walls of epidermis and cortex cells, but never entered 

vascular tissues. Using histochemical stains we could show 

plant defence reactions, e.g. papillae, lignitubers and other 

cell wall appositions, but these never prevented root 

colonization. Nematophagous fungi grew extensively 

especially in monocotyledon plants producing abundant 

mycelia, conidia and chlamydospores (P. chlamydosporia). 

Whether this endophytic growth induces systemic 

resistance in plants is yet unknown, but worth further 

investigations. The ability to colonize plant roots may also 

be a survival strategy of these fungi and could explain soil 

suppressiveness to plant-parasitic nematodes in nature. 

251 - Holocarpic biflagellate parasites of nematodes 

including Chlamydomyzium and Myzocytiopsis 

S.L. Glockling 

Northern Illinois University, NIU, DeKalb, Illinois, IL 

60115-2861, U.S.A. - E-mail: sally@glockling.com 

In comparison to related straminipilous fungi that are easily 

cultured, the obligate nematode parasites are a little known 

group that have been considerably understudied. Because 

of their host-dependent nature and the paucity in 

phylogenetically significant data, their position within the 

peronosporomycetes, and their relationship to one another, 

has yet to be established. The nematode parasites produce a 

holocarpic thallus inside the bodies of their bactiverous 

nematode hosts. The infective agents are either motile 

biflagellate zoospores or adhesive aplanospores, which 

attach to the nematode cuticle and penetrate with a germ 

tube to initiate infection. This presentation looks at the 

main diagnostic characteristics of Chlamydomyzium and 

Myzocytiopsis and at the diversity within the genera. 

Development of the thallus into sporangia or into resting


spores is demonstrated using both light microscopy and 

transmission electron microscopy (TEM), and the process 

of zoospore release is followed in a video sequence. A 

phylogenetic tree (mitochondrial coxII gene) of 

representative peronosporomycetes has been compiled 

(Hudspeth & Hudspeth) and includes a few species of 

nematode parasites. 

252 - What makes a species? Redundancy, 

recombination, and reproductive isolation 

S.G. Oliver 

School of Biological Sciences, University of Manchester, 

2.205 Stopford Building, Oxford Road, Manchester M13 

9PT, U.K. - E-mail: steve.oliver@man.ac.uk 

The Saccharomyces "sensu stricto" group of yeast mate 

with S. cerevisiae, but produce hybrids that yield spores of 

very low viability, such that speciation seems to have 

arisen via reproductive isolation. Chromosomal 

rearrangements might act as a post-zygotic isolating 

mechanism. We studied the distribution of chromosomal 

translocations in 5 Saccharomyces "sensu stricto" species 

(S. paradoxus, S. bayanus, S. cariocanus, S. mikatae and S. 

kudriavzevii) by comparing them to the S. cerevisiae 

genome. No correlation was found between the sequencedbased 

phylogeny of these species and the presence of 

translocations. Instead, bursts of rearrangements are seen 

between closely related species, while more distant ones 

have co-linear genomes. Thus, chromosomal 

rearrangements are not the primary cause of yeast 

speciation, but may intensify reproductive isolation once a 

species has arisen by another route. We are using a novel 

molecular approach to generate yeast strains containing 

specific chromosomal translocations. Strains of S. 

cerevisiae have been constructed that contain the 

translocations found in other Saccharomyces "sensu 

stricto" species. These are then mated, both to wild-type S. 

cerevisiae and to the "sensu stricto" species whose genome 

configuration they mimic. The data show that 

translocations make a significant contribution to the postzygotic 

isolation of species and indicate a mechanism for 

genome duplication via allopolyploidy. 

253 - Genus relationships in the Saccharomycetales 

from multigene analyses 

C.P. Kurtzman 

National Center for Agricultural Utilization Research, 

1815 N. University Street, Peoria, IL 61604, U.S.A. - Email: 

kurtzman@ncaur.usda.gov 

As with most fungi, circumscription of yeast genera is 

based primarily on phenotype. Gene sequence 

comparisons, such as from 18S or 26S rDNAs, have shown 

that many presently defined yeast genera are not 

monophyletic, but single gene analyses seldom provide 

sufficient resolution to unambiguously circumscribe 

genera. Analysis of the ca. 80 known species of the 

Saccharomyces clade from EF-1, RNA polymerase II, 

cytochrome oxidase II and actin, as well as from 18S, 26S 

and mitochondrial small subunit rDNAs, individually gave 

congruent terminal lineages and, when analyzed in 

combination, provided strong genus-level support. 

Intergeneric relationships are less well resolved making 

family assignments uncertain. A comparison of species in 

the phylogenetically distant Stephanoascus/Blastobotrys 

clade gave similar results. Diagnostic phenotypes were 

recognized for many of the phylogenetically defined 

genera, but for some genera, there appeared to be no 

unifying morphological or physiological characters. 

254 - Molecular approaches to the re-appraisal of 

species diversity within the ascomycete genus Taphrina 

A. Fonseca * & M.G. Rodrigues 

Centro de Recursos Microbiológicos (CREM), Secção 

Autónoma de Biotecnologia, Faculdade de Ciências e 

Tecnologia, Universidade Nova de Lisboa, Quinta da 

Torre, 2829-516 Caparica, Portugal. - E-mail: 

amrf@mail.fct.unl.pt 

The dimorphic ascomycete genus Taphrina Fries 

comprises nearly 100 species recognised by their mycelial 

states parasitic on different vascular plants. Whereas the 

filamentous state occurs exclusively in plant tissue, the 

yeast state is saprobic and can be cultured on artificial 

media. Taphrina species are differentiated mainly on the 

basis of host range and geographic distribution, type and 

site of infection, and morphology of the sexual stage in the 

infected tissue. However, there has been little progress in 

the systematics of the genus in recent years mainly due to 

the scarcity of molecular studies and of available cultures. 

Here we report on the molecular characterisation of 

Taphrina spp. obtained from culture collections (yeast 

states) in order to address the following topics: (i) is 

Taphrina a well defined genus?; (ii) are species defined on 

the basis of phenotypic criteria genetically distinct?; (iii) 

does host specialisation underlie speciation in Taphrina?; 

(iv) do Taphrina spp. occur naturally as saprobic yeast 

forms?; (v) can molecular methods help in accurately 

diagnosing the different diseases caused by Taphrina 

species? The molecular methods used comprised PCRfingerprinting 

using single primers for microsatellite 

regions (MSP-PCR), and sequencing of two ca. 600 bp 

long nuclear rDNA regions: the 5' end of the 26S rRNA 

gene (D1/D2 domains) and the Internal Transcribed Spacer 

(ITS) regions (including the 5.8S rRNA gene). 



255 - Biogeography of floricolous yeasts: is everything 

everywhere? 

M.A. Lachance 

Department of Biology, University of Western Ontario, 

London, Ontario N6A 5B7, Canada. - E-mail: 

lachance@uwo.ca 

The old dogma of microbial ecology, 'Everything is 

everywhere, the environment selects,' has had a profound 

influence on the study of yeast biodiversity. One outcome 

has been a widespread neglect of the habitat as a significant 

element of species descriptions. The traditional paradigm 

implies that a global 'yeast seed bank' is available to fill 

any niche that is made available. Biogeographic theory, on 

the other hand, predicts that species diversity should be 

higher in the tropics, lower in isolated localities, and 

proportional to the size of contiguous landmasses. To test 

these opposing models, the yeast communities of 

ephemeral flowers were studied in various Pacific islands 

and several sites in Australia, Brazil, Costa Rica, the 

southern United States, and the eastern Nearctic region. 

These yeasts are vectored and maintained by insects such 

as bees and beetles. The yeast species composition is 

greatly but not exclusively affected by the nature of the 

vector insect species, and much less so by the plant species. 

Most biogeographic factors have a significant influence 

when not confounded by human interference. Different 

yeast species have different ranges on the global scale: 

some can be viewed as cosmopolitan and others as 

endemic. The emerging pattern is that indeed, the 

environment selects. However, geography plays a major 

role. The notion that 'everything is everywhere,' as least at 

it applies to floricolous ascomycetous yeasts, is misleading. 

256 - Molecular ecology of basidiomycetous yeasts in 

tropical marine habitats 

J.W. Fell 

University of Miami, 4600 Rickenbacker Causeway, Key 

Biscayne, Fl, U.S.A. - E-mail: jfell@rsmas.miami.edu 

The current model of yeast systematics includes ^ 1000 

species in >100 genera, numbers that are rapidly increasing 

with the discovery of new species and the description of 

new genera due to increased knowledge of the biology and 

phylogenetics of these unicellular ascomycetous and 

basidiomycetous fungi. Estimates indicate that this number 

of species may only represent 1% of the species in nature. 

Yeasts in marine environments are widespread from the 

tropics to polar regions and intertidal habitats to the deep 

sea floor. The ecological role of yeasts ranges from species 

that are host specific saprophytes to species with diverse 

habitats and appetites. Our specific knowledge of these 

roles is, however, meager. A major reason has been the 

lack of ability to specifically identify these organisms to 

the species level. The advent of molecular techniques has 

provided the necessary cure to the systematic problems. 

82 


Research in our laboratory with the basidiomycetous yeasts 

has centered on the ribosomal DNA, exploring the ITS, 

D1D2 portion at the 5' end of the large subunit, and the IGS 

regions. Based on these data bases, the phylogenetic 

position and identity of yeasts can be determined. The 

sequence differences between strains and species has 

spawned methods for the rapid identification of culturable 

and uncultured species directly from the environment. 

These methods will be discussed. 

257 - Systematics of the human pathogen Cryptococcus 

neoformans in the genomics era 

T. Boekhout 1* , B. Theelen 1 , M. Diaz 2 , J.W. Fell 2 , K.J. 

Kwon-Chung 3 , M.T. Barreto de Oliveira 4 , L. Trilles 5 , M. 

Lazera 5 , R. Falk 6 , I. Polacheck 6 & W. Meyer 7 

1 Centraalbureau voor Schimmelcultures, Uppsalalaan 8, 

3584 CT Utrecht, The Netherlands. - 2 Rosenstiel School of 

Marine and Atmospheric Sciences, Key Biscayne, Florida, 

U.S.A. - 3 Laboratory of Clinical Investigation, NIH, 

Bethesda, Maryland, U.S.A. - 4 Instituto de Ciências 

Biomédicas, Universidade de São Paulo, São Paulo, 

Brazil. - 5 Mediacal Mycology Laboratory-HEC-FIOCRUZ, 

Rio de Janeiro, Brazil. - 6 Hadassah Medical Center, Kyriat 

Hadasah, Jerusalem, Israel. - 7 Molecular Mycology 

Laboratory, Centre for Infectious Diseases and 

Microbiology, University of Sydney, Sydney, Australia. - Email: 

boekhout@cbs.knaw.nl 

Cryptococcus neoformans is a basidiomycetous yeast 

causing life-threatening infections in immunocompromised 

hosts. The species is known in both the asexual and sexual 

states. Since long two varieties C. neoformans var. 

neoformans and var. gattii were known. The observation of 

mating and gene flow resulted in the description of a 

separate genus Filobasidiella. A third variety C. 

neoformans var. grubii (= serotype A) was described 

recently. Molecular studies on the IGS and ITS of the 

rDNA, the mtLrRNA, URA5, laccase and phospholipase 

genes, as well as AFLP and PCR-fingerprinting showed 

that the three varieties belong to different phylogenetic 

lineages and may represent species. Novel genotypes could 

be distinguished, thus further questioning the species 

boundaries. The biological species concept was tested in an 

analysis of a mating between variety neoformans and 

variety gattii. Most of the descendants possessed the 

genotypes of either parental isolate. Gene flow could not be 

demonstrated under natural circumstances. Therefore, we 

proposed that both varieties represent at least two species. 

Hybrid serotype AD strains of C. neoformans originated 

from different parental strains. The rare serotype A MATa 

allele occurs in part of these hybrids. An analysis of 

virulence related phenotypic traits and antifungal 

susceptibility revealed that virulence and susceptibility 

varied widely. Fluconazole resistance was observed in 

some environmental isolates, suggesting an innate 

resistance.


258 - Functional genomics in Candida albicans for drug 

target discovery 

R. Calderone 

Georgetown University Medical Center, Department of 

Microbiology & Immunology, Washington, DC, U.S.A. 

Current anti-fungals used to treat human diseases are few 

in number or invariably toxic. Most new anti-fungals are 

chemically modified versions of existing compounds that 

act against the same fungal targets [ergosterol synthesis, for 

instance]. The complete sequence of the Candida albicans 

genome has made it possible to utilize new strategies in 

order to identify targets that can be exploited in drug 

discovery. How does one select those genes [ORFs] that 

are the most appropriate for target validation? Most of the 

important ORFs are decided so by mostly 'common sense' 

thinking, including, first, the target should be represented 

in a number [if not all] of the human fungal pathogens but 

not in the human genome, or, if present, there should be 

sufficient differences that can be exploited. Secondly, the 

target should be vital to the infection process. Third, the 

function of the gene product should be at least partially 

understood so that appropriate assays can be established. 

Fourth, the gene product should be essential for growth or 

viability of the fungus, although virulence factors that are 

not essential for growth may be candidates. The spectrum 

of a candidate target is limited in definition since, except 

for the C. albicans genome, sequences for other human 

pathogens are non-existent or incomplete. In this 

symposium, topics related to the identification of 

functional gene homologues, target identification and 

validation will be presented. 

259 - Carbon dioxide exchange, diffusion resistances 

and water content in lichens: Laboratory and field 

T.G.A. Green 1* & O.L. Lange 2 

1 Waikato University, Hamilton, New Zealand. - 

2 Universitaet Wuerzburg, Wuerzburg, Germany. - E-mail: 

greentga@waikato.ac.nz 

Jumelle in 1892, first commented on depressed net 

photosynthesis at high thallus water contents and Stocker, 

in 1927, the first to infer that it was due to diffusion 

limitations. The topic attracted attention after 

demonstrations of its occurrence by Kershaw in the 1970s. 

There followed a period of contention as to what actually 

caused the depression. Two possibilities were proposed, 

depression through increased respiration or through 

increased diffusion resistances for carbon dioxide. 

Evidence, including chlorophyll fluorescence analysing 

activity of the photosynthetic apparatus and helox for direct 

determination of resistances, has consistently supported the 

changed diffusion resistances theory. This evidence will be 

summarised as well as field results that demonstrate the 

ecological importance of the depression in carbon gain. 

Although first found and described in the laboratory, 

suprasaturation depression of NP is not an experimental 

artefact but an important ecological feature of many lichen 

species in many habitats. As an example, averaged over a 

total year, net photosynthesis of Leconora muralis was 

heavily reduced through suprasaturation during 38.5% of 

the time when photosynthesis was possible. Despite these 

confirmations of the role of diffusion resistances there is 

still little agreement as to where these diffusion pathways 

are actually located. An interpretation of the thallus will be 

presented based on standard water potential water vapour 

considerations. 

260 - Hydrophobins in lichen-forming asco- and 

basidiomycetes 

R. Honegger 1* , S. Scherrer 2 & M.L. Trembley 3 

1 University of Zürich, Institute of Plant Biology, 

Zollikerstr. 107, CH-8008 Zürich, Switzerland. - 

2 University of Minnesota, 100 Ecology Building, 1987 

Upper Buford Circle, St. Paul, MN 55108, U.S.A. - 3 (no 

institution), Gempenstrasse 18, CH-4053 Basel, 

Switzerland. - E-mail: rohonegg@botinst.unizh.ch 

Main building blocks of heteromerous lichen thalli are 

hydrophilic, conglutinate pseudoparenchyma (peripheral 

cortex) and loosely interwoven plectenchyma built up by 

aerial hyphae with hydrophobic wall surfaces (medullary 

and photobiont layers). Wall surface hydrophobicity 

prevents the thalline interior from getting waterlogged at 

high levels of hydration and facilitates gas exchange 1 . This 

wall surface hydrophobicity was shown to be primarily due 

to class 1 hydrophobins. Several hydrophobins were 

biochemically and genetically characterised: one from the 

haploid vegetative thallus of each of 4 Xanthoria spp. 

(XPH1/parietina, XEH1/ectaneoides, XFH1/flammea, 

XTH1/turbinata) 2, 4 and three from the dikaryotic, 

lichenized basidiocarp of Dictyonema glabratum (DGH1, 

DGH2, DGH3) 5 . With in situ hybridisation techniques 

hydrophobin gene expression was located in medullary 

hyphae of X. parietina 3 or in the photobiont layer, the 

lower stratum and the boundary layer to the hydrophilic 

tomentum in D. glabratum, respectively 6 . An antibody 

raised against the recombinant DGH1 bound on ultrathin 

sections to the electron dense outermost wall layer of 

hyphae in the photobiont layer where rodlets had been 

resolved in freeze-etch preparations 6 . 1 Honegger (2001) 

THE MYCOTA IX: 165-188; 2 Scherrer et al. (2000) 

FGBI 30:81-93; 3 Scherrer et al. (2002) New Phytol. 

154:175-184; 4 Scherrer & Honegger, unpubl.; 5 Trembley 

et al. (2002a) FGBI 35:247-259; 6 Trembley et al. (2002b) 

New Phytol. 154:185-196. 



261 - Rolling stones in a harsh environment. 

Morphology and ecophysiology of globular erratic 

lichens 

L.G. Sancho 1* , B. Schroeter 2 & A. Pintado 1 

1 Universidad Complutense, Dpto. Biologia Vegetal II, Fac. 

Farmacia, Ciudad Universitaria, 28040 Madrid, Spain. - 

2 University of Kiel, Botanisches Institut, Olshausenstr. 40, 

24098 Kiel, Germany. - E-mail: sancholg@farm.ucm.es 

Erratic lichens, growing free without any attachment to the 

substrate, are widely distributed in many biomes all over 

the world. Xanthoparmelia and Aspicilia, have the largest 

number of vagrant taxa world-wide. Habitat supporting 

vagrant lichens is typically windswept, semi-arid, cold in 

winter and sparsely vegetated. In Eurasia, these habitat 

conditions correspond to the steppes and semi-deserts and 

account for almost one-fourth of the continent. Vagrant 

lichens of these regions typically show short and compact 

lobes or branches, thick scleroplectenchymatous tissues 

and low chlorophyll content. Accordingly, the thalli of all 

the investigated species showed very low water-holding 

capacity and low Amax. Studies among and within 

populations of vagrant Aspicilia species, both from the 

Iberian Peninsula and from Kazachstan, allowed us to 

consider the significance of thallus size for water economy 

and CO2 exchange. We found that in these compact 

globular lichens size can be a limiting factor for net 

photosynthesis, since the mass of the fungi increases 

volumetrically while the algal layer is more or less 

restricted to the surface. However, the highly specialized 

thallus structure prevents increased diffusion resistances 

for the transport of CO2, even at the highest level of water 

content. Moreover, the capacity for photosynthetic activity 

remained the same for the entire surface of these globular 

thalli, even though they were partially buried in the soil for 

several weeks. 

262 - Fungal compounds as sun screens for symbiotic 

algae in lichens 

Y. Gauslaa 


Agricultural University of Norway, P.O.Box 5014, 1432 

Ås, Norway. - E-mail: yngvar.gauslaa@ibn.nlh.no 

Lichens, like leaves of plants, have a non-photosynthetic 

tissue located as a filter between the incident solar radiation 

and the photosynthetically active cells. Unlike the 

predominantly green members of the plant kingdom, many 

lichens are coloured by cortical secondary compounds, e.g. 

the orange parietin, the yellow usnic acid and the brownblack 

melanins. Since many compounds can be extracted in 

a non-destructive way by 100% acetone, their screening 

role can be directly assessed. The upper cortex of lichens 

screens a substantial part of the irradiance, particularly 

UVB, even in the absence of extractable secondary 

compounds. However, secondary compounds modify the 

84 


transmittance spectra further by reducing short 

wavelengths more than longer wavelengths. For instance, 

an acetone-rinsed air-dry Nephroma arcticum cortex with 

the usnic acid removed, transmits 1% of UVB, which is ten 

times the transmittance before usnic acid extraction (0.1%). 

Whereas usnic acid only has a potential in screening UV 

radiation, parietin and melanin can additionally protect 

lichens against excessive violet-blue, and all visible 

irradiation, respectively. For some species, the protection is 

not always sufficient. The heat- and high-irradiance 

susceptible Lobaria pulmonaria is caught between the risk 

of being damaged by excess irradiances due to a pale, 

highly transmitting cortex, or by high temperatures due to a 

melanic, highly radiation-absorbing cortex in sun-exposed 

habitats. 

263 - Structural and functional aspects of water uptake 

and photosynthesis in lichens at subzero temperatures 

B. Schroeter 1* , B. Frey 2 & C. Scheidegger 2 

1 Botanisches Institut, Universität Kiel, Olshausenstr. 40, 

D-24098 Kiel, Germany. - 2 Swiss Federal Institute for 

Forest, Snow and Landscape Research, CH-8903 

Birmensdorf, Switzerland. - E-mail: bschroeter@bot.unikiel.de 

It is well known that lichens are able to photosynthesize at 

subzero temperatures. This ability might be of high 

ecological significance in cold environments such as 

Antarctica where snow is the major source for hydration. 

Due to their poikilohydrous nature water relations in the 

lichen thallus are strongly correlated with the ambient 

water regime. At subzero temperatures the lichen thallus is 

partly dehydrated due to the negative ambient water 

potential. We present data from a combined structural and 

physiological study on water relations and metabolic 

activity in green algal and cyanobacterial lichens at subzero 

temperatures. Structural investigations were carried out 

with LTSEM techniques, CO2 gas exchange and 

chlorophyll a fluorescence indicated the status of the 

photosynthetic apparatus of the photobiont of the lichens. 

At subzero temperatures hydrated thalli were kept in 

equilibrium with water saturated air or dry thalli were 

allowed to rehydrate from snow. There was no major 

difference in water relations and activation at subzero 

temperatures in lichens from cold and temperate 

environments. However, a major difference was found 

between green algal and cyanobacterial lichens: while 

green algal lichens were able to reactivate PSII through 

water uptake from snow in cyanolichens this did not lead to 

measureable reactivation of the photosynthesis. This 

difference may contribute to the almost complete absence 

of cyanobacterial lichens in continental Antarctica.


264 - Live-cell imaging of fungal hyphae at high spatial 

resolution 

N.D. Read * , P.C. Hickey, S.R. Swift & H.A. Atkinson 

Fungal Cell Biology Group, Institute of Cell and Molecular 

Biology, University of Edinburgh, Rutherford Building, 

Edinburgh EH9 3JH, U.K. - E-mail: Nick@fungalcell.org 

A revolutionary new perspective of the cell biology of 

fungal hyphae is arising as a result of using live-cell 

imaging techniques to analyse organelle and molecular 

dynamics at high spatial resolution. A primary objective of 

the work in our group is to perform live-cell imaging at the 

highest possible spatial resolution that can be routinely 

obtained. For this purpose we have been primarily using 

laser scanning confocal microscopy although more recently 

we have also started to evaluate multiphoton microscopy 

and spinning disc confocal microscopy for imaging living 

fungal hyphae. A major focus of our research is in 

understanding the mechanisms and regulation underlying 

hyphal tip growth. Laser scanning confocal microscopy has 

enabled us to image living hyphae without significantly 

compromising hyphal growth. Much of this work has 

concentrated on studying vesicle trafficking and organelle 

dynamics in living hyphae using a range of vital 

fluorescent dyes (e.g. FM4-64, FM1-43, and Rhodamine 

123) or recombinant GFP specifically targeted to these 

organelles. Organelles that we are routinely imaging in 

living hyphae include nuclei, mitochondria, ER and 

vacuoles. We are also imaging the dynamic localisation of 

various proteins fused to GFP. Examples of movies from 

our live-cell imaging studies can be found at the following 

website: http://www.fungalcell.org/. 

265 - Measurement and interpretation of biomechanical 

parameters relevant to fungal development 

N.P. Money 

Department of Botany, Miami University, Oxford, Ohio 

45056, U.S.A. - E-mail: moneynp@muohio.edu 

Without biomechanical experimentation, mycologists have 

no hope of understanding mechanisms of fungal growth, 

reproduction, or pathogenesis. Having said this, it must be 

tearfully-acknowledged that most biologists view the study 

of fungal mechanics as an eccentric pursuit whose findings 

can be ignored without grievous consequences. There are 

two major reasons for this impression, and for the 

consequent lack of biomechanical research on fungi: (i) an 

illegitimate conviction that scientific inquiry restricted to 

the molecular level can answer all of the important 

questions, and (ii) a legitimate fear that methods for 

measuring minuscule forces from microscopic cells can be 

very difficult. If successful, this presentation will alleviate 

both problems by offering a balanced appraisal of the value 

and challenges of biomechanical investigation. Methods for 

determining cellular turgor pressure and the invasive 

pressures exerted by hyphal apices will be discussed, along 

with recent efforts to explore the mechanical performance 

of multicellular fruiting bodies. 

266 - Analysis of calcium signalling in living hyphae 

expressing a codon-optimised aequorin gene 

A. Zelter 1* , O. Kozlova-Zwinderman 1 , O. Yarden 2 & N.D. 

Read 1 

1 

Institute of Cell and Molecular Biology, Rutherford 

Building, University of Edinburgh, Edinburgh, EH9 

2 

3JHUK, U.K. - Dept. Of Plant Pathology and 

Microbiology, Hebrew University of Jerusalem, Rehovot 

76100, Israel. - E-mail: azelter@holyrood.ed.ac.uk 

Analysis of the Neurospora genome indicates that that the 

calcium signalling machinery is more complex than that 

found in budding yeast. However, calcium signalling is 

little understood in filamentous fungi largely because easy 

and routine methods for calcium measurement in living 

hyphae have previously been unavailable. We have 

developed the recombinant aequorin method for this 

purpose. High levels of expression of the calcium-sensitive 

aequorin photoprotein have been obtained in a range of 

filamentous fungi, including Neurospora crassa, 

Aspergillus nidulans, A. niger and A. awamori, by codon 

optimisation. Three external stimuli (mechanical 

perturbation, hypo-osmotic shock and high external 

calcium) transiently increase cytosolic free calcium. The 

calcium-signatures associated with each of these 

physiological treatments have been quantified and are 

highly reproducible under any one set of conditions. Each 

of the three stimuli generates a unique calcium-signature 

suggesting that the involvement of different calciummediated 

signal transduction pathways. This is supported 

by results using calcium modulators. However, we have 

also found that various parameters (e.g. amplitude and full 

width half maximum) of calcium-signatures are dependent 

on temperature and whether the fungus is grown on solid or 

in liquid medium. Future directions for research on calcium 

signalling using aequorin will be described. 

267 - Dynamics of organelles and the cytoskeleton in the 

plant pathogen Ustilago maydis 

G. Steinberg 

MPI for Terrestrial Microbiology, D-35043 Marburg, 

Germany. - E-mail: Gero.Steinberg@mailer.unimarburg.de 

In contrast to vertebrates, fungi are surrounded by a rigid 

cell wall that limits motility of fungal cells. However, 

substrate invasion and pathogenic development of fungi 

require migration of the cell that is achieved by hyphal tip 

growth. This process is based on delivery of growth 

supplies to the expanding apex of the hypha involves the 

cytoskeleton and associated motor molecules. Recently, the 

use of advanced light microscopy methods has enabled us 



to monitor dynamics of organelles and the cytoskeleton 

itself in Neurospora crassa and Ustilago maydis. These 

experiments have revealed remarkable levels of motility 

within the fungal cell and it has become obvious that 

intracellular motions are essential for spatial organization, 

growth and morphogenesis of fungi. 

268 - Using light and electron microscopy to explore 

hyphal cytoplasmic order, behavior and mutation mode 

of action 

R.W. Roberson 1* & S. Bartnicki-García 2 

1 

Arizona State University, Department of Plant Biology 

2 

Tempe, Arizona 85287-1601, U.S.A. - Centro de 

Investigación Científica y de Educación Superior de 

Ensenada, 22830, Baja California, Mexico. - E-mail: 

robert.roberson@asu.edu 

To assess the impact of single mutations on cell 

morphogenesis, it is essential to explore the effect of the 

mutation on cytoplasmic organization and behavior. Video 

microscopy provides a real time record of organelle 

behavior and morphological changes in living cells. Such 

images can provide clues about the mode of action of a 

mutation that can be tested by transmission electron 

microscopy of sectioned specimens. One good example is 

in the study of the ro-1 and nudA mutations in hyphae of 

Neurospora crassa and Aspergillus nidulans, respectively. 

These genes encode subunits of cytoplasmic dynein. In 

wild-type cells a well-defined Spitzenkörper (Spk) 

dominated the cytoplasm of the hyphal apex. Vesicles 

exhibited active motility in subapical regions. Hyphae 

contained abundant microtubules (MT) that were mostly 

aligned parallel to the growing axis of the cell. 

Mitochondria and nuclei maintained a near constant 

position in the advancing cytoplasm. Dynein deficiency 

causes disruption of MT organization and function. Beside 

the overall perturbation to cytoplasmic organization and 

organelle motility, these mutations disrupt the organization 

and stability of the Spk, which, in turn, leads to severe 

reduction in growth rate and altered morphology. The 

combined use of light and electron microscopy has lead to 

a more complete understanding of MT disruption and other 

cytoplasmic phenotypes that result from dynein deficiency. 

269 - Four-dimensional laser scanning microscopy of 

fungal plant pathogens expressing fluorescent proteins 

R.J. Howard 1* , T.M. Bourett 1 , J.A. Sweigard 2 , K.J. 

Czymmek 3 & K.E. Duncan 1 

1 

DuPont Crop Genetics, Exp Stn, Wilmington, DE 19880- 

2 

0402, U.S.A. - DuPont Crop Genetics, Delaware 

Technology Pk, Newark, DE 19716, U.S.A. - 3 University of 

Delaware, Department of Biological Sci, Newark, DE 

19716, U.S.A. - E-mail: richard.j.howard@usa.dupont.com 

86 


The three-dimensional mapping of host-pathogen cell 

interfaces over time, or 4D analysis, represents a 

challenging but important prerequisite for understanding 

pathogenesis. We generated fluorescent transformants of 

two different pathogens, Fusarium verticillioides and 

Magnaporthe grisea, and used them for real-time imaging 

by confocal and multi-photon microscopy. Driven by 

strong constitutive fungal promotors, expression of spectral 

variants of green fluorescent protein, as well as the recently 

identified reef coral fluorescent proteins derived from 

several Anthozoa species, had no detectable effect on either 

growth rates or abilities to cause disease. Cytoplasmtargeting 

of fluorescent proteins, coupled with fast image 

capture rates, allowed discrimination of many subcellular 

organelles by differential exclusion and facilitated 

monitoring of rapid changes in permeability of the nuclear 

envelope. Alternatively, use of fluorescent markers as 

fusion proteins, for example with tubulin, made it possible 

to image specific proteins/structures/organelles during cell 

growth, development, and pharmacological treatment. The 

intense brightness of some strains expressing fluorescent 

proteins in the cytoplasm permitted documentation of 

pathogen cells during invasion of plant host tissues. 

AmCyan and ZsGreen reef coral fluorescent proteins were 

sufficiently excited at 855 and 880 nm, respectively, to 

facilitate time-resolved in planta imaging by two-photon 

microscopy. 

270 - Studying cell biology of arbuscular mycorrhizal 

fungi by combined high resolution biochemical, 

molecular biology and microscopy techniques 

B. Bago 1* , P.E. Pfeffer 2 , W. Zipfel 3 & Y. Shachar-Hill 4 

1 CSIC, CIDE, Cami de la Marjal s/n, Albal (Valencia), 

Spain. - 2 USDA, ERRC, 600 E. Mermaid Ln. 19038 

Wyndmoor, PA, U.S.A. - 3 Cornell University, Clarck Hall, 

Cornell University, Ithaca NM, U.S.A. - 4 NMSU, Dept. 

Chemistry and Biochemistry, NMSU, 88033 Las Cruces 

NM, U.S.A. - E-mail: berta.bago@uv.es 

Glomeromycotan or arbuscular mycorrhizal (AM) fungi 

are widespread mutualistic symbionts whose taxonomic 

position, ecological distribution, reproductive strategies, 

not to mention cell biology is still poorly understood. At 

the basis of this lack of knowledge is probably the fact that 

these fungi are obligate biotrophs, i.e. they cannot complete 

their life cycle unless they have successfully colonized a 

host plant. Unsuccessful attempts of culturing AM fungi 

axenically have been made in the last forty years, this 

greatly hindering progress in our knowledge of these 

ecologically important fungi. Moreover, the difficulties to 

study the intraradical fungal phase withouth disturbing 

symbiosis functioning, and the extraradical mycelium 

while maintaining intact their soil-growing hyphae requires 

of non-destructive, in situ techniques. In this presentation 

we will briefly review recent advances in our understaning 

of the cellular biology of these symbiotic fungi. This has 

been possible by combining techniques such as monoxenic 

AM cultures, NMR spectroscopy, image analysis and 

multiphoton microscopy.


271 - Integrating scientific research and information 

systems 

* 

G. Hagedorn , M. Burhenne, M. Gliech, T. Gräfenhan & 

M. Weiss 





The GLOPP project (Global Information System for the 

Biodiversity of Plant Pathogenic Fungi, 

http://www.glopp.net/ ) started in the summer of 2000 and 

will continue until spring 2003. It aims to provide a unified 

view of data about fungi parasitizing higher plants. The 

information system will integrate data on host plant 

specificity, geographical distribution, and a core set of 

descriptive characters that allow interactive online 

identification. European and especially German pathogens 

are treated with priority to guarantee high data quality and 

a certain completeness. We plan to continue our work 

within the framework of international, collaborative 

successor projects. - The GLOPP system is presented as a 

highly useful exercise combining basic organismic research 

with research in biodiversity informatics. The system is 

presented from the perspective of (a) system development 

and information modeling, (b) contributing research 

scientists, and (c) users in applied fields like plant 

pathology extension services. Ultimately such efforts will 

be successfull only if they are as valuable to content 

generators as they are to information consumers. 

Contributing researchers should be able to work more 

efficiently within the information system than otherwise. 

This is currently not the case, but the progress made is 

substantial (see http://www.diversitycampus.net/ ). 

272 - Fundamentals of biodiversity research or: What 

we still do not know! 


Botanical Institute of the University of Frankfurt, 



Our knowledge of fungal biodiversity is limited not only 

because of numerous undiscovered species, but we also 

have many unresolved taxonomic and systematic questions 

(incomplete descriptions, bad type material, unclear 

synonymy), the morphology of many species is not well 

documented, we ignore host ranges of many species, areas 

of distribution, numerous aspects of life cycles, and effects 

of interactions with other organisms. For the 

documentation of existing knowledge of fungal diversity in 

the internet, we include morphological descriptions in a 

database. Therefore, we first need a list of morphological 

characters and character states, which requires a reasonable 

selection of adequate descriptive vocabulary. The 

description of a species in literature might be satisfying at 

first sight, its inclusion into the database, however, in 

addition to problems of 'translation' due to the use of 

different terminology, usually shows that the description is 

far from complete. Often, the germination of spores and 

other morphological details are not well documented. 

During the input of data into the database we are forced to 

ask numerous questions, of which many usually can not be 

easily answered. Only by field work we obtain fresh 

material which allows us to elaborate complete sets of 

character states for the database, and contribute to our 

knowledge on morphology, host range, distribution, and 

ecology of the fungi. 

273 - Connecting traditional and modern data sources: 

Towards an integrated view on the biodiversity of 

powdery mildews (Erysiphales) 

1* 1 2 

C. Kainz , D. Triebel & U. Braun 

1 Botanische Staatssammlung München, Mycology Dept., 

Menzinger Strasse 67, D-80638 Muenchen, Germany. - 

2 Martin-Luther-Universität Halle, Institut für Geobotanik 

und Botanischer Garten, Neuwerk 21, D-06099 Halle, 

Germany. - E-mail: kainz@botanik.biologie.unimuenchen.de 

The Erysiphales form a well delimited order of 

phytopathogenic ascomycetes with about 650 taxa 

occurring worldwide. As they represent a group of strongly 

related fungi, which is relatively well-investigated, they are 

selected to realise an integrated view on biodiversity data 

of different kinds. Within the project, descriptive, 

ecological, distributional and molecular data are collected 

and linked with data available from traditional and modern 

data sources. Information is gained from the material of 

two major collections of Erysiphales in the herbaria of 

Halle and München (label and specimen data including 

sequence data) as well as from literature (descriptive data). 

For the storage and maintenance of collected data the 

database modules of DiversityWorkbench, the onlineinformation 

system LIAS and the sequence data 

management system ARB are used. Subsequent analysis of 

data from the various sources is achieved via export to 

various applications. The employment of new database 

systems and informatics tools will simplify scientific 

studies on the correlation and evolution of characters both 

in a distributional and historical context. 

274 - Reconsidered species and genus boundaries: How 

molecular data influence the content of the database 

M. Goeker * , A. Riethmueller & F. Oberwinkler 

Lehrstuhl Spezielle Botanik/Mykologie, Botanisches 

Institut, Universitaet Tuebingen, Auf der Morgenstelle 1, 

72076 Tuebingen, Germany. 

The GLOPP (Global Information System for the 

Biodiversity of Plant Pathogenic Fungi, 



http://www.glopp.net/) project does not aim at a simple 

compilation of data, so critical appraisal of the data found 

in taxonomic literature is necessary. As a rule, this entails 

experimental re-examination of the published data. 

Unfortunately, the morphological data so far used for 

Oomycete taxonomy are often ambiguous. This is 

especially important where only few morphological 

characters are available, as it is the case in taxon 

delimitation in most plant parasitic Oomycete groups. Even 

the total number of species in Peronosporales and 

Sclerosporales stated in the literature can vary very much 

depending on the underlying species concept used by 

different authors. To cope with these uncertainties, the 

combination of morphological with modern molecular 

methods is required. The content of the database will 

depend to a high degree on the results from this approach. 

275 - Discovering the crunches: the use of the GLOPP 

databases in rust systematics, ecology and biogeography 

R. Berndt 

University of Tübingen, Systematic Botany and Mycology, 

Botanical Institute, Auf der Morgenstelle 1, D-72076 

Tübingen, Germany. - E-mail: reinhard.berndt@unituebingen.de 

The knowledge about rust fungi (Uredinales) is still patchy: 

there are some well-studied pests of economically 

important plants, and there is the bulk of species growing 

on hosts with no obvious economic importance. The latter 

group of rusts has received much less attention 

accordingly. Another striking disparity of knowledge exists 

between tropical and subtropical areas and extratropical 

regions. Whereas floras or monographs on rust fungi are 

widely available in extratropical regions, the student of 

tropical rusts most often has to confine to scattered original 

work that may not be handy or accessible. In our project 

we aim to compile a database on rust fungi with a focus on 

tropical and subtropical species. The database is geared to 

(1) facilitate the determination of rust fungi and to provide 

up-to-date taxonomic information (2) to supply data on the 

family affiliation of the host plants and on the known 

geographic range of the fungi in question. Additional 

information on relevant literature and personal 

observations are also included. The present database allows 

to retrieve data that are not readily available on the 

internet: - host family-rust fungus search - geographical 

search - synonymy of rust fungi - complex queries with 

several parameters Such options enable the user to reduce 

the number of possible determinations of a given rust 

fungus and to find hidden coincidences and contradictions 

in regard of host spectra, systematics, and geographical 

distribution. 

88 


276 - Phytopathogenic fungi from South Africa 

P.W. Crous 

Department of Plant Pathology, University of Stellenbosch, 

P. Bag X1, Matieland 7602, South Africa. - E-mail: 

pwc@sun.ac.za 

The appointment of I.B. Pole-Evans as mycologist of the 

former Transvaal Department of Agriculture in 1905 

heralded the beginning of phytomycology in South Africa. 

As diseases of wild and cultivated plants became more 

well-known, several lists were published of these 

pathogens. The early lists were prepared by Doidge (1924) 

and Doidge and Bottomley (1931). The most significant 

one, however, was published by Doidge et al. (1953), 

which was primarily based on information contained in her 

voluminous Index of South African Fungi and Lichens to 

the end of 1945 (Doidge, 1950). In later years this list was 

updated again, and several new records added in a series of 

three science bulletins published by Gorter (1977, 1981, 

1982). These data formed the basis for the book, 

Phytopathogenic fungi from South Africa, which was later 

published by Crous et al. (2000). An abbreviated version of 

these data can be viewed on the USDA web site in 

Beltsville. Members of the Southern African Society for 

Plant Pathology required a more adaptable database, 

however, that could easily be updated and changed to 

fulfill the needs to the local plant pathological community. 

For this reason a new database template was developed that 

would support a CD-Rom and web-driven version of the 

database, incorporating illustrations, notes, all local and 

relevant publications, culture collection and DNA sequence 

data. 

277 - The polypores of the world - a useful database or 

are we still in the alpha phase? 

L. Ryvarden 

Botany Department, Biological Institute, P. O. Box 1045, 

Blindern, N-0316 Oslo, Norway. - E-mail: 

leif.ryvarden@bio.uio.no 

Over a period of 30 years mycotas of East Africa, North 

America, Europe and North East Asia have been published, 

while those of New Zealand and the neotropics are in 

manuscript and will be published shortly. The generic keys 

in these books will this autumn be available on the Internet 

for free downloading. In 1987 360 poroid species were 

registered in Europe, and since then, i.e. over 15 years, 

about 5 species have been added, some of them probably 

tropical imports. 12 species have been described as new, 

almost all of them based on splitting of previously to 

widely accepted species, such as Antrodiella semisupina. 

This indicates clearly that the alpha phase is finished in 

Europe. The situation is similar in North America and 

North East Asia. In tropical Africa 320 species are 

registered excluding critical genera like Ganoderma and 

Phellinus while at least about 20 species are ready to be


described based on recent collections. Still the Congo-basin 

is strongly under collected and will certainly yield a good 

number of new species, especially those with resupinate 

basidiocarps. Thus, there is still some time before the 

alpha-phase can be said to be finished in this continent. In 

the neotropics the situation is similar and recent intensive 

collecting in the Amazonas basin has revealed a number of 

new species, clearly indicating that we still not have a clear 

picture of the biodiversity in this group. 

278 - Taking your data online: Database experiences at 

the USDA Systematic Botany and Mycology 

Laboratory 

D.F. Farr 

USDA, Systematic Botany and Mycology Laboratory, 

BARC-West, Rm. 304, Bldg. 011A, Beltsville MD 20705, 

U.S.A. - E-mail: davef@nt.ars-grin.gov 

As the Web becomes increasingly pervasive as a primary 

source of information, one might suggest that if the 

information is not on the Web it does not exist. A primary 

role of systematics is to provide a basic framework that 

society can use to communicate about the natural world. 

While there are many potential users of systematic 

information, these data are most readily available to those 

with access to the large libraries associated with 

universities and research institutions. As systematists, we 

need not only to develop and synthesize information, but to 

ensure that it is available to those who need it. The web is a 

visual and interactive medium, which opens up many 

exciting ways for systematists to distribute their 

knowledge. Greater thought should be given to providing 

information on the Web if we want to remain a viable part 

of the information age. At the same time it is important to 

understand the differences between the Web and print 

media and to develop a presentation formats that organize 

and deliver content is a usable way. To assist systematists 

in facilitating their use of the Web, an online tutorial has 

been developed. This tutorial explores the techniques for 

presenting and relating the descriptive text, images, 

specimen data and keys that make up the systematists' 

publication. It is hoped that these examples will stimulate 

systematists to include Web publication as one of their 

research products. 

279 - Foliicolous diversity: the Caribbean example 

M. Rodríguez Hernández 

National Botanic Garden, Carretera del Rocío km 3.5, 

Calabazar, Boyeros, CP 19230 C.Habana, Cuba, Cuba. - 

E-mail: hajb@ceniai.inf.cu 

Flora and vegetation in the Caribbean Islands have a great 

diversity and richness, therefore, it is a nice place for fungi, 

particularly for foliicolous, leaf-surface fungi. As treated 

here, they are those which have the mycelium and fruit 

bodies on the leaf surface and are in contact with the leaf 

and the atmosphere, and subject to the influence of both. 

The Caribbean is a good region for leaf-surface fungi 

because: • There are more than 20 fungal families and 

hundreds of species • There are more than 1,000 host plants 

belonging to 125 families They might be divided, since an 

ecological or practical point of view, in 3 main groups: 

Saprobic, Parasite and Hyperparasite. Interesting examples 

may be shown: • Trichomerium grandisporum: it has a 

world-wide distribution and has been found on more than 

50 host plants belonging to more than 20 families. It grows 

since cultivated areas at sea level to rainforests • Meliola 

opuntiae: on Cactaceae in dry places, in Cuba and Jamaica 

with only 2 records • Meliola orchidacearum: on 

Orchidaceae in humid vegetations, only in Cuba and 

Dominican Republic with 3 records Some outstanding 

examples among host plants are: Mangifera indica, 

Syzygium jambos, Rondeletia canellaefolia, Sideroxylon 

jubilla, Acrosynanthus trachyphyllus, associated with many 

saprobic species, or Cupania americana and Ocotea 

leucoxylon with parasitic ones. The present paper offers a 

list of leaf-surface fungi in the Caribbean and the related 

host plants. 

280 - The ecology of foliicolous fungi 

G.S. Gilbert 1 & D.R. Reynolds 2* 

1 University of California, Environmental Studies, 1156 

High St., Santa Cruz, CA 95064, U.S.A. - 2 Natural History 

Museum, 900 Exposition Blvd., Los Angeles, California 

90007, U.S.A. - E-mail: dreynold@nhm.org 

Foliicolous fungi require leaves to make a living in a forest, 

yet remarkably little is known of such fungi where most 

leaves are available - in the forest canopy. The 3dimensional 

structure of diverse tropical forests creates 

strong vertical gradients and horizontal patchiness in 

suitable environmental conditions and host availability for 

leaf-surface fungi. Using canopy cranes in Australian and 

Panamanian moist tropical forests, we have explored the 

roles of host plant availability, fungal dissemination, and 

microclimate in determining the spatial distribution and 

diversity of foliicolous fungi. Fungi were found in all strata 

of the forest at Cape Tribulation, Australia, but the more 

than 30 genera of non-lichenized leaf-surface fungi were 

much less common on exposed canopy leaves than in the 

understory. Understory environmental conditions were 

darker, relative humidity greater, and temperature variation 

less extreme, all factors that may promote fungal growth. 

For three focal plant species, the density of hyphal growth 

on the leaf surface was greater in the understory than in the 

canopy. Nevertheless, within the understory in both 

Australia and Panama, the probability of fungal 

colonization increased with increase light availability. The 

types of fungi found varied greatly among plant hosts. The 

spatial pattern of fungal colonization in the understory was 

not strongly associated with presence of fungi in the 

canopy. 



281 - Amber fossils of sooty moulds 

J. Rikkinen 

Department of Applied Biology, P.O. Box 27, FIN-0014, 

University of Helsinki, Finland. - E-mail: 

jrikkine@mappi.helsinki.fi 

Fungi are unlikely candidates for fossilization and thus 

relatively few fungal fossils have been described. However, 

perfectly preserved amber fossils have shown that many 

modern genera of fungi, and possibly even species, were 

already present in the Tertiary. For example, several well 

preserved specimens of sooty moulds were recently found 

from European amber dating back to 22-54 million years 

ago. The fossils represent fragments of superficial subicula 

composed of brown moniliform hyphae with markedly 

tapering distal ends. The subglobose cells are quite 

identical to those of extant Metacapnodium 

(Metacapnodiaceae, Dothideales) species. Also the 

production of two distinctive conidial states supports a 

placement in this extant genus. The fossils demonstrate that 

the hyphae of some sooty moulds have remained 

unchanged for tens of millions of years. As there is little 

reason to believe that the fossilizations would have been 

immediately preceded by a period of more rapid evolution, 

the initial divergence of the fungi must have happened in 

the distant past, probably in the Cretaceous. The fossils 

also indicate that hyphal morphology and conidial states 

can be given considerable classificatory significance in this 

group of fungi. Eventually, after more detailed 

phylogenetic hypotheses have been generated for different 

groups of sooty moulds, the available fossils can become 

invaluable for timing branching events and calibrating 

molecular clocks. 

282 - Molecular systematics of some sooty molds 

J.L. Nicklin 

Biological Sciences, Birkbeck, University of London, Malet 

Street, London, WC1E 7HX, U.K. - E-mail: 

j.nicklin@bbk.ac.uk 

Sooty moulds are a community of several hundred darkly 

pigmented mitosporic and ascosporic species of fungi, 

living on tropical plant surfaces in association with 

honeydew from insect infestations. All phases of the life 

cycle of these fungi retain a melanised cell wall creating a 

sooty layer on leaf surfaces. Mitospores of many of the 

species are formed on a variety of pycnidial or coremial 

structures, all of which appear to be adaptations to the leaf 

surface environment that maximise spore dispersal after 

wetting. This guild of fungi was extensively studied on 

citrus and other tropical plant species during the late 19th 

C, and at that time was given the common epithet 

Capnodium citri (Farlow, Underwood). It is now realised 

that this an ambiguous name for a polymorphic group. The 

systematics of these fungi and their interactions with plants 

and aphids is of interest as the economic effects of sooty 

90 


mould colonisation of citrus, pecan and other 

horticulturally important plants begins to become apparent. 

The taxonomic status of some of the sooty moulds has 

recently received some clarification based on 

morphological and physiological characters. However, the 

complexity of the associations mean that identification 

based on morphological and biochemical features is 

unsatisfactory and molecular approaches reported in this 

work, including RAPD, microsatellite fingerprinting and 

ITS sequence analysis, provides better data for taxonomic 

and cladistic analysis. 

283 - Foliicolous ascomycetes in the canopy of a lowland 

rainforest in the Orinoco Basin, Venezuela (Surumoni 

Crane Project) 

P. Otto 1* & D.R. Reynolds 2 

1 University of Leipzig, Institute of Botany, Systematic 

Botany, Johannisallee 21-23, D-04103 Leipzig, Germany. - 

2 Natural History Museum, Research and Collections, 900 

Exposition Boulevard, Los Angeles, California 90007, 

U.S.A. - E-mail: otto@rz.uni-leipzig.de 

The fungi were investigated from the forest floor to the 

upper canopy (till 35 m height) in an area of 1.5 ha of an 

evergreen forest using a mobile crane system. Data are 

available from 1996 to 1998 for rain and dry season and for 

leaves of different ages. The studied fungi, teleomorphs 

and selected characteristic anamorphs, belong to saprobic, 

plant pathogenic or fungal pathogenic species and were 

collected from trees of 25 plant families. Fungi of 15 

genera in 12 families were identified in the canopy. Typical 

genera for tree crowns are e.g. Atichia, Brefeldiella, 

Brooksia, Microthyrium, and Myriangiella. The most 

common genus is Micropeltis, which occur on all tree 

species with a sufficiently thick leaf cuticle and produce 

first fruitbodies on leaves of an age of about 2 months. The 

fungal diversity (including saprobic fungi) is very different 

for individual tree species. Trees particularly in the 

Annonaceae and Caesalpiniaceae have a high fungal 

species number. Diversity and quantity decrease from the 

lower and medium canopy to the top. Probably 

unfavourable climatic conditions (till about 35 °C and 40% 

rel. humidity) are responsible for it. This assumption is 

supported by the fact that in the crane site at the end of rain 

season foliicolous ascomycetes are best developed. Some 

of these have a preference for the undersurface of leaves, 

e.g. Schizothyrium, which is a further indication of the 

strong influence of temperature and humidity on leaf 

inhabiting fungi.


284 - Foliicolous lichens: Evolution and ecology of an 

unusual growth habit 

R. Lücking 

Botany Department, The Field Museum, 1400 S Lake Shore 

Drive, Chicago, IL 60605-2496, U.S.A. - E-mail: 

rlucking@fieldmuseum.org 

Foliicolous lichens inhabit leaves of vascular plants in 

tropical rainforests. They have to adapt to the short 

longevity of their substratum, as leaves are mostly shed 

after 12 to 36 months. Furthermore, they must not interfere 

with leaf functions of gas exchange and photosynthesis. In 

spite of these limitations, foliicolous lichens evolved a 

surprising diversity and unique phenotypic features. Fifty 

species can be found on a small leaf, and 300 taxa within a 

single rainforest spot. Following their needs for rapid 

dispersal in a semi-aquatic environment, foliicolous lichens 

developed particular, convergent dispersal strategies, such 

as disc-shaped isidia (vegetative), derived conidiomata 

(asexual), and facultative parasitism on other lichens 

(sexual reproduction). Based on different evolutionary 

models of foliicolous lichen growth, and using phenotypebased 

phylogenetic analyses of selected groups, we tested 

whether individual features fulfill the criteria of 'key 

innovations' and led to subsequent radiation. It is concluded 

that radiation occurred only when sexual and asexual 

reproduction were maintained as principal dispersal modes 

within a clade, and the nature of radiation, whether specific 

or generic, depended on the inherent plasticity of the 

putative 'key innovation'. Thus, clades characterized by 

facultative parasitism or derived conidiomata show a high 

degree of radiation, while speciation is virtually absent in 

clades with specialized vegetative dispersal organs. 

285 - The phylogeny of Roccella (Roccellaceae) with 

emphasis on the Macaronesian species 

Å. Dahlkild 1* , M. Källersjö 2 & A. Tehler 3 

1 

Botaniska Institutionen, Stockholms Universitet, 106 91 

Stockholm, Sweden. - 2 Molekylärsystematiska laboratoriet, 

Naturhistoriska Riksmuseet, Box 50007, 104 05 Stockholm, 

Sweden. - 

3 Sektionen för kryptogambotanik, 

Naturhistoriska Riksmuseet, Box 50007, 104 05 Stockholm, 

Sweden. - E-mail: Asa.Dahlkild@botan.su.se 

This paper presents a phylogenetic analysis of the 

lichenized fungal genus Roccella (Roccellaceae, 

Arthoniales), focussing on the Macaronesian species. The 

ITS rDNA region, including the 5.8S region, ITS1, and 

ITS2, was sequenced for 14 Roccella species, including 

two putative species pairs. The sequence information was 

analyzed under a range of alignment parameters, and with 

gaps coded as missing data. The genus Roccella is found 

not to be monophyletic, since two species, Roccella 

hypomeca and Roccella portentosa, are more closely 

related to another genus, Roccellina. Dirina is the sister 

group of the core Roccella, as found in earlier studies. The 

Macaronesian species form a monophyletic group in some, 

but not in all analyses. Our study also indicates that the two 

putative species pairs Roccella allorgei/Roccella africana 

and Roccella tinctoria/Roccella canariensis each should be 

treated as conspecific. 

286 - The Sphaerophorus globosus species complex - 

morphology re-interpreted with molecular phylogeny 

M. Wedin 1* & F. Högnabba 2 

1 Dept. of Ecology and Environmental Science, Umeå 

University, SE-901 87 Umeå, Sweden. - 2 Dept. of Ecology 

and Systematics, University of Helsinki, P.O. Box 47, FIN- 

00014 Helsinki, Finland. - E-mail: mats.wedin@eg.umu.se 

The Sphaerophorus globosus species complex 

(Lecanorales, lichenised Ascomycota) shows a surprisingly 

large morphological variation, and several relatively 

distinct morphotypes can be distinguished over the 

distribution area. We have sampled populations from most 

areas where the species complex occurs, with a particular 

focus on the populations in the North American Pacific 

Northwest, where the morphological variation is very large 

in the group. In this study, we utilise a multi-gene based 

maximum-parsimony approach (nITS+LSU rDNA, mtSSU 

rDNA, ß-tubulin, actin) to investigate the phylogeny of this 

complex. If the different morphotypes constitute distinct 

monophyletic groups in the multi-gene phylogenies, this 

would support the hypothesis that the morphotypes are 

different phylogenetic species. Monophyletic groupings 

corresponding to geographical origin would, on the other 

hand, support the alternative hypothesis that the different 

morphologies are likely to be caused by the 

environmentally induced phenotypic variation, rather than 

by common descent. The results point at the presence of 

several distinct phylogenetic species in the group. 

287 - Phylogeny of Ostropales and Gyalectales - 

evidence from molecular and ontogenetical data 

F. Kauff 1* , F. Lutzoni 2 & B. Büdel 1 

1 University of Kaiserslautern, FB Biologie, Abt. Allg. 

Botanik, Postfach 3049, 67653 Kaiserslautern, Germany. - 

2 Duke University, Dept. of Botany, Box 90338, Durham, 

NC 27708, U.S.A. - E-mail: kauff@rhrk.uni-kl.de 

Despite various morphological and anatomical similarities, 

the two orders Gyalectales (lichenized ascomycetes) and 

Ostropales (lichenized and non-lichenized ascomycetes) 

have been considered to be distantly related to each other, 

and their position within the Ascomycota was unsettled. To 

estimate relationships within these groups and their 

respective phylogenenetic placement within the 

Ascomycota, we investigated the ascoma ontegeny and 

analyzed DNA sequences from the SSU and LSU nrDNA 

using Max. Parsimony, Max. Likelihood, and Bayesian 

statistics with Markov chain Monte Carlo algorithms. We 



report here that the Ostropales, in their current 

circumscription, are paraphyletic, and that the Ostropales 

s.l. include the Gyalectales and the Trapeliaceae. Dimerella 

and Coenogonium are congeneric, and Petractis 

thelotremella and P. hypoleuca are reunited with members 

of the genus Gyalecta. The ontogeny of the ascomata is 

variable within the Ostropales and can be used to further 

subdivide different lineages within this large and 

challenging order. Ascoma ontogeny, apothecial anatomy 

and molecular data support the separation of Coenogonium 

(incl. Dimerella) as family Coenogoniaceae from the 

Gyalectaceae. In addition to requiring less computational 

time, Bayesian inference of phylogeny recovered the same 

topology as a conventional heuristic search using Max. 

Likelihood as the optimization criterion, and seems 

superior to bootstrapping in estimating support for short 

internal branches. 

288 - Molecular evaluation of the generic concepts in 

the Pertusariaceae (lichenized ascomycota) 

I. Schmitt * & H.T. Lumbsch 

FB9/Botanik, Universität Essen, Universitätsstr. 5, 45117 

Essen, Germany. - E-mail: imke.schmitt@uni-essen.de 

The Pertusariaceae is a rather large and diverse family of 

crustose lichens. Its members are characterized by 

hemiangiocarpous ascoma development, thick-walled 

amyloid asci and relatively large ascospores. The group 

boasts a great diversity of secondary lichen compounds. 

Because several taxa are quite heterogeneous, 

morphologically and anatomically variable and often 

sterile, there has been a lot of discussion on the taxonomy 

of the family. In the present study we analysed nuclear 

LSU, mitochondrial SSU and LSU sequence data of 45 

species to re-evaluate the generic concepts in the 

Pertusariaceae. We ran maximum parsimony and Bayesian 

(MCMC) analyses and tested for monophyly of the genus 

Pertusaria. Our results suggest that the genus Pertusaria is 

polyphyletic comprising the Pertusaria s.str.-, the 

Monomurata- and Varicellaria-groups. The Monomuratagroup 

is a sister taxon to Ochrolechia; both are closely 

related to the Varicellaria-group. The genera Loxosporopsis 

and Melanaria are included in Pertusaria s.str. in the 

molecular genealogies. Morphological, anatomical and 

chemical characters are re-evaluated in the light of the 

molecular study. 

289 - Character evolution in Phacopsis inferred from 

nrDNA sequences 

D. Persoh 

Lehrstuhl Pflanzensystematik, Universität Bayreuth, 

Universitätsstraße 30 - NW I, D-95440 Bayreuth, 

Germany. - E-mail: derek.persoh@uni-bayreuth.de 

92 


Phylogenetic analyses of SSU nrDNA sequence data of 

two species of the lichenicolous genus Phacopsis (P. 

huuskonenii and P. oxyspora) revealed their membership of 

the lichen family Parmeliaceae. Studies of the ITS nrDNA 

of the family additionally included Phacopsis vulpina. 

While the monophyletic origin of the three lichenicolous 

species could not be rejected by the 'approximately 

unbiased test', phylogenetic trees calculated with 

parsimony and likelihood algorithms indicate no closer 

relationship among the investigated species of Phacopsis. 

Based on these facts, the evolution and adaptive value of 

selected morphological characters within the Parmeliaceae 

in the enlarged concept is discussed. Aside from 

characteristics of asco- and pycnospores, the 'cupulate' 

hypothecial layer is notable because it is present in 

Parmeliacean taxa and in P. oxyspora, but not in P. vulpina 

and P. huuskonenii. Furthermore, the hypothetical ways of 

thallus reduction and delichenisation are addressed. The 

loss of lichenisation (in the sense of a mutualistic 

interaction between a fungal and an algal partner) does 

most likely not occur in members of the genus Phacopsis. 

290 - Evaluation of morphological characters in the 

genus Usnea with the help of molecular data 

K. Articus 1* , M. Grube 2 , M. Wedin 3 & L. Tibell 1 

1 EBC, Systematic Botany, Norbyvagen 18D, SE-75236 

Uppsala, Sweden. - 2 Institut für Botanik, Karl-Franzens- 

Universität Graz, Holteigasse 6, 8010 Graz, Austria. - 

3 Department of Ecology and Environmental Science, Umeå 

University, SE-901 87 Umeå, Sweden. - E-mail: 

kristina.articus@ebc.uu.se 

A molecular phylogeny of a group of Usnea species was 

constructed utilising two independent gene loci, nITS-LSU 

rDNA and ß-tubulin. This phylogeny was used to 1) test if 

the traditionally used morphological species-characters 

actually characterize monophyletic groups, and 2) to 

analyse the species delimitation of the closely related 

pendant Usnea barbata, U. chaetophora, U. filipendula and 

U. rigida. The morphological characters most commonly 

used for species identification were mapped onto the 

molecular tree to assess their usefulness for characterizing 

monophyletic groups. Other species included were: Usnea 

articulata, U. ceratina, U. florida, U. hirta, U. longissima, 

U. subfloridana and U. wasmuthii. The species in focus 

form three monophyletic groups. Usnea barbata and U. 

rigida form one group with intermixed specimens. Most of 

the Usnea chaetophora specimens form a well supported 

group. The third group contains all U. filipendula 

specimens and several U. barbata and U. chaetophora 

specimens. There are some morphological characters that 

support these groups (such as branch shape and branching 

type in the U. chaetophora-group, and isidiomorph and 

isidia occurrence in the U. filipendula-group). Many 

character, though, e.g. the shape and number or papillae 

and fibrils, are inconsistent with the molecular phylogeny.


291 - A new look at the Pezizales 

D.H. Pfister 




Recent molecular phylogenetic studies have shown that the 

Pezizales are positioned at the base of the Euascomycota 

tree. These studies have confirmed the phylogenetic 

importance of a series of characters, such as ascus 

construction, ascus cytology and septal pore configuration, 

that have been used both at ordinal and family levels. 

Within the order molecular phylogenetic studies also have 

provided useful information on relationships among taxa. 

At least three major lineages have been recognized which 

include families and/or groups previously delimited based 

on morphological studies. I will review the characters 

currently in general use at family and genus level and 

suggest areas for further investigation. It is important at 

this juncture in the development of the classification of the 

Pezizales to direct attention to studies of morphology, life 

histories, ecology, population dynamics and biogeography. 

This is essential to the development of a useful and 

biologically informative classification. The idea that the 

Pezizales, with more than 1,000 described species, are 

relatively well known taxonomically will be discussed in 

light of recent field studies and those showing genetic 

diversity within even morphologically well characterized 

taxa. 

292 - A closer look at truffle phylogeny 

R. Percudani 1* , A. Rubini 2 , F. Paolocci 2 & S. Ottonello 1 

1 Dipartimento di Biochimica e Biologia molecolare, Parco 

area delle scienze, 43100 Parma, Italy. - 2 Istituo di 

ricerche sul Miglioramento Genetico delle Piante 

Foraggere, CNR, via Madonna Alta, 130 06128 Perugia, 

Italy. - E-mail: riccardo.percudani@unipr.it 

Small subunit ribosomal RNA (18S rRNA) sequence 

analysis has proven effective in resolving phylogenetic 

relationships among pezizalean families. However, due to 

the limited number of informative sites, a low statistical 

support is usually obtained for branch topologies at the 

intra-family and intrageneric level. Even though 18S rRNA 

topology in Tuberaceae has been corroborated by a parallel 

analysis of rRNA internal transcribed spacer regions (ITS), 

an extended sampling of species was required for a robust 

assessment of some unexpected relationships evidenced 

among Tuber and between Tuber and related genera. 

Moreover, the lack of convenient outgroup sequences 

prevented a correct rooting of the truffle phylogenetic tree. 

The results of a study based on a comprehensive set of 

Tuber, Choiromyces and Paradoxa ITS sequences and 

suitable outgroups such as Dingleya and Redellomyces will 

be presented. Comparisons of molecular phylogenies with 

morphological character trees and the convenience of a 

standard encoding system of truffle descriptors (TuberKey, 

http://www.truffle.org/tuberkey) will also be discussed. 

293 - Phylogeny and biogeography of Morchella 

K. O'Donnell 

USDA-ARS-NCAUR, 1815 N. University Street, Peoria, IL 

61604, U.S.A. - E-mail: kodonnell@mail.ncaur.usda.gov 

True morels classified in the genus Morchella are among 

the most highly prized and widely collected macrofungi by 

mycophiles. In order to infer their phylogeny, 

biogeography and the evolution of reproductive mode, 

multigene genealogies were constructed to investigate 

species limits within this systematically challenging genus. 

294 - Progress toward a phylogenetic classification of 

the Pyronemataceae 

B. Perry 



bperry@oeb.harvard.edu 

Of the families of the Pezizales the Pyronemataceae 

(including Otideaceae) remains the least well studied. The 

family has been considered a default family for pezizalean 

taxa with uninucleate spores and iodine negative asci, 

which lack distinguishing anatomical characters by which 

they could be segregated into putative natural families. 

Standard treatments of the Pyronemataceae include taxa 

with a wide diversity of both morphological features and 

nutritional modes. Recent molecular phylogenetic studies 

indicate that the Pyronemataceae is part of a lineage 

composed of Sarcoscyphaceae, Sarcosomataceae, 

Ascodesmidaceae, and Glaziellaceae. Within these 

analyses however, the Pyronemataceae is only poorly 

sampled. The goal of this investigation is to generate a 

multiple gene phylogeny of the Pyronemataceae and 

closely related taxa using sequence data from three 

unlinked nuclear loci (large subunit ribosomal DNA, ßtubulin 

and RNA polymerase II) to infer the evolutionary 

relationships of the family and the genera it contains. Initial 

phylogenetic analyses of a large subunit ribosomal data set 

will be presented and discussed. 



295 - Phylogenetics in the Pezizaceae / Ascobolaceae 

lineage 

K. Hansen * & K.F. LoBuglio 



khansen@oeb.harvard.edu 

Together Pezizaceae and Ascobolaceae appear to be a wellseparated 

lineage within the Pezizales supported by nuclear 

rDNA sequence data, iodine positive asci (unique to these 

families within the Pezizales), uninucleate spores, presence 

of an internal, circular indentation of the ascus apical 

apparatus, and some similarities in the type of ascus 

septum pore plugging. The family boundaries, however, 

remain questionable and will be addressed here, 

particularly with regard to the placement of Iodophanus 

and Thecotheus. Delimitation and relationships among 

genera within the Pezizaceae are highly controversial. Our 

previous analyses of 90 partial nuclear LSU rDNA 

sequences suggest that the circumscription of the large 

genus Peziza and other genera of the Pezizaceae, e.g. 

Plicaria, Amylascus, Scabropezia, Hydnotryopsis and 

Sarcosphaera should be re-evaluated. Peziza is composed 

of at least 6 major lineages, most of which include other 

genera of the Pezizaceae. In this study we will investigate 

the evolutionary relationships within the Pezizaceae using 

analyses of an expanded LSU rDNA data set, in 

combination with analyses of sequences from a portion of 

the protein coding nuclear genes β-tubulin and RPB2 for 

subsets of the taxa. A phylogenetically based classification 

of Pezizaceae is in progress. 

296 - Ophiostomatoid fungi associated with the Eastern 

Himalayan spruce bark beetle Ips schmutzenhoferi and 

other bark beetles in Bhutan 

T. Kirisits 1* , M.J. Wingfield 2 & D.B. Chhetri 3 

1 Institute of Forest Entomology, Forest Pathology and 

Forest Protection (IFFF), Universität für Bodenkultur 

Wien, Hasenauerstrasse 38, A-1190 Vienna, Austria. - 

2 Forestry and Agricultural Biotechnology Institute (FABI), 

Faculty of Biological and Agricultural Sciences, University 

of Pretoria, Pretoria 0002, South Africa. - 3 Renewable 

Natural Resources Research Centre (RNR-RC) Yusipang, 

P. O. Box 212, Thimphu, Bhutan. - E-mail: 

kirisits@edv1.boku.ac.at 

The bark beetle Ips schmutzenhoferi infests Eastern 

Himalayan spruce (Picea spinulosa) and Himalayan blue 

pine (Pinus wallichiana) and has been known to cause 

destructive outbreaks in conifer forests of Bhutan, Eastern 

Himalayas. We have recently conducted a preliminary 

investigation of the assemblage of ophiostomatoid fungi 

associated with I. schmutzenhoferi. The mycobiota of two 

other Himalayan bark beetles (Polygraphus sp. and 

Dryocoetes sp) was also considered. Nine, two and four 

ophiostomatoid fungi were found to be associated with I. 

94 


schmutzenhoferi, Polygraphus sp. and Dryocoetes sp., 

respectively. In total, 13 ophiostomatoid fungi, including 1 

Ceratocystis sp., 2 Ceratocystiopsis spp., 6 Ophiostoma 

spp., 3 Leptographium spp. and 1 Pesotum species were 

collected. The spectrum of fungi associated with I. 

schmutzenhoferi included Ceratocystis sp., 

Ceratocystiopsis minuta, 2 Leptographium species (sp. 1 

and sp. 2), Pesotum sp., Ophiostoma cf. ainoae, O. cf. 

cucullatum, Ophiostoma sp. 1 and Ophiostoma piceae. 

Polygraphus sp. was associated with Ceratocystiopsis sp. 

and Ophiostoma sp. 3, while Leptographium sp. 3, 

Ophiostoma sp. 1, Ophiostoma sp. 2 and O. piceae were 

associated with the Dryocoetes sp. The majority of these 

fungi are suspected to belong to hitherto unknown taxa and 

investigations on their taxonomic placement are 

continuing. This study has improved our knowledge on the 

occurrence and taxonomy of ophiostomatoid fungi and 

their vectors in the Himalayas. 

297 - Taxonomy, host relationships and breeding 

systems in Ophiostoma minus and O. pseudotsugae 

J.F. Webber 1* & C. Gorton 2 

1 Forestry Commission Research Agency, Alice Holt Lodge, 

Farnham, Surrey, GU10 4LH, U.K. - 2 RHS, Wisley, 

Woking, Surrey, GU23 6QB, U.K. - E-mail: 

joan.webber@forestry.gsi.gov.uk 

Ophiostoma minus is widely recognised both as a bluestain 

fungus causing discoloration of harvested lumber and as a 

pathogen of standing pines. On pine, O. minus is usually 

associated with bark beetle vectors: with Tomicus 

piniperda in Europe and with Dendroctonus frontalis in the 

USA. There is another Ophiostoma species, O. 

pseudotsugae, that is morphologically very similar to O. 

minus and so the two species have been considered to be 

synonomous. However, analysis of behavioural traits, 

molecular markers and breeding systems have 

demonstrated that O. minus and O. pseudotsugae are 

separate biological species. Moreover, it is also apparent 

that isolates currently identified as O. minus, comprise two 

distinct taxa that can be distinguished on the basis of their 

mating behaviour: one is homothallic and the other it is 

heterothallic. The extent to which these taxa should be 

considered as separate species is considered. 

298 - Hitch-hikers with invasive Tetropium fuscum 

(Fabr.) (Coleoptera: Cerambycidae) in Atlantic Canada 

K. Jacobs 1* , K.A. Seifert 1 , K.J. Harrison 2 , G. Smith 2 & T. 

Kirisits 3 


and Agrifood Canada, 960 Carling Avenue, Ottawa, K1A 

0C6, Canada. - 2 Natural Resources Canada, Canadian 

Forest Service, Atlantic Forestry Centre, Fredericton, E3B 

5P7, Canada. - 3 Institute of Forest Entomology, Forest 

Pathology and Forest Protection, Universität für


Bodenkultur Wien, Hasenauerstrasse 38, A-1190 Vienna, 

Austria. - E-mail: karin.jacobs@fabi.up.ac.za 

Species of Ophiostoma are well-known as causal agents of 

blue-stain in living trees and lumber, and as serious 

pathogens, and occur in close association with insects, 

especially bark beetles, which often function as vectors of 

these fungi. In 1999, dying red spruce (Picea rubens) in a 

small area of the Halifax Regional Municipality in Nova 

Scotia, Atlantic Canada were infested by Tetropium 

fuscum. In Europe, T. fuscum is usually a secondary pest of 

Norway spruce and rarely attacks healthy trees. In this area 

of Nova Scotia, T. fuscum is infesting and killing red 

spruce, whose native range is restricted to northeastern 

North America. Several ophiostomatoid fungi occur with 

this beetle. The aims of this study were to identify the fungi 

associated with T. fuscum in Halifax and compare them 

with ophiostomatoid fungi associated with this insect in 

Europe. Fungal associates of T. cinnamopterum 

(indigenous to and transcontinental in Canada) were also 

studied and compared to those of T. fuscum. Species were 

identified based on morphological and molecular 

characters. Ophiostoma tetropii is the most common 

species associated with T. fuscum in Europe and Canada, 

probably introduced into Atlantic Canada by this insect. O. 

tetropii was only isolated from trees initially attacked by T. 

fuscum. Other species isolated from trees attacked by both 

T. fuscum and T. cinnamopterum include O. piceae, a 

secondary colonizer of conifer sapwood, and the rarely 

reported Pesotum fragrans. 

299 - Surviving life in the fast lane: sex and clonality in 

Ophiostoma novo-ulmi 

C.M. Brasier * & J.F. Webber 

Forest Research Agency, Farnham, Surrey GU10 4LH, 

U.K. - E-mail: clive.brasier@forestry.gsi.gov.uk 

On several continents the Dutch elm disease pathogen 

Ophiostoma novo-ulmi is the fungal equivalent of an 

adventurous tramp. Having left its original home or centre 

of origin and 'jumped aboard a moving express train' (a 

spreading series of Dutch elm disease epidemics) it has had 

to cope with a many new and challenging events, processes 

and opportunities. 'Positive' opportunities include less 

distraction from competition between genotypes (a 

consequence of founder effects and selection), escape from 

natural enemies, an abundance of beetle vectors combined 

with a highly susceptible host, easier access to the 

saprotrophic phase and easier access to the vectors. 

'Negative' changes include increased risks from threats 

such as virus infections and sibling species and, potentially, 

a reduced opportunity for genetic variation if the 

environment changes. The impact these lifestyle changes 

appear to have had on the pathogens fitness, behaviour and 

population structure will be discussed, with special 

reference to sexual and asexual reproduction, clonality, the 

influence of fungal viruses and the role of outcrossing and 

horizontal gene transfer. 

300 - Hosts, insects, sex, aleurioconidia and the 

evolution of Ceratocystis 

T.C. Harrington * , A.E. Paulin-Mahady & J. Steimel 

Iowa State University, Department of Plant Pathology, 

Ames, Iowa 50011, U.S.A. - E-mail: tcharrin@iastate.edu 

Phylogenetic analyses of rDNA sequences and a portion of 

the MAT-2 gene show Ceratocystis to be monophyletic and 

most closely related to Gondwanamyces and more distant 

to the Microascales. Five major clades are found within 

Ceratocystis. The C. fimbriata complex and C. albofundus 

form a basal clade. Species in the C. paradoxa clade are 

pathogens on monocots. The Thielaviopsis basicola clade 

includes four soilborne, asexual species. The C. 

coerulescens clade lacks aleurioconidia and includes a 

subclade adapted to the Pinaceae. The remainder of the 

species group loosely in the C. moniliformis clade, which 

includes C. adiposa, C. fagacearum, and three asexual 

symbionts of ambrosia beetles: Ambrosiella xylebori, A. 

hartigii, and A. ferruginea. Characters found in the basal C. 

fimbriata clade and in one or more of the other clades were 

likely present in the most recent ancestor of Ceratocystis. 

The hypothesized ancestor was a wound-colonizer of 

woody dicots, insect-dispersed, and homothallic 

(unidirectional mating type switching); had hat-shaped 

ascospores accumulating at the tip of long-necked 

perithecia; and formed both endoconidia (Chalara-like) 

and aleurioconidia (pigmented, thick-walled spores that can 

survive in insect frass and soil). Shifts to heterothallism, 

loss of sexual reproduction, loss of aleurioconidia, 

adaptations to monocots or the Pinaceae, and bark beetle or 

ambrosia beetle symbiosis appear to have occurred in one 

or more lineages of Ceratocystis. 

301 - Relationships amongst Ceratocystis polonica, 

Ceratocystis laricicola and their bark beetle vectors in 

Europe and Asia 

M.J. Wingfield 1* , M. Marin 1 , T. Kirisits 2 , O. Preisig 1 & 

B.D. Wingfield 1 

1 Forestry and Agricultural Biotechnology Institute (FABI), 

University of Pretoria, Pretoria, South Africa. - 2 Institute 

of Forest Entomology, Forest Pathology and Forest 

Protection, Universität für Bodenkultur Wien, 

Hasenauerstrasse 38, 1190 Vienna, Austria. - E-mail: 

mike.wingfield@fabi.up.ac.za 

The pathogenic blue-stain fungus Ceratocystis polonica 

occurs on various spruce species (Picea spp.) and is 

associated with the bark beetles Ips typographus, Ips 

amitinus and Ips duplicatus in Europe and Ips typographus 

f. japonicus in Japan. Ceratocystis laricicola is the 

dominant pathogenic fungus vectored by the bark beetle 

Ips cembrae that infests larch (Larix spp.) in Europe and 

Japan. These two fungi are morphologically identical. 

However, recent studies have shown that they can be 

clearly separated using a number of molecular characters 



including isozymes and sequences of the ITS region of the 

ribosomal DNA operon, the beta-tubulin gene and the 

HMG box of the MAT-2 gene. Our recent data also show 

that the Japanese isolates of C. laricicola represent a 

distinct taxon. This is consistent with DNA-based 

comparisons of its vectors, which are presently also viewed 

as two separate species occurring in Europe and Asia. 

Reciprocal inoculations in a number of experiments 

including our own in central Europe, have shown that C. 

polonica is preferentially pathogenic to spruce. In contrast, 

C. laricicola is more pathogenic to larch than to spruce. 

These and other studies comparing populations of isolates 

using microsatellite markers, show that C. polonica and C. 

laricicola represent discrete biological entities. They have 

evidently evolved closely with their respective insect 

vectors, on different host trees. 

302 - Pleurotus species as medicinal mushrooms in 

health care and nutrition 

O.S. Isikhuemhen 1* , G. Goktepe 2 & J.V. Kumar 3 

1 Department of Natural Resources & Environmental 

Design, NC A&T State University, 1601 East Market 

Street, Greensboro, NC 27411, U.S.A. - 2 Department of 

Human Environment& Family Science, NC A&T State 

University, 1601 East Market Street, Greensboro, NC 

27411, U.S.A. - 3 Department of Chemistry, NC A&T State 

University, 1601 East Market Street, Greensboro, NC 

27411, U.S.A. - E-mail: omon@ncat.edu 

Pleurotus mushrooms, popularly called oyster mushrooms 

are distributed all over the world. Modern scientific 

analysis has shown that Pleurotus mushrooms are rich in 

proteins (especially those containing essential amino 

acids), vitamins, essential fatty acids and minerals. In past 

three decades in the United States of America, attention is 

increasingly been drawn to the studies of oyster 

mushrooms for its nutritional values and medicinal 

benefits. There are reports on biologically active 

compounds, lectin and different polysaccharides isolated 

from oyster mushrooms that have been tested for their 

medicinal effects. Polysaccharides from different oyster 

mushroom have been shown to have 50-95% reduction in 

cancel cell growth in vitro and animal models in some 

cases. The scientific and clinical studies of the medicinal 

properties of oyster mushrooms are still limited. Generally, 

the problem of standardization and a uniform approach to 

the studies on medicinal benefits of mushrooms also apply 

to oyster mushrooms. This report is aimed at bringing to 

light the traditional uses of oyster mushrooms and the 

current state of knowledge on the scientific studies of 

oyster mushrooms in food and medicine. The limitation in 

methodology and approach to scientific studies and 

application of oyster mushroom in nutrition and health care 

delivery, as well as solutions and better approach to this 

subject are discussed. 

96 


303 - Pleurotus sporocarps - a hypocholesterolemic 

nutraceutical? 

N. Gunde-Cimerman 1* & A. Plemenitas 2 

1 University of Ljubljana, Biotechnical Faculty, Biology 

Dept., Veèna pot 111, SI-1000 Ljubljana, Slovenia. - 

2 University of Ljubljana, MedicalFaculty, Institute of 

Biochemistry, Vrazov trg 2, SI-1000 Ljubljana, Slovenia. - 

E-mail: nina.gunde-cimerman@uni-lj.si 

Coronary artery diseases and hypercholesterolaemia are the 

main cause of morbidity in developed countries. The initial 

step in their prevention and treatment is the modification of 

the nutritional regime (diet low in fats and saturated fatty 

acids; rich in crude fibers). The oyster mushrooms are a 

good source of nonstarchy carbohydrates, have a very high 

dietary fiber content and a low caloric value. Therefore 

their hypolipidaemic efficiency was for a long time 

attributed only to the interaction of crude fibers and 

chitosan with bile acids in the small intestine, causing 

increased excretion and reduced enterohepatal circulation 

of bile acids and accelerating cholesterol catabolism by a 

feedback mechanism. Later it was discovered that addition 

of dried oyster mushroom to high cholesterol diet 

effectively reduces cholesterol accumulations in the serum 

and liver of rats, redistributes cholesterol in favour of high 

density lipoprotein (HLDL) cholesterol, reduces production 

of very low density lipoprotein (VLDL) and low density 

lipoprotein (LDL) cholesterol. It also reduces cholesterol 

absorption and the activity in liver of the major rate 

limiting enzyme in cholesterol biosynthesis pathway, 3hydroxy-3-methylglutary 

CoA (HMG CoA) reductase. By 

the combined action of crude fibers and specific inhibition 

of HMG CoA reductase, a considerable reduction in VLDL 

and LDL cholesterol levels can be explained. 

304 - Antioxidant activity of several basidiomycete 

mushrooms cultures 

S.M. Badalyan 1* & A.V. Gasparyan 2 

1 Yerevan State University, Department of Botany, 375025, 

1 Aleg Manoogian St., Armenia. - 2 State Microbial 

Depository Center, National Academy of Sciences Of 

Armenia, 378510, Abovian city, Armenia. - E-mail: 

badalians@infocom.am 

The active metabolites of medicinal basidiomycete 

mushrooms have different medicinal and preventive 

properties. However, the data concerning their antioxidant 

activity (AOA) is rare. The ability of mushroom-derived 

compounds to prevent oxidative damage was observed 

within cultures mostly from the genera Agaricus, Coprinus 

and Ganoderma. The cultural liquid (CL) was proposed for 

preventing cancer, nephritis, hypertension, and for delaying 

senility. The higher peroxidase inhibition potential of 

fungal melanin pigments was also reported. Mycelial 

melanin is promising for elaboration of new biotechcompounds 

with AOA. In our experiment 17 mushroom


cultures from 13 genera (Coprinus, Coriolus, Flammulina, 

Lentinula, Lepista, Marasmius, Hypholoma, Pholiota, 

Pleurotus, Schizophyllum, Stropharia, Suillus and 

Volvariella) were tested on AOA. The CL and mycelium 

suspension (MS) obtained on days 7 and 14 haven't 

possessed AOA. The all samples showed different degree 

of activity on 21 day. The CL samples were more active 

than MSs. The highest AOA in CL of Pholiota alnicola 

(44%) and in MS of Volvariella bombycina and Lepista 

personata (26%) has been revealed. No correlation 

between AOA and ecological belonging of tested species 

has been observed. Further AOA-screening, study of 

mechanism of action and dosa-effect correlations of active 

compounds will assist in obtaining new biopreparations or 

mushroom dietary supplements with antioxidant effect. 

305 - Mushrooms and weight loss: a randomised, 

double blind study 

B. Donatini * & I. Le Blaye 

Mycoceutics - 40 rue du Dr Roux, 51350 Cormontreuil, 

France. - E-mail: donatini@wanadoo.fr 

We investigated the impact of mycelium shiitake or 

Agaricus bisporus, when associated with fibers, on body 

weight. Material and methods Biscuits A were prepared 

with 6% of mushroom powder (70% of Shiitake mycelia 

grown on wheat and 30% of fibers) and Biscuits B 3% only 

(70% of dried Agaricus bisporus and 30% of fibers). The 

micture was finalised according to the patent 

PCT/FR01/00577. Consumers were randomized. Each had 

to eat 5 biscuits (50g) per day for 4 weeks. Body weight, 

fat index, food intake, physical activity and global caloric 

balance (caloric intake - physical activity) were compared. 

Results 38 consumers were included: 19 (18 women) in 

group A and 19 (14 women) in group B. 26 consumers 

completed the form on food intake and physical activity. 

Table 1: Populations A and B were comparable 

Group A Group B 

Mean age 30 ±10 35 ±10 

Body weight (kg) 67 ±10 71 ±11 

% of fat 30 ±6 32 ±10 

For group A, caloric balance - for 4 weeks - was equal to 

+998 kcal and +1562 for group B. 

Table 2: Difference for body weight and fat weight after 4 

weeks. 

Group A Group B 

Body weight difference -0.2 ±1.1* -1.3 ±1.3* 

Fat weight difference +0.3 ±1.0* -1.4 ±2.1* 

*p


The inhibiting effect of mycelial or fruit body extracts or 

liquid culture filtrates from Lentinus edodes (Berk) Sing. 

and other medicinal mushrooms has been tested on 

aflatoxin production by Aspergillus parasiticus Speare. The 

mushrooms tested were: Lentinus edodes, Trametes 

versicolor (L.:Fr) Pil., Auricularia auricula-judae (Bull) 

Wettstein and Pleurotus ostreatus (Jocquin ex Fr.) 

Kummer. The aflatoxin producer strain was Aspergillus 

parasiticus NRRL 2999. The medicinal mushrooms were 

inoculated on Potato Dextrose Broth (PDB, Difco) and 

incubated at 25 °C till they have reached plateau. The 

cultures were than filtered, and the culture filtrates were 

lyophized and tested at the concentration of 2% w/v in the 

presence of A. parasiticus conidia and incubated at 28 °C 

up to 6 days. The fruit bodies and spent compost of L. 

edodes were water extracted and assayed in the same way. 

The extracts from mycelia and liquid culture filtrate were 

also partially purified and the obtained fractions tested on 

the aflatoxin production. The results obtained showed that 

not purified lyophilised or liquid filtrates, fruit bodies and 

spent compost extracts from L. edodes inhibited aflatoxin 

production of 85-90% without interfering with A. 

parasiticus growth. Among the partially purified extracts 

some showed a strong inhibiting effect both on fungal 

growth and aflatoxin production (T. versicolor), while 

others inhibited only the aflatoxin production (A. 

auricularia-judae). 

309 - Botanical antifungal drug from lichen metabolites 

fight fungal infections 

S.K. Shahi 1* , M. Patra 1 , A. Dikshit 1 & D.K. Upreti 2 

1 Biological Product Lab, Botany Department, University of 

Allahabad, Post Box. 2026, Kuchery PO- 211002, 

Allahabad, India. - 2 Lichen Laboratory, National Botanical 

Research Institute, Rana Pratap Marg, 226 001, Lucknow, 

India. - E-mail: shahiindia@rediffmail.com 

During antifungal screening of some lichen metabolites 

(LM), tested at different concentrations against human 

pathogenic fungi (dermatophytes) Epidermophyton 

floccosum, Microsporum audouinii, M. nanum, M. canis, 

M. gypseum, Trichophyton mentagrophytes, T. rubrum, T. 

violaceum and T. tonsurance. The lichen metabolites of 

Usnea longissima Ach. was found to be most effective. The 

minimum inhibitory concentration of the LM was found to 

be 30 µl ml -1 at which LM showed fungistatic action. The 

minimum fungicidal concentrations (MCCs) of the LM 

were found to be 50 µl ml -1 against human pathogenic 

fungi. The LM at 50 µl ml -1 showed heavy doses of 

inoculum potential. Moreover, LM did not exhibit any 

adverse effect on mammalian skin up to 10% 

concentration. Further, the LM based ointment of Usnea 

longissima was trialed clinicaly in MLN Medical college, 

Allahabad. 30 patients were selected, showing positive 

potassium hydroxide (KOH) results at the start of the trial. 

At the end of medication, 30.0% of patients recovered 

complete cure, 45.0% showed significant improvement 

from the disease. No KOH negative cases of relapse were 

observed when patients were reexamined after two month 

following the end of treatment. The ointment was found 

cost effective and absence of any adverse effects. The 

98 


commercial utilization of the ointment could be determined 

after undergoing successful multicentre clinical trial. 

Which is in progress. 

310 - Mycotoxins as ecotypic characters in Fusarium 

species 

U. Thrane 

Mycology Group, BioCentrum-DTU, Technical University 

of Denmark, Søltofts Plads 221, DK-2800 Kgs. Lyngby, 

Denmark. - E-mail: ut@biocentrum.dtu.dk 

Fusarium species can be found in many different 

ecological niches all over the world, including natural as 

well as man-made niches. Furthermore, it is well-known 

that every Fusarium species is able to produce several 

biological active metabolites, of which some are known as 

mycotoxins. However, this does not imply that every 

Fusarium mycotoxin can be found everywhere, as not all 

Fusarium species occur everywhere. Each ecological niche 

has its own associated set of Fusarium species, which can 

growth on the given substrate under the given conditions. 

Taking an ecological approach a link between Fusarium 

species, their mycotoxins, and the ecological niche will be 

demonstrated, indicating that known mycotoxins may have 

more functions than being toxic to humans and livestock. 

The presentation will focus on the profile of produced 

compounds originating from more metabolite families and 

not on individual compounds of the trichothecenes, 

zearalenones, fumonisins, moniliformin, and other well 

known Fusarium metabolites. The holistic interpretation of 

metabolite profiles is launched as a fruitful platform for 

future discussions on fungal chemistry and toxicology 

towards a better understanding of fungal ecology. 

311 - Ecology and genetics of loline alkaloid expression 

in grass endophyte defensive mutualisms 

H.H. Wilkinson 1* , B.L. Kutil 1 , M.J. Spiering 2 , C.D. Moon 2 

& C.L. Schardl 2 

1 Texas A&M University, Department of Plant Pathology 

and Microbiology 2132 TAMU College Station, Texas 

77845-2132, U.S.A. - 2 University of Kentucky, Department 

of Plant Pathology S-305 Agricultural Sciences Bldg. 

Lexington, KY 40546-0091, U.S.A. - E-mail: 

heatherw@ppserver.tamu.edu 

Loline alkaloids (saturated 1-aminopyrrolizidine alkaloids 

with an oxygen bridge) are fungal secondary metabolites 

often present in grasses symbiotic with endophytes in the 

genera Epichloë and Neotyphodium. Endophytes producing 

loline alkaloids provide grass hosts with enhanced 

protection from herbivory, drought and pathogens. 

Epichloë festucae has proven to be an excellent genetic 

model system for ecological and genetic studies of lolines. 

In an earlier study, segregation analysis supported a single 

locus (LOL) contributing to a naturally occurring lolines


expression polymorphism in E. festucae. Responses of two 

aphid species (Rhopalosiphum padi and Schizaphis 

graminum) to meadow fescue symbiota containing E. 

festucae progeny segregating for LOL (Lol+ vs Lol-) 

clearly implicate lolines as agents of protection against 

insects. Successful map-based cloning has resulted in 

identification of a >40kb gene cluster unique to the Lol+ 

parent. Subsequent sequencing of this region in E. festucae 

(and related gene clusters in Neotyphodium uncinatum; see 

poster by Spiering et al.) reveals relationships of the likely 

loline alkaloid synthesis genes with fungal genes known 

for synthesis of amino acids, polyamines and other 

secondary metabolites. 

312 - Inherent fitness advantage associated with 

progression along the AF/ST polyketide biosynthesis 

pathway in Aspergillus 

H.H. Wilkinson 1* , A. Ramaswamy 1 , S.C. Sim 1 & N.P. 

Keller 2 

1 Texas A&M University, Department of Plant Pathology 

and Microbiology 2132 TAMU College Station, Texas 

77845-2132, U.S.A. - 2 University of Wisconsin, Department 

of Plant Pathology, Madison, Wisconsin 53706, U.S.A. - Email: 

heatherw@ppserver.tamu.edu 

Sterigmatocytin (ST) is a polyketide-derived Aspergillus 

mycotoxin and penultimate precursor of aflatoxin (AF). 

Although the specific functions of many ST cluster genes 

have been elucidated with regard to their role in the ST/AF 

biosynthesis, the specific function of ST/AF for the fungi 

that produce them is not known. We are attempting to 

identify the fitness costs and/or benefits of ST expression. 

Isogenic and prototrophic, A. nidulans mutants interrupted 

at different genes in the ST biosynthesis cluster (delta-aflR, 

delta-stcJ, delta-stcE, delta-stcU) and the wild type (ST) 

were generated. These genotypes were grown on petri 

plates containing glucose minimal media or on live. 

Relative fitness was determined as a difference in the 

number of conidia harvested after incubation for seven 

days at 37°C.Two light conditions were used in these 

fitness tests. In both experiments completing the ST 

pathway was beneficial to fungal fitness. Rather than being 

attributable to the impact of a knockout mutation, the tight 

relationship between fitness ranking and position of each 

gene product in the biochemical pathway support a direct 

effect of these mutations on conidiation. These data suggest 

that ST (and presumably AF) production contributes to the 

fitness of Aspergillus under the environmental conditions 

tested. 

313 - Biology of secondary metabolites 

J.D. Miller 

Chemistry, Carleton University, Ottawa, Ontario K1S 5B6, 

Canada. - E-mail: david_miller@carleton.ca 

There are many perspectives on secondary metabolites of 

the marine fungi, conifer endophytes and agricultural fungi 

I have studied. The genes that code for secondary 

metabolites seem largely to be clustered together implying 

that their acquisition was not incremental but chaotic. Their 

role in determining population structure focuses on the 

benefit to species. Necrotrophic plant pathogens produce 

virulence factors that also discourage the consumption of 

the affected plant tissues by animals. Fungi that produce 

toxins that affect animal competitors typically produce 

several families of toxins. Some toxin families many be 

more or less active against different potential herbivores. 

Others result in additive toxicity with the important 

consequence that acquisition of resistance to the toxin is 

improbable. Despite this high degree of evolution, the 

production of secondary metabolites by filamentous fungi 

must first be thought of in terms of the biochemistry of the 

terminal few cells of the mycelium. It has been known 

since the invention of the microscope that it is only these 

cells that are active, with of course the most active being 

the hyphal tip. Control of the pathways of secondary 

metabolism reflects the regulatory processes of glycolysis, 

the citric acid cycle and electron transport. As nutrients 

become unavailable to the active cells, accumulation of 

acetate, citric acid cycle products and some amino acids 

become available for the production of secondary 

metabolites. 

314 - Liard Hotsprings hypsothermal relict 

commmunity: thermal impacts on fungi and plants in a 

boreal forest ecosystem 

R.S. Winder 1* , A. Ceska 2 & O. Ceska 2 

1 Natural Resources Canada / Canadian Forest Service, 

Pacific Forestry Centre, 506 W. Burnside Rd., Victoria, 

B.C. V8Z 1M5, Canada. - 2 Botanical consultant, 1809 

Penhurst Rd. Victoria, B.C. V8N 2N6, Canada. - E-mail: 

rwinder@pfc.forestry.ca 

The response of plants and fungi to temperature gradients 

was studied in Liard Hot Springs Provincial Park in the 

boreal forest of British Columbia. Within 50 m of the hot 

springs, the forest was characterized by the proliferation of 

understory shrubs such as bitter cherry (Prunus 

emarginata) and bunchberry (Cornus canadensis), and 

hardwood tree species such as balsam poplar (Populus 

balsamifera) and paper birch (Betula papyrifera). By 

contrast, the forest 150 m from the springs was occupied 

primarily by white spruce (Picea glauca). Basal area of 

tree stems declined with proximity to the springs and 

increasing proportion of hardwoods; basal area 50 m from 

the springs was half of the basal area 150 m from the 

springs. Diversity of macrofungal fruiting bodies also 

declined from an average of 33 species 150 m from the 

springs, to an average of 13 species 50 m from the springs. 

Fungi found in the warmer zones included species such as 

Humaria hemispherica and Mycena delicatula. 

Hygrophorus spp., Marasmius spp., and others were found 

in the colder zones, while Cortinarius spp., Inocybe spp., 

and others were found throughout the gradients. Plant 

diversity and the proportion of mycorrhizal species were 

unchanged with respect to distance from the springs. While 



further study would be needed to extrapolate this 

information into general trends, boreal hot springs appear 

to offer unique opportunity to study the potential impacts 

of climate change on boreal fungi and plants. 

315 - Biomass of ectomycorrhizal mycelia at different 

soil depths and along nutrient gradients in the boreal 

forest 

H. Wallander * , L.-O. Nilsson, S. Mahmood & S. Erland 

Department of Microbial Ecology, Lund University, 

Ecology Building, 223 62 Lund, Sweden. - E-mail: 

Hakan.Wallander@mbioekol.lu.se 

Nutrient uptake by forest trees is greatly dependent on 

ectomycorrhizal (EcM) fungi. We quantified the 

production of EcM mycelia in the field by the use of ingrowth 

mesh bags. Mesh bags were placed at different soil 

depths (5-15-30 cm) in spruce and mixed spruce/deciduous 

forests in southern Sweden and in the humus layer along a 

natural nutrient gradient in northern Sweden. Spruce stands 

produced 590 kg EcM biomass ha -1 while mixed forests 

produced 420 kg ha -1 . The delta-13C value of mycelia 

collected from mesh bags was similar to values of EcM 

fruitbodies, and it was not influenced by soil depth, 

indicating that the mycelia were of EcM origin. The delta- 

13C value in mycelia from mixed forests suggested that the 

mycelia received more carbon from spruce trees than from 

oak trees. The production of EcM mycelia decreased with 

soil depth. The decrease was more accentuated in mixed 

stands compared to pure spruce stands. In the natural 

nutrient gradient (low N and low pH changed gradually to 

high N and high pH) the production of EcM biomass 

decreased when moving from the nutrient poor to the 

nutrient rich end. We identified some of the EcM species 

that colonized the mesh bags with PCR/RFLP analysis of 

ribosomal DNA. We found a low level of similarity 

between EcM species found on root tips outside the mesh 

bags and EcM species found as mycelia or rhizomorphs 

inside the mesh bags. Fast growing species such as Paxillus 

involutus were common in mesh bags but rare on root tips. 

316 - New perspectives on the ecological distribution of 

epiphytic hair lichens (Bryoria) in northern forests 

T. Goward 

Enlichened Consulting Ltd., Edgewood Blue, Box 131, 

Clearwater, B.C. V0E 1N0, Canada. - E-mail: 

tgoward@interchange.ubc.ca 

Caribou biologists working in the high-elevation conifer 

forests of inland British Columbia have long observed that 

hair lichens in the genus Bryoria attain much heavier 

loadings in oldgrowth stands than in adjacent younger 

stands. In an attempt to account for this phenomenon, I 

examined Bryoria species composition in regenerating 

forests along successional and within-stand vertical 

100 


gradients. Bryoria appears to enter regenerating stands in 

two phases. In the first phase, to about 70-100 years, B. 

fuscescens and B. glabra dominate throughout the canopy. 

These are small, sorediate lichens with a relatively high 

tolerance for prolonged wetting. After ca. 100 years, these 

species are increasingly replaced, at least in the upper 

canopy, by the non-sorediate B. fremontii and B. 

pseudofuscescens. These are large, relatively xerophytic 

hair lichens favoured by open, well-ventilated conditions. 

B. fremontii and B. pseudofuscescens have indeterminate 

growth; once established in the upper canopy, they tend to 

fragment, thereby being continuously 'parachuted' into the 

lower canopy. It is the gradual accumulation of these nonsorediate 

species at all levels of the canopy that ultimately 

leads to an elevated Bryoria biomass in oldgrowth forests. 

The ability of B. fremontii and B. pseudofuscescens to 

persist in the lower canopy does vary considerably from 

year to year, depending on the occurrence of 'die-back' 

events associated with prolonged dampness. 

317 - Implications of fungal translocation for nutrient 

cycling in boreal forest ecosystems 

B.D. Lindahl 

Dept. of Forest Mycology and Pathology, SLU, Box 7026, 

SE-750 07 Uppsala, Sweden. - E-mail: 

bjorn.lindahl@mykopat.slu.se 

Most current models of nutrient cycling have been 

developed with a vague concept of 'microorganisms' in 

mind, generally relating to bacteria. In boreal forest 

ecosystems, fungi probably play a more important role than 

bacteria in the decomposition of organic matter. Most fungi 

differ from bacteria in that they are multicellular and able 

to translocate resources throughout their mycelia. Current 

nitrogen cycling models are centred around mineralisation; 

as substrates with a low C/N-ratio are degraded, nitrogencontaining 

organic compounds are thought to be used as an 

energy source, leaving ammonium as an undesired 

biproduct that is released. Fungi may, however, degrade 

substrates with a low energy content, using carbohydrates 

that are translocated from external sources such as woody 

debris, litter or living roots in the case of mycorrhizal 

fungi. Obtained nutrients may be translocated back in the 

opposite direction to support colonisation and degradation 

of the nitrogen-poor wood or litter, or to support a 

mycorrhizal host plant. Translocation of resources thus 

enables fungi to utilise resources of various qualities more 

efficiently, conserving nutrients within their mycelia 

instead of exuding them. In light of the capacity of boreal 

forest plants to access organic forms of nitrogen via their 

mycorrhizal associates, it is likely that a significant fraction 

of the nitrogen recycling from organic matter back to plants 

takes place without the nitrogen ever being mineralised.


318 - The natural abundance of 15 N in mat-forming 

lichens 

C.J. Ellis 1* , P.D. Crittenden 1 & C.M. Scrimgeour 2 

1 School of Life and Environmental Sciences, University of 

Nottingham, Nottingham, NG7 2RD, U.K. - 2 Scottish Crop 

Research Institute, Invergowrie, Dundee, DD2 5DA, U.K. - 

E-mail: christopher.ellis@nottingham.ac.uk 

Natural abundance of 15 N and [N] was studied in thalli of 

mat-forming lichens collected from tundra and heathland 

sites in the Northern and Southern Hemispheres. The study 

includes samples of British Cladonia portentosa from sites 

in regions of high and low N-loading and in heathland 

growing on peat and independently of the soil substratum, 

in a canopy of prostrate gorse. In the mat-forming lichens 

examined, a non-random pattern in [N] and δ 15 N was 

characterised by a minimum in δ 15 N, which occurred most 

frequently at 40-60 mm below the thallus apex. [N] 

increased above this point, towards the apex, though 

remained invariably low towards the thallus base. We 

discuss the significance of the pattern in [N] and δ 15 N for 

current theories describing the uptake and recycling of 

nitrogen by mat-forming lichens in oligotrophic habitats. 

Our data are incompatible with the suggested uptake of soil 

organic nitrogen depleted in 15 N, though are consistent with 

possible internal recycling and the development of a 

structural necromass. The study emphasises the internal 

fractionation of nitrogen isotopes and provides a caveat 

against the assumption that values of δ 15 N provide an 

unequivocal indicator of source-sink relationships in 

nitrogen cycling. 

319 - Role of environmental conditions and birch tree 

genotype in endophyte-herbivore interactions 

K. Saikkonen 

University of Turku, Section of Ecology, Department of 

Biology, 20014 Turku, Finland. - E-mail: karisaik@utu.fi 

We compared phenotypic and genetic frequency 

correlations for two endophytic fungal genera (Fusicladium 

and Melanconium) and bich rust (Melampsoridium 

betulinum) with the performance of six invertebrate 

herbivores growing on the same half-sib progenies of 

mountain birches (Betula pubescens ssp. czerepanovii) in 

two environments over a 3-year period. We found little 

support for causal association between fungal frequencies 

and performance of herbivore species. Instead, genetic 

correlations, particularly between autumnal moth (Epirrita 

autumnata) and birch rust, suggest that herbivore 

performance may be affected by (1) genetic differences in 

plant quality for fungi and herbivores, or (2) genetic 

differences in responses to environmental conditions. 

Genetic analysis (RAMS-PCR) of Venturia ditricha 

(anamorph Fusicladium betulae) revealed that (1) host 

genotypes and environment influence the probability of 

infection by particular endophyte genotypes, (2) genetic 

variation correlated negatively with infection frequencies 

of the fungus, and (3) the susceptibility of the host to a 

particular endophyte genotype may change when 

environmental conditions are changed (environment-host 

genotype interaction). Our results propose that the 

seemingly direct interactions between herbivores and fungi 

may actually indicate genetic differences in plant quality 

for fungi and herbivores or responses to environmental 

conditions. 

320 - Specificity of fungal-algal interactions on leaves 

M. Grube 1* , E. Baloch 1 & R. Luecking 2 

1 Institute of Botany, Holteigase 6, A-8010 Graz, Austria. - 

2 The Field Museum of Natural History, 1400 South Lake 

Shore Drive, Illinois 60605-2496, U.S.A. - E-mail: 

martin.grube@uni-graz.at 

Several types of algae are adapted to the growth on living 

leaves, coccoid green algae and members of 

Trentepohliales being most common and also serving as 

photobionts of lichen-forming fungi. Initials of these 

lichens need to form composite thalli with appropriate 

algae in a relatively short time. Therefore, particular 

strategies are pursued to ensure efficient dispersal and 

colonialization of leaves. Specific growth patterns of 

trentepohlioid algae, found in both lichenized and nonlichenized 

forms, suggest that foliicolous mycobionts have 

a high specificity for their photobionts. Adjacent freeliving 

Phycopeltis colonies belonging to different 

morphotypes are not accepted as photobionts by the same 

fungus. Different ecological preferences of photobionts 

apparently shape the diversity of foliicolous lichens found 

in a given microhabitat. A particular strategy regarding 

photobiont selection of lichens is found in lichenized, 

foliicolous species of Chroodiscus, which grow 

facultatively parasitic on lichens of the genus Porina. C. 

australiensis and C. coccineus show a high degree of host 

specificity: C. australiensis parasitizes Porina mirabilis, 

whereas C. coccineus is specifically found on P. 

subepiphylla. In genera of lichens, which include both 

lichenized and non-lichenized, lichenicolous species (e.g. 

Arthonia), the latter take advantage of the host photobionts. 

321 - Fungal endophytes in a neotropical forest: 

ecological, systematic, and physiological insights 

A.E. Arnold 1* , L.C. Mejía 2 , E. Rojas 2 , D.A. Kyllo 2 & E.A. 

Herre 2 

1 University of Arizona, Department of Ecology and 

Evolutionary Biology, 1041 E. Lowell, BSW 310, Tucson, 

AZ 85721, U.S.A. - 2 Smithsonian Tropical Research 

Institute, Apartado 2072, Balboa, Panama. - E-mail: 

betsya@email.arizona.edu 

Fungal endophytes associated with woody angiosperms are 

abundant and diverse; yet especially in tropical forests, 



they are poorly known. In particular, the scale of endophyte 

diversity, and the nature of endophyte-host interactions, are 

not well understood. Drawing from extensive field studies 

at scales ranging from individual leaves to disparate sites 

across lowland Panama, I will discuss evidence for spatial 

structure, temporal variability, host affinity, and horizontal 

transmission among tropical forest endophytes. Drawing 

from experimental results, I will describe a potential 

mechanism for host affinity based on defensive chemistry 

of host leaves. Using molecular sequence data (nrDNA: 

ITS1, ITS2, 5.8s), I will show that tropical endophytes are 

diverse at both low and high taxonomic levels, and will 

discuss phylogenetic patterns of diversity. Finally, I will 

discuss roles of endophytes in mediating host defense 

against foliar pathogens, and will assess general costs and 

benefits of endophyte infection in tropical woody 

angiosperms. 

322 - Effect of agricultural management on diversity of 

root endophytes: the role of dark septate endophytes 

E.M. Wilberforce 1* , G.W. Griffith 1 , L. Boddy 2 & R. 

Griffiths 3 



3DA, Wales, U.K. - 2 School of Biosciences, Cardiff 

University, PO Box 915, Cardiff CF10 3TL, Wales, U.K. - 

3 National Botanic Garden of Wales, Middleton Hall, 

Llanarthne, Carmarthenshire, SA32 8HG, Wales, U.K. - Email: 

emw96@aber.ac.uk 

Three mesotrophic grassland sites of similar physical 

characteristics but differing management histories were 

chosen to test the hypothesis that agricultural disturbance 

has a deleterious effect on the diversity of fungi inhabiting 

plant roots and the prevalence of potentially pathogenic 

species (e.g. Fusarium spp.). Species abundance data were 

collected for fungi isolated from surface sterilised root 

samples (>40 taxa). Shannon and Brillouin indices of 

diversity, TWINSPAN and detrended correspondence 

analysis were applied to the community data. Quantitative 

ordination separated the samples by site showing that the 

communities differed in fields of contrasting management. 

Species presence and absence appeared to be affected 

seasonally; site differences were manifested in relative 

abundance. Diversity did not appear to vary by site, but a 

methodological explanation for this is proposed. Sterile 

dark septate endophytes (DSE) were shown among the 

most abundant groups at all sites, and diversity among 

these fungi was investigated using a PCR-based approach. 

It is suggested that DSE, in addition to the overall diversity 

of root-colonising species, may be implicated in relating 

plant root communities and plant health. Microcosm 

systems have been used to study the dynamics of root 

colonization by DSE, as well as Fusarium spp. 

102 


323 - Rock inhabiting fungi and lichen photobionts: 

symbiotic or antibiotic interactions? 

A. Gorbushina 1* , A. Beck 2 & A. Schulte 1 

1 Geomicrobiology, ICBM, Oldenburg University, P.O.Box 

2503, D-26111 Oldenburg, Germany. - 2 Lehrstuhl für 

Pflanzensystematik, Universität Bayreuth, Universitätsstr. 

31, D-95447 Bayreuth, Germany. - E-mail: 

anna.gorbushina@uni-oldenburg.de 

Phototrophs are considered to be the first land colonisers, 

although this ability is often supported by mutualistic 

associations with fungi. Furthermore bare rock surfaces are 

frequently dominated not by phototrophs, but by free-living 

and symbiotic ascomycetes. On desert rock surfaces 

lichens often yield and in the harshest environments only 

microcolonial fungi (MCF) are present. MCF - highly 

stress-tolerant free-living organisms - could represent the 

remnants of symbiotic associations, which implies their 

capability to interact with photobionts. Four typical MCF 

were cultivated with 4 lichen photobionts isolated from 

similar desert locations. The relations between the partners 

were investigated by electron microscopy and histological 

methods. After several months of cultivation a structure 

involving both partners has been developed. Photobiont 

cells were not changing size, but fungal branching was 

more expressed in the vicinity of green algal cells. 

Histological analysis of interwoven colonies exhibits 

spatial adjustment of the partners. After continued 

cultivation algae lost vitality. Only one MCF strain enabled 

survival of the algae tested. The absence of antibiotic 

influences in the first stage of the experiment hints to an 

universal ability of MCF to form unstable mutualistic 

relations with lichen photobionts. The later loss of 

mutualistic balance may be connected to the diverse 

taxonomic position of the MCF strains in question. 

324 - Cyanobiont diversity in ecological lichen guilds 

J. Rikkinen 

Department of Applied Biology, PO Box 27, 00014 


jouko.rikkinen@helsinki.fi 

Many cyanolichen species are organized into guilds around 

groups of phylogenetically related cyanobacteria. In a 

study of old-growth associated cyanolichens in central 

Finland all bipartite epiphytes were found to house closely 

related Nostoc strains and most of the thalli contained one 

specific strain. While the cyanobionts of all these lichens 

were closely related, the lichen-forming fungi were 

necessarily not. Moreover, some related terricolous species 

associated with a different group of Nostoc symbionts and 

thus belonged to a different ecological guild. A comparison 

with cyanolichens from North America and East Asia 

revealed concurrent patterns. Thus, a similar specificity 

was evident on all spatial scales ranging from single tree 

trunks to global distributions. All lichen-forming fungi


within a specific guild can potentially share cyanobionts 

with each other. Further studies of guild structure will give 

us a better understanding of the dispersal ecologies and 

conservation requirements of old-growth associated 

epiphytes and other cyanolichens. 

325 - Grass endophyte host specificity and evolutionary 

implications 

C.L. Schardl * , K.D. Craven & C.D. Moon 

University of Kentucky, S-305 ASCN, Lexington, Kentucky 


The Epichloë (anamorph = Neotyphodium) species are 

fungal endophytes of grasses, and span an evolutionary 

continuum from antagonistic to mutualistic symbionts. The 

antagonistic endophytes are sexual, horizontally 

transmitted (contagious), and suppress host seed 

production; the more mutualistic can transmit vertically in 

host seeds. Some vertically transmissible species are sexual 

and capable of occasional contagiously spread; others are 

strictly seed-borne and asexual. In general, each Epichloë/ 

Neotyphodium species occurs in a single host genus or 

tribe. However, one of the more antagonistic species, 

Epichloë typhina, can infect grasses in at least three diverse 

tribes. The high genetic diversity in E. typhina compared to 

other Epichloë species suggests that speciation has been 

relatively slow in E. typhina. This may be due to its 

dependence on horizontal transmission, because 

diversification by sexual recombination may enhance the 

potential of E. typhina to adapt to the diversity of available 

hosts. Among the seed-borne sexual species, host 

specialization appears to help drive speciation, such that 

there is good correspondence between the phylogenetic 

relationships among some Epichloë species and the 

phylogeny of their respective hosts. Curiously, the strictly 

seed-borne endophytes do not exhibit co-phylogeny with 

their hosts. Instead, they usually seem to have arisen by 

transfers between host tribes, and often have undergone 

interspecific hybridization. 

326 - Exceptionally high nucleotide substitution rate 

differences between lichenized Omphalina species and 

their symbiotic green algae Coccomyxa 

S. Zoller * & F. Lutzoni 

Duke University, Dept. of Biology, Durham, NC 27708, 

U.S.A. - E-mail: szoller@duke.edu 

To test the hypothesis of a low rate of genetic change in the 

inhabitant of mutualistic symbiotic systems, we 

investigated substitution rates in the nuclear ribosomal 

spacer region of the Omphalina/Coccomyxa lichens. 

Thallus fragments of six lichenized Omphalina species 

were collected in Greenland, Iceland, and in Eastern 

Canada. The average number of substitutions per 100 sites 

in the Omphalina ITS1 portion was 25.7, 2.8 in the 5.8S 

portion, and 25.5 in the ITS2 portion. The corresponding 

substitution rates for Coccomyxa were 1.6 (ITS1), 0.8 

(5.8S) and 1.0 (ITS2). On average, rates in ITS1 were 23.4 

times higher in Omphalina compared to Coccomyxa, 3.6 

times higher for 5.8S, and 29.9 times higher in ITS2. This 

finding is in accordance with the hypothesis that the lichen 

inhabitants have lower rates of genetic change when 

compared to their exhabitants. A comparison of rates 

between lichenized and free living sister species revealed 

that the rate differences observed between the mycobiont 

and photobiont is explained in part by an accelerated rate in 

the lichenized fungi. No shifts in rates were detected 

between free living and lichenized algae. Based on the 

extremely low variation among Coccomyxa ITS sequences 

and on morphological evidence (Friedl, pers. comm.), we 

believe that the Coccomyxa isolates belong to a single 

species. 

327 - Copper toxicity in Saccharomyces cerevisiae 

S.V. Avery 

University of Nottingham, School of Life and 

Environmental Sciences, University Park, Nottingham NG7 

2RD, U.K. - E-mail: Simon.Avery@nottingham.ac.uk 

Copper is an essential metal, but can also exert toxicity. 

One proposed mechanism of Cu toxicity arises from its 

capacity to promote free radical generation. Several Cu 

resistance mechanisms have been described in 

Saccharomyces cerevisiae, including metallothionein 

(Cup1) and Cu,Zn superoxide dismutase (Sod1). These 

products confer resistance by diminishing free Cu in cells, 

though they also have antioxidant activity. To examine the 

roles of such factors in the Cu resistance of growing cells, 

we asked the question: when only ^50% of cells in a 

genetically-homogeneous yeast culture are killed by Cu, 

what dictates which cells die or survive? Such phenotypic 

heterogeneity is well known, but is poorly understood even 

though it can be critical for the fitness of organisms. Using 

flow cytometry to sort Cu-sensitive/-resistant cells within 

isogenic cultures, it was shown that heterogeneity was 

partly dependent on cell cycle stage. Other potential 

contributory factors include cell age, cell size, rhythms and 

stochastic variation. The actions of various potential Cu 

resistance genes in heterogeneous Cu sensitivity were 

examined. One antioxidant gene was identified that was 

responsible for cell cycle-dependent Cu resistance. The 

same gene may also underpin heterogeneity imparted by 

cell age, cell size etc. The study highlights the specific 

factors that can account for the variable Cu resistances of 

cells in non-manipulated natural yeast populations. 



328 - Fungal influence on metal mobility: mechanisms 

and relevance to environment and biotechnology 

G.M. Gadd * , E.P. Burford, M. Fomina, F.A. Harper & H. 

Jacobs 

Division of Environmental and Applied Biology, Biological 

Sciences Institute, School of Life Sciences, University of 

Dundee, Dundee, DD4 1HN, Scotland, U.K. - E-mail: 

g.m.gadd@dundee.ac.uk 

In the terrestrial environment, fungi are of importance as 

decomposer organisms and plant symbionts (mycorrhizas), 

playing important roles in carbon and many other 

biogeochemical cycles. For example, the ability of fungi to 

solubilize insoluble metal phosphates is important for plant 

and microbial nutrition. Metals and their derivatives can 

interact with fungi in various ways depending on the metal 

species, organism and environment, while fungal metabolic 

activities can also influence speciation and mobility. 

Certain mechanisms may mobilize metals into forms 

available for cellular uptake and leaching from the system, 

e.g. complexation with organic acids, other metabolites and 

siderophores. Metals may also be immobilized by, e.g. 

sorption onto cell components, exopolymers, transport and 

intra- and extracellular sequestration or precipitation. The 

relative importance of such apparently opposing 

phenomena of solubilization and immobilization are key 

components of biogeochemical cycles for toxic metals, 

whether indigenous or introduced into a given location, and 

fundamental determinants of fungal growth, physiology 

and morphogenesis. Furthermore, several processes are of 

relevance to environmental bioremediation. This 

contribution seeks to highlight selected physico-chemical 

and biochemical mechanisms by which fungi can interact 

with and transform toxic metal species between soluble and 

insoluble forms, and to draw attention to the environmental 

significance of these processes. 

329 - Molecular mechanisms induced upon cadmium 

exposure in the ectomycorrhizal fungus Paxillus 

involutus 

M. Courbot 1* , C. Jacob 1 , P. Leroy 2 , A. Brun 1 & M. Chalot 3 

1 Université Henri Poincaré Nancy I, Faculté des Sciences 

et Techniques, UMR INRA-UHP 1136 'Interactions 

Arbres/Micro-organismes', BP 239, 54506 Vandoeuvre-les- 

Nancy Cedex, France. - 2 Laboratoire 'Thiols et fonctions 

cellulaires' - Faculté de Pharmacie, BP 403, 54001 Nancy 

Cedex, France. - 3 Centre INRA-Nancy, UMR INRA-UHP 

1136 'Interactions Arbres/Micro-organismes', 54280 

Champenoux, France. - E-mail: mikael.courbot@scbioluhp.nancy.fr 

The response of mycorrhizal fungi to toxic metals is of 

importance in view of their interest in the reclamation of 

polluted sites and their importance in tree growth. 

However, the molecular mechanisms underlying their 

response towards metals remain poorly understood. We 

104 


have used the suppression substractive hybridization (SSH) 

procedure combined to cDNA array hybridization, as well 

as direct cloning to isolate genes which expression is 

modulated by cadmium (Cd) in Paxillus involutus (Pi). 

More particularly, we are investigating antioxidant and 

detoxification enzymes such as those involved in free 

oxygen radical scavenging, in glutathione and chelating 

agents biosynthesis. The increasing activity of the Pi 

superoxide dismutase as well as the fact that it can 

functionally substitute for the E. coli SODs under Cd stress 

suggests that this enzyme is involved in the cellular 

response of Pi to Cd. The transcript levels of gglutamylcysteine 

synthetase, GSH synthetase, 

metallothioneins, thioredoxins and tyrosinase were 

measured. Of these, metallothioneins and tyrosinase were 

shown to be highly regulated by Cd. The intracellular pools 

of amino acids, glutathione, gamma-glutamate-cysteine and 

different phytochelatins in Pi were also subjected to 

changes under Cd exposure. The isolation of genes and 

proteins involved in the response to cadmium-induced 

stress open new perspectives in the understanding of 

molecular mechanisms that promote tolerance in 

mycorrhizal fungi. 

330 - Interactions of ericoid mycorrhizal and soil fungi 

with heavy metals: implications for tolerance and 

bioremediation 

E. Martino 1* , S. Cerminara 1 , P. Bonfante 2 & S. Perotto 1 

1 Dipartimento di Biologia Vegetale, Università di Torino, 

Viale Mattioli 25, 10125, Torino, Italy. - 2 Istituto per la 

Protezione delle Piante del CNR, Sezione di Torino, Viale 

Mattioli 25, 10125, Torino, Italy. - E-mail: 

elena.martino@unito.it 

Ericoid mycorrhizal fungi increase the ability of their host 

plants to colonize soils polluted with toxic metals, although 

the underlying mechanisms are unclear. We investigated 

the molecular and cellular responses of ericoid mycorrhizal 

fungi to heavy metals, and found a novel range of 

extracellular proteins being produced in the presence of 

different soluble metal compounds. Some of these proteins 

were identified by N-terminus sequencing as antioxidant 

enzymes (e.g. superoxide dismutase, SOD). SOD enzymes 

play a pivotal role in stress responses and are involved in 

metal tolerance. We also found that ericoid mycorrhizal 

fungi can solubilize insoluble metal compounds thanks to 

the release of organic acids. Solubilization activity was not 

the same for all the strains tested, with isolates from 

unpolluted soils being more efficient in the solubilization 

process than those from metal polluted soils. We are also 

exploring the potential of ericoid mycorrhizal and other 

soil fungi to mobilize iron from a very specific insoluble 

substrate: asbestos fibers. These fibers contain variable 

amounts of iron, one of the most relevant chemical features 

in the mechanism of asbestos carcinogenicity. Iron 

extraction from asbestos may thus decrease their cytotoxic 

potential. Our results suggest that some fungal species can 

remove significant amounts of iron from the fibers, thus 

opening new perspectives for the decontamination of 

asbestos-contaminated sites.


331 - A zinc tolerant ectomycorrhizal fungus protects 

zinc-stressed pine seedlings 

K. Adriaensen * , J. Vangronsveld & J.V. Colpaert 

Limburgs Universitair Centrum, Laboratory of 

Environmental Biology, Universitaire campus, 3590 

Diepenbeek, Belgium. - E-mail: 

kristin.adriaensen@luc.ac.be 

The effect of sublethal zinc concentrations on pines 

mycorrhizal with either a Zn-adapted or a Zn-sensitive 

Suillus bovinus isolate was studied. A dose response 

experiment was performed. During the zinc treatments the 

nutrient uptake capacity (NH4 + and P i) was measured in a 

semi-hydroponic environment. The Zn sensitive plantfungus 

combination already failed to sustain nutrient 

acquisition of the pines at 38 µM Zn. While nutrient uptake 

in plants inoculated with the Zn tolerant S. bovinus was not 

affected when exposed to the same Zn levels. Subsequently 

plants mycorrhizal with the tolerant isolate captured more 

Pi and were less iron deficient as plants mycorrhizal with 

the sensitive fungus. Plant biomass was not yet affected, 

but excess Zn significantly reduced the biomass of the Zn 

sensitive fungus. These results show that the genetic 

adaptation for increased Zn tolerance in Suillus bovinus is 

required for its own survival and to maintain nutrient 

acquisition in pines exposed to high Zn concentrations. 

332 - Arbuscular mycorrhizal fungi and stress by heavy 

metals and salts 

H. Bothe * , U. Hildebrandt & F. Ouziad 

Botanical Institute, University of Cologne, D-50923 Koeln, 

Germany. - E-mail: hermann.bothe@uni-koeln.de 

Plants from heavy metal soils can be colonized by 

arbuscular mycorrhizal fungi. A Glomus intraradices 

isolate has been obtained from the roots of the zinc violet, 

Viola calaminaria which confers heavy metal tolerances to 

diverse plant in diverse heavy metal soils. The genes which 

are differentially expressed in AMF colonized tomato roots 

are currently studied in our laboratory. The genus Thlaspi 

(pennycress) of the Brassicaceae contains heavy metal 

hyperaccumulating species. Some of them have now been 

found to be strongly colonized by AMF. The fungi 

colonizing Thlaspi in Nature have now been identified. 

Many plants of salt marshes are also good mycorrhizal 

plants. Up to 80% of all spores isolated from diverse saline 

habitats in Germany and Hungary belong to one single 

species, Glomus geosporum, as shown by molecular 

characterization. The role of this fungus in conferring salt 

resistance to plants will be discussed. 

333 - Gene order evolution and paleopolyploidy in 

hemiascomycete yeasts 

S. Wong 1 , G. Butler 2 & K.H. Wolfe 1* 

1 University of Dublin - Trinity College, Dept. of Genetics, 

Dublin 2, Ireland. - 2 University College Dublin, Dept. of 

Biochemistry, Dublin 4, Ireland. - E-mail: khwolfe@tcd.ie 

The wealth of comparative genomics data from yeast 

species allows the molecular evolution of these eukaryotes 

to be studied in great detail. We used 'proximity plots' to 

visualize gene order information from 14 

hemiascomycetes, including the recent Génolevures 

survey, to Saccharomyces cerevisiae. Contrary to the 

original reports we find that the Génolevures data strongly 

support the hypothesis that S. cerevisiae is a degenerate 

polyploid. Using gene order information alone 70% of the 

S. cerevisiae genome can be mapped into 'sister' regions 

that tile together with almost no overlap. This map 

confirms and extends the map of sister regions that we 

constructed previously using duplicated genes, an 

independent source of information. Combining gene order 

and gene duplication data assigns essentially the whole 

genome into sister regions, the largest gap being only 36 

genes long. The 16 centromere regions of S. cerevisiae 

form eight pairs, indicating that an ancestor with eight 

chromosomes underwent complete doubling; alternatives 

such as segmental duplications can be ruled out. Gene 

arrangements in Kluyveromyces lactis and four other 

species agree quantitatively with what would be expected if 

they diverged from S. cerevisiae before its 

polyploidization. In contrast, S. exiguus, S. servazzii and 

Candida glabrata show higher levels of gene adjacency 

conservation, and more cases of imperfect conservation, 

suggesting that they split from the S. cerevisiae lineage 

after polyploidization. 

334 - A major role for gene duplication and gene loss in 

the evolution of synteny and redundancy in yeast 

G. Fischer * , I. Lafontaine, E. Talla & B. Dujon 

Institut Pasteur, Structure and Dynamics of Genomes Dpt, 

Génétique Moléculaire des Levures, 25 rue du Dr Roux, 

75724 Paris cedex, France. 

Based on Génolevures, a recent large-scale sequencing 

program of 13 yeast species (FEBS Letter, special issue, 

487(1), 2000), a comparative sequence study showed that 

90% of the synteny breakpoints characterized between S. 

cerevisiae and S. uvarum corresponded to micro-synteny 

rearrangements. So, as little as 10% of all the synteny 

breakpoints are attributable to gross rearrangements such as 

reciprocal translocations. Micro-synteny breakpoints 

resulted from ancestral duplications of one (or few) gene(s) 

onto two different chromosomes followed by the 

differential loss of the two copies between the two 

genomes. In few cases, traces of the ancient presence of 

one duplicate in the corresponding S. cerevisiae intergenic 



regions were identified (Fischer et al., 2001, Genome Res, 

11, 2009-2019). These traces were called relics rather than 

pseudogenes because of the high number of mutations 

accumulated in these sequences that erased all 

characteristics of an ORF. A genome-wide search for relics 

in S. cerevisiae reveals the presence of at least 130 

intergenic regions showing weak homology to functional 

ORFs. The combined chromosomal localization of the 

relics, the members of the multigene families and the transchromosomal 

series identified during the Génolevures 

program reveal some overlap between the different 

duplicated regions showing that at least part of the 

redundancy in the yeast genome results from a dynamic 

equilibrium between gene duplications and gene losses. 

335 - Comparison of genomic data of the cotton 

pathogenic fungus Ashbya gossypii with Saccharomyces 

cerevisiae 

P. Philippsen 1* , F. Dietrich 1 , S. Brachat 1 , S. Voegeli 1 , A. 

Lerch 1 , K. Gates 2 & T. Gaffney 2 

1 Institute of Applied Microbiology, Biozentrum, University 

of Basel, Klingelbergstrasse 50/70, CH-4056 Basel, 

Switzerland. - 2 Syngenta, 3054 Cornwallis Road Reaserch 

Triangle Park, NC 27709, U.S.A. 

Ashbya gossypii and Saccharomyces cerevisiae are very 

different ascomycetes with respect to growth form and 

habitat but their genomes share remarkable similarities. 

The comparison at the nearly complete map of the 4800 A. 

gossypii ORFs with the map of the 6200 S. cerevisiae 

ORFs reveals the following results: 1. For 96% of the A. 

gossypii ORFs homologs are found in S. cerevisiae. 2. The 

majority of these homologs are present in S. cerevisiae as 

duplicated segments with up to 50 genes displaying relaxed 

synteny with the A. gossypii gene order. 3. Most duplicated 

S. cerevisiae ORFs are present as single copy ORFs in A. 

gossypii, e.g. RAS1/RAS2, TOR1/TOR2, MYO2/MYO4, 

CLB1/CLB2 and many others. Several of these 'ancient 

twin ORFs' code for functionally different proteins in S. 

cerevisiae and it is an open question which function is 

encoded by the single A. gossypii ORF. 4. At least 300 

functional S. cerevisiae ORFs are not present, or no longer 

present in A. gossypii. 5. Homologs for several of the S. 

cerevisiae ORFs annotated as questionable were detected at 

syntenic positions in A. gossypii. However for most of 

these ORFs no homolog was found in A. gossypii. 6. The A. 

gossypii genome contains many gene families, which are 

also, present in the S. cerevisiae genome, but often with 

fewer members. Implications of these results with respect 

to the evolution of both genomes will be discussed. 

336 - Evolution of the yeast genome 

J. Piskur 

Section for Molecular Biology, BioCentrum-DTU, 

Technical University of Denmark, Building 301, DK-2800 

Lyngby, Denmark. - E-mail: imjp@pop.dtu.dk 

106 


The genetic material has often been rearranged during the 

evolutionary history of various organisms. For example, 

gene duplications and rearrangements of the gene order 

have been particularly frequent. The most plausible way to 

deduce the molecular mechanisms, which are responsible 

for these changes, is to compare the genomes of closely 

related contemporary species. Because yeast species are 

easy to manipulate in the laboratory and the yeast genomes 

are relatively small, these organisms represent ideal models 

to understand the molecular evolution of eukaryotic 

genomes. In collaboration with laboratories from France 

and U.S., several Saccharomyces species were recently 

analysed for the structure of their mitochondrial and 

nuclear genomes. Several mitochondrial DNA molecules 

were mapped for their genes and a couple of the 

mitochondrial genomes was recently totally sequenced to 

get an idea about the origin of the present mitochondrial 

gene order. A number of nuclear genes, homologous to the 

duplicated S. cerevisiae genes, were analysed for their 

phylogenetic relationship to deduce the timing of the 

duplication, as well as the gene differentiation event(s). 

337 - PHOREST: a web based tool for comparative 

analysis of EST data 

D.G. Ahrén 1* , C. Troein 2 , T. Johansson 1 & A. Tunlid 1 

1 Microbial Ecology, Lund University, Ecology Building, 

223 62 Lund, Sweden. - 2 Theoretical Physics, Complex 

systems, Lund University, Sölvegatan 14 A, 223 62 Lund, 

Sweden. - E-mail: dag.ahren@mbioekol.lu.se 

Large-scale sequencing of cDNAs prepared from specific 

tissue or material prepared under different growth 

conditions has proven to be an efficient way for gene 

discovery. For such applications, we present PHOREST, a 

web-based tool for managing, analyzing and comparing 

various collections of expressed sequence tags (ESTs). 

PHOREST is specifically designed to support EST data 

projects and for successively receiving and incorporating 

data as project grows. After entry of sequence data a search 

for homologous sequence information in publicly available 

databases is automatically conducted, followed by 

assembly of ESTs/clones into contigs. After manual 

annotation of contigs the redundancy and distribution of 

transcripts/contigs into functional categories can be 

analyzed. Several projects/databases can be managed in 

parallel and overall assembly and normalization of data 

display transcripts/contigs being differentially regulated in 

the different projects can be compared. Access can be set 

for multiple users to search and annotate the same dataset 

without interference. The tool has been used for two EST 

projects comparing three different stages of fungal growth 

and infection. The projects are comparing stages in a 

nematode-trapping fungus, Monacrosporium haptotylum 

and a mycorrhizal fungus, Paxillus involutus.


338 - Genetic Mapping of Phytophthora cinnamomi with 

Microsatellite Fragment Length Polymorphisms 

(MFLP) 

M.P. Dobrowolski 1* , G.E.St.J. Hardy 1 , I.C. Tommerup 2 , 

B.L. Shearer 3 , I. Colquhoun 4 & P.A. O'Brien 1 

1 School of Biological Sciences and Biotechnology, 

Murdoch University, Murdoch, WA, 6150, Australia. - 

2 Forestry and Forest Products, CSIRO Perth, PO Box 5, 

Wembley, WA, 6913, Australia. - 3 Science and Information 

Division, Department of Conservation and Land 

Management, Como, WA, 6152, Australia. - 4 Alcoa World 

Alumina Australia, Environmental Department, PO Box 

252, Applecross, WA, 6153, Australia. - E-mail: 

dobrowol@murdoch.edu.au 

Phytophthora cinnamomi is a major oomycete pathogen of 

an extensive range of mainly woody plants. We are 

conducting genetic mapping of P. cinnamomi to localise 

genes involved in pathogenicity. Because we are restricted 

to analysing mapping populations of F1 hybrid progeny in 

this diploid species, choosing an informative (i.e. highly 

polymorphic) marker system for genetic mapping is 

critical. Microsatellite loci are ideal markers for this 

purpose. Their high polymorphism can be harnessed 

without prior DNA sequence information of the subject 

organism by a method called microsatellite fragment length 

polymorphism (MLFP) (1). MFLP are analysed in a very 

similar way to amplified fragment length polymorphisms 

(AFLP). In our analyses of four F1 mapping populations of 

P. cinnamomi progeny, we have detected between 1 and 4 

informative loci per primer combination. We are in the 

process of constructing a genetic map of the P. cinnamomi 

genome with this data. 1. Yang, H., Sweetingham, M. W., 

Cowling, W. A., and Smith, P. M. C. (2001) DNA 

fingerprinting based on microsatellite-anchored fragment 

length polymorphisms, and isolation of sequence-specific 

PCR markers in lupin (Lupinus angustifolius L.). 

Molecular Breeding 7:203-209. 

339 - Introduction to the theory of metabolic (modelling 

and) control. Application to the citric acid production 

by Aspergillus niger 

N.V. Torres 

Universidad de La Laguna, Dpto. Bioquímica y B.M. Avda. 

Asfco. Fco. Sánchez s/n 38206 La Laguna Tenerife Islas 

Canarias, Spain. - E-mail: ntorres@ull.es 

Analysis, control (and optimization) of biochemical 

pathways requires mathematical modelling frameworks 

that are able to integrate different types of data and to 

capture the essence of complex systems. In this talk we will 

introduce a modelling approach that provides such a 

framework. It begins with the derivation of models from 

basic concepts of metabolic modelling; introduces the 

different types of system analysis, namely the stability, 

sensitivity (or control) and dynamics analysis; illustrate the 

parameter estimation techniques and finally, apply them to 

the citric acid production by A. niger. Once the model has 

been successfully tested and fine-tuned, we are provided 

with a full, quantitative and unambiguous description of the 

pathway behaviour, including its steady state control 

structure and dynamics. This description can be used for 

tasks that are otherwise difficult or impossible to execute. 

One of such tasks is the optimization of the pathway with 

respect to criteria that are not given by nature but defined 

by human demand. But this is another history. 

340 - Carbon metabolism in Aspergillus and Penicillium 

J. Nielsen 

Center for Process Biotechnology, Technical University of 

Denmark, Building 223, DK-2800 Kgs. Lyngby, Denmark. 

- E-mail: jn@biocentrum.dtu.dk 

Filamentous fungi belonging to the genera Aspergillus and 

Penicillium are used extensively in the fermentation 

industry for production of a variety of products. These 

products fall in many different industrial sectors: 1) food 

additives, e.g. citric acid; 2) antibiotics, e.g. penicillin; 3) 

high-value pharmaceuticals, e.g. statins; and 4) industrial 

enzymes, e.g. amylases and xylanases. In the bioprocess 

industry there is a trend towards applying specific 

microbial strains for production of many different products. 

Hereby optimisation of the function of these specific 

strains can be harnessed for the production of many 

different products. These plug-bugs include A. oryzae and 

A. niger, which are both used extensively for production of 

industrial enzymes, where a strong and tailor made 

promoter structure is used to drive the production of the 

enzyme of interest. Recently it has also been demonstrated 

that P. chrysogenum can be used as plug-bug for 

production of different b-lactams, e.g. penicillins and 

adipoyl-7-ADCA. With the development of plug-bugs it is 

of significant industrial interest to obtain fundamental 

insight into the carbon metabolism of these organisms, as 

well as to engineer the strains in order to redirect the 

carbon fluxes towards the product of interest. In this 

presentation several different techniques for analysis of the 

central carbon metabolism in Aspergillus and Penicillium 

will be presented. 

341 - Regulation of catabolic fluxes and the energetics 

of adaptive responses in Saccharomyces cerevisiae 

F.I.C. Mensonides 1 , G.P.M.A. Hardy 2 , H.F. Tabak 2 , J. 

Blom 3 & M.J. Teixeira de Mattos 1* 

1 Swammerdam Institute for Life Sciences, Dept Microbiol., 

University of Amsterdam, Nieuwe Achtergracht 166, 1018 

WV Amsterdam, The Netherlands. - 2 Laboratory of 

Biochemistry, Academic Medical Center, University of 

Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The 

Netherlands. - 3 Swammerdam Institute for Life Sciences, 

Micro-Array Division, University of Amsterdam, Kruislaan 



308, 1098 SM Amsterdam, The Netherlands. - E-mail: 

teixeira@science.uva.nl 

The metabolic network of the living cell is endowed with 

an amazing capacity to cope with different environmental 

conditions. This capacity is due to a variety of signal 

transducing and regulatory systems designed to tune the 

cell's metabolic machinery to prevailing physico-chemical 

conditions e.g. nutrient availability, temperature, 

osmolarity and the nature of the energy source. Although 

the mechanisms that underly various adaptive responses 

may differ in many respects, they seem to have in common 

that the energetic and redox state of the cell is crucial to 

their functioning on the one hand and that they affect 

energy and redox catabolism on the other. In this context, it 

will be shown that the extent to which exponentially 

growing cells can withstand a heat shock is related to the 

ATP pools and to their potential to adapt the internal 

trehalose and glycogen metabolism. Further, a quantitative 

flux analysis of strains modified in the levels of proteins 

(hap4p and hexokinase 2) that take part in general glucose 

repression regulatory pathways, will illustrate the role of 

these pathways in the bioenergetics of yeast growth and 

maintenance. Finally, we will present an example of the 

transient transcriptional and physiological events which are 

invoked when steady state glucose-limited chemostat 

cultures are challenged with oleate as energy source. Here 

again, the initial adaptive response (prior to peroxisome 

biosynthesis) is gouverned by redox and energetic 

imbalance. 

342 - Uptake and catabolism of glucose 

M. Mattey 

University of Strathclyde, 204, George Street, Glasgow G1 

1XW, Scotland, U.K. - E-mail: m.mattey@strath.ac.uk 

Fungi are chemoheterotrophs found in a variety of habitats 

and their life cycles are similarly varied. Strategies adopted 

by fungi vary from typical 'r' strategists, with a high 

reproductive rate and a short life cycle, to 'K-'strategists 

which use resources efficiently and reproduce more slowly. 

Whatever their environmental niche most fungi can use 

glucose and do so with the same basic biochemistry. The 

uptake of glucose uses members of the major facilitator 

superfamily which mediate the thermodynamically 

downhill movement of glucose across the cell membrane. 

Free glucose within the cell is maintained at a low level by 

hexokinase, which also confines the subsequent 

metabolism to the cell by phosphorylation. Glycolysis is 

the most common pathway for the degradation of glucose. 

Although the basic biochemistry is the same in most cases, 

the regulation of the enzymes differs for different 

strategies. Faced with a surfeit of glucose, a situation that 

may occur naturally or in industrial processes, fungi grow 

rapidly, but this may not be enough to use all the glucose 

that enters the cell by simple diffusion. The rate of 

diffusion can exceed the maximum uptake rate of 

facilitated transport. When this happens facilitated 

transport appears to shut down by an unknown mechanism, 

and free internal glucose levels rise. In the opposite 

situation, with oligotrophic media, the problem becomes 

108 


one of how to maintain internal metabolite levels when the 

entry rate is less than the flux. 

343 - Galactose metabolism in Trichoderma and 

Aspergillus: properties, regulation and identification of 

a second, reductive pathway 

B. Seiboth 1 , L. Karaffa 2 & C.P. Kubicek 1* 

1 Section Microbial Biochemistry and Gene Technology, 

Institute of Chemical Engineering, TU Wien, 

Getreidemarkt 9/166, A-1060 Wien, Austria. - 2 Department 

of Microbiology and Biotechnology, Faculty of Sciences, 

University of Debrecen, H-4010, P.O.Box 63, Debrecen, 

Hungary. - E-mail: ckubicek@mail.zserv.tuwien.ac.at 

D-Galactose metabolism via the Leloir pathway is a 

ubiquitous trait in pro- and eukaryotic cells. Its metabolic 

regulation has been extensively studied in yeast 

(Saccharomyces, Kluyveromyces) but not in filamentous 

fungi. A more detailed knowledge on the latter would be 

worthwhile, as lactose (1,4-0-β-D-galactopyranosyl-Dglucose) 

arising from whey - represents a renewable carbon 

source for several fungal fermentations, notably cellulase 

production by Hypocrea jecorina (anamorph Trichoderma 

reesei), but an effective exploitation of lactose for the 

biotechnical utlization is still hampered by its slow 

metabolism and a lack of basic knowledge on its utilization 

in H. jecorina and filamentous fungi in general. We 

therefore have functionally characterized the genes for the 

Leloir pathway in H. jecorina. Here we will report that 

they differ from the yeast counterparts with respect to 

genomic organization, protein structure and genetic 

regulation. We will also provide evidence for the 

involvement of the Leloir pathway in cellulase induction 

by lactose. Further, we will provide evidence for the 

existence of a second, so far unknown pathway of Dgalactose 

breakdown in H. jecorina and Aspergillus 

nidulans which involves a NADPH-aldose reductase 

dependent reduction to D-dulcitol, and its subsequent 

oxidation by 2-arabinitol dehydrogenase to D-tagatose.

IMC7 Friday August 16th Lectures 

344 - The practicalities of integrating anamorph and 

teleomorph taxonomies 

G.J. Samuels 1 & M. Reblova 2* 

1 U.S. Dept. of Agriculture, Systematic Botany and 

Mycology Lab., 10300 Baltimore Ave., Beltsville, MD 

20705-2530, U.S.A. - 2 Institute of Botany, Academy of 

Sciences, CZ - 25243 Pruhonice, Czech Republic. - E-mail: 

reblova@ibot.cas.cz 

Article 59 of the International Code of Botanical 

Nomenclature enshrines a system that biases naming of 

pleomorphic fungi in favor of the teleomorph. The system 

maintains artificial separation of teleomorph and anamorph 

taxonomy resulting in loss of information and confusion in 

understanding species. DNA sequencing has eroded the 

significance of the teleomorph to taxonomy by integrating 

species and genera based on anamorph characters into the 

'botanical' system. That all fungi may undergo outcrossing 

of some sort has further eroded the importance of the 

sexual morph in the life-cycle. Three scenarios are 

anticipated if Art. 59 is dropped. I. Retain primacy of the 

teleomorph but permit description of species for which no 

teleomorph is known as botanical species; permit inclusion 

of genera for which no teleomorph is known as botanical 

genera. In case of priority conflict, the oldest type based 

upon teleomorph material will determine the correct name. 

II. Permit names based on teleomorph material compete 

with names based on anamorph material strictly on the 

basis of priority. III. Selectively give priority to teleomorph 

or anamorph names based on defined criteria (e.g. common 

usage). The advantages and disadvantages of the scenarios 

are discussed. The authors conclude that despite its 

shortcomings, the current system is so firmly established 

that change based on any of the three scenarios presented 

would result in an unacceptably high level of disruption. 

345 - Anamorphs, teleomorphs and users of names 

M.E. Palm 1* & P.M. Kirk 2 

1 USDA/APHIS, Systematic Botany and Mycology 

Laboratory, Beltsville, Maryland 20705-2350, U.S.A. - 

2 CABI Bioscience, Bakeham Lane, Egham, Surrey TW20 

9TY, U.K. - E-mail: mary@nt.ars-grin.gov 

The eventual abandonment of Article 59 is inevitable. The 

separate naming of anamorphs has provided a practical 

solution in some groups of fungi to the challenge of 

pleomorphism. Phylogenetic placement of anamorph taxa 

increasingly is possible as a result of data from molecular 

phylogenetic studies combined with morphological 

approaches. Systematists can now craft an alternative to the 

dual naming system, one which should be the least 

disruptive to communication and information retrieval. A 

practical strategy would be to consider groups on a case by 

case basis as monographic studies using morphological and 

molecular approaches are conducted. Harmonization and 

revision of the Code should be a gradual process in order to 

avoid total chaos and in order to learn of the pitfalls and 

problems that could not be predicted, as they come to light. 

Several case studies will be presented along with an 

analysis of nomenclatural options. 

346 - Dual nomenclature and classification in higher 

fungi: Six procedures to resolve it, with proposals to 

emend Art. 59 

G.L. Hennebert 

University of Louvain (professor emeritus), 32 Rue de 

l'Elevage, 1340 Ottignies-LLN, Belgium. - E-mail: 

hennebert@mbla.ucl.ac.be 

Since the symposium, 'The Fungal Holomorph', the 

Deuteromycetes are considered group of form-taxa, the 

deuteromycetes (decapitalized), rather than a (capitalized) 

fungal Class. Integrated classification implies unification of 

the two parallel nomenclatures. The fundamentals of dual 

nomenclature lie in the distinction of Linnean or botanical 

nomenclature based on botanical (holomorphic) 

typification and anatomical nomenclature based on 

anatomical (anamorphic and teleomorphic) typification. 

Article 59 assigns holomorphic application to teleomorphic 

types, but denies it to anamorphic types, which are 

restricted to anamorphic application. The nature of the type 

of a name is thus distinguished from its application. Any 

process for nomenclature integration must envisage a 

change of anamorphic to holomorphic type applications, ie. 

the change of form-names into botanical names, and the 

suppression of alternate names. This can be achieved using 

six different procedures, from conservative to 

revolutionary, depending on the extent of the changes in 

type application, extent of retroactivity, the taxonomic 

ranks affected, and the choice of preserved name amongst 

alternate names of pleomorphic and pleoanamorphic fungi. 

Preservation and suppression of names, already implicit in 

Art. 15.2, cannot be processed by conservation and 

rejection, because they must be revisable when the organic 

connections between correlated names are disproven. See 

Hennebert & Gams on http://www.cbs.knaw.nl/ for full 

text. 

347 - The history of Article 59 and anamorph 

definitions - and a new twist: synteleomorphic names 

S.A. Redhead 

ECORC, Agriculture & Agri-Food Canada, Bldg. 49, CEF, 

AAFC, Ottawa, Ontario K1A 0C6, Canada. - E-mail: 

redheads@em.agr.ca 

The Linnean system of classification dates back to 1753 

(Species plantarum). Less obvious to young scientists is 

knowledge that the starting dates for fungal systematics 

used to be 1753, 1801 (Persoon's Synopsis), or 1821 (Fries' 

Systema) depending upon fungal groups. The latter two 

authors' named publications currently serve as Sanctioning 



works. Lost in time are when the Codes of nomenclature 

were written, when pleomorphic fungi were allowed 

multiple names (i.e. Art. 49bis in 1910), or why? Murkier 

are the dates when it was commonly accepted that fungi 

had sexual and asexual morphs, or when teleomorphs of 

Basidiomycota could be distinguished from Ascomycota or 

from anamorphs. When were lichens and Zygomycota 

added as exceptions to the article on pleomorphic fungi, 

and why? While DNA sequence-based technology paves 

the way for merging anamorph typified names with 

teleomorph typified names, another nomenclatural and 

taxonomic issue emerges, that of merged taxa typified by 

grossly dissimilar teleomorphs (e.g. false truffles within 

agaric or bolete genera and visa versa). At the generic 

level, the nomenclatural problems presented by anamorph- 

versus teleomorph-typified names mirrors the problems of 

teleomorph versus teleomorph name conflicts that now 

spawn ideologies leading to creation of a PhyloCode. 

Future changes to the Botanical Code should consider these 

two conflicts jointly and not separately because they are the 

results of data generated by the same technology. 

348 - Consequences of deletion of Article 59 for 

economically important fungi 

P.F. Cannon 1* & K.A. Seifert 2 

1 CABI Bioscience, Bakeham Lane, Egham, Surrey TW20 

9TY, U.K. - 2 Eastern Cereal and Oilseed Research Centre, 

Agriculture & Agri-Food Canada, 960 Carling Avenue, 

Ottawa, Ontario K1A 0C6, Canada. - E-mail: 

p.cannon@cabi.org 

Modification of the wording of Art. 59 has been the 

preferred avenue of change for governing the application of 

anamorph names. We will examine the consequences of 

complete deletion of this article from the ICBN, and the 

consequent loss of sanctioned status for teleomorph-based 

holomorph names at all taxonomic ranks. In this scenario, 

the oldest name of a species would be available for the 

actual name in combination with the oldest generic name, 

in line with priority, irrespective of what morphs were 

described in the protologue. We consider this essential for 

many groups of economically important fungi, where the 

number of described anamorph taxa often greatly exceeds 

the number of teleomorphic-holomorphs. For example, the 

ratio of anamorph:teleomorph species is about 10:1 in the 

Glomerellaceae, 5:1 in the Pleosporaceae, 3:1 in the 

Trichocomaceae, 2:1 in the Chaetosphaeriaceae and 1.5:1 

in the Hypocreales, Tubeufiaceae, and Microascaceae. This 

approach will result in far fewer changes in binomials than 

the alternative of retaining teleomorph priority. These 

changes will have negligable effects on the names of 

families or higher taxa. Since 969 of 1361 genera of 

hyphomycetes and 196 of 602 genera of coelomycetes 

currently lack known teleomorph connections, deletion of 

Art. 59 would protect the status of these names even if 

teleomorphs are eventually discovered. 

110 


349 - Has dual nomenclature for fungi run its course? 

The Article 59 debate. Arguments for retaining Article 

59 

W. Gams 

Centraalbureau voor Schimmelcultures, P.O. Box 85167 


Gams@cbs.knaw.nl 

Fungal taxonomy is mainly morphology-based. 

Understanding the morphology of a fungus is part of 

understanding its function. Many modern studies illustrate 

the indispensability of morphology. Replacing 

identification methods by molecular routines is a form of 

reductionism to be opposed. The users' needs must not be 

ignored. Permitting separate names for the morphs of 

pleomorphic higher fungi conflicts with the principle of 

'one species - one name'. No higher-rank classification of 

anamorphic fungi is recognized and integration of 

anamorph taxa in the teleomorph system is essential, but 

anamorph morphology is inadequate to do this. Retaining a 

separate anamorph nomenclature (particularly at the 

generic level) has advantages in morphological 

identification. The provisions of Art. 59 have grown 

through time to satisfy the needs of identifying 

mycologists. In many cases, the anamorph absolutely 

dominates in culture and often also in nature; it often is 

more differentiated morphologically than the teleomorph. 

Complete integration of teleomorph and anamorph 

nomenclature with all its consequences could work only if 

a 1:1 relationship of anamorph and teleomorph genera were 

possible. If the correlation between anamorph and 

teleomorph species is not 1:1, retaining both systems is of 

particular advantage. Abandoning Art. 59 immediately will 

cause nomenclatural chaos; any solution will require very 

complicated procedures (see detailed texts on CBS 

website! ). 

350 - Has dual nomenclature for fungi run its course? 

The Article 59 debate. Arguments to reject dual 

nomenclature 

D.L. Hawksworth 1* , P.M. Kirk 2 & M.L. Berbee 3 

1 Departmento de Biologia Vegetal II, Facultad de 

Farmacia, Universidad Complutense, Plaza de Ramon y 

Cajal, Ciudad Universitaria, E-20840 Madrid, Spain. - 

2 CABI Bioscience, Bakeham Ln, Egham, Surrey TW20 9TY, 

U.K. - 3 Department of Botany, University of British 

Columbia, 6270 University Blvd, Vancouver, BC V6T 1Z4, 

Canada. - E-mail: davidh@ucmail.ucm.es 

Dual nomenclature, the practise of giving separate 

scientific names to teleomorphs and anamorphs of 

pleomorphic fungi, predates the governance of fungal 

nomenclature by formal Codes. While there may be some 

practical benefits of having identification systems and 

separate names for the anamorphic fungi that are often 

collected or cultured independently of any teleomorph,


having two names confuses non-mycologists and can cause 

problems in communication. There has always been 

discomfort at the existence of an artificial taxonomic 

system for anamorphs that imperfectly parallels the 

supposedly phylogenetic holomorphic system. Calls for the 

abolition of dual nomenclature have become more frequent 

with the advent of molecular systematics, which allows all 

fungi to be included in a unified phylogeny irrespective of 

the morphology expressed. Dual nomenclature for some 

groups of fungi is governed by Article 59 of the 

International Code of Botanical Nomenclature, but this 

does not apply to all and no such provision exists for any 

other kinds of organisms. We will argue for the eventual 

rejection of formalized dual nomenclature and a major 

revision of Article 59. 

351 - Biogeography and species complexes in some 

corticioid fungi 

N. Hallenberg * & R.H. Nilsson 

Botanical Institute, Box 461, SE-405 30 Gothenburg, 

Sweden. - E-mail: nils.hallenberg@systbot.gu.se 

Species diversity has by tradition been based on a 

morphological species concept. As shown in numerous 

studies such morpho-species among corticioid 

basidiomycetes are frequently complex, consisting of 

morphologically indistinguishable biological species. 

Phylogenetic analyses based on ITS sequences support the 

biological species concept. Moreover, from such analyses it 

may be possible to estimate genetic divergence within and 

between geographical regions as well as evolutionary 

relationships between biological species. For a few species 

an extensive sampling has been done, originating from 

temperate areas around the world. From datasets in 

Hyphoderma praetermissum and Hyphoderma setigerum it 

is shown that representatives from some of the different 

biological species have very wide distribution areas at 

least. The different groups revealed in a complex are 

distinctly separated in their sequence divergence but, on the 

other hand, more closely related to each other than to 

morphologically distinguished relatives. Homothallic 

isolates group together with heterothallic ones, indicating 

that homothallism must be looked upon as a life strategy 

which repeatedly have come into existence within certain 

populations. 

352 - Genetic and molecular evidence for vicariance 

events in the hymenomycetes 

K.W. Hughes 1* , R.H. Petersen 1 & S.A. Redhead 2 

1 Dept. of Botany, University of Tennessee, Knoxville, TN, 

USA 37996-1100, U.S.A. - 2 Agriculture and Agri-Food 

Canada, Ottawa ON K1A 0C6, Canada. - E-mail: 

khughes@utk.edu 

Many Northern Hemisphere basidiomycete fungi have 

intercontinental distributions (North America, Europe and 

Asia). For the taxa we have studied, most collections retain 

the ability to interbreed in vitro throughout their 

geographical range (e.g., Auriscalpium vulgare, 

Flammulina velutipes, F. rossica, Panellus stypticus, 

Lentinellus castoreus, Clavicorona pyxidata, Collybia 

cookei, C. cirrhata and C. tuberosa). We have examined 

the genetic relationships among collections of selected taxa 

from widely separated geographical regions based on 

ribosomal ITS DNA sequences. For Panellus stypticus, 

Flammulina velutipes and F. rossica, the data suggest that 

western NA and Asian populations are most closely 

related, perhaps via Beringia. Eastern NA populations of 

these species are clearly distinct but their origins are 

obscure, perhaps derived from western NA populations, 

perhaps from Europe. For Clavicorona pyxidata, the data 

suggest that collections from eastern NA and Europe are 

most closely related, possibly through a North Atlantic land 

bridge open during the Eocene. For species with both 

Northern and Southern Hemisphere distributions, migration 

may have occurred via the Malaysian Archipelago 

(Panellus stypticus) or via South and Central America 

(possibly Lentinellus castoreus). The finding of distinct but 

contaxic fungal populations across broad geographical 

areas suggests that fungal species may be very old. 

353 - Natural biogeography and movement by humans 

of host-specialized forms in the Ceratocystis fimbriata 

complex 

T.C. Harrington * & C.J. Baker 

Iowa State University, Department of Plant Pathology, 

Ames, 50011, Iowa, U.S.A. - E-mail: tcharrin@iastate.edu 

There is little morphological diversity but a wide range of 

hosts in the wilt pathogen Ceratocystis fimbriata. 

Sequences of rDNA and the MAT-2 gene have shown a 

surprising amount of variation among more than 200 

isolates studied. Phylogenetic analyses group all isolates 

into three major geographic clades, each comprised of hostspecialized 

lineages. The center of diversity of the complex 

is found in the USA, and the North American clade (NAC) 

includes lineages on Prunus, Quercus, Populus and Carya. 

The Asian clade (AC) includes a Ficus lineage and a 

Colocasia lineage. The Latin American clade (LAC) 

appears indigenous from southeastern USA to South 

America. The LAC has a wide range of hosts, many of 

which are exotic crop plants, but it is less genetically 

diverse and appears younger than either the AC or NAC. 

Host specialization was demonstrated in LAC populations 

on Ipomoea, Platanus, the family Araceae, Coffea, 

Theobroma, Gmelina, Eucalyptus, and Mangifera. The 

LAC appears to be comprised of relatively small and 

geographically isolated populations in the process of hostspecialization 

and speciation. Genetic isolation of LAC 

populations may be due to limited geographic dispersal by 

insects. However, there is genetic and historical evidence 

for dissemination by humans of C. fimbriata strains on 

plant materials: Colocasia (AC) to Hawaii, Populus (NAC) 

to Poland, and LAC forms on Ipomoea to Asia, Syngonium 

to Hawaii, Platanus to Europe, and Theobroma to Brazil. 



354 - Disjunct distributions of some tropical agarics and 

boletes 

R.E. Halling 

Institute of Systematic Botany, New York Botanical 

Garden, Bronx, NY 10458-5126, U.S.A. - E-mail: 

rhalling@nybg.org 

The agarics and boletes in Neotropical forests exhibit 

several different patterns of origin and distribution. The 

patterns and diversity are directly related to the types of 

habitats in which these fungi occur and have been 

influenced by geological history, native and introduced 

phanerogamic vegetation, climate and, ultimately, human 

impact. Specifically, the aforementioned factors affect the 

distribution of those agarics and boletes that occur 

naturally in the montane oak forests of Latin America. 

Documentation and personal observations of 

ectomycorrhizal associations and distributions during the 

last two decades have revealed some distinctive patterns: 

(1) relictual disjunct distributions; (2) generic and specific 

distributions along a cline; (3) local endemism; (4) high 

generic similarity; (5) low species similarity; and (6) high 

levels of macromycete diversity. 

355 - Biogeography and possible origin of Armillaria 

species 

M.P.A. Coetzee 1* , B.D. Wingfield 1 , P. Bloomer 2 & M.J. 

Wingfield 1 

1 Department of Genetics, Forestry and Agricultural 

Biotechnology Institute (FABI), University of Pretoria, 

Pretoria, 0002, South Africa. - 2 Department of Genetics. 

University of Pretoria, Pretoria, 0002, South Africa. - Email: 

martin.coetzee@fabi.up.ac.za 

Armillaria (Fr:Fr.) Staude is a globally distributed plant 

pathogenic basidiomycete that causes root rot on a variety 

of hosts. The phylogeny of Armillaria species from the 

Northern Hemisphere and Australasia has been relatively 

well studied in recent years. However, very little is known 

regarding the phylogenetic relationships between species 

from the Northern and Southern Hemispheres. In a series 

of studies, we have attempted to elucidate these 

relationships and to develop a hypothesis regarding the 

origin of Armillaria species. Isolates that we have studied 

included those from Africa, Asia, Australia, New Zealand, 

South America, North America and Europe. Phylogenetic 

analyses were based on distance analysis using DNA 

sequences from the large subunit (LSU) ribosomal RNA 

operon. Resulting phylogenetic trees separate species from 

the Southern and the Northern Hemisphere into two 

strongly supported monophyletic clades. Armillaria species 

that grouped within the Southern Hemisphere clade showed 

a higher interspecific diversity than the Northern 

Hemisphere species. Results suggest that the Southern 

Hemisphere Armillaria group is older than its Northern 

112 


counterpart. Furthermore, our results lead us to believe that 

Armillaria probably has a Gondwanean origin. 

356 - Phylogeography of the widely disjunct lichen 

Cavernularia hultenii 

C. Printzen 1* , S. Ekman 1 & T. Tønsberg 2 

1 Botanisk Institutt, Universitetet i Bergen, Allégaten 41, 

5018 Bergen, Norway. - 2 Botanisk Museum, Universitetet i 

Bergen, Allégaten 41, 5018 Bergen, Norway. - E-mail: 

christian.printzen@bot.uib.no 

Up to now, almost nothing is known about the population 

structure and history of lichenized ascomycetes. The largescale 

infraspecific disjunctions displayed by many species 

have alternatively been explained by high evolutionary age 

or exceptional dispersal abilities of the species. In order to 

infer pleistocene and holocene population history of the 

widely disjunct lichen Cavernularia hultenii, we sequenced 

the ITS and part of the IGS region of 300 samples from 62 

populations across the species' range. While four ancestral 

haplotypes are found in all areas, none of the observed 

descendant haplotypes is present in more than one of the 

part ranges. Although this is evidence for a past 

fragmentation event, nested clade analysis fails to 

unambiguously infer allopatric fragmentation due to the 

shallow gene genealogy and widespread ancestral 

haplotypes. Long-distance dispersal is not inferred either. 

Mismatch distributions yield evidence for exponential 

population growth, probably in connection with postglacial 

invasion of C. hultenii into formerly glaciated areas of 

Western North America. The presence of a southern and at 

least one northern glacial refugium in south Central Alaska 

is inferred from the data. Nested clade analysis does not 

infer range expansion from our dataset either. This and the 

failure to infer fragmentation is explained by slow action of 

genetic drift, which has not completely removed ancestral 

haplotypes from the post-fragmentation and post-expansion 

areas. 

357 - Biogeography of mushrooms: Finding Wallace's 

line for the fungi 

R. Vilgalys * , J.-M. Moncalvo & T.Y. James 

Dept. of Biology, Duke University, Durham, NC 27707, 

U.S.A. - E-mail: fungi@duke.edu 

The combined study of phylogeny and biogeography 

provides a framework for understanding the relationship 

among different components of speciation, including 

population genetic structure, isolation mechanisms, 

vicariance, morphological evolution, and ecological 

adaption. Examples of fungal groups with rich 

phylogeographic histories include Pleurotus, Ganoderma, 

Schizophyllum, and Xeromphalina. In these groups, 

patterns of genetic variation among geographically isolated 

populations indicate that speciation is primarily allopatric


in mode. However, gene flow between widely isolated 

locations is also evident, and repeated dispersal to distant 

provenances may provide opportunities for 

biodiversification through repeated rounds of allopatric 

speciation. Older lineages may also be more likely to have 

broader distributions than recently evolved ones. This 

presentation will review recent advances in understanding 

diversification in fungi through phylogeographic studies. 

358 - In vitro fruiting of Cordyceps militaris 

J.M. Sung * & B. Shrestha 

Kangwon National University, Hyoja 2 Dong, Chuncheon, 

200-701, Korea. - E-mail: jmsung@kangwon.ac.kr 

Every year, 200-400 specimens of C. militaris are collected 

mainly from Kangwon Province of Korea and their isolates 

preserved in EFCC, KNU, Korea. Cordyceps species are 

highly regarded as medicinal mushrooms in East Asia, 

including Korea. Stromata of C. militaris could be 

produced in vitro when its liquid suspensions were 

inoculated in brown rice medium supplemented with 

silkworm pupae and incubated in 20-25 °C under light and 

high humidity conditions. But, the isolates showed unstable 

variation in fruiting. Most of the isolates failed to produce 

stromata or produced only few deformed stromata. Other 

isolates, which produced good fruiting, could not produce 

the same quality of fruiting when their subcultures were 

used, while other few isolates produced good fruiting when 

their subcultures were used. This was the main problem in 

commercial cultivation of C. militaris. In order to 

understand the fruiting character of C. militaris, several 

single ascospore strains were crossed with each other and 

observed for their fruiting. Out of eighteen strains, ten 

strains produced stromata profusely with the remaining 

eight strains in all the combinations, but not among 

themselves and vice versa. When two opposite strains were 

repeatedly inoculated together in large scale, they produced 

good quality of fruiting continuously. This paper intends to 

report that C. militaris is a bipolar heterothallic fungus and 

crossing between two opposite types is a good method of 

fruit body production. 

359 - Breeding of mushroom by using cell fusion 

techniques 

T. Morinaga 

School of Bioresources, Hiroshima Prefectural University, 

Nanatsuka 562, Shobara-city, Hiroshima Prefecture 727- 

0023, Japan. - E-mail: tmorina@bio.hiroshima-pu.ac.jp 

Twenty-two homokaryons were isolated from one 

commercial fruiting body of P. ostreatus. Then, all 

homokaryons were crossed each other. In crossing tests of 

homokaryotic mycelia, only two sexual phenomena were 

observed: clamp formation and "Barrage" reaction, in 

contrast with Schizophyllum commune (Raper 1966). We 

determined the mating type of P. ostreatus by this two 

reactions. As a result, P. ostreatus was determined to have 

a bifactorial incompatibility; KM 4 (A1B1), KM 7 (A2B2), 

KM 18 (A1B2) and KM 22 (A2B1). All compatible 

dikaryons formed their fruiting bodies on commercial 

sawdust media. By means of UV-irradiation to protoplasts 

of these homokaryons, five strains of auxotrophic mutant 

were obtained. The ratio of regeneration to mycelia from 

protoplasts of homokaryons was about 0.1%. Reversion 

rate of each mutant to prototroph was calculated from the 

number of colonies that appeared on the complete medium 

and that on the minimal medium against the number of 

protoplasts or regenerated protoplasts. And then, protoplast 

fusion between strain KM 4-1 (wet-) and KM 4-34 (ade-) 

was done in the same mating type. Fifteen strains which 

could grow on the minimal medium and possessed one 

nucleus per each cell obseved by fluorescent microscopy, 

were obtained with this fusion treatment. These strains 

were inoculated to MYA medium containing methyl - 1 - 

(butylcarbamoyl) - 2 - benzimidazole carbomate 

(benomyl). Benomyl is known as a regent of 

haploidization. Two strains (KM D-4-1 and KM D-4-2) 

occurred sector on this medium. Therefore, these strains 

were found to have one diploid nucleus per each cell. On 

the other hand, the other diploid strains having the opposite 

mating type were made by the same cell fusion technique. 

After getting the strains, crossing were done among the 

diploid strains. Finally, tetraploid mycelium were gotten. 

The growth rate of these strains became about two times of 

diloid strains and their fruiting also became faster than 

normal commercial strains. 

360 - The genus Pleurotus as a bioremediation tool 

E. Sanjust 

Cattedra di Chimica Biologica, Università di Cagliari, 

Complesso Universitario, 09042 Monserrato (CA), Italy. - 

E-mail: sanjust@unica.it 

The genus Pleurotus belongs to the white rot fungi, capable 

of rapid growth on lignocellulosics and with a preference 

for lignin. It lacks lignin peroxidase, but actively excretes 

other oxidative enzymes upon proper induction. Oxidised 

products arising from extracellular enzyme action are 

recycled by mycelium-bound reductases; enzymatic and/or 

non-enzymatic re-oxidation of such reduced molecules 

results in hydrogen peroxide production in the growth 

media. Some details of this particular mechanism have 

been recently elucidated whereas others are under study. 

Pleurotus acts as a biochemical hydrogen peroxide factory 

by means of quinone/polyphenol and benzyl 

alcohol/benzaldehyde redox cycles. The bleaching power 

of hydrogen peroxide as well as its production in 

autoxidation reactions could be enhanced by the ability of 

the fungus to raise the pH of culture media when stressed 

by xenobiotics. Hydrogen peroxide and excreted enzymes 

co-operate in the breakdown of a wide range of different 

molecules. Therefore the fungus efficiently grows on a 

variety of substrates. Chemicals that are toxic for other 

fungi are bleached by Pleurotus just out of the hyphae and 

cannot accumulate within the fungal cells. So safe and 

valuable sporocarps could be obtained when starting from 



some substrates that are unsuitable for direct use as human 

food and/or animal feed, while concomitantly destroying 

hazardous substances they eventually contain, and 

operating an efficient bioremediation. 

361 - Development of functional foods by use of 

basidiomycetal strains 

J.S. Lee 1* , H.C. Ha 1 , S.Y. Kim 1 , H.J. Kim 1 & K.B. Park 2 

1 Dept.of Bioindustry,Yeungnam University, 214-1 

Daedong,Gyungsan Gyungbuk, Korea. - 2 Haechandle Co., 

284-3 Yongmundong Seogu Daejeon, Korea. - E-mail: 

jslee@yumail.ac.kr 

In this study, it was attempted to investigate the 

hepatoprotective effects of two fractions of methanol 

extracts, larger molecular fraction(above 20,000 MW) and 

smaller molecular fraction(under 20,000 MW), obtained 

from waxy brown rice cultured with Phellinus igniarius. 

After administering these sample fractions to rats orally for 

2 consecutive weeks, rats were treated with CCL4. The 

determination of enzyme activities in sera of treated rats as 

well as biochemical analysis of liver were conducted and 

compared with those data from control group. Serum GOT 

and GPT of rats increased highly in the control group 

treated with CCL4 only. The treatment group fed with 

larger molecular fraction of the methanol extracts obtained 

from waxy brown rice cultured with P. igniarius showed a 

strong suppression on the GOT and GPT augmentation. In 

the group treated with only CCL4, all the activities of ALP, 

LDH, γ-GTP, SDH increased considerably compared to the 

normal rats without CCL4 treatment. However, both the 

groups fed with methanol extracts from brown rice cultured 

with P. igniarus prevented the increase in activities of 

these enzymes, almost to the level of normal rats without 

CCL4 treatment in some of the data. The larger molecular 

fraction demonstrated higher effectiveness in general. 

362 - High incidence of mycotoxin zearalenone in stored 

wheat from Mazandaran Province, north of Iran 

M.T. Hedayati * & A.R. Dalili 

Faculty of Health, Amir Mazandarani Boul. Vesal Shirazi 

St. Post code 48167, Sari, Iran. - E-mail: 

hedayaty2001@yahoo.co.uk 

In Iran, wheat is one of the main cereals for human 

consumption. Wheat grains containing zearalenone can 

cause genital problem and reproductive disorders in 

domestic animals. In human, episodes of breast 

enlargement in young boys and sexual precocity have been 

reported. This toxin may therefore have a role in hormonal 

balance and mammary cancer in regions with high 

zearalenone ingestion. In this study the incidence of 

zearalenone in wheat samples from Mazandaran 

warehouses were investigated. A total of 118 wheat 

samples (1 kg) were randomly collected from all 12 

114 


warehouses of Mazandaran province. Competitive 

microtitre plate enzyme immunoassay for the quantitative 

determination of zearalenone was used according the 

manufacturer instructions (Sigma-Aldrich 

Laborchemikalien GmbH). Zearalenone was found in 81% 

of samples and 64.4% of samples contained >200 µg/kg of 

this toxin. The minimum levels of zearalenone were 29 

µg/kg. All of the warehouses were positive for the presence 

of zearalenone. 50-100% of samples in each of warehouse 

had >200 µg/kg contamination of zearalenone. Our results 

showed the high percentage and levels of contamination of 

wheat samples with zearalenone. With regard to proposed 

standards given by the Standard Administration of Iran, 

under which the content of zearalenone should not exceed 

200 ppb in food products, 64.4% of samples meet these 

criteria. 

363 - Mycelial growth of Ganoderma lucidum and 

Pleurotus ostreatus on the selenium-enriched media 

M. Stajic 1* , I. Brceski 2 , M. Kezman 1 , S. Duletic-Lausevic 1 

& J. Vukojevic 1 

1 Faculty of Biology; University of Belgrade, Takovska 43; 

11000 Belgrade, Yugoslavia. - 2 Faculty of Chemistry; 

University of Belgrade, Studentski trg 3; 11000 Belgrade, 

Yugoslavia. - E-mail: stajicm@hotmail.com 

Ganoderma lucidum and Pleurotus ostreatus are 

mushrooms of high pharmacological value. 119 different 

kinds of triterpenoids and various polysaccharides from G. 

lucidum fruiting bodies have shown antitumor and 

immunostimulating effects. Polysaccharides isolated from 

P. ostreatus have shown antitumor, immunomodulating, 

antiviral, antibacterial activities. Selenium is one of the 

essential oligoelements, and it is indispensable for cell 

functions. Selenium adding in nutrition has been shown to 

have anticancer and antimutagenic effects. The aim of our 

research was to investigate the influence of selenium added 

in media on the growth rate and biomass changes of 

mycelia and them ability to accumulate this microelement. 

The medium used for analyzing the growth rate of the 

mycelia was Malt agar. The same submerged medium was 

used for measuring mycelial biomass and the accumulated 

selenium. The concentrations of selenium, used in a form 

of sodium selenite, were the following: 0.3 mg/l, 0.7 mg/l, 

1 mg/l and 1.3 mg/l. The accumulation of selenium in 

mycelia was measured using the Atomic Absorption 

Spectrophotometer. The analyzed concentrations of 

selenium did not show any influence to the growth rate of 

both species mycelia in comparison to the control. 

Different concentrations of selenium showed influence on 

mycelial biomass in submerged culture. The results of 

selenium accumulation in mycelia have shown higher 

concentration in samples grown on selenium containing 

media.


364 - Antiprotozoal activity and mitogen effect of edible 

medicinal mushroom Lentinus edodes (Berk.) Sing. 

(Shiitake) 

S.M. Badalyan 1* & S.H. Sisakyan 2 

1 Yerevan State University, Department of Botany, 375025, 

1 Aleg Manoogian St., Armenia. - 2 Yerevan State Medical 

University, Department of Medical Biology and Genetics, 

375025, 2 Koryun St., Armenia. - E-mail: 

badalians@infocom.am 

The Lentinus edodes is a worldwide-cultivated mushroom 

of high nutritional value and excellent pharmacological 

properties. It has been used in traditional medicine of Asian 

countries as elixirium ad longam vitam. In nowadays, 

different forms of dietary supplements with curative and 

preventive properties are obtained from L. edodes. The 

mitogen and antiprotozoal activities of L. edodes have not 

been investigated sufficiently. As part of our Medicinal 

Mushrooms Pharmacological Activity screening program, 

the presented results concern the study of antiprotozoal 

activity (APA) and mitogen effect (MGE) of L. edodes 

against Paramecium caudatum. The 25-day-old mycelium 

of L. edodes cultured on liquid malt-extract medium was 

used. The mycelium extract (ME, 0.03%, 0.03 ml) and 

cultural filtrate (CF, 0.03 and 0.06 ml; not diluted and 

diluted by Lozinskii solution in ratio 2:1, 1:1, v/v) were 

tested. All ME and CF samples during 76 hours showed 

100% APA. The fastest effect was observed after 2 hours 

under the influence of 0.03 ml CF without dilution. The 

highest APA with 21% MGE showed 0.06 ml CF (2:1, 

v/v). The strongest MGE (221%) possessed ME. The 

revealed physiological activities of L. edodes allow us to 

conclude that its exo- and endo-metabolites can be used for 

formulation of different bio-preparations, with 

antiprotozoal and mitogen (wound-healing) activities. 

Experiments including pathogenic test-organisms 

Entameoba histolytica and Balantidium coli are in 

progress. 

365 - Knowledge and utilisation of edible mushrooms 

by local populations of the rain forest of south 

Cameroon 

T.W. Kuyper 1* , J.F.W. Van Dijk 2 & N.A. Onguene 3 

1 Wageningen University, Soil Quality, P.O. Box 8005, 

6700 EC Wageningen, The Netherlands. - 2 Wageningen 

University,, P.O. BOX 342, 6700 AH Wageningen, The 

Netherlands. - 3 Institute of Agricultural Research for 

Development (IRAD), P.O. BOX 2123, Nkolbisson 

(Yaounde), Cameroon. - E-mail: 

THOM.KUYPER@BB.BENP.WAU.NL 

Indigenous knowledge on edible fungi and their utilisation 

by local populations in the rain forest zone of southern 

Cameroon were investigated. Members of both major 

tribes, Bantu and Bagyeli (Pygmy), were interviewed. 

Mushroom usage was monitored on a daily basis for more 

than a year. Mushroom knowledge among both Bantu and 

Bagyeli is very high. More than 50 vernacular names were 

retrieved. At present, about 35 species have been identified 

to species level. Bantu sample most mushrooms in 

secondary forests, and their mushroom diet consists mainly 

of saprotrophic fungi. Bagyeli collect mushrooms more 

frequently in the primary rain forest, and ectomycorrhizal 

and termitophilous fungi constitute a larger part of their 

diet. Some species are also used in traditional medicine. 

Toxic mushrooms were not mentioned by local 

populations. Some differences in mushroom collection and 

consumption do exist between Bantu and Bagyeli, but 

actual rates of consumption are low for both groups, with 

resp. 1.4 and 1.1 kg fresh mushrooms per person per year. 

Consumption is much lower than in other parts of central 

and eastern Africa. Suggestions are given how the apparent 

discrepancy between widespread and extensive mushroom 

knowledge and rather infrequent mushroom use can be 

explained. 

366 - Evolutionary and co-evolutionary genomics in 

Saccharomyces cerevisiae 

C. Zeyl 

Wake Forest University, Department of Biology, P.O. Box 

7325, Winston-Salem, NC 27109, U.S.A. - E-mail: 

zeylcw@wfu.edu 

I used laboratory populations of budding yeast maintained 

by daily transfers to compare rates of adaptation by haploid 

and diploid populations over 2000 generations, and tested 

whether adaptation occurred through many mutations or by 

a few major mutations. As predicted by recent theoretical 

work, haploids adapted more quickly than diploids, but lost 

this advantage when the experiment was repeated with 

much smaller population sizes. Genetic analysis of one 

haploid evolved genotype indicates that its 72% fitness 

increase is attributable to approximately 4 mutations. 

Adaptive changes are being further characterized by 

hybridization of genomic DNA and cDNA to microarrays. 

Despite the artificial simplicity of the environment, 

ecological diversity evolved from a single ancestral 

genotype in at least one population within 1000 

generations. Two genotypes with contrasting growth curves 

are maintained by negative frequency-dependent selection 

(competitive advantage when rare). Interesting interactions 

also occur between the mitochondrial and nuclear genomes 

within cells. The population of mitochondrial 

chromosomes within a cell experiences conflicting 

selective pressures at different levels: selection among cells 

favors mitochondria that contribute to cellular fitness by 

encoding respiratory proficiency, while selection among 

mitochondrial chromosomes within cells favors less 

mutualistic mutants that encode no respiratory function but 

replicate faster than wild-types. 



367 - Selfish genes, sex and adaptation in yeast 

M.R. Goddard 

Centre for Population Biology, Imperial College at 

Silwood Park, Ascot, U.K. - E-mail: m.goddard@ic.ac.uk 

Most biologists agree that evolution occurs because those 

members of a population best adapted to the current 

environment will flourish at the expense of those less well 

adapted. But what makes certain members of a population 

better adapted? What is the link between genotype and 

phenotype? How does this adaptation arise? The short 

answer is by chance, random genetic mutations or novel 

gene combinations will confer a greater fitness. But what 

sort of mutations (e.g. duplications/point mutations), how 

many mutations, does one mutation always win out or are 

there many equally good solutions to a problem? I will 

describe an experiment with asexual yeast which looks at 

some of the mechanisms of adaptation to a saline 

environment. The vast majority of organisms are sexual to 

some degree, so how does sex affect adaptation? I will 

describe a second experiment which tests the function of 

sex - does it, as originally suggested, allow adaptation to 

occur more rapidly? Lastly, even though sex has presumed 

advantages, I will discuss some of the more unfortunate 

consequences of sex - namely how it allows selfish or 

parasitic genes to prosper. 

368 - Mutation and selection in the yeast Saccharomyces 

cerevisiae 

R. Korona 

Institute of Environmental Biology, Jagiellonian 

University, Gronostajowa 3, 30-387 Krakow, Poland. - Email: 

korona@eko.uj.edu.pl 

Fitness of organisms is to a large extent determined by 

quantitative traits, such as growth rate. The study of 

mutation affecting such traits is impeded by the fact that 

individual alleles often cause small and recessive effects. 

Fungal experimental systems, such as laboratory 

populations of baker's yeast, provide unparalleled 

opportunities for detection of new alleles. One reason is 

that mutations can be studied both in haploids and diploids, 

the other is the ease of propagation and growth rate assays. 

We screened for mutations that appeared in diploids and 

therefore were initially masked by wild-types but then, 

after tetrad dissection analysis, were revealed as growth 

defective haploids. We found that a deleterious or lethal 

mutation occurs spontaneously at a rate of about one per 

thousand of diploid genome replications. This seemingly 

slow pace of mutation rate in unicellular organisms with 

small genomes becomes substantial when extrapolated for 

species that have several times larger genomes and many 

cell replications per sexual generation. The collected 

mutations were subsequently assayed for their dominance 

and epistatic interactions (the latter only for non-lethals). 

Heterozygous effects of lethals and non-lethals were very 

116 


small and equal on average which means that the former 

were much better masked. Non-lethal mutations in 

heterozygous loci interacted mostly multiplicatively 

suggesting that neither their synergism nor antagonism 

were strong. 

369 - Evolution of azole resistance in yeasts: Genetics 

and genomics 

L.E. Cowen * , L.M. Kohn & J.B. Anderson 

Department of Botany, University of Toronto, 3359 

Mississauga Road North, Mississauga, Ontario, L5L 1C6, 

Canada. - E-mail: lcowen@utm.utoronto.ca 

We established twelve replicate experimental populations 

of Candida albicans to study the dynamics and 

mechanisms of adaptation to the azole antifungal agents. 

The experimental populations were founded from a single 

drug-sensitive cell; six populations were evolved with 

inhibitory concentrations of fluconazole and six were 

evolved without drug, over 330 generations. While all 

populations evolved with fluconazole adapted to the 

presence of drug, they followed strikingly different 

trajectories. The experimental populations also diverged in 

fitness. We measured changes in genome-wide gene 

expression that became entrenched during adaptation and 

persisted in the absence of drug using DNA microarrays. 

Cluster analysis of the 301 genes with significantly 

modulated expression identified three patterns of 

adaptation to drug. One pattern was unique to one 

population and included upregulation of the ABC 

transporter gene, CDR2. A second pattern occurred at a late 

stage of adaptation in three populations; for two of these 

populations profiled earlier in their evolution, a different 

pattern was observed at an early stage of adaptation. The 

succession of early- and late-stage patterns of gene 

expression, both of which include upregulation of the 

major facilitator gene, MDR1, must represent a common 

program of adaptation to this antifungal drug. We compare 

these results with the dynamics and mechanisms of evolved 

fluconazole resistance in experimental populations of 

Saccharomyces cerevisiae. 

370 - Predicting the evolutionary potential of plant 

pathogenic fungi: a model framework 

B.A. McDonald * & C.C. Linde 

Institute of Plant Science, ETH Zurich, Universitatstrasse 

2, Zurich CH-8049, Switzerland. - E-mail: 

bruce.mcdonald@ipw.agrl.ethz.ch 

Plant pathogenic fungi are notorious for evolving rapidly in 

agroecosystems. Hence they offer good models for studies 

in experimental population genetics and evolution. 

Interactions among five population genetic forces 

(mutation, migration, selection, drift, recombination) 

ultimately affect the evolutionary potential of fungal


populations. We propose a model framework for predicting 

the evolutionary potential of a range of plant pathogenic 

fungi based on assessment of the life history traits 

underlying the individual population genetic forces. The 

proposed 'evolutionary risk' model is flexible and provides 

a quantitative prediction of the expected relative rate of 

fungal evolution in experimental studies. The components 

of the model can be tested individually and in combination. 

We believe this model will be useful for formulating 

specific hypotheses and designing experiments regarding 

evolution and population genetics for a wide range of 

fungi. A preliminary test of the model based on the 

observed rate of breakdown of resistance genes in 35 plant 

pathosystems showed a significant correlation with 

predicted rates of evolution. This suggests that the model 

may be useful to predict the relative rates at which 

pathogenic fungi will evolve virulence. We hypothesize 

that the model will also be useful for predicting the rate at 

which fungi develop resistance to fungicides. 

371 - Molecular mechanisms involved in the evolution 

of the modern yeast genomes, proteomes and 

metabolomes 

J. Piskur * , R.B. Langkjaer, K. Moeller, Z. Gojkovic & W. 

Knecht 

Section for Molecular Biology, BioCentrum-DTU, 

Technical University of Denmark, Building 301, DK-2800 

Lyngby, Denmark. - E-mail: imjp@pop.dtu.dk 

Saccharomyces cerevisiae is one of the most important 

industrial and laboratory model organisms. It can 

successfully grow in the almost complete absence of 

oxygen (anaerobic yeast), it utilises sugars via fermentation 

even at aerobic conditions (aerobic-fermenting yeast), it 

constantly produces respiration-deficient mitochondrial 

mutants (petite-positive yeast) and the genome shows large 

segmental duplications (post-genome-duplication yeast). 

Recently, several related yeasts have been studied to get an 

idea about the origin of the modern traits found in the 

contemporary S. cerevisiae species. It is likely, that the 

progenitor of the modern Saccharomyces yeasts, as well as 

other related genera, was an aerobic and petite-negative 

yeast, which was not very prone to aerobic alcoholic 

fermentation. In the next step, some of the yeast lineages 

progressively decreased their dependence on oxygen by 

reshaping several of their metabolic pathways. For 

example, as soon as the fermentation pathway could 

provide enough energy for growth, the mitochondrial 

genome became dispensable, and several biochemical 

pathways, as de novo pyrimidine biosynthesis, could 

become independent on the integrity of the respiratory 

chain. A progressive independence of these pathways from 

the presence of oxygen has promoted the anaerobic life 

style. The origin of these gross metabolic changes might 

have coincided with extensive chromosome reorganisation 

and the origin of the large segmental genome duplications. 

372 - Pathogenicity factors involved in fungus-fungus 

interactions and biotechnological applications for 

disease control 

M. Lorito * , M. Ruocco, R. Ciliento, D. Piacenti, V. Scala, 

S. Lanzuise, A. Zoina, F. Scala & S. Woo 

Dept. ARBOPAVE- Plant Pathology, University of Naples, 

Via Università, 100, 80055 Portici (NA), Italy. - E-mail: 

lorito@unina.it 

Fungus-fungus interactions play an important role in 

biological processes that affect crop production and food 

quality, as well as animal and human health. They may 

serve as model systems useful in the understanding of 

various microbe-other organisms interactions. Further, the 

mechanisms involved represent sources of genes, 

promoters and compounds which have interesting 

biological properties. However, we are still quite far away 

from having a substantial understanding of these 

interactions. Our group, studies the Trichoderma-host 

system, which is based on several and complex 

mechanisms able to protect crops from pathogens and 

stimulate plant growth. By applying a variety of 

biochemical, genetics and molecular techniques, including 

the use of targeted gene disruption and transgenic 

expression of Trichoderma genes, we have identified 

pathogenicity factors required by the mycoparasite to 

attack the host and behave as a biocontrol agents. A few 

chitinases and glucanases, within the extensive and 

redundant lytic enzyme system of Trichoderma spp., have 

been found to contribute substantially to the antagonistic 

ability of these fungi. In vivo testing of knock-out mutants 

having one or two different ORFs interrupted, have 

indicated that Trichoderma uses different sets of genes 

depending on the strain and the host fungus. In addition, 

gene disruption affected expression of other biocontrol 

genes, and produced unexpected and potentially useful 

effects. 

373 - Pathogenicity factors in nematode-trapping fungi 

A. Tunlid 

Department of Ecology, Ecology Building, SE 223 62 

Lund, Sweden. - E-mail: anders.tunlid@mbioekol.lu.se 

The nematode-trapping fungi comprise a rather large group 

of soil living fungi that can infect nematodes by forming 

special morphological structures (traps) like mycelial 

networks, adhesive knobs or constricting rings. Following 

the development of the traps, the fungi infect the 

nematodes through a sequence of events: attachment to the 

host surface, penetration, followed by invasion and 

digestion of the host tissue. Among the pathogenicity 

factors that have been studied in some detail in these fungi 

are extracellular serine-proteases and lectins. To identify 

additional enzymes, toxins etc. that are involve in the 

killing and paralyzing of infected nematodes, we have 

started an EST sequencing project in the fungus 



Monacrosporium haptotylum which captures nematodes 

with the aid of adhesive knobs. In this fungus, functionally 

intact trap cells can be isolated from the mycelium. Three 

different cDNA libraries are analyzed representing 

transcripts of mycelium, knobs and knobs infecting C. 

elegans. We have so far sequenced and analyzed ca 5.000 

transcripts. Annotation of the transcripts displaying high or 

moderate similarities to sequences in Genbank indicate 

major metabolic differences between the saprophytic 

mycelium, the infection structures (knob) and the fungus 

when infecting C. elegans. 

374 - Gene structure and evolution of fungal secondary 

metabolites involved in plant disease 

J.D. Walton 

Michigan State University, DOE-Plant Research Lab, E. 

Lansing MI 48824, U.S.A. - E-mail: walton@msu.edu 

In filamentous fungi, the genes necessary for the 

biosynthesis of a particular secondary metabolite are 

typically clustered. Examples include penicillin, 

trichothecene, gibberellin, aflatoxin, lovastatin, and HCtoxin. 

Because known genetic mechanisms would tend to 

break up clusters, their prevalence and persistence suggests 

that clustering confers selective advantage either to the 

organism or to the cluster itself. Since housekeeping 

pathways tend not to be clustered in fungi, the selective 

advantage is probably related to the unique properties of 

secondary metabolite pathways, for example, their weak 

selective advantage. In the plant pathogenic fungus 

Cochliobolus carbonum, the genes for biosynthesis of the 

cyclic peptide HC-toxin are clustered within a 600-kb 

region of DNA called the 'TOX2' locus. TOX2 comprises 

at least seven genes, encoding a non-ribosomal peptide 

synthetase, a putative HC-toxin efflux carrier, at least three 

other structural biosynthetic genes, and a pathway-specific 

transcription factor. In all known HC-toxin producing 

isolates, these genes are all in multiple functional copies. 

All of the genes are completely absent from toxin nonproducing 

isolates, suggesting that acquisition of the HCtoxin 

biosynthetic genes occurred after speciation of C. 

carbonum, perhaps by horizontal gene transfer. The 

organization of the TOX2 genes within and between 

isolates suggests that duplication and translocation 

occurred subsequent to the acquisition of the TOX2 genes. 

375 - Glycosylation of cell wall proteins: importance in 

pathogenicity of Candida albicans 

N.A.R. Gow 

University of Aberdeen, Institute of Medical Sciences, 

Aberdeen AB25 2ZD, U.K. - E-mail: n.gow@abdn.ac.uk 

Candida albicans is the most common agent of serious 

human infection caused by a fungus. Cell wall 

mannoproteins act as the immediate point of contact 

118 


between the fungus and host surface epithelia, endothelia 

and phagocytic cells. Therefore, glycosylation of cell wall 

proteins has a direct impact on the outcome of host-fungus 

interactions. We are exploring the biological roles of the O- 

and N-linked mannan in via the analysis of strains in which 

a range of genes encoding mannosyl transferases have been 

disrupted. Periodate oxidation of mannan on the Candida 

cell surface or deletion of either MNT1 or MNT2, which are 

involved in O-glycosylation, results in strains with reduced 

adherence and attenuation of virulence. Mutations in Nglycosylation 

have multiple phenotypes including changes 

in drug sensitivity, adhesion and in some cases changes in 

morphogenesis. Mutants with deletions in the MNN4 gene 

are almost devoid in phosphomannan, which has been 

implicated in recognition of C. albicans by macrophages. 

However, mnn4 mutants were still able to be taken up by 

macrophages suggesting that this epitope is not a key 

mediator of macrophage recognition. Analysis of 

glycosylation mutants to date establishes that the 

carbohydrate epitopes of mannoproteins play key roles in 

aspects of C. albicans related to its pathogenesis including 

adherence, cell wall permeability, drug sensitivity, 

virulence, and yeast-hypha morphogenesis. 

376 - Genomic analysis of pathogenicity factors of the 

insect pathogen Metarhizium anisopliae 

F.M. Freimoser 1* , S. Screen 2 , G. Hu 1 & R.J. St. Leger 1 

1 University of Maryland, 4112 Plant Sciences Bldg., 

College Park, MD, 20742, U.S.A. - 2 Monsanto Company, 

St. Louis, Missouri, U.S.A. - E-mail: ff34@umail.umd.edu 

The ascomycete Metarhizium anisopliae attacks many pest 

insects and is widely used as a mycoinsecticide. It has been 

studied extensively at the genetic and biochemical level 

and serves as a model insect pathogenic fungus. M. 

anisopliae secretes a large number of enzymes with 

potential targets in the insect host including proteases, 

chitinases, lipases, esterases as well as toxic secondary 

metabolites. ESTs and microarrays offer unprecedented 

opportunities for the identification and characterization of 

these complex fungal responses to host-related signals. 

ESTs of M. anisopliae were obtained under growth 

conditions that optimize production of secreted proteins. 

The EST sequences revealed many potential virulence 

factors and categories of biologically active molecules 

previously unreported in Metarhizium and sometimes other 

fungi. The largest and most diverse category were the 

proteases representing more than 30% of all transcripts. To 

analyze expression of all discovered genes and in particular 

potential pathogenicity determinants cDNA microarrys 

were made. Several clusters of genes had distinct 

expression patterns and were specifically up- or downregulated 

during growth on insect cuticle. For example 

many proteases were differentially expressed in different 

media and at various time-points. These results 

demonstrate the suitability of such genomic approaches for 

understanding pathogenicity as a whole and identifying and 

characterizing pathogenicity factors in M. anisopliae.


377 - Fungal biodiversity in Colombian Amazon forests: 

species composition, ecology and function 

A.E. Franco Molano 1* , C.A. Lopez Quintero 1 & T. 

Boekhout 2 

1 

Dept. de Biologia, Universidad de Antioquia, Apartado 

Aereo 1226, Medellin, Colombia. - 

2 

TROPENBOS, 

Amsterdam, The Netherlands. - E-mail: 

afranco@catios.udea.edu.co 

This work constitutes an effort to establish the role of fungi 

during regeneration processes of tropical lowland forests, 

and was performed at the plots of Tropenbos Colombia in 

Araracuara and Peña Roja, in central Caquetá of 

Amazonian Colombia. The plots represent different 

regeneration stages and primary forests and have been 

investigated for a long time at Tropenbos Colombia. Our 

project generated new data on the occurrence of macro- 

and microfungi along a regeneration gradient in Colombian 

Amazon lowland forests. Functional inferences suggest a 

considerable role for the putative ectomycorrhizal fungi 

and litter decomposing fungi in the nutrient cycling of 

these forests. Decomposition of leaf litter can be relatively 

fast, but seems to be influenced by the environmental 

conditions. Shortly after trees are cut down, many woodinhabiting 

fungi appear, which may occur as endophytes in 

healthy trees. The formation of fruit bodies may be a result 

of the stress imposed by the death of the hosts. After the 

felled trees are burnt, soil borne microfungal populations 

are dominated by heat resistant fungi. Many new taxa of 

macro- and microfungi have been found, and await formal 

description. New ideas about the role of microfungal 

populations in nutrient cycling led to a new research 

proposal to study the role of the microbial interface 

connecting the roots of the trees with the environment in 

secondary and primary forests. 

378 - Diversity and complexity in Ascomycota from 

Venezuela 

T. Iturriaga 1* & D.W. Minter 2 

1 

Universidad Simón Bolívar, Dpto. Biología Organismos, 

Aptdo. 89000, Sartenejas, Baruta, Edo. Miranda, 

Venezuela. - 

2 

CAB International Mycological Inst., 

Bakeham Line, Egham, Surrey TW20 9TY, England, U.K. - 

E-mail: titurri@usb.ve 

Since the beginning of the 20th century and up to 1950, 

there was a remarkable interest of the US towards Latin 

America. The US Department of Agriculture sent UStrained 

mycologists to conduct inventories on the natural 

resources of several Latin American countries, and as a 

result, neotropical mycological knowledge widened. That 

was the period when Hans Sydow, and Chardon and Toro 

published the results of their expeditions to Venezuela. 

More recently, there has been an emphasis in fungal 

systematics, an interest towards basic research, and more 

participation of local mycologists, all of which have helped 

improved knowledge on fungi in general. Programs such as 

Flora Neotropica and Flora Amazonica Project, have 

contributed toward an increased knowledge of fungi of the 

neotropics. R. W. G. Dennis, has been a major contributor 

towards neotropical fungal knowledge, specifically in 

regards to ascomycetes. In his 'Fungus Flora of Venezuela 

and Adjacent Countries', 1970, he treated approximately 

1346 species of Ascomycetes. This talk will present 

ascomycete collectors in Venezuela over the last 50 years, 

as well as numbers of publications dealing with neotropical 

ascomycetes in major mycological journals and congress 

presentations. in this same area. Knowledge of the total 

numbers of Ascomycetes for some neotropical areas will 

be compared with other groups of fungi from the same 

geographical regions. 

379 - Ectomycorrhizal fungi associated with Dicymbe 

(Caesalpiniaceae) in the Pakaraima Mountains of 

Guyana 

T.W. Henkel 

Department of Biological Sciences, Humboldt State 

University, Arcata, California 95521, U.S.A. - E-mail: 

twh5@humboldt.edu 

Mycological explorations in the Pakaraima Mountains of 

western Guyana have unearthed unique forests dominated 

by ectomycorrhizal (EM) tree species of the genus 

Dicymbe Spruce ex Benth. (Caesalpiniaceae, tribe 

Amherstieae). This remote region of tropical South 

America is situated on the central Guiana Shield and is 

characterized by highly oligotrophic soils. While EM 

associations are still poorly known from South American 

rain forests, the discovery of EM Dicymbe forests extends 

the known range of EM associations substantially from 

previously documented sites in the central Brazilian 

Amazon and southern Venezuela. Species of Dicymbe 

reach extreme levels of dominance (> 90% of stand basal 

area) in a mosaic of primary forests in the Pakaraima 

Mountains. Dicymbe stands are sharply demarcated from 

surrounding mixed rain forests lacking in EM trees, and 

harbor a rich EM mycota. Collecting efforts have currently 

recorded > 150 species/morphospecies of EM fungi 

occurring in close association with Dicymbe species. These 

fungi are primarily basidiomycetes representing most of the 

commonly accepted EM families, including Boletaceae, 

Russulaceae, Amanitaceae, Cantharellaceae, 

Cortinariaceae, and Paxillaceae. Putative EM ascomycetes 

include Elaphomyces spp. and Pseudotulostoma volvata 

O.K. Mill. & T.W. Henkel, a new taxon in the 

Elaphomycetaceae. Ongoing systematic work on these 

fungi will be discussed, with a focus on new and unusual 

taxa. 



380 - Polypore diversity in Costa Rica 

J. Carranza 1* & A. Ruiz-Boyer 2 

1 Escuela de Biología, Universidad de Costa Rica, San 

Pedro, Costa Rica. - 2 Departamento de Historia Natural, 

Museo Nacional, San José, Costa Rica. - E-mail: 

julietac@biologia.ucr.ac.cr 

The systematic study of polypore fungi in Costa Rica 

started with the collections by Dr. Josiah L. Lowe in 1963. 

Recent studies have contributed to the knowledge of this 

group in Costa Rica. Currently, 81 genera and 284 species 

of pore fungi have been reported for Costa Rica. It has 

been calculated that the total amount of species will be 

around 350-400. Most of the species reported are 

cosmopolitan, and are distributed through out the country. 

Nevertheless, the highest diversity is found from sea level 

to 900 m with less taxonomic diversity on high elevations 

(2000-3000 m). 

381 - Biodiversity of Russula in Costa Rica: a first 

account 

B. Buyck 

Muséum national d'histoire naturelle, 12 rue Buffon, 75005 

Paris, France. - E-mail: buyck@mnhn.fr 

Information on Russula in Costa Rica is scarce and very 

fragmentary notwithstanding the fact that many hundreds 

of specimens have been collected, especially in the last ten 

years. Only very few new Russula's have been described 

from Costa Rica so far, but it is clear that the genus is 

highly divers and is characterized by an important degree 

of endemism. As can be expected, the Costa Rican taxa - at 

least in the more temperate mountain areas - reveal strong 

similarities with the Russula's from eastern North America. 

Unfortunately however, even in North America the 

knowledge of Russula remains rather superficial. With the 

exception of some subsections, there exists no complete, 

modern, taxonomic study of American Russula that can 

serve as an adequate basis for a revision of the genus in 

Costa Rica. To illustrate the above, some examples of 

biogeographically or otherwise interesting Russula's will be 

presented in more detail. The relation between species 

diversity, host distribution and general ecology will be 

addressed and compared with observations from other 

continents. 

120 


382 - Diversity of Costa Rican wood-inhabiting 

pyrenomycetes and loculoascomycetes 

S.M. Huhndorf * & F.A. Fernandez 

Field Museum of Natural History, 1400 S. Lake Shore Dr., 

Chicago, IL 60605, U.S.A. - E-mail: shuhndorf@fmnh.org 

Ascomycetes constitute the largest known group of fungi 

worldwide, occurring in all ecosystems and geographic 

areas as saprobes, pathogens and endophytes. Almost all 

historical reports of pyrenomycetes and loculoascomycetes 

for Costa Rica pertain to leaf-inhabiting fungi. Woodinhabiting 

taxa in these groups are virtually unknown - one 

of the only reports listed seven species in three genera in 

one family. A total of 531 specimens were collected from 

1996 to 2001. The specimens were distributed among 213 

species in 45 families. Species in six families were 

frequently encountered: 26% of the fungi collected were in 

the Chaetosphaeriaceae, 20% in the Lasiosphaeriaceae, 

12% in the Annulatascaceae s.l., 7% in the Xylariaceae, 7% 

in the Melanommataceae, 6% in the Hypocreaceae, 6% in 

the Platystomaceae and 6% in the Tubeufiaceae. In 

addition to our own collections, specimens have been 

examined from parataxonomists collecting in 5 

conservation areas in 2001 and 2002. Parataxonomists 

collected 1133 specimens of ascomycetes on trunks and 

455 specimens from plots. Field identifications reported 

specimens (% of trunk or plot collections) in the following 

groups: Xylariales (32% trunk; 35% plot), 

Lasiosphaeriaceae (13% trunk; 8% plot), Hypocreales (8% 

trunk; 7% plot), Melanommataceae (8% trunk; 4% plot), 

Platystomaceae (4% trunk; 2% plot) and others. 

Comparative data from collecting trips to lowland French 

Guiana and Puerto Rico show proportions of taxa similar to 

those for Costa Rica. 

383 - Diversity of neotropical smut fungi 


Botanical Institute of the University of Frankfurt, 



After ten years of field work in neotropical countries, 

morphological investigations, and study of the literature, a 

monograph on neotropical smut fungi (Ustilaginomycetes, 

Basidiomycota) is accepted for publication in Flora 

Neotropica. 227 species in 34 genera are described. Among 

these 17 species and 5 genera were described as new taxa 

and ca. 34 new combinations were necessary. Further 

interesting results were obtained concerning the diversity 

of soral morphology, germination of teliospores, host 

ranges, and geographical distribution of the different 

species of smut fungi. However, much work remains to be 

done before we might have a rather complete 

understanding of the existing diversity of smut fungi in the 

neotropics. An overview of the diversity of the known 

neotropical species of smut fungi will be presented.


384 - The importance of fungi 

A.J.S. Whalley 

School of Biomolecular Sciences, Liverpool John Moores 

Univeristy, Byrom Street, Liverpool, L3 3AF, U.K. - Email: 

bmsawhal@livjm.ac.uk 

Why study fungi? Politicians and grant awarding bodies 

focus on the applied, the popular or the trendy. Fungi sadly 

are rarely seen to fit into these categories. It is however our 

duty as mycologists to demonstrate that fungi are one of 

the Worlds greatest living resources and estimates of 1.5 

million species or more are becoming increasingly likely. 

Their track record for the benefit of man is without 

question. Where would we be without penicillins, 

cyclosporins cephalosporins and other important 

pharmaceutical products? At least 80 percent of the worlds 

plants have mycorrhizal associations without which they 

would grow poorly or not at all. We could be justified in 

saying no fungi no world! These of course are beneficial 

activities but their global significance as pathogens of 

economic plants and, more recently though HIV, man 

should not be underestimated. Imagine the political 

reaction if fungi were the perfect bio-weapons! This 

symposium follows on from Vancouver where the then 

President of IMA stated stand up and be proud to be a 

mycologist because if you dont there will be few left in ten 

years time. Eight years on is a good time to reflect and 

prepare for the future. Mycology is in our hands and it is 

our responsibility to make the subject exciting, emphasize 

its relevance and to woo the appropriate individuals and 

organizations. 

385 - Awareness to Public 

R. Watling 

Caledonian Mycological Enterprises, Crelah, 26 

Blinkbonny Ave, Edinburgh, EH4 3HU, Scotland, U.K. - Email: 

caledonianmyc@compyserve.com 

Promoting fungi at the grass roots is essential for the 

development of a healthy appreciation of the role fungi 

play and as mycologists its our duty to the community to 

inform. The paper will present case-studies in which 

pubic/mycological interface has been forged. It is 

necessary to inject interest in fungi at a range of levels 

from schools to adult education programmes. There are 

more people in Britain than ever before collecting and 

eating wild fungi and keen to know about their quarry and 

many natural historians who wish to know about their 

natural heritage but lack the knowledge and resources or 

where to look for such information. These are very 

receptive audiences. Children have strong influences on 

their adults and they are the adults of the future. The 

informed public is a useful ally in that it understands wellpresented 

media coverage, lobby on behalf of mycology 

and in addition includes a volunteer army happy to spend 

long hours collecting, recording and monitoring. In contrast 

many others, including politicians, consider fungi of 

nuisance value or even deadly and rarely think (or know) 

of the benefits. Celebrities can help to disseminate the 

message. If mycology is to be supported by Government 

agencies then they and voters require to know how 

important fungi are to everyday life and well-being of our 

planet. Mycologists are the people who know about fungi 

so it is up to us; if all fails we cannot blame anyone else!. 

386 - Integration of mycology with other disciplines 

N.A.R. Gow 

Univerity of Aberdeen, Insitute of Medical Sciences, 

Aberdeen AB25 2ZD, U.K. - E-mail: n.gow@abdn.ac.uk 

One vision of mycology is that our discipline is 

undervalued and underfunded. The accuracy of such a view 

is open to debate but reiteration of this negative view 

undermines the value and potential of mycology. Other 

disciplines also feel disadvantaged and have equal cause 

for concern about the future. Therefore, it may seem 

perverse or naïve to present on overtly positive vision of 

mycology in the current climate, but there are many 

opportunities for the modern era of mycology. The 

mycological remit is broad with highly studies model 

systems taking their place amongst the most important and 

fashionable and systems in which to explore basic 

biological processes. Genomics and post-genomic data 

bases and technologies are being developed by cell and 

molecular biologists, bacteriologists, parasitologists and 

botanists that will have tremendous utility in the analysis of 

fungi. The fungi are rich in organisms with critical roles to 

play in ecology, biotechnology, medicine, agriculture and 

other aspects of human endeavour, and remain one of the 

few relatively unexplored corners of global biodiversity. 

With the right tools, a multidisciplinary approach and an 

open mind, mycology can be seen to be at the beginning of 

an exciting and highly competitive era. This talk will 

accentuate this naively positive view with selected 

examples of how mycology is already evolving in this new 

era. 

387 - Educational aspects of promoting mycology 

D. Moore 

The University of Manchester, School of Biological 

Sciences, 1.800 Stopford Building, Manchester M13 9PT, 

U.K. 

From the standpoint of recent experiences, I will discuss 

the need for active promotion of mycology at all 

educational levels. However strongly, and however loudly, 

the case for the importance of fungi is put, we face an 

uphill struggle to have mycology included in general 

educational courses. Inclusion as a 'specialist study of a 

taxonomic group' is feasible in college and university 

courses, but there is little sympathy for the view that a 



course on 'biology' (at any level) should naturally include a 

major input dealing with fungi. The traditional division of 

biology into animal biology and plant biology still rules - 

from the school curriculum to the university degree 

programme. Changing this blinkered (and essentially 

ignorant) view of the natural world is likely to take a long 

time. We have to create an understanding that fungi form 

an essential and normal part of the biology that surrounds 

us, and I believe that we need to start with school, and even 

pre-school, children if we are to achieve this change in 

attitude. 

388 - Mycology in Thailand 

P. Sihanonth 1 , L. Manoch 2 , S. Rodtong 3* , E. Sangvichien 4 

& N. Jonglaekha 5 


Bangkok 10330, Thailand. - 2 Department of Plant 

Pathology, Kasetsart University, 10900, Bangkok, 

Thailand. - 3 Faculty of Science, Suranaree University of 

Technology, Nakhon Ratchasima, 30000, Thailand. - 

4 Faculty of Science, Ramkhamhaeng University, Bangkok, 

10240, Thailand. - 5 Department of Plant Pathology, 

Faculty of Agriculture, Chiang Mai University, Chiang 

Mai, 50200, Thailand. - E-mail: sureelak@ccs.sut.ac.th 

Thailand traditionally has a strong base in plant pathology 

and this is continuing. However in the last ten years 

mycology has received increasing attention and support in 

both the university and government sectors. Mycology, 

however, is not seen by the younger generation as a 

commercial subject and considerable effort in the 

university sector is required to switch on perspective 

students. The British Council has recently approved a 

higher education link programme between our universities 

and Liverpool John Moores University linking with the 

British Mycological Society aimed at stimulating the 

teaching of mycology through workshops and teaching 

manuals. BIOTEC as a government funded organization is 

running a number of research programmes mainly targeted 

at applied aspects of mycology. This presentation will 

review the development of mycology in Thailand through 

to the present day and evaluate the current situation and 

future prospects. 

389 - Conservation of the fungal diversity in Cuba 

J.M. Portales 

Instituto de Ecología y Sistemática, Carretera de Varona 

Km. 3.5, Capdevila, Boyeros, AP 8029, Ciudad de la 

Habana 10800, Cuba. - E-mail: botanica.ies@ama.cu 

One of the main objectives of Darwin Initiative project, 

'Fungi of the Caribbean' was to produce national 

conservation strategies for fungi in Cuba. The strategy is 

based on the analysis of the available information in the 

computerised databases of the project and in the Cuban 

122 


mycological literature. This proposal constitutes the first 

document that in our country, in the Caribbean region - and 

possibly in Latin America - approaches the specific 

problem of the fungal diversity conservation and one of the 

few ones that exist in the world. The document is 

structured in two fundamental parts: in the first one we 

offer the most current state of knowledge of Cuban 

mycobiota; while in the second part we develop the 

strategy and action plan in correspondence with the 'holes' 

identified in this study. The goals of this proposition 

belong together with those traced in the Cuban National 

Strategy of Biodiversity. However, the objectives and 

actions answer the particularities that the rational use and 

conservation of this important group of organisms has. The 

strategy and action plan highlights those areas where basic 

information about fungi are lacking, and discusses the best 

way to prepare 'red data list' for the fungi. The question of 

what scientific society would be most suitable to represent 

mycology in Cuba is also addressed, as well as the 

importance that has the habitats preservation for the 

conservation of the fungal diversity. 

390 - Conservation of macrofungi in the Pacific 

Northwest United States 

T.A. Dreisbach 1* , M.A. Castellano 1 , T. O'Dell 2 & R. 

Molina 1 

1 USDA Forest Service, Pacific Northwest Research 

Station, Forestry Sciences Laboratory, 3200 SW Jefferson 

Way, Corvallis, Oregon 97331, U.S.A. - 2 Grand Staircase- 

Escalante National Monument, 190 E. Center, Kanab, 

Utah 84741, U.S.A. - E-mail: tdreisbach@fs.fed.us 

The Northwest Forest Plan was developed in 1994 to 

conserve biodiversity and species viability by maintaining 

habitat in the forests of the Pacific Northwest United 

States. More than 400 species of concern are listed in the 

official Record of Decision, including mammals, birds, 

amphibians, mollusks, arthropods, plants, lichens, mosses, 

and fungi. Of the 400 species, the largest proportion (more 

than 50%) is fungi. The goal of the Forest Mycology Team 

of the USDA Forest Service, Pacific Northwest Research 

Station, Corvallis, Oregon, is to provide scientific 

knowledge on the biology and functional diversity of forest 

fungi and to apply this knowledge to fungal conservation as 

well as forest ecosystem health and sustained productivity. 

Information regarding taxonomy, species distribution and 

occurrences, population dynamics, and environmental 

variables are essential to improve strategies for species 

conservation. We are developing a number of tools to assist 

land managers and scientists in this endeavor, including: 

survey techniques, population analysis methods, and 

habitat-based models for predicting species presence and 

potential habitat. Using the 'most noble polypore' 

Bridgeoporus nobilissimus as a case study, we illustrate the 

concepts, issues, and management approaches relating to 

conservation of rare forest fungi on public lands in the 

United States.


391 - The IUCN threat categories as a challenge for 

mapping fungi 

B. Senn-Irlet * & S. Egli 

WSL, Swiss Federal Research Institute, CH- 8903 

Birmensdorf, Switzerland. - E-mail: beatrice.senn@wsl.ch 

IUCN criteria for Red lists (2000) demand mapping on an 

even base and on a fine scale for all taxa studied. To 

overcome the weakness of classical mapping projects, i.e. 

spatial, temporal, and taxonomical inconsistencies three 

new approaches were tested. The first approach includes 

sampling on transects at stratified randomly chosen forest 

plots of the Swiss National Forest inventory by trained 

mycologists. The second approach is directed towards 

volunteers with observations at randomly selected grid 

points in all habitat types. Modelling with spatially explicit 

models and expert models as the third approach are chosen 

to simulate the potential distribution and to define indicator 

values for each species. A comparison of the different 

approaches shows a similar abundance pattern for most 

agarics and boletes and contradicts objections towards 

classical mapping approaches. Species distribution in all 

approaches follow the same pattern, a Weibull distribution 

with few abundant and many rare species. To overcome 

problems of scale for the range estimation the link with 

habitat types and their simulation models is essential. Subdatasets 

allow to test size-dependant effects on species 

richness and conservation value. For some habitat types 

such as bog vegetation area alone seems not to be a very 

important criterion for fungi. 

392 - Fungi as indicator organisms for endangered 

forest ecosystems 

E. Parmasto 

Institute of Zoology and Botany, Estonian Agricultural 

University, 181 Riia St., 51014 Tartu, Estonia. - E-mail: 

e.parmasto@zbi.ee 

Indicator fungi are species, easily identifiable by 

paramycologists, which co-exist with rare and Red Data 

Book organisms. A 'classical' inventory of the 

Scandinavian forests used only 5 rare species earlier, but 

109 species and 16 large species groups of 'signal-species' 

were added later in Sweden. In the first case, many 

valuable habitats may be omitted; in the second case, good 

knowledge of fungi is needed by surveyors. In Finland, 33 

species are used as indicators, in Estonia, 51 species and 5 

species groups (genera). Selection of indicators should be 

based on a significant statistical correlation between their 

presence and general species richness. Such a study would 

be extremely time-consuming and has not yet been carried 

out anywhere. Lists of indicators have been compiled 

intuitively on the basis of observations by experienced 

mycologists. There are many examples of the existence of 

such a dependence. In an old forest of 19 ha protected since 

1924 in Estonia, 23 indicator species but also half (97) of 

the Estonian polypores and 140 other wood-rotting fungi 

have been found. Use of indicators is an arbitrary tool but 

useful in demonstrating the natural value of sites to nonspecialists. 

In Estonia, mycologists participated in the 

project 'Estonian Forest Conservation Area Network' 

(1999-2001; 51 new selected conservation areas); 

paramycologists have found indicator fungi at 175 sites 

studied during the inventory of Woodland Key Habitats 

(2000-2002). 

393 - The impact of environmental pollution on 

macrofungi 

A.J. Termorshuizen 

Marijkeweg 22, 6709 PG Wageningen, The Netherlands. - 

E-mail: aad.termorshuizen@biob.dpw.wau.nl 

An update of the literature on the impact of environmental 

pollution on the mycoflora will be presented. Much 

emphasis has been laid on nitrogen immissions, and the 

question is raised which other types of pollution may 

affect, positively or negatively, the quality of the 

mycoflora. Since the first reports on effects of air pollution 

on the mycoflora appeared, several types of air pollution 

have drastically been changed. The question will be 

addressed whether these changes have affected the 

mycoflora. Finally, an update will be presented on the 

mechanisms involved in pollution affecting the mycoflora. 

394 - Effects of forest management on wood-inhabiting 

fungi in Scandinavian boreal forests 

J.N. Stokland 1* , K.-H. Larsson 2 & L. Ryvarden 3 

1 Norwegian institute of land inventory, P.O.Box 115, 1430 

Ås, Norway. - 2 Göteborg University, Department of 

Botany, P.O. Box 461, SE 405 30 Göteborg, Sweden. - 

3 University of Oslo, Box 1045 Blindern, N-0316 Oslo, 

Norway. - E-mail: jns@nijos.no 

The species composition of wood-inhabiting fungi was 

investigated in 90 managed and 34 unmanaged spruce and 

pine forests in SE Norway. Altogether, we found 56 

Polypore and 234 Corticoid species. Comparisons of logs 

with similar properties (standardized tree species, decay 

class and dimension class) showed a significant reduction 

in species number per log in managed spruce forests 

compared to unmanaged forests, but there was no such 

effect in pine forests. The species number per log in 

managed spruce forests was 90-45% of that in the 

unmanaged forests, and the reduction was highest on logs 

of large dimensions. A closer inspection revealed that 

species preferring medium and very decayed spruce logs 

were disfavored in managed stands, whereas species on 

early decay classes and decay generalists were unaffected. 

Similarly, species preferring large spruce logs were 

disfavored in managed forests. Forest management had 

strongest impact on low-frequent (rare) species in the 



spruce forests (more than 50% reduction), whereas 

common species were modestly affected. The Corticoid 

species seemed more adversely affected than the 

Polypores. These results indicate that many woodinhabiting 

species in spruce forests are adversely affected 

by temporal (continuity) gaps in the supply of dead wood, 

whereas the species in pine forests seem more adapted to 

forest disturbances. This implies that forest management 

practices should secure sites with a continuous supply of 

dead wood in spruce forests. 

395 - Conceptual problems of Ecological Continuity and 

its bioindicators 

B. Nordén 

Botanical Inst. Göteborg University, Box 461, 405 30 

Göteborg, Sweden. - E-mail: bjorn.norden@systbot.gu.se 

Old, undisturbed forest stands may be important for 

biodiversity through their content of microhabitats or for 

the long periods available for colonisation. The term 

Ecological Continuity (EC) has been used to ascribe value 

to old forest stands, but its use may lead to underestimation 

of the importance of forest dynamics and dispersal. If bioindicators 

of EC are to be used, species with low dispersal 

capacity should be chosen. However, many lichens and 

other fungi seem to have a patch-tracking lifestyle, and to 

merely indicate specific microhabitats. This is exemplified 

with data from two papers on wood-inhabiting ascomycetes 

and basidiomycetes in temperate deciduous woodland. In 

the first paper, a negative correlation between species 

richness and stand age in hazel woodland was reported. 

The number of Red List species was not affected by stand 

age. In the second paper, fine woody debris (FWD; diam 1- 

10 cm) was found to be very important for species richness 

in oak dominated woodland. This finding is discussed in 

relation to the required time for microhabitat formation. 

75% of the ascomycetes were found only on FWD, 

compared to 35% for basidiomycetes. The volume of CWD 

(coarse woody debris; diam > 10 cm) and FWD, and the 

number of Red List species on CWD and FWD were 

similar. Indicators of EC should be sought among 

microhabitats (logs), terrestrial molluscs, vascular forest 

plants, bryophytes and lichenized fungi which are 

'perennial stayers', and perhaps ground-floor fungi. 

396 - Cortinarius subgenus Myxacium: phylogeny and 

distribution in North America 

M.T. Seidl 

University of Washington, Dept. of Botany, Box 351330, 

Seattle, WA, U.S.A. - E-mail: seidl@u.washington.edu 

Subgenus Myxacium is a relatively small group within 

Cortinarius, the largest genus of agarics. The subgenus in 

North America presently comprises 3-4 sections with 15-25 

species. A review of names used for Myxacium taxa will be 

124 


presented. Distribution maps will be shown for selected 

taxa within this region. To begin understanding 

relationships within Myxacium phylogenetic analyses were 

performed using nuclear DNA sequences from the internal 

transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA 

gene on 23 Cortinarius taxa representing 7 subgenera, and 

later on 42 taxa from the Northern Hemisphere. Evidence 

from these studies suggests the subgenus Myxacium is 

polyphyletic as currently recognized. A summary of 

current knowledge of phylogenetic relationships within 

Myxacium will be discussed together with individual 

species and their probable taxonomic placement. 

397 - Multiple-gene phylogenies indicate a recent 

radiation of Cortinarius 

U. Peintner 1* , J.-M. Moncalvo 2 , R. Vilgalys 2 & M. Moser 1 

1 University Innsbruck, Technikerstr. 25, A-6020 Innsbruck, 

Austria. - 2 Duke University, P.O. Box 90338, U.S.A. - Email: 

Ursula.Peintner@uibk.ac.at 

The main obstacle for a broadly acceptable classification of 

Cortinarius (Agaricales, Basidiomycetes) is the huge 

number of species, and the lack of good, non-transitional 

characters. In this study, we tested suitability and power of 

three protein coding genes in combination with nuclear 

rDNA to resolve deeper relationships within Cortinarius. 

We also compare the resolving power of increased taxon 

sampling versus increased character sampling. To address 

these questions, portions of ATPase subunit 6 (ATP6), 

elongation factor 1-alpha (EF1-alpha), NADH-ubiquinone 

oxidoreductase chain 5 (NADH5) and nuclear rDNA (ITS 

and LSU) were sequenced for 18-132 taxa, respectively. 

Sequences of 44 species were combined totalling 4955 bp 

of DNA. Datasets were analyzed separately and combined 

with Maximum Likelihood and Maximum Parsimony with 

different weighting schemes. Ribosomal DNA and proteincoding 

genes failed to provide adequate discriminatory 

power for deeper relationships. The combined gene 

genealogy yielded the best resolution and thus provides the 

most robust initial hypothesis of phylogenetic relationships 

within Cortinarius. We suggest that the difficulties to 

resolve the evolutionary relationships of Cortinarius are 

linked to two sequential rapid radiation events in the 

evolution of this ectomycorrhizal mushroom. Thus, 

unresolved branching orders during these time periods may 

represent an accurate representation of the evolutionary 

history of Cortinarius. 

398 - Taxonomy of genus Cortinarius based on 

phylogeny obtained from nuclear rDNA sequences 

K. Høiland 1* , K. Røberg 1 , A. Holst-Jensen 2 & K.T. 

Hansen 3 

1 Department of Biology, University of Oslo, P.O. Box 

1066, N-0316 Oslo, Norway. - 2 National Veterinary 

Institute, P.O. Box 8156 Dep., N-0033 Oslo, Norway. - 

3 Norwegian Polar Institute, P.O. Box 720, N-9171


Longyearbyen, Norway. - E-mail: 

klaus.hoiland@bio.uio.no 

ITS and LSU rDNA sequences of 48 species (ITS) and a 

subset of 29 species (LSU) of Cortinarius from the 

Northern Hemisphere were studied. Gymnopilus sapineus 

was the outgroup taxon. Phylogeny from the ITS data was 

studied by: maximum parsimony followed by successive 

weighting (MPsw), maximum likelihood analysis (ML), 

and neighbour joining (NJ). In all analyses, the subgenera 

Telamonia and Dermocybe were monophyletic. MPsw also 

rendered subgenus Phlegmacium as monophyletic if we 

accept Rozites caperatus, C. limonius (formerly subgenus 

Leprocybe), and subgenus Myxacium section Colliniti 

belonging to this subgenus. A tentative taxonomy of 

Cortinarius is: (1) subgenus Dermocybe with its 'classical' 

inventory. (2) subgenus Telamonia comprising the 'old' 

Telamonia species (minus C. obtusus) together with 

species formerly placed in Sericeocybe or Leprocybe. (3) 

subgenus Phlegmacium, also comprising some species 

from Leprocybe and Myxacium, and the genus Rozites. (4) 

subgenus Cortinarius, a paraphyletic group positioned at 

the bottom of the phylogenetic tree consisting of 

miscellaneous species from the 'old' subgenera. The LSU 

sequence data had few variable sites, and the phylogeny 

was not so obvious as from ITS. MPsw gave following 

indications: Telamonia is monophyletic, Rozites is inside 

Cortinarius, C. cinnabarinus belongs to Telamonia, C. 

obtusus falls outside Telamonia. The analysis could not 

support Phlegmacium as monophyletic. Dermocybe 

appeared paraphyletic on the LSU tree. 

399 - Evolution of subgenus Phlegmacium in light of its 

pigment chemistry - a case of 'trivialization' and loss of 

character diversity? 

T.E. Brandrud 

Norwegian institute for nature research (NINA), P.O.Box 

736 Sentrum, N-0105 Oslo, Norway. - E-mail: 

tor.brandrud@ninaosl.ninaniku.no 

Cortinarius subgenus Phlegmacium includes many 

spectacular, bright-coloured species, especially in sect. 

Fulvi ss. lat. (incl. sect. Scauri ss. Mos. p.p.). Section Fulvi 

has more than 60 different anthraquinonic pigments, 

mainly of the atrovirin, flavomannin and phlegmacin 

series. These pigment series are probably developed 

independently, from different precursors, and occur in 

different species. It is thus likely that sect. Fulvi is a 

polyphyletic group, even though it is morphologically 

homogeneous based on numerical character analyses. 

According to chemical pathways, the atrovirin-containing 

species (subsect. Atrovirentes), and the ones with the least 

methylated flavomannins and phlegmacins (subsect. 

Splendentes and Rufoolivacei) are the most primitive. 

These species are characterised by (i) high pigment 

concentrations, (ii) high diversity of pigments, and (iii) 

easy oxidized compounds. The supposed primitive pigment 

characters are co-occurring with extreme 'phlegmacioid' 

features such as (i) bright colours, (ii) strongly glutinous 

surfaces, (iii) marginate bulb, (iv) citriform, coarsely 

verrucose spores, and (v) lack of brown encrusting-parietal 

pigments. The supposed derived species possess more 

'trivial', 'telamonioid' characters such as brown colours and 

a clavate stem. It seems thus that the ongoing evolutionary 

trend in Phlegmacium is a kind depauperation of pigment 

diversity, going from spectacular to more trivial looking 

organisms. 

400 - Molecular systematics in the genus Inocybe 

(Agaricales) 

P. B. Matheny 1* , Y. J. Liu 2 , J. F. Ammirati 1 & B.D. Hall 2 

1 University of Washington, Departments of Botany, Box 

351330, Seattle, WA 98195, U.S.A. - 2 University of 

Washington, Departments of Botany and Genetics, Box 

355325, Seattle, WA 98195, U.S.A. - E-mail: 

matheny@u.washington.edu 

Three genes are employed to infer the phylogeny of 

Inocybe, a genus of ectomycorrhizal agarics distributed 

world-wide but particularly diverse in temperate regions of 

the northern and southern hemispheres. Partial DNA 

sequences of RPB1, RPB2, and nuclear large subunit 

ribosomal RNA (nLSU) all indicate Inocybe is 

monophyletic and composed of several lineages. Combined 

RPB1 and RPB2 DNA sequences and amino acid data 

reveal the I. calamistrata group (sect. Cervicolores) 

occupies the most basal branch in the clade and that 

subgenus Mallocybe, which includes the I. dulcamara 

group, is monophyletic. However, these results are not 

supported by all data sets. Members of the I. rimosa group 

(sect. Rimosae) appear polyphyletic based on RPB1 and 

nLSU data. Strong support is shown in all instances for the 

monophyly and derived status of species with metuloid 

cystidia, but nodulose-spored taxa are polyphyletic. 

Examples of cystidiate clades include the I. lanuginosa 

group and I. geophylla group. Several novel relationships 

have also been detected: (1) a southern hemisphere 

mallocyboid clade including the tropical West African I. 

aureoplumosa and undescribed taxa from Australia, 

including a secotioid form; and (2) a monophyletic group 

of several species with nodulose spores from neotropical 

Dicymbe-dominated forests in Guyana. RPB1 and nLSU 

data suggest the saprophytic genus, Phaeomarasmius, is 

indeed sister to Inocybe, although these two data sets are 

incongruent for some taxa. 

401 - Distribution of Cortinarius in the world flora and 

their relations to phanerogams. An update 

M. Moser 

University of Innsbruck, Technikerstr. 25, A-6020 

Innsbruck, Austria. - E-mail: Meinhard.Moser@uibk.ac.at 

The genus Cortinarius was for a long time regarded as 

being restricted to the temperate Northern and Southern 

Hemisphere associated with Pinaceae, Fagales, Eucalyptus, 

Tilia and Dryas. The work of Favre initiated the interest in 



alpine and arctic mycology and numerous Cortinarii have 

been described from the Northern Hemisphere associated 

with Salix and Dryas. The increased exploration of tropical 

fungi have brought to light quite numerous ectomycorrhizal 

taxa but the number of Cortinarii remain low. Few species 

are known from Congo, the Amazon area and South India 

associated with leguminous and sapotaceous genera or 

Dipterocarpaceae. The distribution of some special groups 

like subgenus Cortinarius, subgenus Leprocybe sensu 

stricto, the Cortinarius orellanus group, subgenus 

Dermocybe, subgenus Phlegmacium subsection 

Coleopodes, the Cortinarius salor group and others are 

discussed. 

402 - Species and speciation in the Hebeloma 

crustuliniforme complex 

D.K. Aanen 1* & T.W. Kuyper 2 

1 Zoological Institute, University of Copenhagen, 

universitetsparken 15, dk-2100 Copenhagen, Denmark. - 

2 Soil Biology and Biological Soil Quality Group, 

University of Wageningen, PO Box 8005, 6700 EC 

Wageningen, The Netherlands. - E-mail: dkaanen@zi.ku.dk 

Within the genus Hebeloma species have traditionally been 

defined typologically by their morphological differences. 

In many cases this has not lead to uncontroversial species 

delimitations. An extreme example is the Hebeloma 

crustuliniforme complex, where a number of species have 

been described, but where the borderlines between these 

species have been disputed. Here we used sexual 

intercompatibility to define species within this species 

complex. We found a minimum of 20 intercompatibility 

groups (biological species). A phylogeny including species 

of the other main groups of the genus Hebeloma shows that 

the Hebeloma crustuliniforme complex consists of two 

well-supported clades that together possibly do not form a 

monophyletic group. Speciation is the process of becoming 

a species. Under the biological species concept, speciation 

can thus be defined as the origin of sexual incompatibility. 

Within a monophyletic group consisting of nine closely 

related ICGs belonging to the Hebeloma crustuliniforme 

complex, four ICGs had varying levels of partial 

incompatibility (within groups 100% compatibility, 

between 0.4, 8.3 and 15.0%). We estimated phylogenies of 

different individuals of these ICGs to reconstruct the 

evolution of intercompatibility. Our results indicate that 

generalizations between the level of intercompatibility and 

genetic divergence within this species complex can not be 

made. 

403 - Correlations between classical systematics and 

molecular phylogenetic hypotheses in the genus 

Cortinarius 

S. Garnica 1* , M. Weiß 1 , B. Oertel 2 & F. Oberwinkler 1 

1 Universität Tübingen, Botanisches Institut, Lehrstuhl 

Spezielle Botanik und Mykologie, Auf der Morgenstelle 1, 

126 


D-72076 Tübingen, Germany. - 2 Universität Bonn, Institut 

für Obst- und Gemüsebau, Auf dem Hügel 6, D-53121 

Bonn-Endenich, Germany. - E-mail: 

sigisfredo.garnica@uni-tuebingen.de 

A wide spectrum of Cortinarius species from South 

America, Australia, Asia and Europa were studied by 

means of comparative morphology and anatomy, scanning 

electron microscopy of basidiospores, and molecular 

phylogenetic methods. Sequencing of both the ITS region 

and the D1/D2 region of the nuclear coded large ribosomal 

subunit (LSU) allowed estimation of phylogenetic 

relationships both within the genus Cortinarius and also 

between Cortinarius and closely related taxa. We present 

results of our molecular phylogenetic analyses and discuss 

them in relation to classical taxonomic concepts in 

Cortinarius and our own macro- and microscopical 

observations. 

404 - A molecular phylogeny of Gymnopilus, using 

ribosomal ITS DNA sequence data 

D.A. Orlovich 1* , A.-M.B. Oliver 1 & B.J. Rees 2 

1 Department of Botany, University of Otago, PO Box 56, 

Dunedin, New Zealand. - 2 School of Biological Science, 

University of New South Wales, Sydney, NSW 2052, 

Australia. - E-mail: david.orlovich@botany.otago.ac.nz 

Gymnopilus P. Karsten is a genus of wood-rotting 

basidiomycetes with over 150 species distributed 

worldwide. Gymnopilus produces a rusty-brown spore print 

and has rough spore ornamentation, commonly without a 

plage. The genus has recently been revised in Australia and 

there are several detailed accounts of the genus for other 

regions including Norway, North America and Great 

Britain. In conjunction with an ongoing revision of New 

Zealand species, we are sequencing the internal transcribed 

spacer region of nuclear ribosomal DNA for phylogenetic 

analyses using parsimony and maximum likelihood. We 

will present a phylogeny of the genus incorporating both 

southern and northern hemisphere species. There is 

considerable similarity between the New Zealand and 

Australian Gymnopilus flora, and several species pairs exist 

that span the northern and southern hemispheres. 

405 - Myconet 

O.E. Eriksson 

Department of Ecology and Environmental Science, Umeå 

University, SE-90187 Umeå, Sweden. 

Myconet MYCONET is an international project intended 

for the development of a natural classification of the 

Ascomycota. Relevant literature is screened and all new 

data that may necessitate changes in the classification are 

presented in the series Notes on ascomycete systematics (at 

present c. 3500 Notes). An Outline of the Ascomycota is


continuously updated. Decision on changes is taken by 

eight Boards of experts, viz. for Ascomycota (higher taxa), 

Taphrinomycotina / Saccharomycotina, Arthoniomycetes / 

Lecanoromycetes, Chaetothyriomycetes / 

Dothideomycetes, Eurotiomycetes, Leotiomycetes, 

Pezizomycetes, and Sordariomycetes. Results are published 

in the electronic journal Myconet and in hard copies of the 

same journal. 

406 - FungalWeb 

L. Lange 

Molecular Biotechnology, Novozymes A/S, Denmark. 

The FungalWeb has been established to make a 

mycological web site that can provide the most current and 

accurate fungal taxonomy. The users it is designed for 

range from professional mycologists in academia and 

industry to advanced high school students. What has been 

done is to provide a classification for all fungal telemorph 

genera, based on a combination of phenotypic and genetic 

data. The FungalWeb as it is available today is the result of 

a joined effort of a number of leading experts, together 

covering all the fungal taxonomic groups. The planned next 

steps for improving of the web site are the following: 

Extend the web site to cover the genera Aspergillus and 

Penicillium to species level. This will be done summer 

2002, building on contribution of data from Samson, 

Frisvad and Pitt. Extend the web site to as far as possible 

also include and accommodate implacement of the 

anamorphs. This will be done based on guidance and data 

from K. Seifert. Planned implemetation fall 2002. In a 

second generation FungalWeb it is planned to establish 

links to all major mycological web sites and databases, 

including pictures, secondary metabolites, pathogenesis, 

ecology, industrial use etc. Further, FungalWeb needs 

resources for scientific and technical maintenance and 

development. Attempts will be made to have this secured 

on coordination with the GBIF initiative. 

407 - LIAS - A global information system for lichenized 

and non-lichenized ascomycetes 

D. Triebel 

Botanische Staatssammlung Muenchen, Menzinger Strasse 

67 D-80638 Muenchen, Germany. - E-mail: 

triebel@botanik.biologie.uni-muenchen.de 

The LIAS project was initiated in 1995. It is a multiauthored 

information system for collecting and distributing 

descriptive and other biodiversity data on lichens and nonlichenized 

ascomycetes. Its goal is to provide a working 

space for cooperation and collaboration of experts 

(http://www.lias.net/). The data of the two core databases 

are stored in DiversityDescriptions (= DELTAAccess) and 

made available through several web interfaces. Using 

HTML data submission forms automatically generated 

from the LIAS databases, sets of descriptive data can be 

established and interactively revised over the internet. 

Taxon names are linked to various types of information (e. 

g. more than 700 partly multi-stated descriptive characters), 

allowing mycologists and lichenologists to co-operate 

online and add their data to a common system for multiple 

usage (identification keys, database generated natural 

language descriptions, revisions etc.; with public or 

restricted access). Currently more than 60 mycologists and 

lichenologists world-wide are contributing as authors or 

revisers. LIAS is co-operating with several flora and 

database projects (e. g. the Sonoran Lichen Flora Project, 

the Checklists of Lichens Project) and the Internet Portal 

'Mycology.Net'. It will serve as Global Species Database 

for lichens in context with the Species2000 initiative and 

the EuroCat project. 

408 - Recent Literature on Lichens 

E. Timdal 

Botanical Museum, University of Oslo, Sars' gate 1, N- 

0562 Oslo, Norway. - E-mail: einar.timdal@nhm.uio.no 

"Recent Literature on Lichens" (RLL) is a series published 

in The Bryologist since 1951. It aims at listing all recently 

published papers in lichenology, with a complete 

bibliographic reference, keywords, and abstract - including 

mention of all new scientific names and combinations. The 

authors are William L. Culberson (in the period 1951- 

1978), Robert S. Egan (1979-1991), and Theodore L. 

Esslinger (from 1991). The lists have been computerized 

by Egan and Esslinger, and the database is put on the Web 

by the present author 

(http://www.nhm.uio.no/botanisk/lav/RLL/RLL.HTM). 

Together with "Mattick's Literature Index", originally a 

card index in Berlin computerized and expanded by Harrie 

J.M. Sipman, the data set now goes back to 1532, and 

consists of more than 32,000 records. Members of 

Myconet's Lecanoromycetes Board may use RLL as a tool 

when scanning for taxonomic papers for review. 

409 - Saccharomycotina and Taphrinomycotina 

C.P. Kurtzman 

Microbial Genomics Bioprocessing Research Unit 

National Center for Agricultural Utilization Research, 

ARS/USD Peoria, Illinois 61604, U.S.A. 

Gene sequence comparisons have shown that presently 

described, genera, families and orders in the 

Saccharomycotina and Taphrinomycotina are often not 

phylogenetically circumscribed. Single gene analyses are 

generally inadequate for genus circumscription, but for the 

Saccharomycetales genera seem to be strongly resolved in 

datasets comprised of around 5,000 nucleotides. Family 

groupings in such analyses are generally only weakly 

supported. Consequently robust molecular phylogenies of 



higher level classification may require 20-50 gene 

sequences. 

410 - Lecanoromycetes 

G. Rambold 

Lehrstuhl für Pflanzensystematik, Universität Bayreuth, 

Universitätsstrasse 30 - NW 1, D-95447, Bayreuth, 

Germany. 

In the current Myconet classification, lichenized 

discomycetes are mainly placed in the two classes 

Arthoniomycetes and Lecanoromycetes with 6 orders, 

about 64 families, 72 Arthoniomycetes and 494 

Lecanoromycetes genera. The class Arthoniomycetes with 

one order is represented by nearly 400 known species, the 

class Lecanoromycetes might well comprize ca. 6.300 

species. Following the Myconet classification, the order 

Lecanorales s. str. is segregated into four suborders, 

whereas some other units formerly treated at suborder rank, 

have been (re-)assigned the status of orders. In the last 

years there were main activities in molecular phylogenetic 

research concerning both classes. From 2930 (partial) 18S 

rDNA sequences actually existing for Pezizomycotina, ca. 

1,6% refer to Arthoniomycetes and ca. 10% to 

Lecanoromycetes. For Arthoniomycetes additional 78 ITS 

sequences (ca. 1.2%) have been published aside from a 

negligible number of 26S and mitochondrial sequence data. 

For Lecanoromycetes, nearly 900 ITS sequences, 250 

sequences of 26S and a small number of mitochondrial 

sequence data are available in GenBank. To estimate the 

support of existing classifications of the two ascomycete 

classes by published molecular data, topologies of 18Strees 

of 20 publications are evaluated and compared with 

phylogenies calculated from 600 high quality sequences of 

1600 bp minimum length. Supplementary information from 

ITS- and LSU-based phylogenies is respected in the 

considerations. 

411 - Chaetothyriomycetes and Dothideomycetes 

H.T. Lumbsch 

Fachbereich 9/Botanik, Universität Essen, 

Universitätsstraße 5 D-45117 Essen, Germany. 

Major results of recent studies, mainly based on molecular 

methods, concerning the phylogeny of these two classes of 

ascolocularous fungi are presented and discussed. This 

includes the inclusion of lichenized fungi (e.g. 

Verrucariales) in the Chaetothyriomycetes and their 

relationships to other ascomycetes. Recent studies on the 

Dothideomycetes mainly dealed with the subdivision of 

this large class of fungi into natural lineages. Results and 

prospects for further is discussed using some examples. 

128 


412 - Eurotiomycetes 

R.S. Currah 

Department of Botany, University of Alberta, b414 

Biological Sciences Centre Edmonton, Alberta T6G 2E9, 

Canada. 

The morphological signature of the Eurotiomycetes 

embodies fungi that generally produce spherical, 

evanescent, eight-spored asci within more or less globose 

ascomata. Ascospores are minute, single-celled and hyaline 

or very pale in colour. DNA sequence analyses underline 

the convergent nature of this signature and have been the 

basis of arguments to align taxa with the typical 

eurotiomycete signature with other classes (e.g., 

Cryptendoxyla, Myxotrichum). Similarly, DNA analyses 

have been used to align taxa with quite anomalous 

morphology within the Eurotiomycetes (e.g., 

Pseudotulostoma). Understanding the nature of the 

ecological pressures creating these morphological variants 

will help us rationalize the taxonomic journeys of taxa 

among the classes of the Ascomycota. 

413 - Leotiomycetes 

H.-O. Baral 1 & R. Galán 2* 

1 Blaihofstr. 42, D-72074 Tuebingen, Germany. - 

2 Department of Plant Biology, University of Alcalá 28871 

Alcalá de Henares, Madrid, Spain. 

Molecular sequence data have suggested a more or less 

severe rearrangement of some genera or species within the 

Helotiales, or even their removal from the order to which 

they have previously been placed according to 

morphological characters. Whether these modern results 

can be accepted in every case remains to be seen. 

Morphological studies are based on a high number of 

species and specimens, while molecular studies often still 

concern only one or a few species per genus. Some 

morphologies are so complex in structure that a 

polyphyletic origin advocated by molecular data in some 

cases seems quite impossible to imagine. Furthermore, 

identification of species is still problematic in the 

Ascomycetes, considering the fact that a large number of 

taxa still remains to be described, followingly erroneous 

specific or even generic names may have been attached to 

specimens from which molecular data derive. Nobody has 

control over correct determination when using data from 

GenBank. The potential of descriptive light-microscopical 

morphology is far from being fully exhausted. This 

especially concerns the various characters of the living 

fungus (vital taxonomy) which offer a huge amount of 

stable information on both generic and specific levels, but 

also concerns exact studies of apical apparatus, croziers 

etc. Such carefully gained morphological results often 

coincide with those of the sequence data, especially if a 

combination of characters is used for the generic concepts.


414 - Pezizomycetes 

K. Hansen 


Cambridge, MA02138, U.S.A. 

Molecular phylogenetic studies in the Pezizomycetes (with 

single order Pezizales) have over the past five years 

revealed new information on relationships between 

families and genera. Pezizales currently includes 14 

families, and is composed of at least 3 distinct lineages: A) 

Ascobolaceae and Pezizaceae, B) a Discinaceae- 

Morchellaceae clade, a Helvellaceae-Tuberaceae- 

Karstenellaceae clade and a monotypic Rhizinaceae clade, 

C) Ascodesmidaceae, Glaziellaceae, Pyronemataceae, 

Sarcoscyphaceae and Sarcosomataceae. The placement of 

the monotypic family Carbomycetaceae is unknown. 

Lineage C is the most heterogeneous lineage and the 

delimitations of all included families are problematic. To 

resolve family level relationships within lineage C it will 

be necessary to evaluate the largest and presumed 

polyphyletic Pyronemataceae in the context of closely 

related taxa (Ascodesmidaceae, Glaziellaceae, 

Sarcoscyphaceae and Sarcosomataceae). The 

Pyronemataceae has been a 'default' family for pezizalean 

taxa with uninucleate spores and iodine negative asci, 

which lack unifying anatomical characters by which they 

could be segregated into putative natural families. The 

evolutionary relationships of Ascodesmis and Glaziella, 

and the justification of the families Ascodesmidaceae and 

Glaziellaceae, or their circumscription, is still uncertain. 

Lineages A and B appears to be well-separated 

evolutionary lineages and the delimitation of families 

within these has to a large extent been clarified. 

415 - Sordariomycetes 

T. Læssøe 

Department of Mycology, Botanical Institute, Oester 

Farimagsgade 2D, DK-1353 Copenhagen K, Denmark. 

Current classification of higher taxa in the class will be 

discussed. The genera currently accepted within the 

Xylariaceae will be given brief comments as to their 

current status and various problems will be raised. Several 

if not most of the so called core genera of the family can 

with the present knowledge not be considered 

monophyletic. 

416 - Laboulbeniomycetes, Spathulosporomycetes and 

Ascomycota inc. sed. 

O.E. Eriksson 


University, S-90187 Umeå, Sweden. - E-mail: 

ove.eriksson@eg.umu.se 

The two classes Laboulbeniomycetes and 

Spathulosporomycetes are briefly presented and 

Ascomycota with uncertain affinities are discussed. 

417 - Integrating biological data within an evolutionary 

framework 

I. Carbone 

Center for Integrated Fungal Research (CIFR), North 

Carolina State University, Box 7567 - Partners II Building, 

Raleigh, NC 27695-7567, U.S.A. 

The rapidly expanding field of bioinformatics is providing 

researchers with many tools and techniques for building, 

updating, and accessing biological data. With the 

exponentially increasing amounts of data from SNPs, gene 

expression profiling, proteomics, clinical and 

epidemiological studies, the focus is now on developing the 

conceptual and practical tools for understanding and 

exploring biological data to its fullest potential. One of the 

biggest challenges facing researchers is the move away 

from studying one gene or one molecular data set at a time 

to looking at multiple genes or multiple genetic and 

phenotypic data sets simultaneously, in a holistic fashion. 

The integration of multiple molecular and phenotypic data 

sets is accelerating the pace of biological discoveries and 

our overall understanding of biological systems. 

Computational methods are being developed to allow 

researchers to make the critical link between genotype and 

phenotype and in the process distinguish between 

associated and causative genetic variation. Evolutionary 

theory is at the core of these computational algorithms and 

is providing the conceptual bridge between bioinformatics 

and biological inference. 

418 - The utility of haplotypes in evolutionary analysis 

A.G. Clark 

Penn State University, Dpet Biology/University Park, PA, 

U.S.A. - E-mail: c92@psu.edu 

Genetic variation in natural populations is organized on 

chromosomes in linked sets that often exhibit some 

correlated structure or linkage disequilibrium. When 

considering sets of adjacent polymorphic sites, it is often of 



use to note that only a subset of possible arrangements 

occur. These 'haplotypes' have proven highly useful for 

making a variety of inferences about population structure, 

migration, ages of polymorphisms, and departures from 

neutral evolution. An overview of the methods for 

inference of haplotypes from genotype data will be 

presented, along with illustrations of the applications of 

haplotype analysis in populations of Drosophila and other 

organisms. 

419 - Inferring ancestry from DNA sequences; modeling 

and inference with the coalescent process 

R.C. Griffiths 

University of Oxford, Department of Statistics, 1 South 

Parks Road, Oxford, OX1 3TG, U.K. - E-mail: 

griff@stats.ox.ac.uk 

A unique gene tree describing the mutation history of a 

sample of DNA sequences can be constructed as a perfect 

phylogeny under an assumption of non-recurrent point 

mutations. There is much interest in thinking of the data as 

a tree, rather than just sequences. This talk will describe, 

with examples, how coalescent trees and gene trees are 

used to model and infer ancestry of a sample of DNA 

sequences. 

420 - Demographic inferences from molecular data 

using Bayesian and maximum likelihood methods 

R. Nielsen 

Department of Biometrics, Cornell University, 439 Warren 

Hall, Ithaca, NY 14853-7801, U.S.A. - E-mail: 

rn28@cornell.edu 

A popular approach for inferring demographic parameters, 

such as levels of gene flow between populations, or rates of 

population growth, is to collect molecular data and to use 

this data to estimate demographic parameters based on 

specific population models. In the past there have been few 

attempts to test these models and to develop methods for 

choosing between alternative models. In this talk I will 

present some recent methods for hypothesis testing and 

model choice using molecular population genetic data. In 

particular I will investigate models for estimating levels of 

gene flow and population divergence times. 

421 - Progress and pitfalls in aflatoxin studies 

D.M. Wilson 

University of Georgia, Coastal Plain Station, P. O. Box 

748, 109 Plant Science Drive, Tifton, GA 31793, U.S.A. - 

E-mail: dwilson@tifton.cpes.peachnet.edu 

130 


Aflatoxin contamination of foods and feeds is a global 

problem which is most severe in the warmer portions of the 

globe. Aflatoxin research is difficult because the fungi that 

produce the aflatoxins are not aggressive pathogens and 

often occur as saprophytes in feedstuffs. Aflatoxin 

contamination can occur before harvest, during harvesting 

and drying and in storage. The environmental conditions 

favoring contamination before harvest generally includes 

late season drought and high temperatures. The 

relationships between damage and contamination begin in 

the field and progress throughout the marketing channels 

with moisture content being the most critical post harvest 

factor. There are different marketing regulations in the 

USA and European Union. The market for the product 

sometimes dictates the marketing and management 

strategies. Countries with few or unenforced regulations 

sometimes have highly contaminated products in the 

marketplace. Therefore, management strategies must be 

tailored to the climate and the country. There are at present 

no completely effective management tools to eliminate 

aflatoxin contamination from the human food chain. 

422 - Fumonisin exposure in different populations and 

reduction of fumonisin by the process of 

nixtamalization 

M. Dombrink-Kurtzman 

Mycotoxin Research, NCAUR, USDA, ARS, 1815 N. 

University Street, Peoria, IL 61604, U.S.A. - E-mail: 

dombrink@ncaur.usda.gov 

In countries where a major part of the daily diet is maize, 

the presence of fumonisins in maize products represents an 

emerging health concern. The U.S. Food and Drug 

Administration has issued recommended guidance levels 

for maize. Although unequivocal harm to humans has not 

been demonstrated, fumonisins have been shown to cause 

diseases in horses and pigs and to produce tumors in 

laboratory rats. Exposure to fumonisins is highest in South 

Africa and China due to high intake of maize and the 

potential for environmental conditions favoring mycotoxin 

production. Fumonisins, mycotoxins produced by 

Fusarium verticillioides (synonym F. moniliforme) and F. 

proliferatum, can be present in normal-appearing maize. 

The alkaline-cooking process, nixtamalization, is used to 

produce many maize products in Mexico, Central America 

and the United States. Research had shown that masa and 

tortillas from Mexico and the United States contained 

fumonisin B1. To determine the fate of fumonisins during 

processing, maize naturally contaminated with fumonisins 

underwent nixtamalization for production of tortillas. 

Material was analyzed at each step for fumonisins, weights 

and moisture contents so that mass balance determinations 

could be calculated. Less than twenty percent of the 

fumonisins remained in the tortillas. Nixtamalization 

appears to be a way to reduce significantly fumonisins 

occurring in maize products.


423 - Human exposure to ochratoxin A: Analytical 

methods for determining ochratoxin A in wine and beer 

and as a biomarker in human urine 

M. Pascale * & A. Visconti 

Istituto di Scienze delle Produzioni Alimentari, CNR, V.le 

L. Einaudi, 51 - 70125 Bari, Italy. - E-mail: 

m.pascale@area.ba.cnr.it 

Ochratoxin A (OA), a mycotoxin widely distributed in 

various foodstuffs and beverages, has been shown to be 

nephrotoxic, hepatotoxic, teratogenic and immunotoxic to 

several animal species and to cause kidney and liver 

tumours in mice and rats. IARC has classified OA as a 

possible carcinogen to humans (Group 2B). The 

widespread occurrence of OA in human blood provides 

evidence of a continuous human exposure to this 

mycotoxin. Wine and beer are products widely consumed 

by adult individuals and, due to the high frequency of 

contamination with OA, they may represents a serious 

source of daily OA intake for human. A new analytical 

method for the determination of OA in wine and beer has 

been recently developed by the authors and validated by an 

inter-laboratory study. The method, based on the use of 

immunoaffinity columns and HPLC, has been recently 

adopted by the AOAC International, CEN (European 

Committee for Standardisation) and OIV (Office 

International de la Vigne et du Vin) as official method. In a 

recent study carried out in the UK, a good correlation 

between OA consumption through the diet and OA 

concentration in urine was observed suggesting that urine 

could provide a useful marker of OA intake. A rapid and 

accurate method, based on immunoaffinity columns and 

HPLC, to quantify OA at pg/ml levels in urine has been 

developed by the authors. The method can be used as a 

rapid and non-invasive tool to assess human and animal 

exposure to OA in epidemiological studies. 

424 - Changes in concentration of mycotoxins during 

storage of wheat 

B. Birzele 

Institute for Plant Diseases, Dept. of Agricultural and 

Food Microbiology, University of Bonn, Meckenheimer 

Allee 168, 53115 Bonn, Germany. - E-mail: b.birzele@unibonn.de 

Species of the genera Fusarium, Penicillium and 

Aspergillus are well-known for their occurrence on grains 

and their ability to produce mycotoxins. Whereas Fusarium 

invade and damage grains predominantly on the field, they 

are increasingly displaced by Penicillium and Aspergillus 

during storage. Fusarium species produce a variety of 

mycotoxins, of which deoxynivalenol (DON), a Btrichothecene, 

is detected most frequently and in highest 

concentrations. DON is mainly produced on the field by F. 

graminearum and F. culmorum. Ochratoxin A (OTA), 

which is one of the mycotoxins produced by Aspergillus 

and Penicillium species, does not normally occur before 

harvest and is therefore considered to be a storage toxin. 

Due to its carcinogenicity and nephrotoxicity it is regarded 

to be the toxicologically most important mycotoxin 

occuring during storage of wheat. It was the purpose of the 

studies presented to investigate the influence of different 

suboptimal storage conditions on the viability and 

mycotoxin production of Fusarium and 

Penicillium/Aspergillus species and to deduce information 

about their inter-relationship during suboptimal storage. 

For that reason, DON and OTA contents were quantified 

and set into relation to the frequency of fungal isolation 

and biomass. 

425 - Penicillium species in food leftovers intended for 

feed 

I. Skaar * & M. Torp 

National Veterinary Institute, Department of Food and 

Feed hygiene, P.O.Box 8156 Dep. 0033 Oslo, Norway. - Email: 

ida.skaar@vetinst.no 

Swill feeding was widespread in Norway until feeding of 

unsterilized swill was prohibited due to the high risk for 

spreading of contagious animal diseases. However, there 

has been increasing interests for utilisation of food 

leftovers for animal feed, especially for pigs and fur 

animals. The authorities want to avoid food leftovers on 

waste disposal sites and at the same time utilize this 

resource in production of an economic feed. Plants 

producing liquid feed from food have been established 

several places in Norway. Food leftovers from large-scale 

households and food industry are actual resources. Since 

food leftovers have variable composition, a high number of 

mould species and mycotoxins may be expected. In a 

descriptive study of the fungal flora of food leftovers 

intended for feed, altogether 89 samples of food waste 

were subjected to mycological examinations. Genus 

Penicillium was demonstrated in 88% of the samples with 

mean count 5,5 ×10 5 KDE/g. Altogether 31 Penicillium 

species were demonstrated, of which 8 were demonstrated 

in ≥10% of the samples. The species most frequently 

demonstrated were P. crustosum, P. roqueforti, P. 

viridicatum, P.brevicompactum, P. aurantiogriseum, P. 

chrysogenum, P. expansum and P. echinulatum. Genus 

Penicillium does produce a high number of mycotoxins, 

which makes it difficult to choose indicator-toxins for feed 

based on food leftovers. Further studies on toxin 

production in food leftovers are in the process. 

426 - Mutualism and antagonism in bark beetle-fungusmite 

interactions 

K.D. Klepzig 1* , R.W. Hofstetter 2 , M.P. Ayres 2 & J.C. 

Moser 1 

1 USDA Forest Service, 2500 Shreveport Hwy, Pineville, LA 

71360, U.S.A. - 2 Dartmouth College, Gilman Hall, 

Hanover, NH 03755, U.S.A. - E-mail: kklepzig@fs.fed.us 



Bark beetles present an economically and ecologically 

significant model for the study of multilevel symbiotic 

interactions. The southern pine beetle - Dendroctonus 

frontalis - is associated with both mutualistic and 

antagonistic fungi. The bluestain fungus - Ophiostoma 

minus - may initially assist the beetle in killing its tree host. 

However, as beetle larvae begin developing, it competes 

with the beetle for host tissue. The beetle also carries two 

fungi in specialized mycangia. These fungi 

(Ceratocystiopsis ranaculosus and Entomocorticium sp. A) 

are nutritional mutualists of the beetle. These mycangial 

fungi are able to grow and fructify within the host tree, 

providing nitrogen rich tissue for the beetles to feed upon - 

unless they are interfered with and outcompeted by O. 

minus. All three fungi interact in both positive and negative 

fashion with phoretic mites, which are carried into the tree 

by the beetle. These mites in turn vector and feed upon two 

of the beetle associated fungi (O. minus and C. 

ranaculosus). Though O. minus usually outcompetes the 

mycangial fungi, abiotic factors (such as water potential) 

may alter these interactions. In addition, the fungi are not 

of equal benefit to the beetle, or the mites. We review 

recent research into the biology and ecology of this 

complex system, pose hypotheses as to the effects of these 

interactions on the population dynamics of this destructive 

beetle, and consider the implications to the field of fungal 

symbioses. 

427 - Epidemiology of southern pine beetle-fungal 

interactions 

R.W. Hofstetter 1* , K.D. Klepzig 2 , J.C. Moser 2 & M.P. 

Ayres 1 

1 Dartmouth College, 202 Gilman Hall, Hanover NH 

03755, U.S.A. - 2 USDA Forest Service, 2500 Shreveport 

Hwy, Pineville LA 71360, U.S.A. - E-mail: 

rwh@dartmouth.edu 

The effects of microbes on the dynamics of animal 

populations may be due to direct interaction between 

associates and hosts, or may be indirectly mediated by 

other organisms within the community. We studied a 

fungus (Ophiostoma minus) that is, at various times and 

stages, an antagonist and competitor of the southern pine 

beetle (Dendroctonus frontalis), a mutualist of several 

mites associated with the beetle, and a facultative pathogen 

of the pines that play host to this beetle. We hypothesized 

that the population dynamics of D. frontalis are influenced 

by negative feedback through community interactions 

involving other (mutualistic mycangial) fungi and O. 

minus, and mites that transport and feed upon the 

associated fungi. We found O. minus abundance within 

bark to be positively related to mite abundance but 

negatively related to D. frontalis survival. The abundance 

of O. minus gradually increased as D. frontalis infestations 

progressed through time. Changes in O. minus abundance 

were more correlated with mite abundance than with the 

prevalence of O. minus on attacking beetles. Factors that 

affect the population dynamics and behavior of these mites, 

and the growth of O. minus in pines, likely play an 

important role in the population dynamics of the southern 

pine beetle. 

132 


428 - The consequences of oligophily in bark beetlefungus 

associations 

D.L. Six 

School of Forestry, University of Montana, Missoula, MT, 

U.S.A. - E-mail: six@forestry.umt.edu 

Fungi are ubiquitous associates of bark beetles (Coleoptera: 

Scolytidae). Investigations have revealed a diverse array of 

association types ranging from mutualism to antagonism. 

Monophilic (one symbiont) and polyphilic (many 

symbionts) associations appear to be rare, while 

oligiophilic (two or a few symbionts) associations are 

common. Oligophily is likely to have several important 

consequences for both the host beetle and the symbiotic 

fungi. For any given association, fungal associates vary 

greatly in their effects on host beetle fitness, and therefore, 

are likely to differ in their influence on host population 

dynamics. Additionally, interactions, including competition 

for hosts, among multiple fungal associates of a beetle 

species may determine the relative abundance of each in a 

population, which, in turn, may also influence host 

population dynamics. Oligophily may be a less desirable 

state than monophily in associations that involve at least 

one mutualistic fungus. In such associations, for at least 

some beetles in a population, the mutualist can be 

displaced by less beneficial associates or by detrimental 

fungi, resulting in lower overall fitness. Some associates 

may actually be cheaters in the system,conferring few to no 

benefits to the host. While cheaters are potentially 

detrimental, beetles may be unable to develop effective 

means of avoiding cheaters without also negatively 

impacting beneficial associates. 

429 - Relationships among hosts, horntails, and fungi in 

Japan 

M. Tabata 

Shikoku Research Center, Forestry & Forest Products 

Research Institute, 2-915 Asakura-nishi, Kochi 780-8077, 

Japan. - E-mail: butter@ffpri.affrc.go.jp 

Wood discoloration of Chamaecyparis obtusa (hinoki) and 

Cryptomeria japonica (sugi) trees by the fungus associated 

with horntails, Urocerus japonicus and U. antennatus was 

studied in Japan. Wood discoloration in hinoki and sugi 

caused by the horntails and the fungus was found in several 

prefectures. The emergence periods of U. japonicus and U. 

antennatus were from the beginning of July to the middle 

of October and from the middle of May to the middle of 

August, respectively. Cultures isolated from the mycangia 

of adult females of horntails showed the same cultural 

characteristics. Basidiocarps found on felled logs of hinoki 

and sugi were identified as Amylostereum laevigatum based 

on morphological characteristics. The cultures from the 

basidiocarps had the same cultural characteristics as those 

from the mycangia of horntails. Two mycangial isolates 

produced on basidiocarps on the stem segments of sugi by


artificial inoculation and were identified as A. laevigatum. 

Isolates from the basidiocarps and the mycangia of 

horntails had similar ITS and peroxidase A sequences. The 

wood of all inoculated trees showed discoloration, with no 

difference in shape and pattern of discoloration among the 

two isolates from the basidiocarps of A. laevigatum and 

two from the mycangia of horntails. The inoculated fungi 

were reisolated from the areas of discoloration in the 

inoculated trees. This is the first report on wood 

discoloration of hinoki and sugi caused by the horntails and 

A. laevigatum. 

430 - Relationships amongst the fungal symbionts of 

Siricid woodwasps and their spread in the Southern 

Hemisphere 

B. Slippers 1* , T.A. Coutinho 1 , B.D. Wingfield 2 & M.J. 

Wingfield 1 

1 

Forestry and Agricultural Biotechnology Institute (FABI), 

Dept of Microbiology and Plant Pathology, University of 

2 

Pretoria, Pretoria, South Africa. - Forestry and 

Agricultural Biotechnology Insititute (FABI), Dept of 

Genetics, University of Pretoria, Pretoria, South Africa. - 

E-mail: bernard.slippers@fabi.up.ac.za 

The Basidiomycetes genus Amylostereum is best known for 

its symbiosis with siricid wood wasps. In this study, the 

phylogenetic relationships between Amylostereum spp. and 

other Basidiomycetes were investigated, using sequence 

data of the nuc-IGS and mt-SSU rDNA regions. A. 

areolatum was more distantly related to the three other 

species of Amylostereum, than they were to each other. A. 

ferreum and A. laevigatum were the most closely related 

species. These data support hypotheses relating to the 

mating behaviour and ecology of Amylostereum spp. 

Furthermore, contrary to previous suggestions, the 

Amylostereum spp. were more closely related to 

Echinodontium tinctorium (Echinodontiaceae) than to the 

Stereaceae. Apart from phylogenetic studies, vegetative 

compatibility and PCR-RFLP analyses were also used to 

study the spread of the introduced S. noctilio-A. areolatum 

complex in the Southern Hemisphere. Isolates of A. 

areolatum from South Africa and South America were 

found to represent a single VCG. PCR-RFLP profiles from 

nuc-IGS region showed that all isolates from the Southern 

Hemisphere shared the same profile, which differed from 

that of other populations. This genetically uniform 

population of A. areolatum has emerged from its obligate 

relationship with S. noctilio, which disperses only asexual 

fungal propagules. These results suggest a single or limited 

introduction of the A. areolatum-S. noctilio into the 

Southern Hemisphere. 

431 - Interspecific combative interactions - an overview 

L. Boddy * & D.P. Donnelly 

Cardiff School of Biosciences, Cardiff University, PO Box 

915, Cardiff CF10 3TL, U.K. - E-mail: 

boddyl@cardiff.ac.uk 

Competition by fungi for nutrients in dead organic 

resources is effectively brought about by competition for 

space. Such competition can be divided into primary 

resource capture (obtaining uncolonized resources) and 

secondary resource capture (combat to obtain resources 

already colonized by other fungi). Combat occurs not only 

in organic resources but also when mycelia grow out of the 

resource, e.g.into soil, in search of new resources. 

Combative mechanisms include antagonism at a distance, 

mycoparasitism, hyphal interference and gross mycelial 

contact. Outcome of interactions can be deadlock, where 

neither species gains territory from the other, or 

replacement, where one mycelium partially or completely 

wrests territory from the other. A hierarchy of combative 

abitlity can be discerned amongst fungi that inhabit a 

particular resource, but within this hierarchy there is 

intransitivity and outcomes can vary depending on abioic 

variables and the presence of other organisms. Interactions 

can dramatically alter mycelial function even when the 

outcome is deadlock. The review will be illustrated largely 

with examples of wood decay fungi and of wood decay 

fungi interacting with ectomycorrhizal mycelium. 

432 - Interactions among wood-inhabiting fungi: 

implications for fungal succession and biocontrol 

J. Stenlid * , L. Holmer & A. Iakovlev 

Dept Forest Mycology and Pathology, Swedish University 

of Agricultural Sciences, Box 7026, S- 750 07 Uppsala, 

Sweden. - E-mail: Jan.Stenlid@mykopat.slu.se 

Interactions among wood inhabiting fungi are important for 

succession and have also implications for biocontrol. Early 

establishment secures resources for the mycelium while 

late arrival to a substrate necessitates strategies for entering 

an already established decay community. Mycelial 

parasitism and replacement in wood has proven to be one 

important driving force in succession in wood. 

Replacement involves developmental changes in the 

mycelia and can often be facilitated by production of 

antimicrobial secondary substances. Other key factors for 

succession include differences in ability to handle 

recalcitrant compounds, differences in growth rate, 

differences in responses to gaseous regime and water 

potential etc. We will describe experiments indicating both 

species specific and general replacement patterns, 

metabolite production, and gene expression during 

interaction between various wood inhabiting fungi. Special 

reference will be given to the possibilities to find 

biocontrol against economically important root rot fungi 

such as Heterobasidion annosum. 



433 - Primary decayers - a key to understanding biodiversity 

in decaying wood? 

J. Heilmann-Clausen 

Danish Forest and Landscape Research Institute, 

Hørsholm Kongevej 11, 2970 Hørsholm, Denmark. - Email: 

jhc@kvl.dk 

Primary decayers, here defined as fungi initiating and 

causing extensive wood decay, have in several cases been 

found to be important determinants of subsequent decay 

development and species composition in decaying wood. 

Based on own research this concept of decay pathways is 

reviewed, and it is discussed if such pathways are well 

defined and whether they are shaped by interspecific 

interactions, by passive facilitation, or both. Further, the 

importance of decay pathways for biodiversity in decaying 

wood is considered, and it is discussed whether decay 

pathways are important to consider in the development of 

management guidelines aiming to preserve and restore 

biodiversity in forests. 

434 - Tritrophic relationships in the phyllosphere: 

fungal parasites and hyperparasites in action 

L. Kiss 

Plant Protection Institute, Hungarian Academy of 

Sciences, H-1525 Budapest, P.O. Box 102., Hungary. - Email: 

LKISS@NKI.HU 

Traditionally, the interactions between plant parasitic fungi 

and host plants are regarded as closed, two-species 

systems. However, both parasites and their hosts are, in 

fact, components of complex multitrophic interactions in 

which parasitic fungi are often attacked and killed by 

hyperparasites or other antagonists. Parasites, by definition, 

have a negative effect on host fitness, so hyperparasitism 

should be favourable for plants infected with parasites. 

However, studies on the possible role of hyperparasites in 

the natural control of plant parasites are missing from the 

literature. There are a few quantitative studies even on the 

natural occurrence of hyperparasitism that represents only 

the first step towards evaluating the impact of 

hyperparasites on host fungal and plant populations in 

nature. This paper synthesizes the current knowledge on 

structural, physiological and evolutionary aspects of natural 

host-parasite-hyperparasite relationships. A case study is 

also presented in which the effects of Ampelomyces 

hyperparasites on the fitness of powdery mildew infected 

Lycium halimifolium plants were studied by measuring the 

chlorophyll content of the healthy and infected leaves with 

and without hyperparasites in the field. 

134 


435 - Understanding the communicating mycelium - 

Translocation, past present and future 

S. Olsson 

Dept of Ecology, Royal Veterinary and Agricultural 

University, Thorvalsensvej 40, DK-1871 Frederiksberg C, 

Denmark. - E-mail: stefan.olsson@ecol.kvl.dk 

A fungal mycelium with its cytoplasmic continuum is a 

single organism. The mycelium is thus a network of 

communicating daughter nuclei. Communication consists 

of both signalling and sharing of material resources, like 

nutrients and energy. The study of nutrient and organelle 

translocation has a long history in mycology and some 

classical works will be briefly presented. The difference 

between large mycelia and the unicellular organisms in 

need for resourse redistribution are mainly two. 1. Need for 

fast high capacity nutrient translocation: Four different 

mechanism has been suggested, simple diffusion, simple 

diffusion + active uptake, active translocation by 

cytoplasmic movements and pressure driven bulk flow 

through vessel hyphae. There appear to be large differences 

between fungal mycelia of different species in their ability 

to translocate nutrients which might reflect the mechanisms 

employed. 2. Need for nutrient storage: To be able to 

scavenge nutrients quickly from the environment for 

redistribution there has to be efficient uptake systems and 

somewhere to store what has been taken up. Nutrients are 

taken up as small molecules but their storage have to be in 

osmotically neutral form. Great advances have been made 

in understanding the mechanisms for nutrient reallocation 

in mycelia but there are still much to do. Long distance 

signalling as well as nutrient storage needs attention in 

future research to be able to understand the physiology of 

mycelial organisms. 

436 - Bidirectional transport - an insurmountable 

obstacle to the use of tracer isotopes in quantitative 

translocation studies? 

B.D. Lindahl 

Dept. of Forest Mycology and Pathology, SLU, Box 7026, 

SE-75007, Uppsala, Sweden. - E-mail: 

bjorn.lindahl@mykopat.slu.se 

Tracer isotopes have been used in a long range of studies to 

demonstrate translocation of various substances through 

fungal mycelia. In a microcosm experiment, translocation 

was studied in rhizomorphs of the wood rotting fungus 

Hypholoma fasciculare. Non-destructive electronic 

autoradiography was used in combination with the two 

radioactive phosphorus isotopes 32P and 33P to show that 

phosphorus was transported in two directions 

simultaneously. This experiment confirms earlier studies 

suggesting that phosphorus, and most likely a range of 

other substances, are transported by mechanisms based on 

circulation throughout the mycelium rather than direct 

unidirectional transport from sources to sinks. The net


translocation between two parts of the mycelium cannot be 

quantified by adding a tracer isotope at the supposed source 

only, as the transport in the opposite direction must also be 

assessed. Even when tracer isotopes are added at both the 

source and the sink, conclusions about the net translocation 

are difficult to draw, as the tracer concentration in relation 

to the concentration of the studied analogue (the specific 

activity), also has to be known at both sites. Tracer isotopes 

may thus not be very useful as tools in quantitative 

translocation studies. The more straightforward method of 

destructive serial harvests followed by determination of 

total pool sizes is likely to be a better alternative. 

437 - In vivo imaging of nutrient dynamics in woodland 

fungi 

M. Tlalka * , S.C. Watkinson, P.R. Darrah & M.D. Fricker 

Plant Sciences, Oxford University, South Parks Road, 

Oxford OX1 3RB, England, U.K. - E-mail: 

monika.tlalka@plants.ox.ac.uk 

Nitrogen translocation by fungi, although a key ecosystem 

process, is poorly understood because current tools to study 

it have limited spatial and temporal resolution. In a novel 

technique to image transport of an amino-acid analogue in 

realistic microcosms, P. velutina was grown on a 

scintillation screen, and transport of 

14 C-AIB was 

quantitatively imaged with a photon-counting camera. 

Fourier analysis of transport showed a rapidly propagated 

pulsatile component, with period ranging from 18.3h at 19 

°C to 11.6h at 25.5 °C. Velocity of transport (23 mm h -1 ) 

was significantly faster than diffusion. Pulses in the 

inoculum and the mycelium were asymmetric and 

complementary, with the mycelium photon signal 

mirroring that from the inoculum. This can be mimicked by 

a model that includes exchange of amino acids in 

outgrowing mycelium with a storage pool in the inoculum 

mycelium. The vacuoles may both store amino acids and 

act as a transport pathway. Confocal microscopy showed a 

dynamic vacuolar network in which bulk movement of 

vacuoles and transient contacts between them via tubules 

were seen. FRAP and FLIP techniques showed an 

exchange of content between the vacuoles. By combining 

amino acid tracking in mycelium with confocal imaging, 

observed global nitrogen movements in the mycelial 

system can be explained in terms of cellular and metabolic 

events. 

438 - The septal pore cap as a key structure in cell to 

cell transport 

W.H. Müller 1* , B.M. Humbel 1 , B. Koster 1 , T.P. van der 

Krift 1 , A.J. Verkleij 1 , A.C. van Aelst 2 , R.C. Montijn 3 , J. 

Stalpers 4 , K.G.A. van Driel 4 & T. Boekhout 4 

1 MCB-EMSA, Utrecht University, Padualaan 8 NL-3584 

CH Utrecht, The Netherlands. - 2 PCB, Wageningen 

University, Arboretumlaan 4, NL-6703 BD Wageningen, 

The Netherlands. - 3 TNO Food and Nutrition Research 

Institute, Utrechtse weg 48, NL-3700 AJ Zeist, The 

Netherlands. - 4 Centraalbureau voor Schimmelcultures, 

Uppsalalaan 8, NL-3584 CT Utrecht, The Netherlands. - Email: 

W.H.Muller@bio.uu.nl 

The function of the septal pore cap (SPC) in 

basidiomycetes remains largely unknown. The connection 

of the SPC to the endoplasmic reticulum, and its position 

close to the orifice of the dolipore septum, may suggest 

several functions of the SPC. These may include a role in 

the transport of subcellular structures from one cell to 

another; a sieve function; and a barrier function during cell 

ageing, stress or cell lysis. While the application of 

tomography is a prerequisite for the fine analysis of the 

SPC, its use in combination with the conventional methods 

may complete the overall view. Interpreting the data 

obtained after the different preparation methods for 

electron microscopy may lead to a better understanding of 

the SPC structure. We suggest that the SPC in Rhizoctonia 

solani plays a key role in intercellular transport of for 

example mitochondria and ribosomes in the hyphal 

filaments. Moreover, the SPC may be involved in plugging 

the orifice of the septal pore channel. This plug-formation 

is not yet understood, but likely a signaling pathway exists 

to coordinate the assembly of plug-material at the orifice. 

Possibly, this pathway starts with an initial cytoplasmic 

signal that indicates that the orifice needs to be plugged, 

then the concerted action of the endoplasmic reticulum, the 

SPC and the filamentous network would finally result in 

the formation of a plug. Once plugged, no intercellular 

transport is possible. Most likely this plugging process is 

reversible. 

439 - Armillaria air pores and rhizomorphs really do 

conduct oxygen 

M. Pareek 1 , A.E. Ashford 1 & W.G. Allaway 2* 

1 University of New South Wales, Sydney, NSW 2052, 

Australia. - 2 The University of Sydney, NSW 2006, 

Australia. - E-mail: allaway@bio.usyd.edu.au 

Armillaria luteobubalina Watling & Kile produced 'air 

pores' at the origin of rhizomorphs in cultures on agar. Air 

pores grew upwards into the air from the colony surface, 

attaining a height of about 7 mm. When mature they were 

pigmented like the surface of the colony. Rhizomorphs 

originated below the air pores, grew down into the agar and 

then turned horizontally beneath the agar surface, 

eventually growing submerged in the agar to near the edge 

of the Petri dish. Air pores consisted of an aggregation of 

hyphae intertwined to form a cylinder. The pigmented layer 

at the surface of the colony itself extended into the base of 

the air pores, where it was elevated into a mound inside the 

base of the air-pore. Beneath the whole of the pigmented 

layer of the colony there was a region of loose hyphae with 

extensive gas space between them. This gas space extended 

into the base of the air pores and was continuous with the 

central gas canal of the rhizomorphs. This gas space was 

also continuous with the internal spaces of the air pore (and 

atmosphere) through gaps in the pigmented layer in its 

basal region. Conductance to oxygen of agar blocks with 

air pores was measured with oxygen electrodes: a 



substantial oxygen conductance averaging about 7 x 10-10 

m3 s-1 was observed. We conclude that the air pores 

conduct oxygen into the gas space below the pigmented 

mycelium of the colony, where the rhizomorphs - which 

can also conduct oxygen - originate. 

440 - Control of motility in tubular vacuole systems in 

fungi 

A.E. Ashford 1* , L. Cole 2 , D. Davies 1 & G. Hyde 1 

1 University of New South Wales, Sydney NSW 2052, 

Australia. - 2 The University of Sydney, NSW 2006, 

Australia. - E-mail: a.ashford@unsw.edu.au 

Tubular vacuole systems occur in mycorrhizal, 

saprotrophic and pathogenic fungi. In an Australian 

Pisolithus they consist of two distinct components: less 

mobile large compartments linked to the plasma membrane 

variously interconnected with motile tubular elements. 

These systems are implicated in endosomal processing, 

intra- and inter-cellular transport, storage, and enhanced 

interaction with the cytoplasm. In mycorrhizal associations 

they may facilitate transport of elements between nutrient 

absorbing hyphal tips and the interface with the plant 

partner. Recent evidence for and against this hypothesis 

will be evaluated. Tubule formation is a widespread 

phenomenon in organelle systems of eukaryote cells and 

offers an alternative to transport via vesicles. Vacuolar 

tubule formation is a regulated process that is modulated by 

both external and internal conditions and in Pisolithus is 

dependent on microtubules but apparently not on 

filamentous actin. Results with other inhibitors support a 

role for GTP-binding proteins in regulation of vacuolar 

tubule formation and the data are consistent with 

involvement of a dynamin-like GTPase. 

441 - The use of genomic repeated sequences to 

characterize arbuscular mycorrhizal fungi 

D. van Tuinen * , A. Gollotte, C. Arnould, O. Chatagnier, S. 

Gianinazzi & V. Gianinazzi-Pearson 

UMR 1088 BBCE-IPM INRA/Burgundy University, CMSE- 

INRA, BP 86510 21065 Dijon Cedex, France. - E-mail: 

tuinen@epoisses.inra.fr 

Arbuscular mycorrhizal fungi (Glomales) form a root 

symbiosis with more than 80% of land plant families, 

improving plant mineral uptake and general plant health. 

The different arbuscular mycorrhizal fungal species which 

have been described do not generally show host-plant 

specificity, suggesting a great plasticity of the fungal 

genome. To date, knowledge concerning genome 

organization and structure in these fungi is poor, so 

limiting the understanding of their biology at a molecular 

level. It is however known that arbuscular mycorrhizal 

fungi have a large genome size, and evidence points to the 

presence of a high proportion of repeated sequences. 

136 


Phylogeny analyses and identification of arbuscular 

mycorrhizal fungi has been based on ribosomal or nonribosomal 

repeated sequences, and the former have been 

used to show genome variability between nuclei of a same 

fungal isolate. In order to improve knowledge about 

genome organization in arbuscular mycorrhizal fungi, 

different types of repeated sequences have been 

characterized and their distribution in the fungal genome 

analyzed. Sequences containing elements found in 

transposons have been isolated, and their role in the 

evolution of the genome of these fungi will be discussed. 

Part of this work is supported by the European Project 

GENOMYCA (QLK5-CT-2000-01319; 

http://www.dijon.inra.fr/bbceipm/genomyca/). 

442 - Vegetative incompatibility and genetic diversity 

among geographically different isolates of Glomus 

mosseae 

C. Sbrana 1* , L. Avio 1 , P. Strani 2 , V. Rinaudo 2 & M. 

Giovannetti 2 

1 Istituto di Biologia e Biotecnologia Agraria C.N.R., 

Sezione di Pisa, Via del Borghetto 80, 56124 PISA, Italy. - 

2 Dipartimento di Chimica e Biotecnologie Agrarie, Via del 

Borghetto 80, 56124 PISA, Italy. - E-mail: 

sbranac@agr.unipi.it 

Population studies based on vegetative compatibility tests 

revealed the occurrence of genetically different isolates and 

of vegetative compatibility groups within the same fungal 

species in pathogenic, saprophytic and ectomycorrhizal 

fungi. Successful anastomoses occur widely between 

hyphae belonging to different individually germinated 

spores of the same isolate of arbuscular mycorrhizal (AM) 

fungi, though nothing is known about mycelial 

compatibility between conspecific isolates. We studied 

mycelial compatibility between geographically different 

isolates of the worldwide distributed AM fungal species 

Glomus mosseae. Vegetative compatibility tests performed 

on germlings belonging to the same isolate showed that all 

isolates were capable of self-anastomosing, whereas no 

anastomoses were detected in pairings between germlings 

of different isolates. Microscopic examinations detected 

hyphal incompatibility responses, consisting in apical wall 

thickening, followed by protoplasm withdrawal and septa 

formation, in hyphal swellings produced by the 

approaching hypha on the surface of the contacted one. The 

intraspecific genetic diversity of G. mosseae revealed by 

vegetative compatibility tests was confirmed by total 

protein profiles and ITS-RFLP profiles. Since AM fungi 

lack a tractable genetic system, vegetative compatibility 

tests may represent a powerful tool for the detection of 

genetically different mycelia and for investigating 

population structure of these obligate symbionts.


443 - Investigating the propagation pathway of 

endobacteria in Gigaspora margarita 

V. Bianciotto 1* , A. Genre 1 , P. Jargeat 2 , G. Bécard 2 & P. 

Bonfante 1 

1 Istituto Protezione Piante - Sezione di Torino CNR and 

Dipartimento di Biologia Vegetale Universita' di Torino, 

Viale Mattioli 25 -10125 Torino, Italy. - 2 UMR 5546 

CNRS/Université Paul Sabatier Pôle de Biotechnologie 

Végétale, 24, chemin de Borde-Rouge BP 17 Auzeville 

31326, Castanet-Tolosan, France. - E-mail: 

v.bianciotto@csmt.to.cnr.it 

Intracellular bacteria live in the cytoplasm of many isolates 

of arbuscular mycorrhizal fungi belonging to 

Gigasporaceae. On the basis of 16S rDNA analysis these 

endobacteria were assigned to a new taxon closely related 

to the genera Ralstonia, Pandorea and Burkholderia. To 

understand their propagation pathways through the life 

cycle of Gigaspora margarita (BEG 34 isolate), in vitro 

mycorrhizal cultures of transformed roots were produced. 

The presence of bacteria in spores from filial generations 

have been monitored by using confocal microscopy 

observations together with molecular analysis. We 

demonstrate for the first time the vertical transmission of 

endobacteria from a single 'parental' spore (F0) to filial 

spores through germinating and symbiotic mycelia. A 

quantification of the endobacteria population was carried 

out using 3D volume reconstruction. To verify whether the 

presence of the whole plant could determine differences in 

the transmission of bacteria, a F1 generation of spores was 

produced on clover pots. In contrast with the in vitro 

conditions, all the F1 spores from monosporal inocula were 

free of bacteria, suggesting that bacterial migration is 

controlled by multiple factors. Endobacterial activity was 

investigated by using Acridine Orange. The bacterial 

distribution pattern and activity, closely related to the 

fungal life cycle, reinforces the hypothesis of a strong 

symbiotic association between the two organisms. 

Research supported by the EU Project GENOMYCA. 

444 - Molecular bases of appressorium formation in 

AM fungi 

N. Requena * , E. Serrano & M. Breuninger 

Botanical Institute, University of Tübingen, Auf der 

Morgenstelle 1, 72076 Tübingen, Germany. - E-mail: 

natalia.requena@uni-tuebingen.de 

The appressorium development is the first morphogenetic 

change which precedes the formation of the symbiotic 

association between arbuscular mycorrhizal (AM) fungi 

and their host roots. This event takes place after 

recognition of yet unknown plant signals which trigger the 

developmental decision of abandoning the so-called 

asymbiotic life stage. Upon recognition of those signals the 

fungus changes its straight pattern of hyphal tip growth to 

form a swollen structure that hooks over a rhizodermis cell 

and later develops the penetration peg among two adjacent 

epidermal cells. The physical contact with the host root is 

essential for the appressorium development. To investigate 

the molecular bases for this morphogenesis we took a 

comparative molecular approach and studied the changes 

on gene expression that occurred to the fungus upon 

induction with a host root. We induced appressorium 

development on water agar medium by bringing into 

contact parsley seedlings with germinated spores or 

sporocarps of AM fungi. A time course of appressorium 

development showed that first induction takes place around 

120 h, while at 168 h a plateau in the number of 

appressoria is achieved. We investigated both, early gene 

expression and later gene expression in order to selectively 

search for genes involved in signaling and recognition or 

for genes responsible for structural changes. Results 

concerning our progress in this topic will be presented. 

445 - Extensive tubular vacuoles in arbuscular 

mycorrhizal fungi 

Y. Uetake * & M. Saito 

Natl. Inst. Livestock Grassland Sci., Nishinasuno, Tochigi, 

329-2793, Japan. - E-mail: yuetake@uoguelph.ca 

Hyphae of Gigaspora margarita were stained with Oregon 

Green 488 carboxy-DFFDA that is known to accumulate in 

the lumen of fungal vacuoles, and were observed by laser 

scanning confocal microscopy. Germ tubes had vacuoles 

with one of the following types: A, longitudinally oriented 

bundles of long tubules; B, both tubular and various sizes 

of spherical vacuoles in various proportions; C, a mass of 

spherical vacuoles. Stained germ tubes with cytoplasmic 

streaming always showed A or B types of vacuoles, but 

never C type. Tubular vacuoles were extremely fragile 

when exposed to laser irradiation; many small spheres were 

formed. Bundles of tubular vacuoles were also observed in 

extraradical hyphae and intercellular hyphae of G. 

margarita from co-cultures with onion seedlings. Tubular 

vacuoles were observed also in the germ tubes of G. rosea, 

Glomus leptotichum, Gl. intraradices, Scutellospora 

cerradensis and in hyphae of other members of 

Zygomycota, Rhizopus stolonier, Absidia repens, Mucor 

meguroence, Choanephora infundibulifera, Mortierella 

chlamydospora, Syncephalastrum racemosum, Linderia 

bicolumnata. These results suggest that tubular vacuoles 

are universal in Zygomycota. Ultrastructure of rapid 

freeze-freeze substituted germ tubes of G. margarita 

showed longitudinally oriented profiles of vacuoles 

occupying most of the cell volume. 



446 - Biolistic transformation of AM fungi: Advances 

and perspectives 

R. Bergero 1 , A. Gollotte 2 , P. Mutch 1 , D. Walsh 1 & L.A. 

Harrier 1* 

1 Scottish Agricultural College, Kings Buildings, West 

Mains Road, Edinburgh, Lothian, Scotland, U.K. - 2 UMR 

BBCE-IPM 1088, INRA/CMSE, BP 86510, 21065 Dijon 

Cedex, France. - E-mail: l.harrier@ed.sac.ac.uk 

Arbuscular mycorrhizal (AM) fungi of the unique phylum 

Glomeromycetes are ubiquitous in nature and constitute an 

integral component of terrestrial ecosystems, forming 

symbiotic associations with plant root systems of over 80% 

of all terrestrial plant species. Understanding the root 

interactions of AM fungi is one of the most challenging 

and exciting areas of research. The development of 

transformation strategies for AM fungi has provided a great 

opportunity for the enhancement of our understanding of 

the biology of AM fungi but also for the interactions 

between plant roots and the surrounding soil environment. 

In this presentation we discuss the development of 

transformation strategies for AM fungi by biolistics and 

highlight the areas of this technology which are being 

further developed for the stable transformation of these 

elusive organisms. These include the use of genetic 

reporters, vector construction and transgene persistence. 

We thank the Scottish Executive Environmental Rural 

Affairs Division (SEERAD) and the EU (Project 

GENOMYCA QLRT-2000-01319) for financial support. 

138 


IMC7 Main Congress Theme I: BIODIVERSITY AND CONSERVATION Posters 

451 - Study the effect of different techniques on 

diversity of freshwater hyphomycetes in the river Nile 

(Upper Egypt) 

A. Abdel-Raheem 

South Valley University, Faculty of Science, Botany 

Department, Sohag 82524, Egypt. - E-mail: a.abdelraheem@mailcity.com 

Four different techniques were applied, leaf mapping of 

Eucalptus rostrata; randomly leaf sampling; Millipore 

filtration; spores in foam, for aquatic hyphomycets 

communities study in the river Nile. Triscelophorus 

monosporus, Anguillospora longissima, Flagellospora 

penicilloides and Tetracladium marchalianum were the 

dominant species in the all techniques used. Aquatic 

hyphomycetes diversity was high by using leaf mapping 

(23 species) and randomly leaf sampling (31 species) but it 

was low by using Millipore filtration (11 species) and 

spores in foam (8 species) techniques. 

452 - Do marine fungi in Egypt form a distinct group ? 

M.A. Abdel-Wahab * & H.M. El-Sharouny 

Department of Botany, Faculty of Science at Sohag, South 

Valley University, Botany Dept., Faculty of Science, Sohag 

82524, Egypt. - E-mail: bhhoba@yahoo.com 

Diversity of subtropical marine fungi is comparable to that 

recorded from temperate and tropical ones. Clear line can 

be drawn between marine mycota in temperate and tropics, 

however that is not the case when we compare marine 

mycota of tropical and subtropical sea shores. Mangroves 

in tropical and subtropical locations harbor similar marine 

taxa, though specific groups of fungi are found more 

frequently in tropical (e.g. Aigialus spp., Lophiostoma spp., 

Pyrenographa xylographoides, Acrocordiopsis patlii and 

Quintaria lignatilis) or subtropical (e.g. Lignincola laevis, 

Lulworthia grandispora and Periconia prolifica) 

mangroves. Mangroves in tropics have a higher fungal 

diversity than that in subtropics but several factors may 

account for this. Marine mycota diversity in subtropical 

habitats varies from one location to another. Mangroves 

and sea shores in Egypt are harsh environments (i.e. high 

salinity and temperature accompanied with low humidity) 

surrounded by desert. Frequent species recorded in samples 

collected from the Red Sea coast were different from other 

subtropical habitats (e.g. Swampomyces spp. and 

Kirschsteiniothelia maritima), while frequently recorded 

species from other subtropical locations were completely 

absent (e.g. Lignincola laevis, Halorosellinia oceanica and 

Massarina spp.). Mycota of Avicennia marina and 

Rhizophora sp. in Egypt will be compared with a 

discussion for several ecological parameters. 

453 - Impact of harvest of tree tops and branches of 

Norway spruce on the biodiversity of fungi 

J. Allmér * , A. Dahlberg, J. Stenlid & R. Vasiliauskas 

Department of forest mycology and patholog, SLU, Box 

7026 SE-750 07 Uppsala, Sweden. - E-mail: 

johan.allmer@mykopat.slu.se 

There has been an increased use of logging waste, tree tops 

and branches, in Swedish forestry for biofuel purposes. The 

aims of this PhD-project are 1) to identify and describe the 

fungal communities associated with fine woody debris 

(FWD), 2) to analyse to what degree fungal communities 

overlap in composition of species between FWD, coarse 

wood debris (CWD) and the litter layer, 3) to analyse 

impacts of removal of logging waste on the diversity and 

community structure of these fungi and 4) from a fungal 

perspective, model the carbon availability in a spruce forest 

generation by considering a) the amount and quality influx 

of litter from CWD, FWD, cones, needles and the field 

layer and b) the decomposition rate of these fractions. I will 

present results from the first study, where we have 

identified and compared the fungal species composition in 

branches, tree tops and stumps, seven years after thinning. 

The identification was conducted by analyses of ITS 

sequences both directly from wood samples and from pure 

cultures. As a comparison, sporocarps were monitored on 

the same samples. 

454 - Septoria diversity, biogeography and life history 

T.V. Andrianova 

M.G. Kholodny Institute of Botany, NAS Ukraine, 

Tereshchenkivska, 2, Kiev, 01601, Ukraine. - E-mail: 

tand@darwin.relc.com 

The fungal genus Septoria Sacc. contains about 1000 

names and is a paraphyletic group considered as 

coelomycetous anamorphs of the Dothideales genus 

Mycosphaerella Johanson. Sexual holomorphic species 

with Septoria anamorph phenotypes in the life cycle 

number about 14%. Most Septoria species are strictly 

asexual and very probably had independent evolution. 

Cryptic sexuality for some anamorphic species (S. 

clematidis-rectae, S. convolvuli, S. cruciatae, etc.) may be 

supposed because spermatial synanamorphs are formed in 

pure culture. A morphological continuum among Septoria 

anamorphs with related holomorphs and anaholomorphs 

confirms the aggregative character of the group. Observed 

homology of conidiogenous cells, conidia and conidiomata 

leads to reassessment of these fungi, as has already 

happened with the Cercospora complex of genera. Septoria 

species are with wide geographical distribution. Most 

common are fungi with a broad Eurasia or Europe and 

Middle Asia, Eurasia and North America geographical 

distribution. Sporadic dispersion within the limits of 

temperate-subhumid, temperate-semiarid, and 

submeridional-semiarid natural habitats is inherent to 



anaholomorphs. Continuum dispersion is more typical for 

holomorphs with Septoria anamorphs spread over the 

Eurasian-American region, while constituent teleomorphs 

sometimes are geographically limited with disjunctions in 

arid areas. 

455 - New species of Marasmius (Basidiomycetes, 

Tricholomataceae) from tropical Africa - I 

V. Antonín 

Moravian Museum, Dept.of Botany, Zelny trh 6, CZ-659 37 

Brno, Czech Republic. - E-mail: vantonin@mzm.cz 

Descriptions, drawings of microscopic features and 

photographs of 12 new taxa (11 species and 1 variety) of 

the genus Marasmius s. str. (Basidiomycetes, 

Tricholomataceae) of the sect. Epiphylli, Fusicystides, 

Globulares, Hygrometrici and Neosessiles from tropical 

Africa are presented. Six species (M. albidocremeus, M. 

camerunensis, M. lacteoides, M. muramwyanensis, M. 

kigwenensis, M. tshopoensis) belong to the sect. 

Globulares, two species and one variety (M. minutoides, 

M. minutoides var. angustisporus, M. nyika) to the sect. 

Hygrometrici, one species to the sect. Epiphylli (M. 

foliiphilus), sect. Fusicystides (M. longicystidiatus) and 

sect. Neosessiles (M. bururiensis). Species presented here 

represent a part of results of the monographic studies of the 

genus Marasmius in tropical Africa. 

456 - Macrofungal species proposed for a Tuscan red- 

List (Italy) 

D. Antonini 1* , M. Antonini 1 , A. Laganà 2 & C. Perini 2 

1 Via F. Ferrucci, 626, - 51036 Larciano (PT), Italy. - 2 Dip. 

Scienze Ambientali 'G. Sarfatti' - Università, Via 

P.A.Mattioli, 4 - 53100 Siena, Italy. - E-mail: 

dantonini@tin.it 

Due to the close relationship between fungi and 

environment, conservation of fungal species often depends 

on environmental protection. This is particularly true for 

entities that grow exclusively in certain habitats; if their 

habitat is threatened, so are they. Areas of particular 

phytogeographical interest and/or limited size were listed 

for Tuscany under the 'Bioitaly' project which implements 

the Habitats Directive 92/43; many of these areas were 

surveyed from the mycological point of view in the Tuscan 

mapping project. The mycoflora of some of these areas was 

not systematically monitored until recently. The results for 

the relict mountain mires and autochthonous spruce forest 

in the National Natural Reserve of Campolino in the upper 

Sestaione Valley (pSIC IT5130001), the coastal dunes of 

Burano Lake (pSIC IT5190032) and Diaccia Botrona 

(pSIC IT5190011) are reported here. Information on 

environmental features is given for each of the areas listed, 

together with a list of epigeous macromycetes to be 

included in a future Tuscan Red-List. The IUCN category 

140 

of each fungal species is indicated. The exsiccata of taxa 

proposed for the regional Red-List are preserved in the 

Herbarium Universitatis Senensis (SIENA). 

457 - The new project of mapping macrofungi in 

Tuscany (Italy) 

D. Antonini 1* , M. Antonini 1 , M. Mannini 2 , G. Nocentini 3 & 

C. Perini 4 

1 AGMT, via F. Ferrucci 626 - 51036 Larciano, Italy. - 

2 AGMT, via del Pino, 38 - 56025 Pontedera, Italy. - 

3 ARSIA, via Pietrapiana, 30 - 50121 Firenze, Italy. - 4 Dip. 

Scienze Ambientali 'G.Sarfatti' - Università, via 

P.A.Mattioli, 4 - 53100 Siena, Italy. - E-mail: 

dantonini@tin.it 

The Association of Tuscan Mycology Groups (AGMT) 

was formed in 1993. It currently coordinates 22 mycology 

groups, is active in scientific research and in the field of 

education. In collaboration with the Tuscan Agency for 

Development and Innovation in Agriculture and Forestry 

(ARSIA), the Tuscan Regional Administration, the 

botanical gardens and the universities, AGMT is beginning 

a new programme involving census, mapping and 

conservation of epigeous fungi in Tuscany. The first census 

(1995-98) led to publication of a check-list of about 1000 

taxa. Besides adding to the check-list, the new convention 

will draw up a regional red-list based on IUCN criteria 

(2000). The first part of the programme involves 

bibliographic research to create a Tuscan mycological 

database. The data of the mapping projects presented by 

the Italian Botanical Society (SBI), the Confederatio 

Europeae Micologiae Mediterraneensis (CEMM) and the 

European Council for the Conservation of Fungi (ECCF) 

will then be extrapolated. The database will include data on 

vegetation and substrate, frequency, and chart coordinates 

for each taxon. The exsiccata will be kept in the Herbarium 

Universitatis Senensis (SIENA). Details of the project can 

be viewed on the ARSIA website. 

458 - Fusarium species newly recognized from Japan 

since 1990 

T. Aoki 


National Institute of Agrobiological Sciences, 2-1-2 

Kannondai, Tsukuba, Ibaraki 305-8602, Japan. - E-mail: 

taoki@nias.affrc.go.jp 

Taxonomy of Fusarium in Japan has long been based on 

the Snyder & Hansen's taxonomic system, especially 

among workers in the phytopathology. The taxonomic 

system was really epoch-making and recognized only nine 

species within the giant anamorphic genus based on the 

present sense of Fusarium mycology. Application of 

taxonomic systems contradictory to it, such as 

Wollenweber & Reinking's or Booth's, was also tried in 

Japan, but recognition or description of a new taxon was


scarce. Most of study efforts have, therefore, been 

concentrated to find infraspecific groupings such as formae 

speciales or phytopathogenic races. A substantial 

reinvestigation of Fusarium species has been conducted in 

the last decade, according to the species concept based on 

recent molecular-phylogenetic analyses and precise 

comparative morphological studies. Presence of more than 

40 species of Fusarium in Japan has been until now 

recognized. Among them, F. fractiflexum, F. kyushuense 

and F. nisikadoi were new species and F. circinatum, F. 

concentricum, F. fujikuroi (= F. moniliforme MP-C), F. 

globosum, F. incarnatum (= F. semitectum var. majus), F. 

proliferatum (= F. moniliforme MP-D), F. phyllophylum, 

F. verticillioides (= F. moniliforme s. str. = MP-A) and so 

on were new records from Japan, including renaming. 

459 - Physical and chemical effects of fungi and lichens 

in rock weathering 

C. Ascaso 1 & J. Wierzchos 2* 

1 Centro de Ciencias Medioambientales, CSIC, c/ Serrano 

115 dpdo, 28006 Madrid, Spain. - 2 Universitat de Lleida, 

Servei de Microscòpia Electrònica, c/ Rovira Roure 44, 

25198 Lleida, Spain. - E-mail: jacekw@suic-me.udl.es 

The epilithic lichen thallus frequently contain microdivided 

minerals and biominerals, meanwhile mycobiont hyphae 

invade the rock surface. Within the rocks the protolichens 

and hyphae constitutes an important part of the endolithic 

biomass. The studies of organo-mineral phase constituted 

by minerals and hyphae, algae, cyanobacteria and bacteria 

should be perform by microscopy techniques that do not 

disturb the biofilm structure. These techniques should 

allow investigation of the internal ultrastructure of the 

biotic components, leading to their identification. Such 

techniques should also permit the observation of mineral 

physical changes induced by microorganisms. The 

equipment which allows the carrying out of this work is the 

SEM operating in Backscattered mode (SEM-BSE). In situ 

SEM-BSE (EDS) observation of the organo-mineral phase 

allows detection of the biomobilisation processes and 

minerals biotransformation. In Antarctic extreme 

environment, biologically transformed minerals have been 

observed, and as a result, diagenetic iron hydroxide 

nanocrystals and clays have been found around 

chasmoendolithic hyphae in Beacon sandstone. Also in 

granitic rocks the biominerals, such as calcium oxalate and 

silica deposit have been found close to the fungal cells. In 

the biomobilisation processes as well as in the biomineral 

formation, the fungi are involved leaving a distinct traces 

of their microbial activity. 

460 - Mycorrhizal fungi of seven Paphiopedilum species 

in Thailand 

P. Athipunyakom 1* , L. Manoch 2 & M. Tanticharoen 3 

1 Plant Pathology and Microbiology Division, Department 

of Agriculture, Chatuchuk, Bangkok 10900, Thailand. - 

2 Department of Plant Pathology, Faculty of Agriculture, 

Kasetsart University, Bangkok 10900, Thailand. - 

3 National Center for Engineering and Biotechnology, 

Bangkok 10400, Thailand. - E-mail: 

pornpimon_a@yahoo.com 

Healthy roots from mature plants of seven Paphiopedilum 

species, namely Paphiopedilum concolor, P. concolor var. 

striatum, P. exul, P. godefroyae, P. niveum, P. parashii and 

P. villosum, were collected from various locations in 

Thailand. Mycorrhizal fungi were isolated using a 

modification of the Masuhara and Katsuya method. Root 

pieces were washed with tap water and surface sterilized 

for 5 min in 5% NaOHCl and then rinsed twice in sterile 

distilled water. The root segments were then cut into 

longitudinal sections and the epidermal layer was removed. 

Cortical cells containing pelotons were macerated and 

placed on 1/6 NDY medium and incubated for 3-7 days. 

After the incubation period, hyphal tips were transferred to 

PDA slants and kept as pure cultures. Identification was 

based on morphological characters as observed under a 

light microscope. Nuclei were stained with safranin O 

using the Bandoni method. Three genera and six species of 

mycorrhizal fungi were found as follow: Rhizoctonia 

repens (P. concolor and P.exul), R. ramicola (P. exul), 

Ceratorhiza goodyerae-repentis (P. godefroyae), 

Epulorhiza calendulina (P. concolor var. striatum and P. 

exul), Rhizoctonia sp.1 (P. parashii) and Rhizoctonia sp.2. 

Nuclear staining revealed that all strains were binucleate 

except Rhizoctonia sp.2 (P. niveum) which was 

multinucleate. Pure cultures were maintained on slant 

PDA, tissue paper, soil and liquid paraffin for further 

studies on molecular phylogeny and symbiotic 

germination. 

461 - Canadian Collection of Fungal Cultures 

(CCFC/DAOM 

C. Babcock 

Canadian Collection of Fungal Cultures, Eastern Cereal 

and Oilseed Research Centre, Agriculture and Agri-Food 

Canada, Rm 1015, K.W. Neatby Bldg., Ottawa, Ontario 

K1A 0C6, Canada. - E-mail: babcockc@em.agr.ca 

The culture collection maintains ˜ 11,000 plant pathogenic 

and wood rotting fungal cultures representing ˜ 2,900 

species. The catalogue can be viewed on the World Wide 

WEB: http://sis.agr.gc.ca/brd/ccc. The CCFC is supported 

by the taxonomic expertise of one of the largest groups of 

systematic mycologist in North America, with 

internationally recognized expertise in most major 

taxonomic groups. Research is concentrated on solving 

agricultural problems. The collection originated as an 

amalgamation of individual working collections and now 

serves as a primary Canadian repository for research 

strains. It functions as a gene bank for this microbial 

genetic resource and provides pure cultures to researchers 

in agriculture, forestry, medicine, private industry and 

biotechnology. Authenticates strains have been provided 

for research programs involved in: crop resistance to fungal 

diseases, biological control, diagnostic probe development 

and identification, screening for new metabolites and 



enzyme pathways, post harvest diseases, and mushroom 

spawn. The methods of long term preservation of cultures 

by cryostorage (vapour phase) and lyophilization are aimed 

at longevity and the stabilization of genetic properties. An 

extensive oil collection is also maintained. Isolates are 

available to researchers at a fee. The CCFC is a member of 

the WFCC and abides by all international regulations 

concerning the distribution and transport of biological 

organisms. 

462 - Identification of genetic diversity in the ascochyta 

blight pathogen of chickpea [Ascochyta rabiei (Pass.) 

Lab.] by used RAPD markers 

A. Bagheri 1* , F. Shokoohifar 2 & M.F. Rastegar 1 

1 Ferdowsi University of Mashhad, College of Agriculture; 

P.O.Box:91775-1163, Iran. - 2 Research Center For Plant 

Science, 2 Sardadvar; Bhahar St.; Zip Code:91747484815, 

Iran. 

The poor information about genetic diversity in the 

ascochyta blight fungus is one of the basic difficulties for 

breeding resistance cultivars of chickpea to blight disease 

in Iran. In this study we have employed RAPD marker for 

distinguish genetic variation in DNA of this fungus 

isolates. We have used 12 random primers for comparison 

of 26 selected isolated isolates from 16 Iranian provinces. 

Ten primers have showed polymorphism among isolates. 

The OPK-01 primer defined the highest number of 

polymorphism (11) and the OPK-09 primer defined only 2 

polymorphism bands. On the base of all primers, isolates 

classified into 22 genotypes. The genetic diversity index 

was estimated - and revealed high variation in Iranian 

population of this pathogen. The pair-wise genetic distance 

has estimated for all isolates. This experiment 

demonstrated that RAPD markers have high ability for 

genetic analysis of this pathogen population. The 

comparison of the results of RAPD marker with virulence 

test showed the very low correlation coefficient (0.03) 

between the virulence and genetic matrixes. 

463 - Survey of Phytophthora species in oak forests in 

Austria and Turkey 

Y. Balci * , E. Halmschlager & T. Kirisits 

Inst. of Forest Entomology, Forest Pathology and Forest 

Protection, Hasenauerstraße 38, 1190 Vienna, Austria. - Email: 

balci@edv1.boku.ac.at 

Oak trees are occupying a wide range of environments and 

are of enormous ecological and economical importance. 

Since the early 20th century oak ecosystems have been 

suffering from decline and mortality. Species of 

Phytophthora cause serious disease problems in natural oak 

stands and plantations. During the course of a survey in 

declining and healthy oak stands in Austria and Turkey 

between 1999-2001 ten Phytophthora species were isolated 

142 


from soil samples using the oak leaflet baiting method. In 

total, 86 sites (35 in Austria and 51 in Turkey) were 

investigated. Phytophthora spp. were recovered from 17 

and 38 stand in Austria and Turkey, respectively. The 

assemblage of species included P. quercina, P. citricola, P. 

cinnamomi (A2), P. gonapodyides, P. syringae, P. 

europaea, P. cryptogea, Phytophthora sp.1 and 

Phytophthora sp.2. In addition, P. cambivora (A2) was 

isolated from beech (Fagus sylvatica) trees admixed in oak 

forests in Turkey. The associated oak species were Quercus 

robur, Q. petraea, Q. cerris, Q. pubescens, Q. frainetto, Q. 

hartwissiana and Q. vulcanica. Among the recovered 

species P. quercina showed the widest distribution and P. 

citricola was also common. Isolates resembling P. citricola 

could be separated into four distinct groups due to 

morphological, physiological and cultural characteristics. 

The results of this study suggest that P. quercina, P. 

cinnamomi and P. cambivora are a potential threat to oak 

ecosystems in Austria and Turkey. 

464 - A new species of Ceratocystis discovered on 

Eucalyptus nitens in Australia 

I. Barnes 1* , J. Roux 1 , B.D. Wingfield 2 , M.J. Dudzinski 3 , 

K.M. Old 3 & M.J. Wingfield 1 

1 University of Pretoria, Forestry and Agricultural 

Biotechnology Institute, Department of Microbiology and 

Plant Pathology, 74 Lunnon Road, FABI, University of 

Pretoria, Pretoria, 0002, South Africa. - 2 University of 

Pretoria, FABI, Department of Genetics, 74 Lunnon Road, 

FABI, University of Pretoria, Pretoria, 0002, South Africa. 

- 3 CSIRO Forestry and Forest Products, CSIRO Forestry 

and Forest Products, Canberra, Australia, 2604, Australia. 

- E-mail: irene.barnes@fabi.up.ac.za 

Ceratocystis fimbriata and C. eucalypti are the only two 

species of Ceratocystis reported to occur on Eucalyptus. C. 

fimbriata is recognized as a serious pathogen, especially in 

Uganda, Congo and Brazil, where it causes rapid wilting 

and death of these trees. C. eucalypti is considered native 

to Australia and is not pathogenic. It does, however, 

colonize fresh wounds on Eucalyptus. This study was 

conducted to determine whether Ceratocystis spp., 

including C. eucalypti, would infect artificially induced 

stem wounds on Eucalyptus nitens (Canberra) and 

Eucalyptus globulus (Cann River), Australia. Ten trees 

were wounded in October 2000 at each site and wounds 

were examined a month later. Perithecia, characteristic of 

Ceratocystis spp., were found covering the wounds and 

also in isolations using carrots as bait. Pure cultures were 

obtained by transferring ascospores masses from 

perithecial necks onto 2% MEA plates. The Ceratocystis 

sp. collected from Eucalyptus nitens has hat-shaped 

ascospores, different to those of C. eucalypti but similar to 

those of C. fimbriata. It differs, however, from C. fimbriata 

and all other Ceratocystis spp. in its unique pear-shaped 

perithecial bases. DNA sequence data from the ITS and 

5.8S rRNA operon confirmed that the fungus from 

Eucalyptus nitens in Australia is unique, with C. fimbriata 

and C. albofundus as its closest relatives. The Ceratocystis 

sp. discovered in this study is in the process of being 

described as a new taxon.


465 - Mycorrhizal fungal richness in ponderosa pine 

(Pinus ponderosa Dougl. ex Laws.) plantations in 

Patagonia (Argentina) 

C. Barroetaveña 

Centro de Investigación y Extensión Forestal Andino 

Patagónico, CC. 14 (9200), Esquel, Chubut, Argentina. - 

E-mail: carolina@ciefap.cyt.edu.ar 

In Argentina, there are about 2,000,000 ha of grasslands 

suitable for afforestation with fast growing non-native 

conifers along the piedmont of the Patagonian Andes, and 

ponderosa pine is the most widely planted species. Many of 

this planting sites are under water stress conditions. The 

aim of this study was to survey mycorrhizal species 

richness and their distribution along the precipitation 

gradient in ponderosa pine plantations in Patagonia. Eleven 

plantations, four under a precipitation regime of 1000 mm 

(humid plantations) and six under 400-600 mm (dry 

plantations), were surveyed for two springs and two 

autumns for hypogeous and epigeous fungi. Amphinema 

byssoides was the most widely distributed species, found in 

all plantations. The other species differed in their presence 

according to the humidity gradient. Humid plantations 

presented 9 different taxa, each plantation displaying a 

species richness between 4 to 6. Hebeloma mesophaeum 

was present in all of them, followed by Suillus luteus and 

Rhizopogon rubescens. Dry plantations presented 5 

different taxa, and each plantations displayed a species 

richness between 1 to 4. S. luteus, H. mesophaeum and R. 

rubescens were the most widely distributed. Data showed 

that species richness was strongly dependent on 

precipitation. The principal species, though, were the same, 

but their relative occurrence varied in different conditions. 

466 - Macromycetes diversity of Russia and its 

conservation ex situ in the collection LE (BIN) 

N.V. Belova 

Komarov Botanical Institute RAS, Prof. Popov str., 2. 

St.Petersburg. 197376, Russia. - E-mail: 

cultures@NJ3873.spb.edu 

Modern social and economic conditions in Russia are 

characterized by numerous negative factors and processes 

which influence on biodiversity. Loss and destruction of 

natural locations are the main danger for mycobiota 

diversity. For long-term purposes of macromycetes 

conservation ex situ specialized culture collections serve. 

During the past ten years the main trend of development of 

the Komarov Botanical Institute Basidiomycetes Culture 

Collection - LE(BIN) has been considering as conservation 

and maintaining of macromycetes cultures with emphasis 

on preservation of macromycetes from reserved territories. 

About 250 new macromycetes cultures were isolated 

during field works in Russian reserves such as 'Nizhne- 

Svirsky' (Leningrad region), 'Les na Vorskle' (Belgorod 

region), Island Valaam (Karelia) and others. LE(BIN) 

species representation was increased by new cultures from 

genera Clavicorona, Collybia, Marasmiellus, Marasmius, 

Mycena and Lentinellus. Strain diversity of Collybia, 

Marasmiellus, Marasmius and Mycena was also increased 

essentially. Considerable representation of biological 

species of the genus Lentinellus is maintained in the 

Collection. The genus Clavicorona is presented by six 

strains of the typical species Clavicorona pyxidata. 

Cultures of ectomycorrhizal fungi, rare species and ones 

that were recommended for conservation in Russia are also 

preserved in the Collection LE(BIN). 

467 - Ecological, management, policy and community 

research in British Columbia relevant to the 

commercial harvest of wild forest mushrooms 

S.M. Berch 1* , B. Chapman 2 , J. Dennis 3 , T. Ehlers 4 , S. 

Fredrickson 4 , M. Kranabetter 5 , D. Mitchell 6 , R. 

Outerbridge 7 , M. Peterson 7 & S. Tedder 1 

1 2 

BC Ministry of Forests, Victoria, BC, Canada. - BC 

Ministry of Forests, Williams Lake, BC, Canada. - 3 Pacific 

4 

Forestry Centre, Victoria, BC, Canada. - Tysig 

Consulting, Winlaw, BC, Canada. - 5 BC Ministry of 

Forests, Smithers, BC, Canada. - 6 Royal Roads University, 

Victoria, BC, Canada. - 7 Applied Forest Science Ltd., 

Victoria, BC, Canada. - E-mail: 

shannon.berch@gems7.gov.bc.ca 

Tricholoma magnivelare (Pine Mushroom) and 

Cantharellus formosus (Chanterelle) are commercially 

important wild forest mushrooms in British Columbia. 

Recently, research to integrate these mycological resources 

into forest planning has been carried out. Productive Pine 

Mushroom habitat throughout BC was characterized and 

the common factor found was dry and nutrient poor soil. It 

was then determined in north western and south central BC 

that it is possible to protect and manage Pine Mushroom 

habitat without an unacceptable impact on timber 

harvesting because of the relatively small area of highly 

productive Pine Mushroom habitat and low tree 

productivity of this habitat. On Haida Gwaii, a study 

determined that Chanterelle productivity on sites burned 

prior to regeneration was greater than on similar sites with 

no fire history. In British Columbia over 95% of the 

forestland is publicly owned which creates challenging 

policy and regulatory issues. A study of social and 

economic potential of non-timber forest products (NTFPs) 

on Haida Gwaii discussed many of the issues resulting 

from the annual influx of mushroom pickers. A subsequent 

study assessed how property rights could be used to 

overcome concerns and enhance stewardship. Integrating 

many of these findings and concerns, the Northern 

Vancouver Island NTFP Demonstration Project is 

researching how best to manage non-timber forest 

resources in a way that is environmentally sustainable, 

economically viable and socially equitable. 



468 - The BIOTA Southern Africa project: concepts 

and first results 

R. Berndt, C. Görke * , M. Mennicken, E. Uhlmann & F. 

Oberwinkler 

University of Tuebingen, Spezielle Botanik & Mykologie, 

Auf der Morgenstelle 1, 72076 Tuebingen, Germany. - Email: 

claudia.goerke@uni-tuebingen.de 

The BIOTA (Biodiversity Monitoring Transect Analysis in 

Africa) Southern Africa project is a long term, 

multidisciplinary project that aims at monitoring 

biodiversity along the rainfall gradient from the Cape of 

Good Hope in South Africa to the Angolan border in 

Namibia. The Cape of Good Hope receives 500 mm 

rain/year. The area at the Namibian/South African border is 

extremely arid with 50 mm rain/year, a transition between 

winter and summer rainfall occurs here. Annual rainfall in 

summer increases towards northern Namibia, which 

receives 450 mm/year. In addition to rainfall regimes, 

different land management systems are compared. This 

poster presents first results from a study of Namibian fungi, 

focussing on soil fungi, arbuscular mycorrhizae, and rust 

fungi. Soil fungi were isolated by soil washing; arbuscular 

mycorrhizae were studied with morphological and 

molecular tools. This revealed differences in species 

composition along the Namibian part of the transect. Rust 

fungi are monitored to assess their biodiversity under 

different climatic and management conditions. In Namibia 

rust fungi are collected countrywide to compile a national 

checklist. 

469 - Some Aphyllophorales in mediterranean 

ecosystems 

A. Bernicchia 1* & E. Salerni 2 

1 Dipartimento di Scienze e Tecnologie Agroambientali, 

Universita' di Bologna, Via F. Re 8, 40126 Bologna, Italy. 

- 2 Dipartimento di Scienze Ambientali, Universita' di Siena, 

Via P.A. Mattioli 4, 53100 Siena, Italy. - E-mail: 

abernicc@pop.agrsci.unibo.it 

The knowledge on distribution of Aphyllophorales in 

Mediterranean area has a pre-eminent position both from 

an ecological and taxonomic point of view. This interest is 

essentially due to the peculiarity of the Mediterranean 

vegetation patterns and, consequently, of the woodinhabiting 

fungi. The drop in rainfall during the last years, 

with the consequent increase in aridity, determined 

considerable changes in the vegetation with a short-lived 

fruiting period as first effect. In this context, we report the 

results of researches concerning the Mediterranean 

Aphyllophorales that, in spite of the studies carried out 

during the last years are till now poorly known. Between 

the collected species are worthwhile: Neolentiporus 

squamosellus (Bernicchia & Ryvarden) Bernicchia & 

Ryvarden, growing on burnt Juniperus oxycedrus, in 

Supramonte di Orgosolo (NU) Sardinia. It is the only 

144 


species of Neolentiporus present in the northern 

hemisphere while N. maculatissimus Rajchenb. is growing 

in the southern hemisphere. Echinodontium ryvardenii 

Bernicchia & Piga is the first European collection of 

Echinodontium while all previous collections are North 

American or Asiatic. It must looked upon as a relict of a 

previously larger distribution. Piloporia sajanensis 

(Parmasto) Niemelä is a rare species with a boreal 

distribution, collected in Sardinia on J. oxycedrus. It is 

suggested that the isolated locality could be an Ice Age 

relict from a time when Italy was covered with coniferous 

forests. 

470 - Fungal biodiversity in a regeneration series in 

Colombia Amazonia 

T. Boekhout 1* , A.E. Franco Molano 2 , C.A. Lopez 

Quintero 2 , M. Silvestri 2 , A.M. Cleef 3 & R.C. Summerbell 1 


3584 CT Utrecht, The Netherlands. - 2 Instituto de Biologia, 

Universidad de Antioquía, Medellín, Colombia. - 

3 2Institute for Biodiversity and Ecosystems Dynamics, 

University of Amsterdam, Amsterdam, The Netherlands. - 

E-mail: boekhout@cbs.knaw.nl 

Fungal biodiversity has been investigated in a series of 

secondary forests and two types of primary lowland forest 

in Colombian Amazonia. Functional inferences suggest a 

role for ectomycorrhizal fungi and litter decomposing fungi 

in the nutrient cycling of these forests. Decomposition of 

leaf litter can be relatively fast, but seems to be influenced 

by the environmental conditions. Shortly after trees are cut 

down, many fruitbodies of wood-inhabiting fungi occurred. 

This raises the question whether these fungi may have 

occurred as endophytes in the healthy trees. The formation 

of fruit bodies by these fungi may be a result of the lack of 

stress imposed by the death of the hosts, and the 

availability of a suitable substrate. After the felled trees are 

burnt, soil-borne microfungal populations are dominated by 

heat resistant fungi. Many new taxa of macro- and 

microfungi have been found, and await formal description. 

Analysis of secondary metabolites in soil borne Penicillia 

revealed some clues to explain the observed uptake of iron 

during decomposition of leaf litter. Initial observations 

suggest the presence of c. 15 undescribed species of 

Penicillium. A novel antagonistic interaction between 

bacteria and certain soil borne basidiomyceteous yeasts has 

been observed. Associated research has been initiated to 

establish potential use of fungi by the indigenous 

populations.


471 - Yeasts of the world, an interactive CD-ROM 

T. Boekhout 1* , V. Robert 1 , J. Stalpers 1 , G. Gijswit 2 , C.P. 

Kurtzman 3 , J.W. Fell 4 & I. Roberts 5 


3584 CT Utrecht, The Netherlands. - 2 Expertisecenter 

Taxonomic Indentification, Amsterdam, The Netherlands. - 

3 NRRL-USDA, Peoria, U.S.A. - 4 RSMAS, University of 

Miami, Key Biscayne, U.S.A. - 5 NCYC, Norwich, U.K. - Email: 

boekhout@cbs.knaw.nl 

This CD-ROM presents a complete taxonomic data set of 

all currently accepted yeast species, including 

morphological and physiological data, and ribosomal DNA 

sequences. The interactive software contains modules for 

the comparison and integrated use of physiological, 

sequence and morphological information, facilitating the 

identification of yeasts using complementary data sets. 

Many species are illustrated by microscopic and 

macroscopic images. This product will be useful in a wide 

range of yeast studies throughout the agro-industrial and 

medical sciences. 

472 - Biodiversity of discomycetes associated with 

coniferous trees 

A.V. Bogachova 

Institute of Biology and Soil Science FEBRAS, Stoletya- 

Street, 159 Vladivostok-22, Russia. - E-mail: 

cryptogamy@ibss.dvo.ru 

Ligniphylous discomycetes are developing on the various 

woods. Ones of they have special ferments for destroying 

of a broad-leaves wood. Other has special ferments for 

destroying of conifers. They to some extent show a species 

specialization according to plants. Discomycetes are not 

aggressive wood-destroying fungi. But some species of 

discomycetes produce diseases, death or a depression of a 

developing of plant. These fungi settle on a bark of robust 

trees and provide a saprotrophic cycle of life. It is not get 

up 1,5 meter on a stem of tree. If a resistance power of tree 

slacken, fungus change self cycle of life to parasitic. Some 

species of Discomycetes develop fruit-bodies only on 

coniferous trees. At first they invade the bark of healthy 

trees and live as saprotrophic fungi. If a resistance of trees 

is weakened, these fungi change their way of life and 

become parasitic. When the tree dies, these fungi come 

back to the saprotrophic way of life. In accordance with 

literature data, 43 species of Discomycetes were reported 

on Pinus spp., 37 - on Abies, 18 - on Picea, and 12 - on 

Larix. In coniferous forests, Discomycetes develop on 

attached and fallen needles and branches in the litter. In the 

latter case their number is smaller in comparison with 

wood-destroying species. 

473 - The Biodiversity of Basidiomycota in Thailand 

T. Boonpratuang 1* , P. Ruksawong 1 , N.L. Hywel-Jones 1 & 

T. Flegel 2 

1 BIOTEC, 73/1 Rama VI Rd., Bangkok, Thailand. - 

2 Mahidol University, Rama VI Rd., Bangkok, Thailand. - Email: 

artbp@biotec.or.th 

The biodiversity of Thailand's fungi is poorly known. Until 

1990 less than 500 spp. (of all fungi) had been reliably 

reported. Since 1990 increased effort has been made to 

survey this rich biodiversity. Thailand is entirely tropical 

offering a range of habitats from rain forest in the south to 

dry deciduous forest in the north. Elevations are from sea 

level to over 2500 m (Doi Inthanon). In the last ten years 

ca. 3100 specimens, from 40 orders, 96 families, 326 

genera and 677 spp. have been added to the BIOTEC 

Herbarium. Most accessions were from 1992 to 2001 from 

various parts of Thailand. Within the herbarium there are 

significant deposits from many researchers having either 

short-term or long term associations with young Thai 

researchers. These activities continue to the present. A 

major effort is being made with the marasmioid and 

mycenoid fungi of Thailand but in the course of that work 

other agarics have also been collected. Notable records of 

these for Thailand include: Cintractia amazonia, Fistulina 

hepatica, Crinipellis zonata, Amanita hemibapha, 

Strobilomyces velutipes, Dacryopinax spathularia, 

Clavulinopsis miniata, Irpex flavus, Steccherinum 

rawakense, Craterellus verrucosus, Anthracophyllum 

nigritum and Mutinus bambusinus. 

474 - Marasmioid and mycenoid fungi of Thailand 

T. Boonpratuang 1* , P. Ruksawong 1 , N.L. Hywel-Jones 1 , T. 

Flegel 2 & D.E. Desjardin 3 

1 BIOTEC, 73/1 Rama VI Rd., Bangkok, Thailand. - 

2 Mahidol University, Rama VI Rd., Bangkok, Thailand. - 

3 San Francisco State University, Holloway, U.S.A. - Email: 

artbp@biotec.or.th 

Marasmius (1600+ spp.) and Mycena (1200+ spp.) are the 

most diverse genera of saprobic basidiomycetes. The 

greatest species diversity is in the tropics, although 

hundreds of taxa are found in temperate regions of both 

southern and northern hemispheres. Nearly all species are 

saprotrobic, having important roles in litter decomposition, 

nutrient recycling/retention, pedogenesis etc. A few are 

parasitic on important food crops. Importantly, some form 

antibiotics such as strobilurin. Many species are host- or 

substrate-specific accounting for some of the high diversity 

seen in tropical forest. Over 460 samples were collected 

from various parts of Thailand. Of these, 93% were 

identified to species or morphotaxa while 7% remain 

unidentified. To date, 100 genera and 51 species are 

known. Samples were classified into three major groups of 

fungi; namely marasmioid, mycenoid and other 

macrofungi. Marasmioid fungi were 25% of the collection, 



mycenoid 40% and other agarics 35%. Of the marasmioid 

fungi, 78% were the genus Marasmius. Two new species of 

marasmioid fungi have been named: Incrustocalyptella 

orientalis and Clitopilus chalybescens. New records of 

Marasmius include M. conicopapilatas, M. papuraceus, M. 

purpureostriatus and M. micraster. Of the mycenoid fungi, 

57% were Mycena. Notable new species include: M. 

dermatogloea, M. khonkhem, M. palmicola, M. pseudoseta 

and M. minicoseta. 

475 - The diversity of Ingoldian fungi in Thailand 

N. Boonyeun * , S. Sivichai & N.L. Hywel-Jones 



Technology Development Agency, 113 Pahonyothin Rd.. 

Khlong 1, Khlong Luang, Pathum Thani12120, Thailand. - 

E-mail: nattawut@biotec.or.th 

While much work on Ingoldian fungi is from temperate 

regions knowledge of tropical species is poor. Surveys 

were made in the last 10 yr, especially for Khao Yai 

National Park. Foam samples were collected from 8 sites in 

Thailand. Rivers were ca. 5-10 m with overhanging forest. 

Foam samples were fixed in the field before spotting to 

microscope slides for examination. From all sites, 49 spp. 

were found. A sample from Khao Yai had only 2 spp. 

while one from Bala Hala had 25. This river (Khlong I- 

Gading) supported 28 spp. including 11 not reported 

elsewhere. By contrast, the similar Lumtakhlong (Khao 

Yai) had 11 spp. in total: none were exclusive to this river. 

In the north of Thailand Nam Nao National Park had 10 

spp. recorded but half were exclusive to this site. 

Triscelophorus spp. were present in all streams. Other 

commonly recorded genera included Lunulospora spp. and 

Anguillospora spp. Clavatospora tentacula and 

Brachiosphaera tropicalis were also regularly found. Of 

the 49 spp. reported, 26 were known from a single site. 

Most species were present as single spores in foam 

samples. But species of Anguillospora, Campylospora, 

Flabellospora and Triscelophorus were represented by 

many spores. While these reports were mostly of 

mitosporic forms of Ascomycota a significant find was the 

discovery of Erynia conica and Erynia rhizospora. These 

insect pathogenic Zygomycetes are mostly known from 

temperate regions and poorly surveyed in the tropics. 

476 - Novel characters of New Zealand polypore fungi: 

septate basidiospores and spore print colour 

P.K. Buchanan 1* & L. Ryvarden 2 


Zealand. - 2 Department of Botany, University of Oslo, P.O. 

Box 1045, Blindern, N-0316 Oslo, Norway. - E-mail: 

buchananp@landcareresearch.co.nz 

146 


About 170 species of polypore fungi in 60 genera are 

currently recognised in New Zealand, with over 80% of 

accepted names differing from those used in the most 

recent monograph published in 1965. Half of the genera 

contain a single New Zealand species, and only one genus, 

Australoporus, is considered to be endemic to the region. 

Many collections of polypore fungi in the New Zealand 

Fungal Herbarium (PDD) bear misapplied Northern 

Hemisphere names. Among these collections we recently 

discriminated two new species that have septate 

basidiospores: Polyporus septosporus and Dichomitus 

newhookii. These species are characterised by occasionally 

uni- to multiseptate basidiospores, a feature newly recorded 

among Aphyllophorales, and apparently also unknown 

among agarics. Septation appears to be confined to older 

spores, especially those dispersed from the tubes. Spore 

print colour, considered taxonomically useful for agarics, 

also has application for polypore fungi and provides the 

only useful field character to differentiate two common 

New Zealand species of Ganoderma. 

477 - Endophytic and plant pathogenic fungi from 

terrestrial orchids in Thailand 

K. Busarakam 1 , L. Manoch 1* , C. Chamswarng 1 & P. 

Suwanarit 2 


Kasetsart University, Bangkok, 10900, Thailand. - 

2 Department of Microbiology, Faculty of Science, 

Kasetsart University, Bangkok, 10900, Thailand. - E-mail: 

agrlkm@ku.ac.th 

Healty leaves and roots of two terrestrial orchids, Ludisia 

discolor and Spathoglottis plicata collected from Lopburi 

and Chaing Mai provinces were used to isolated 

endophytic fungi. A modification of Okane's method was 

employed. The samples were washed with tap water and 

the leaves were cut into small pieces of 5x5 mm. Leaf 

pieces and the whole root were surfaced sterilized for 1 

min in 70% EtOH and 2 min in 10% sodium hypochlorite 

and rinsed 2-3 times with sterile distilled water. Leaf and 

root segments were placed on sterile filter paper and 

transferred on 2% malt extract agar and incubated for 2-3 

days. Endophytic fungi found on leaves included 

Gliocladium penicilloides, Colletotrichum coccides, 

Nodulisporium gregarium, Pestalotiopsis guepinii, 

Pestalotiopsis sp. and Xylaria spp., whereas 2 isolates of 

binucleate Rhizoctonia and 1 multinucleate species were 

found from the healthy roots. In a second investigation, 

disease leaves of three terrestrial orchids: Spathoglottis 

affinis, Spathoglottis plicata and Phaius tankervilleae 

showing spot symptoms, were collected from Bangkok, 

Lopburi and Trang Provinces. The tissue transplanting 

method with 10% clorox and potato dextrose agar was 

used. Plant pathogenic fungi found in this study were 

Cladosporium cladosporioides, Colletotrichum 

gloeosporioides, Curvularia eragrostidis, C. lunata, 

Curvularia sp., Cylindrocladium sp., Drechslera 

hawaiiensis, Fusarium oxysporum, Pestalotiopsis guepinii, 

Phomopsis sp. and Phyllosticta sp.


478 - A preliminary study of Polyporales and poroid 

Hymenochaetales from Equatorial Guinea 

J.N. Campoamor * & M.T. Tellería 

Real Jardín Botánico, C.S.I.C., Plaza de Murillo 2, 28014 

Madrid, Spain. - E-mail: campoamor@ma-rjb.csic.es 

Within the scope of the project 'Catalogación y estudio de 

la diversidad vegetal y fúngica del bosque virgen 

continental de Guinea Ecuatorial' the basic aim of this work 

is to elaborate a catalogue of Polyporales and poroid 

Hymenochaetales from the area. We start with the study of 

samplings collected from 1998 to 2000 at different places 

of the territory, in most cases at the 'Monte Alen National 

Park ' (ECOFAC). Up to the date, the material 

corresponding to the genus Amauroderma, Favolus, 

Ganoderma, Lenzites, Loweporus, Microporus, Oxyporus, 

Polyporus, Phellinus, Pycnoporus, Rigidoporus, 

Schizopora and Trametes, has been examinated. 

479 - Additions to the discomycetes of the Dominican 

Republic and the Caribbean Region 

S.A. Cantrell 1* , D.H. Pfister 2 & T. Iturriaga 3 

1 Science & Technology, Universidad del Turabo, PO Box 

3030, Gurabo, PR 00778, U.S.A. - 2 Harvard University 

Herbaria, 22 Divinity Ave., Cambridge, MA 02138, U.S.A. 

- 3 Universidad Simon Bolivar, Apartado Postal 89000, 

Sartenejas, Baruta, Edo. Miranda, Venezuela. - E-mail: 

sharonac@coqui.net 

The discomycetes of the Dominican Republic and the 

Caribbean region are poorly known, especially the 

inoperculate. In a previous presentation made by S. A. 

Cantrell she reported for the Dominican Republic 9 and 12 

new records in the Pezizales and Helotiales, respectively. 

Some of the species were new records for the Greater 

Antilles and the Caribbean. In a recent collecting trip made 

in January 2002, 111 discomycetes were collected divided 

in 22 Pezizales, 81 Helotiales, and 8 Rhytismatales. Some 

new additions within the Pezizales are: Helvella macropus, 

Plectania cf. platensis, Pachyella babingtonii, a beautiful 

black-green Peziza, a burnt-orange Kompsoscypha and an 

orange-yellow Pulvinula. Within the Helotiales, some new 

additions are: Dicephalospora rufo-carnea, 

Podophacidium xanthomelum, Rhizodiscina lignota, 

Hyaloscypha aureliella, Polydesmia cf. dumontii and 

Hyalorbilia inflatula. So far, we know that there are 75 

species of discomycetes in the Dominican Republic, 

divided in 28 Pezizales and 47 Helotiales. Within the 

Pezizales, 24 are new reports for the Domincan Republic, 

10 for the Caribbean region and 5 might represent new 

species. For the Helotiales, all are new reports for the 

Dominican Republic, 10 for the Caribbean region and 15 

might represent new species. 

480 - Mycogeographic affinities of Tabasco (México) 

polypores 

S. Capello 1 , J. Cifuentes 2 , F. García-Pantaleón 3 & V. 

Carballo 1* 

1 División de Ciencias Biológicas. UJAT, Av. Universidad 

s/n, Villarhermosa,Tabasco, Mexico. - 2 Herbario FCME, 

Fac. de Ciencias, UNAM, A.P. 70-399 Coyoacán, C.P. 

04510, México D.F., Mexico. - 3 Departamento de Biología 

Vegetal U de C., Campus Rabanales, Cordoba, Spain. 

A quantitative mycogeographic analysis was made to asses 

the mycological affinities of the southern tropical Tabasco 

State (Mexico) based on the inventory of 61 wood polypore 

species (Cappello, 2001). The Multi-Variate Statistical 

package was used to perform UPGMA groupings. In the 

first part of the analysis Tabasco's polypores were 

compared with polypore listings from the mexican 

phytogeographic regions as defined by Rzedowski (1978) 

and in the second part they were compared with selected 

regions of the world such as Africa Asia, Cuba, Europe, 

Northamerica (divided in regions after Gilbertson & 

Ryvarden, 1986), Central and Southamerica. Interesting 

Tabasco appears somewhat separated when compared with 

the mexican regions and the remaining areas form two 

distinctive groups. Furthermore Tabasco forms a distinctive 

group with SE northamerica, Cuba and southamerica when 

compared with the chosen world areas which in part agrees 

with Guzmán (1973), Ryvarden (1998) and Welden & 

Lemke (1961) findings. 

481 - Epiphytic lichens on different fractions of Norway 

spruce Picea abies 

A. Caruso 

Department of Conservation Biology, Swedish University 

of Agricultural Sciences (SLU), Box 7002, SE-750 07 

Uppsala, Sweden. - E-mail: Alexandro.Caruso@nvb.slu.se 

The use of logging residues as fuel in Sweden has 

increased during the last decades but still only a small part 

of the total wood fuel supply is today being harvested. 

Since there is a potential for a further increase in the 

harvest of logging residues efforts are now being made to 

investigate the effects this may have on the forest flora and 

fauna. My research is focused on fruticose, foliose and 

crustose lichens, both epiphytic and epixylic. As a first step 

it is of great interest to examine the structure of the lichen 

community on a whole-tree basis. Several lichen studies 

have been made on basal trunks and basal branches but tree 

tops and branches higher up in the canopy have been 

poorly investigated. The aim of this study is to describe the 

species composition of epiphytic lichens on different 

fractions of Norway spruce Picea abies, a common tree in 

northern Europe. The study area, Kilsbergen, is located in 

the boreonemoral zone of Sweden northwest of the city of 

Örebro (59°17'N, 15°13'E). In this area I selected ten forest 

stands dominated by Norway spruce and with similar stand 



characteristics. In each stand I then made a whole-tree 

inventory of epiphytic lichens on two felled individuals of 

Norway spruce. 

482 - Filamentous fungi isolated from soil of mangrove 

at the Itamaracá island, state Pernambuco, Brazil 

M.A.Q. Cavalcanti * , I.P.M.W. Costa, M.J.S. Fernandes, 

D.M.M. Lima & Z. Passavante 

Universidade Federal de Pernambuco, Rua Nelson Chaves 

s/n, 50670-420 Recife, Brazil. - E-mail: 

xiliamac@terra.com.br 

Filamentous fungi were isolated from mangrove soils 

collected at the Itamaracá Island, State of Pernambuco, 

Northeast Brazil, using the serial dilution technique. 

Samples (1ml) from the final dilution (1:1000) were 

tranferred to Petri dishes with Sabouraud plus 

chloramphenicol and maintained at room temperature 

(28°C ±1°C), with the growth of colonies followed for 72 

hours. The following taxa were identified: Aspergillus 

niger, A. sydowi, A. terreus, A. ustus, Cladosporium 

sphaerospermum, Eupenicillium brefeldianum, Eurotium 

chevalieri, Penicillium corylophilum, P. lividum, P. 

restrictum, Scopulariopsis sphaerospora, Spegazzinia 

tessarthra, Talaromyces bacillosporus, T. trachyspermus, 

T. flavus and Stilbella sebacea. Scopulariopsis 

sphaerospora and Talaromyces bacillosporus are being 

referred for the first time to Brazil. 

483 - The mycoflora of hot spring soil of northern 

Taiwan 

K.Y Chen * , C.C Liu & D.J Huang 

Department of Biology, Chinese Culture University, Taipei 

112, Taiwan. - E-mail: thermophilic@ms52.url.com.tw 

An investigation on the mycoflora, particularly of 

thermophilic and thermotolerant fungi inhabiting on 

sulfurous hot spring soils of northern Taiwan resulted 12 

taxa viz. Aspergillus fumigatus var. fumigatus (66.85%), A. 

fumigatus var. 1 (green colony) (7.86%), A. fumigatus var. 

2 (brown colony) (4.81%), A. niger (1.14%), unidentified 

Aspergillus sp.(0.045%), Papulospora thermophila 

(2.72%), Chrysosporium sp. (0.18%), Scytalidium 

thermophilum (0.045%), Sporotrichum sp. (0.045%), 

Mycelia Sterilia sp.1 (white colony) (6.63%), M. Sterilia 

sp. 2 (yellow colony) (5.27%) and M. Sterilia sp.3 (gray 

colony) (4.405%). 2202 colonies were isolated from three 

sampling sites, i.e. site 1 (hot springhead), site 2 (2 m from 

site 1) and site 3 (4 m from site 1). Fungal colonies isolated 

as well as species richness in three sites were as follows: 

32.92% in 9 taxa from site 1, 37.87% in 11 taxa from site 2 

and 29.21% in 8 taxa from site 3, respectively. The 

dominant species was A. fumigatus var. fumigatus that was 

isolated year around from three sampling sites. A. 

fumigatus var. 1 appeared from Feb. to Jun. (2000). In 

148 


contrast, A. fumigatus var. 2 was isolated only in Aug. and 

Oct.(1999). Within the sampling range of present hot 

spring niche, there is an evidence of presence of ecotypes 

in A. fumigatus complex. Chrysosporium and Sporotrichum 

were isolated only from the soils without hot water 

treatment but Aspergillus sp. and S. thermophilum were 

isolated only from the soils pre-treated with hot water for 

30 min at 60°C. 

484 - The fluctuation of marine Mastigomycota in 

estuary tidewater of Tsengwen River 

S.-F. Chen 

Chia Nan University of Pharmacy and Science, 60 Erh-Jen 

Road, Section 1, Jen Te, 717, Tainan, Taiwan. 

The occurrence and seasonal fluctuation of marine 

zoosporic fungi were investigated at four stations in the 

estuary tidewater of Tsengwen River, southwestern 

Taiwan. Thirteen species of Thraustochytrids and seven 

species of Chytrids were isolated. The highest species 

abundance (S) recorded in summer was 15. The highest 

species similarity (CC) recorded between the spring and 

winter was 77.78. According to their frequency at 

individual stations, T. aggregatum, T. proliferum, U. 

minuta were ubiquitous, while the rest were common to 

scare. Chytrids was only found at tidewater of stations 1L 

and 1M in summer and autumn. The presence of 

Thraustochytrids and Chytrids revealed fluctuation when 

the saltility descend to zero. 

485 - Two new species of Amanita from Korea 

D.H. Cho 

Woosuk University, Chonju, 565-701, Korea. - E-mail: 

chodh@woosuk.ac.kr 

Two new species of Amanita were collected in central 

areas of Korea from 1997 to 2000. They were identified, 

described and illustrated. One of them, Amanita longstipeta 

has turtle-shaped warts of crust and its base is longer under 

the soil than is above the soil. The other A. aureofarinosa is 

covered with golden yellow farina and annulus is absent. 

Both are solitary in soil with sand. 

486 - Database of Korean mushrooms 

D.H. Cho 1* , W.K. Cho 1 , J.Y. Chung 1 , H.S. Park 2 & B.Y. 

Ahn 2 

1 Woosuk University, Chonju, 565-701, Korea. - 2 KISTI, 

Taejeon, 565-701, Korea. - E-mail: chodh@woosuk.ac.kr


In Korea about 2000 species of mushrooms (higher fungi) 

have been studied from basdiomyctes to ascomycetes. 

Among them, about 1500 species are constructed in 

database. This database contains several items; 

taxonomical system, the application, the ecological 

resources, the geographical distributions. Each of 

mushrooms is illustrated with descriptions, photographies 

and spores. Also, information retrieval system is available 

by using KRISTAL II for query searches on the Web in 

URL http://ruby.kisti.re.kr/^mushroom. 

487 - Macrofungi distribution in a region of the Sierra 

Madre Oriental (México): an approximation of area of 

endemism methods 

J. Cifuentes * , M. Villegas, R. García-Sandoval, G. Vidal, S. 

Sierra-Galván, R. Valenzuela, L. Pérez-Ramírez & E. 

Morales-Torres 

FCME Herbaria, UNAM., Po.Box 70-399, CP 04510, 

Coyoacán, DF, Mexico. - E-mail: 

jcb@hp.fciencias.unam.mx 

Several protocols of areas of endemism methods (Morrone, 

1994, Jardine, 1972, Linder 2001) were applied to a 

mapped list of 220 species of selected groups of 

macrofungi (Stipitate hydnoids, polypores, Amanita, 

Laccaria, Psilocybe and Scleroderma. Taxa were chosen 

both for practical (better known and collected groups) and 

ecological(to represent both saprotrophs and biotrophs) 

reasons. The best result, lesser fragmented areas, was 

obtained with Linder's method of inverse weighting. 

Though three well defined groups were observed they were 

not supported enough to be considered robust. Nevertheless 

when these groupings were contrasted with vegetation and 

altitudinal levels maps it was found an acceptable non 

exact concordance with the last one; one subgroup related 

with lower altitudes is accordingly distributed. Contrary to 

our findings based on the application of parsimony 

methods, previous ecological studies in Mexico had shown 

a relation of macrofungi distribution and vegetation 

(Guzmán and Guzmán-Dávalos 1979, Varela and Cifuentes 

1979, Heredia 1989) but they were based solely on 

similarity list comparisons. Tan & Wu (1986) in China 

rather found that macrofungi distribution was highly 

correlated with temperature which agrees better with our 

results. In spite of limitations to the use of herbaria data to 

apply areas of endemism methods, our results support to 

certain extent their applicability. 

488 - The 609 red listed fungi in Sweden: an analysis of 

their ecological characteristics, distribution and threats 

A. Dahlberg * & H. Croneborg 

Swedish Species Information Centre, P.O.Box 7007, 750 07 

Uppsala, Sweden. - E-mail: 

anders.dahlberg@artdata.slu.se 

Red lists are extremely important instrument in both 

national and international conservation work. The recent 

Swedish Red List is based on an evaluation of factors that 

affect risk of extinction, a system to classifying species into 

threat categories developed by the World Conservation 

Union (IUCN), which is successively replacing the old 

systems around the world. The Red List comprises 609 

species, of which 254 are assessed as threatened, selected 

from an assessment of the 4620 known Swedish 

macromycetes (http://www.artdata.slu.se/home.htm). In 

principle, all available knowledge of each of these fungi 

have been compiled and analysed; life forms, preferences 

of habitats and substrates, distribution within and outside 

Sweden, and threats. The poster will summarize their 

ecological characteristics and identify the major threats. 

The red list can be looked upon as a deficiency analysis of 

habitats and substrates. Among the most severe problem is 

the large scale on going fragmentation of valuable biotopes 

like primeval forest with a high amount of dead wood of 

appropriate quality and unfertilised grassland. Today, an 

alarming number of fungal populations survive only in 

small number of isolated populations. Reference: 

Gärdenfors U (ed) 2000. Red List of Swedish Species. 

ArtDatabanken, SLU Uppsala. 

489 - Polypores on Pinus koraiensis in NE China 

Y.C. Dai 

Institute of Applied Ecology, Academia Sinica, Shenyang 

110016, Wenhua Road 72, China. - E-mail: 

yuchengd@yahoo.com 

Pinus koraiensis is the main coniferous tree in NE China. 

The species of polypores on wood of the tree were 

investigated, and 79 species were recognized. Among them 

23 species are considered as rare species, 31 are occasional 

and 25 are common. 16 species were found only in forest 

reserves, 12 in unprotected forests, and 49 in both forest 

reserves and unprotected forests. 6 species were found on 

dead trees, 28 on fallen trunks, 41 on rotten wood and 4 on 

charred wood. 19 species occur on Pinus koraiensis only, 

and 60 on other substrates as well. 

490 - 'Guide des champignons comestibles du Bénin', a 

tool for valorisation, preservation and sustainable use 

of West-african savanna woodlands 

A. De Kesel 1* , J.T.C. Codjia 2 & S.N. Yorou 3 

1 National Botanic Garden of Belgium, Domein van 

Bouchout, B-1860 Meise, Belgium. - 2 Faculté des Sciences 

Agronomiques, Université d'Abomey-Calavi au Bénin, 01 

BP 526, Cotonou, Bénin. - 3 Laboratoire d'écologie 

Appliquée, Faculté des Sciences Agronomiques, Université 

d'Abomey Calavi au Bénin, 01 BP 526, Cotonou, Bénin. - 

E-mail: dekesel@br.fgov.be 



Wild edible mushrooms are highly appreciated by local 

people in Bénin (West Africa). The majority of edible 

species in Bénin, however, are not cultivatable because 

they are ectomycorrhizal or associated with termites. In 

Bénin, as well as in the neighbouring countries, many 

ecosystems suffer from the influence of human activities. 

This guide is a tool for valorisation, preservation and 

sustainable use of West-african savanna woodlands and 

helps the reader to recognise the place of the fungi, their 

identity and function within the natural world. Aspects 

concerning morphology, ecology and distribution of fungi 

in general, as well as aspects on poisoning, cultivation, 

recording local knowledge, ethnomycology and methods 

for assessing natural productions and valuation of wild 

edible fungi. The guide presents over 70 colour pictures 

and drawings, accompanied by morphological descriptions, 

local names and information on distributions and edibility 

of more than fifty wild species. Everyone interested in 

collecting or studying West-African fungi will find this 

book a useful reference. Those dealing with the sustainable 

use and management of natural resources in tropical Africa, 

will find a multitude of techniques and tools for a better 

management and conservation of natural ecosystems in a 

participative way, i.e. with the participation and for the 

benefit of local people. This guide is to be released by the 

end of June 2002. 

491 - Soil microfungi in a banana (Musa paradisiaca) 

plantation in Tabasco (México) 

M. Del Olmo, J. Cifuentes * , G. Vidal & J.E. Rosique 

FCME Herbaria, UNAM., Po.Box 70-399, CP 04510, 

Coyoacán, DF, Mexico. - E-mail: 

jcb@hp.fciencias.unam.mx 

Soil fungi have been extensively studied all over the world 

(Viaud et al., 2000; Stchigel, et al., 2000) but poorly and 

rather recently in Mexico (Mena et al., 1998; Bills et al., 

2001). Since Mexico has been recognized as one of the 

megadiverse countries (Guzmán, 1998) it is a priority to 

undertake studies increasing our mycobiota knowledge. 

Furthermore micromycetes are increasenly important as a 

source of new bioactive metabolites and enzymes for 

biotechnology (Pointing & Hyde, 2001). Soil samples were 

taken in a banana plantation at the Teapa county, Tabasco 

State (17°32'N, 92°57'E) in a three months intervals during 

August 2001 to May 2002. To isolate fungi plate dilution 

and direct soil techniques on PDA/streptomycin and 

bengala rose added were used. So far a listing of 30 species 

and 15 genera of mostly mitosporic fungi has been 

obtained, which agrees in general with other banana 

plantation soil fungi listings Goos (1960, 1963). 

Abundance and frequency data are provided to discern 

phenological and successional patterns. 

492 - Flora of the macrofungi of South Cameroon 

C. Douanla-Meli * & E. Langer 

150 



Str. 40, Germany. - E-mail: cmeli@yahoo.com 

The macrofungi of South Cameroon have been surveyed 

for the first time including detailed illustrations of the 

micromorphology. In the megatherm forests with high 

precipitation of South Cameroon the Aphyllophorales are 

the most abundand fungal group with 60% of all collected 

material followed by agaricoid fungi (32, 7%). Fungi from 

the Auriculariales make 3% of all collections. Some 

important species are presented including illustrations of 

their macro- and micromorphology. 

493 - Mycorama - an international center of mycology 

in Switzerland 

S. Egli 1* & J. Keller 2 

1 Swiss Federal Research Institute WSL, 8903 Birmensdorf, 

Switzerland. - 2 Association suisse du Mycorama, 2053 

Cernier, Switzerland. - E-mail: simon.egli@wsl.ch 

In 1998 the Swiss Association of the Mycorama (ASM) - 

representing 400 members and 50 mycological societies - 

started with the planification of this project, examining its 

feasibility and finding the necessary funding. The start of 

the construction of the mushroom-shaped building is 

planned for the end of this year, in the Jura region near 

Neuchatel in Western Switzerland. The Mycorama will be 

an international museum presenting the many aspects of 

mycology, between myth and magic, medicine and 

gastronomy, ecology and economy, addressing itself as 

much to the public as to the mycologist. It will include 

facilities for the cultivation of edible species, a demo 

laboratory and a restaurant with typical mushroom dishes. 

494 - Mycorrhizal plants of traditionally managed 

boreal grasslands in Norway 

M. Eriksen 1* , K. Bjureke 2 & S.S. Dhillion 1 

1 Dep. of Biology and Nature Conservation, Agricultural 

University of Norway, P.O. Boks 5014, 1432 Ås, Norway. - 

2 Natural History Museum and Botanical Garden, P.O. 

Boks 1172, Blindern, 0318 Oslo, Norway. - E-mail: 

marit.eriksen@ibn.nlh.no 

This poster reports on the mycorrhizal status of 82 plant 

species growing in traditionally managed grasslands in 

three different locations in the boreal and boreo-nemoral 

vegetation zone in the eastern part of Norway. Seventyfour 

species were found to have arbuscular mycorrhiza 

(AM). To our knowledge, we report AM for the first time 

in Achillea ptarmica, Ajuga pyramidalis, Alchemilla 

glaucescens, Carex brunnescens, Carex pallescens, Crepis 

praemorsa, Hieracium lactucella, Rumex longifolius, 

Scorzonera humilis, Trifolium aureum and Trifolium 

spadiceum. The rare and threatened species Arnica 

montana, S. humilis, C. praemorsa, Gentianella


campestris, Parnassia palustris, T. aureum and T. 

spadiceum, all confined to grasslands, were found to 

possess AM fungi. 

495 - Fungi from the highlands of Eastern Iceland, an 

area at risk because of the proposed Karahnjukar 

hydropower project 

G.G. Eyjolfsdottir 

Icelandic Institute of Natural History, Akureyri Division, 

Hafnarstraeti 97; P.O.Box 180; IS-602 Akureyri, Iceland. - 

E-mail: gge@ni.is 

This study of the macromycota of the highlands (450-650 

m a.s.l.) in Eastern Iceland was a part of research aimed at 

assessing the environmental impact of the proposed 

Karahnjukar hydropower project. Fungi were collected at 

the proposed site of a 57 km 2 , 24 km long reservoir 

Halslon, to be formed by building dams in the river Jökulsa 

a Bru and flooding its channel and surrounding area. 

Collection took place in middle of August 2000, at 24 

transects for vegetation analyses. The fungi were 

photographed and dried in the field and later identified. A 

total of 35 species of macromycetes and four plant parasitic 

micromycetes were identified. Most were typical highland 

fungi, e.g. Omphalina pyxidata, Lactarius pseudouvidus, 

Russula nana, Laccaria laccata, Hebeloma mesophaeum, 

H. kuehneri, Inocybe calamistrata, Cortinarius subtorvus, 

C. alpinus, Lactarius pubescens, Puccinia septentrionalis, 

Exobasidium vaccinii-uliginosi and Rhytisma salicinum, 

found in four or more transects. Those found in one 

transect only were: Anthracoidea elynae, Bovista 

nigrescens, Entoloma sericeum, Galerina clavata, 

Galerina pseudomycenopsis, Geopora arenicola, Helvella 

albella, Hygrocybe turunda, Inocybe geophylla, I. hirtella, 

I. lacera, Laccaria pumila, Lyophyllum decastes and 

Scutellinia patagonica, some of which are more common at 

lower elevation or confined to specific habitats. The known 

mycota of the highlands of Eastern Iceland is compared to 

that of Central Iceland. 

496 - Rapid approach to evaluate persistence of fieldintroduced 

AMF by nested PCR 

M.J. Farmer 1* , D. van Tuinen 1 , B. Zhao 2 , X. Li 3 , G. Feng 3 , 

W.K. Chan 4 , S. Gianinazzi 1 & V. Gianinazzi-Pearson 1 

1 UMR 1088 INRA/Université de Bourgogne BBCE-IPM, 

INRA-CSME, BP 86510 21065 Dijon cedex, France. - 2 Key 

Laboratory of Agromicrobiology (Ministry of Agriculture, 

China) College of Life Science and Technology, Huazhong 

Agricultural University, Wuhan 430070, China. - 

3 Department of Plant Nutrition, College of Natural 

Resources and Environment Sciences. China Agricultural 

University, Beijing 100094, China. - 4 Department of 

Applied Biology & Chemical Technology, Hong Kong 

Polytechnic University, Hunghom, Koloon, Hong Kong, 

China. - E-mail: farmer@epoisses.inra.fr 

In China 74% of agricultural soils (more than 7 million 

hectares) are phosphorus-deficient. Arbuscular mycorrhizal 

fungi (AMF), which form symbiotic associations with plant 

roots, are involved in the maintenance of sustainable soil 

fertility. Colonization of plant roots by such fungi improves 

the availability and uptake of soil phosphate resulting in 

increased plant growth. Within the framework of a 

European Project (1) focussing on the use of AMF 

technology to improve staple food crop production in 

small-scale sustainable agriculture in China, one of our 

objectives is the use of molecular tools to monitor 

persistence, within host roots, of selected AMF inoculated 

onto three target food crops: maize, sweet potato and 

cassava. We have studied different techniques of 

conservation of mycorrhizal root systems and genomic 

DNA (DNAg) extraction. A method of grinding in a Tris 

EDTA buffer with the addition of activated charcoal was 

retained for its simplicity, rapidity and efficiency. The 

large ribosomal subunit of DNAg was amplified by nested 

PCR using eukaryotic universal primers for the first PCR, 

and primers specific for selected AMF(2) inoculated onto 

target crops, for the second. (1) INCO-DEV: 

MYCHINTEC No ICA4-CT-2000-30014. (2) van Tuinen, 

D., Jacquot, E., Zhao, B., Gollotte, A., Gianinazzi-Pearson, 

V. (1998) Characterization of root colonization profiles by 

a microcosm community of arbuscular mycorrhizal fungi 

using 25S rDNA-targeted nested PCR. Molecular Ecology 

7: 879-887. 

497 - Influence of resource size on wood-inhabiting 

fungal communities in a tropical forest 

A. Ferrer 

Smithsonian Tropical Research Institute, Apartado 2072, 

Panama. - E-mail: astridferrer@hotmail.com 

The diversity of ascomycetes and basidiomycetes woodinhabiting 

communities were examined for different 

resource sizes of the host tree Prioria copaifera in five 

low-diversity and high-diversity forests in central and 

eastern Panama. A positive correlation between size of the 

resource and basidiomycetes diversity was found, large 

dead branches and trees had a large number of 

basidiomycetes than small branches or small diameter 

trees. The same significant relationship was found for 

ascomycetes in three of these five sites. A relationship 

between stage of decay and number of fungal species was 

also tested, and more basidiomycetes species were found at 

early stages of decay in three sites. However, this 

relationship was weak and explained less than 10% of the 

deviance in species diversity. Stage of decay did not have 

any effect on the number of ascomycetes species. A field 

experiment was conducted to determine the effect of the 

size of the resource on fungal colonization. Resource size 

was manipulated using different sized wood blocks of 

Prioria, placed in two sites and censured every 3 months 

for 15 months. The size of the blocks did had a positive, 

significant effect on the fungal species number, and the 

most common species were able to fruit in both small and 

large wood blocks. These results suggest that resource size, 

and therefore forest structure, is an important determinant 

of fungal community structure. 



498 - Description of Lactarius indigo and L. deliciosus 

mycorrhizas on nine pine species 

R.E. Flores 1* , M. Honrubia 1 , G. Díaz 1 & M.C. Bran 2 

1 University de Murcia, Depto. Biología Vegetal (Botánica), 

Facultad de Biología. Campus de Espinardo 30100, Spain. 

- 2 Universidad de San Carlos de Guatemala, DIGI/Fac. 

CCQQ y Farmacia, Edif T-12 Ciudad Universitaria. 

01012, Guatemala. - E-mail: rfloresa@yahoo.com 

The main macro and microscopical features of mycorrhizas 

of Lactarius indigo Schw. ex Fr. and Lactarius deliciosus 

Fr. with five species of Mesoamerican pines (Pinus 

ayacahuite, Pinus hartwegii, P. oocarpa, P. pseudostrobus 

and P. rudis) are presented. Also description with 

microscopical and SEM pictures of mycorrhizas of L. 

indigo with Pinus halepensis, P. nigra, P. pinaster and P. 

sylvestris are shown. The work was made in a growth 

chamber and using plastic containers with peat mossvermiculite 

substrate. Inocula of selected strains of 

Lactarius deliciosus and L. indigo were applied directly to 

the radicular systems of the seedlings. The results seems to 

be the first published for Lactarius indigo, a known edible 

species in America, especially in Mexico and Guatemala, 

and also the mycorrhizas of L. deliciosus with 

Mesoamerican pines. These results are very important to 

the production of mycorrhized plants in Central America, 

especially with these edible mushrooms, because they 

could be of use in potentiating reforestation and for 

providing extra income and food to farm people, specially 

in Guatemala, Honduras, El Salvador and Nicaragua. 

499 - Russian species of the genus Alternaria Nees 

Ph.B. Gannibal 

St.Petersburg State University, Universitetskaja nab. 7-9, 

St.Petersburg, 199034, Russia. - E-mail: 

philipp@rsp.spb.org 

Alternaria is a very widespread genus of dematiaceous 

fungi. Many species are serious pathogens of crops. 

Despite on long history of its investigation, there are some 

different opinions about genus taxonomy. Formerly closed 

species were distributed between two genera - Alternaria 

and Macrosporium Fr. In 1969 the name Macrosporium 

was abolished, and the name Alternaria became the only 

(Levkina L.Ì. 1984. Mycol. and Phyt. 18, 1). The genus 

Alternaria was revised several times. There are more than 

100 described species now, but not all descriptions are 

admitted. The paper of Egorova about fungi on Russian Far 

East contains the most complete list of Alternaria species 

in Russia (Egorova L.N. 1999. Mycol. and Phyt. 33, 1). 

There are 43 well-known and 7 insufficiently investigated 

species in this region. They infect 72 plants. A. citri Ell. et 

N. Pierce, A. mali Roberts, A. longipes (Ell. et Everh.) 

listed by Egorova are considered A. alternata (Fr.) Kiessl. 

now (Rotem J. 1994). There are a few papers, which add 

some other species in total list. For instance, there is A. 

152 


saponariae (Peck) Neerg. in the roll of Russian arctic fungi 

(Rus. Arctic Fungi. 1999). Our collections include some of 

these Alternaria species. In two main Russian mycological 

herbariums we reveal at least 5 species, which are absent in 

the list of Alternaria species from Far East. Using prior 

analysis of literature and herbarium material, we can 

conclude that there are about 50 Alternaria species in 

Russia. 

500 - Diversity of ectomycorrhiza in Quercus rubra L. 

stands of different age on reafforested mining sites 

S. Gebhardt * , J. Wöllecke & R.F. Hüttl 

Brandenburg University of Technology, Chair of Soil 

Protection and Recultivation, Postfach 101344, 03013 

Cottbus, Germany. - E-mail: gebhardt@tu-cottbus.de 

Red oaks are planted on 15% of reforested mining sites in 

Lower Lusatia and thus belongs to the main tree species 

used in reclamation. The mining substrates are 

characterized by poor nutrient supply e.g. N and P. Under 

such adverse conditions mycorrhizal fungi may play an 

important role for the vitality and the survive of these trees. 

One aim of this investigation is to charakterize the 

morphotypes of mycorrhizae of Quercus rubra. We want to 

know if the fungi are above all generalists or if the fungi 

are species adapted to these extrem conditions. An other 

aim is to compare the ectomycorrhizal fungi which are 

common with the neophyt Quercus rubra and indigenous 

oak species like Quercus petrea. Soil samples are taken in 

oak stands of different age within an area of 720 m& 2 ;. The 

mycorrhiza are separated by morphological and genetic 

methods. To characterize the extrem soil conditions, the 

samples are analysed of pH, phosphate, nitrate, ammonium, 

C, N, S, water retention and bulk density. First results 

show, that 19 morphotypes of mycorrhizal fungi can be 

separated by their mycorrhizae. Some fungi can be 

identified by their fruitbody, for example Boletus edulis or 

Amanita muscaria. Scleroderma citrinum and Paxillus 

involutus can be found as fruitbody and mycorrhiza. The 

generalists and 'early stage fungi' Cenococcum geophilum 

and Paxillus involutus have an important part in the fungi 

coenoses. 

501 - Aphyllophorales (Basidiomycotina) of the Atlantic 

Rain Forest in Northeast Brazil I. States of Sergipe and 

Alagoas 

T.B. Gibertoni * & M.A.Q. Cavalcanti 

Universidade Federal de Pernambuco, Rua Nelson Chaves 

s/n, 50670-420 Recife, Pernambuco, Brazil. - E-mail: 

tbgibertoni@hotmail.com 

Eight collections of Aphyllophorales were undertaken 

between October 2000 and July 2001 in 3 Atlantic Rain 

Forest remnants in Northeast Brazil. In the State of Sergipe 

4 species of Polyporaceae (Hexagona hydnoides, H.


papyracea, Megasporoporia cavernulosa, Trametes 

caperata); 2 species of Podoscyphaceae (Caripia 

montagnei, Cymatoderma dendriticum) and 1 species of 

Hymenochaetaceae (Phellinus gilvus) were identified. In 

the State of Alagoas 19 species of Polyporaceae 

(Coriolopsis rigida, Daedalea sprucei, Earliella scabrosa, 

#Echinochaete brachyporus, Hexagona hydnoides, H. 

papyracea, #Junghuhnia luteoalba, Lentinus crinitus, 

Lenzites stereoides, Megasporoporia cavernulosa, 

Nigrofomes melanoporus, Physisporinus vitreus, 

Rigidoporus linteus, Pycnoporus sanguineus, Trametes 

caperata, #T. lactinea, T. versicolor, T. villosa, Trichaptum 

abietinum); 10 species of Hymenochaetaceae 

(#Hymenochaete fulva, H. luteo-badia, Phellinus discipes, 

P. gilvus, P. fastuosus, #P. inermis, P. merrillii, #P. 

portoricensis, P. punctatus, P. rimosus); 2 species of 

Ganodermataceae (Ganoderma applanatum, G. lucidum); 2 

species of Podoscyphaceae (Caripia montagnei, 

Cymatoderma dendriticum); 2 species of Stereaceae 

(Lopharia cinerascens, Stereum lobatum); 1 species of 

Corticiaceae (Trechispora farinacea) and 1 species of 

Schizophyllaceae (Schizophyllum commune) were 

identified. All the species are new occurrences to the States 

and those marked with a # represent new occurrences to 

Brazil. 

502 - Aphyllophorales (Basidiomycotina) of Atlantic 

Rain Forest in the Northeast Brazil II. State of 

Pernambuco 


Rua Nelson Chaves s/n, 50670-420 Recife, Pernambuco, 

Brazil. - E-mail: tbgibertoni@hotmail.com 

Twelve collections of Aphyllophorales were undertaken 

between September 2000 and July 2001 in 4 Atlantic Rain 

Forest remnants in Northeast Brazil. In the State of 

Pernambuco 23 species of Polyporaceae (#Coriolopsis 

asper, Fomes fasciatus, Fomitella supina, Hexagona 

apiaria, H. hydnoides, H. papyracea, Lentinus crinitus, 

Lenzites stereoides, Megasporoporia cavernulosa, 

Microporellus obovatus, Nigrofomes melanoporus, 

*Nigroporus vinosus, *Perenniporia martius, Polyporus 

dictyopus, *P. guianensis, P. leprieuri, P. tenuiculus, 

Pycnoporus sanguineus, Rigidoporus microporus, *R. 

ulmarius, Trametes caperata, T. membranacea, 

Trichaptum abietinum); 11 species of Hymenochaetaceae 

(Hymenochaete aspera, H. luteo-badia, #H. pinnatifida, 

*Phellinus calcitratus, P. gilvus, P. fastuosus, P. linteus, P. 

rimosus, P. senex, *P. tropicalis, P. umbrinellus); 5 species 

of Ganodermataceae (Amauroderma omphalodes, *A. 

sprucei, Ganoderma applanatum, G. lucidum, G. 

resinaceum); 3 species of Podoscyphaceae (Caripia 

montagnei, Cymatoderma dendriticum, *Podoscypha 

ravenelii); 3 species of Stereaceae (Lopharia cinerascens, 

Stereum australe, S. lobatum); 1 species of Corticiaceae 

(*Trechispora farinacea); 1 species of Hericiaceae 

(#Stecchericium fistulatum) and 1 species of 

Schizophyllaceae (Schizophyllum commune) were 

identified. The species marked with a # are new 

occurrences to Brazil and those marked with an * represent 

new occurrences to the State of Pernambuco. 

503 - Aphyllophorales (Basidiomycotina) of Atlantic 

Rain Forest in the Northeast Brazil III. States of 

Paraíba and Rio Grande do Norte 


Rua Nelson Chaves s/n, 50670-420 Recife, Pernambuco, 

Brazil. - E-mail: tbgibertoni@hotmail.com 

Seven collections of Aphyllophorales were undertaken 

between October 2000 and May 2001 in 3 Atlantic Rain 

Forest remnants in Northeast Brazil. In the State of Paraíba 

11 species of Polyporaceae (Antrodia albida, Fomes 

fasciatus, Fomitella supina, Hexagona hydnoides, Lentinus 

crinitus, Megasporoporia cavernulosa, Nigrofomes 

melanoporus, Polyporus dictyopus, Pycnoporus 

sanguineus, Trametes caperata, Trichaptum abietinum); 3 

species of Hymenochaetaceae (#Hymenochaete dura, 

Phellinus gilvus, P. merrillii); 2 species of Podoscyphaceae 

(Caripia montagnei, Cymatoderma dendriticum); 2 species 

of Stereaceae (Stereum hirsutum, S. lobatum); 1 species of 

Ganodermataceae (*Ganoderma applanatum) and 1 

species of Schizophyllaceae (Schizophyllum commune) 

were identified. All the species represent new occurrences 

to State of Paraíba, except the one marked with an *. In the 

State of Rio Grande do Norte 7 species of Polyporaceae 

(Fomes fasciatus, Hexagona hydnoides, Lentinus crinitus, 

L. velutinus, Megasporoporia cavernulosa, #Perenniporia 

martius, Trametes caperata); 7 species of 

Hymenochaetaceae (#Hymenochaete fulva, Phellinus 

fastuosus, P. gilvus, #P. maxonii, #P. meleoporus, P. 

rimosus, P. umbrinellus); 1 species of Corticiaceae 

(Trechispora farinacea) and 1 species of Schizophyllaceae 

(Schizophyllum commune) were identified. All the species 

are new occurrences to the State of Rio Grande do Norte. 

The species marked with a # in both States are new 

occurrences to Brazil. 

504 - Floristics of Burroughs Mountain, Mount Rainier 

National Park, Washington State, USA 

K.A. Glew * , C.L. Miller & L. Koepke 

University of Puget Sound, Department of Biology, 1500 N. 

Warner, Tacoma, WA 98416, U.S.A. - E-mail: 

kglew@ups.edu 

Washington State houses three national parks with alpine 

zones, yet limited baseline documentation has been 

undertaken regarding species inventories or descriptive 

works of lichen communities. These areas are subject to 

disturbance from both hikers and climbers. Vascular plant 

surveys have been carried out in limited areas of the parks, 

but do not include lichens. In the summer of 2001 a study 

was conducted on Burroughs Mountain in Mount Rainier 

National Park to focus on an inventory of lichen species 

and to study these sensitive communities. At 14,410 ft 

(4392m) elevation, Mount Rainier reflects alpine 

communities rarely seen in other areas of the state or the 

Pacific Northwest. Burroughs Mountain is made up of 



three plateaus ranging from 7000 to 7500 ft. Although each 

plateau is subject to the same climatic conditions and at 

very similar elevations, the vegetation of each plateau is 

drastically different. Snowmelt conditions and substrate 

appear to be major factors affecting the species of lichens 

found at each location. Analyses include lichen species 

richness, abundance, frequency, and the association of 

macrolichens with plant communities. Data was analyzed 

using PC-ORD and CANOCO. An inventory of lichens 

was also conducted at Camp Muir (10,000ft) to determine 

rare and sensitive lichen species found at this elevation. 

Highly subject to disturbance, this is the base camp for 

climbers hiking to the summit of Mount Rainier. A list of 

rare lichens collected will be presented. 

505 - Contribution to the knowledge of diversity and 

distribution of ascomycetes from Mexico 

M.C. Gonzalez 1* , R.T. Hanlin 2 , M. Ulloa 1 & E. Aguirre 1 

1 Departamento Botanica Instituto de Biologia, UNAM I, 

Mexico City DF 04510, Mexico. - 2 Department of Plant 

Pathology, University of Georgia, Athens Georgia 30602- 

7274, U.S.A. - E-mail: mcgv@ibiologia.unam.mx 

Diversity surveys of Mexican Ascomycetes from terrestrial 

or aquatic environments are limited mostly by a lack of 

necessary resources for mycological biodiversity studies. 

However, currently in Mexico there is interest in 

biodiversity issues, especially in relation to the 

conservation of our natural resources. The importance of 

biodiversity in this country is well recognised, and there is 

a long history of Botanical, Mycological and Zoological 

research which still continues to the present day. The 

current study aims to assess the diversity of Ascomycetes 

from Mexico. The numbers of Mexican Ascomycetes 

recorded from the literature in relation to global estimates 

of fungal diversity are also addressed in this work. The 

Pezizales and Xylariales are the best studied groups, 

probably because they are among the largest Ascomycetes 

and therefore more easily collected. The most explored 

areas in the country are the State of Veracruz and the State 

of Mexico, both located in the central-southeast region of 

the country. The least studied areas are the State of Nayarit, 

the State of Aguascalientes, and the Peninsula of Baja 

California. 

154 


506 - Biodiversity of keratinophyles in Indian hills 

(Shimla, H.P.) and Plain (Agra, U.P) and perforator 

nature of dermatophytes in relation to taxonomy 

N. Govil * & J.N Shrivastava 

Dayalbagh Educational Institute, Microbiology lab, Deptt 

of Botany, faculty of Science, Dayalbagh Educational 

Institute, Dayalbagh, AGRA, India. - E-mail: 

nidhi_dei@yahoo.com 

A comparative biodiversity of keratinophyles reveals that 

hilly area was rich in both species content and relative 

diversity as showing 100% prevalence of keratinophyles 

where as plains shows 80% prevalence as these areas differ 

markedly in prevailing environmental conditions. The 

percentage prevalence of Chrysosporium indicum (26%) 

and Microsporum fulvum (26%) was maximum indicating 

its dominance in hilly area. In plains Trichophyton equinum 

(20%) and T. rubrum (20%) was dominant. Out of seven 

genera of keratinophyles three isolated genera of 

dermatophytes were subjected for their ability to perforate 

hair in vitro considering only their superficial pigmented 

characteristics. Fourteen isolates belonging to three genera 

of Microsporum, Trichophyton and Epidermophyton were 

found to have wide range of flexibility in pathogenecity 

evaluating that they can be distinguished on the basis of 

their perforating nature without identification. M. fulvum 

was found to be best perforator releasing maximum protein 

(5.98 µg/ml). Among the six strains of M. baullardii strain 

6 was found to be non-perforator indicating their nonpathogenic 

nature although similar in microscopic and 

macroscopic morphology with other strains. So it can be 

considered as atypical strain and can achieve a nascent 

taxonomic status. The taxonomic status of all the fourteen 

isolates of three genera and seven species was studied with 

respect to their morphology. 

507 - Screening of keratinophyles isolated from Indian 

hill (Shimla, H.P.): A biotechnological approach 

N. Govil * & J.N. Shrivastava 


of Botany, Faculty of Science, Dayalbagh Educational 



Eighteen keratinophilic species was isolated from Indian 

hills (Shimla, H.P.) belonging to seven genera 

Chrysosporium, Microsporum, Trichophyton, 

Epidermophyton, Emmonsia, Geomyces and Zymonema 

were screened for their ability to degrade keratin (feather) 

in vitro. All test fungi was found positive for producing 

keratinase enzyme, total protein and sulphur containing 

amino acids on keratin degradation. G. pannorum and M. 

fulvum produced maximum keratinase (20.5 Ku/ml and 

14.2 Ku/ml respectively, protein (6.42 µg/ml and 5.84 

µg/ml respectively) indicating the biotechnological 

importance of their pathogenic nature.


508 - Above and below ground community of 

ectomycorrhizal fungi on beech from four small gaps in 

mature natural and managed beech forests 

T. Grebenc 1* , M. Christensen 2 & H. Kraigher 1 

1 

Slovenian Forestry Institute, SI-1000 Ljubljana, Vecna pot 

2, Slovenia. - 

2 

KVL, Unit of Forestry, DK-1958, 

Rolighedsvej 23, Denmark. - E-mail: 

tine.grebenc@gozdis.si 

The mycelium of ectomycorrrhizal (ECM) fungi represents 

a crucial link between forest ground vegetation and biotic 

and abiotic sources of nutrients. Mapping of fruitbodies has 

been shown as unadequate for studies of the distribution of 

the fungal community. We have applied a combined 

method of mapping of fruitbodies, identification of their 

PCR-ITS-RFLP patterns and identification of types of 

ECM after anatomical characteristics. The abundance of 

types of ECM have been studied in four natural or manmade 

canopy gaps in beech forest, two in Denmark and 

two in Slovenia. The fruitbodies were mapped and 

collected in two growing seasons on the comparable 

research plots and the ECM root tips were analysed twice 

in a transect line through each of the plots. In soil samples 

(V=274 ml) all roots were cleaned, nonmycorrhizal, old 

mycorrhizal root tips and morphotypes were separated and 

counted. On each plot from 14 to 33 different species of 

ECM fungi were determined. In the soil samples from all 

four plots 37 different types of ECM were separated, from 

these 31 on beech. 21 of them have been determined to the 

species level. Only ca one half of the ECM species 

determined from mycorrhizal root tips corresponded to the 

fruitbodies from the same plot. Some of the fruitbodies 

were not yet found in ECM. For identification of unknown 

ECM root tips the PCR-ITS-RFLP patterns have been 

compared with the database of RFLP patterns, while 

sequencing of the ITS region in rDNA has also been 

started. 

509 - Biodiversity patterns: kinds of rarity of the 

corticioid fungi (Basidiomycetes) in Tierra del Fuego 

(Argentina) 

A.G. Greslebin 


Patagónico, CC 14, 9200, Esquel, Chubut, Argentina. - Email: 

alina@ciefap.cyt.edu.ar 

Usually fungi do not appear in the lists of endangered 

species. This is probably due to our limited knowledge of 

some ecological features of the species such as the 

vulnerability. The form of rarity, based on the geographic 

distribution, local abundance and habitat specificity, helps 

us identify the species that should be protected and the way 

this protection should be carried out in a certain area. 

During a floristic study of Corticiaceae s.l. of Tierra del 

Fuego the kind of rarity of the species was evaluated. Six 

extensive collecting were made along 3 years, visiting 18 

localities selected in 5 areas. The kind of rarity of each 

species was estimated based on: 1. Frequency measured as 

the number of collecting where the species was recorded. 2. 

Geographic distribution range in Tierra del Fuego 

measured as the number of localities visited where the 

species was recorded. 3. Substrate specificity measured as 

the number of woody substrates where the species was 

recorded. A total of 104 were distributed in kinds of rarity 

as follows: Common (class 0): 15 sp.; slightly rare (classes 

1-2): 26 sp.; rare (classes 3-4): 8 sp.; highly rare (classes 5- 

7): 55 sp. Of 26 endemic species of the area 13 were highly 

rare, 10 were slightly rare and 3 were common. Of 4 

species with an austral distribution 3 are extremely rare and 

1 slightly rare. This estimation of rarity considers the 

situation of the species only in Tierra del Fuego and did not 

aim to be generalized for other areas. 

510 - A comparative account of poisonous mushroom 

biodiversity of Turkey and northeast North America 

F. Gucin 1 & B.A. Bunyard 2* 

1 Fatih University, Biology Dept., Buyukcekmece, 34900 

Istanbul, Turkey. - 2 Ursuline College, Biology Dept., 2550 

Lander Road, Pepper Pike, OH 44124, U.S.A. - E-mail: 

bbunyard@ursuline.edu 

Mushroom poisonings are reported every year in Turkey. 

The primary reason for this is that local persons may be 

unfamiliar with poisonous mushrooms and are unable to 

distinguish between edible and poisonous mushroom 

species. Ninety two taxa of poisonous mushrooms occur in 

Turkey. These are listed here together with their locations 

as well as poisoning syndromes such as: Phalloides 

syndrome (ten species, one variety), Muscarine syndrome 

(nine species), Pantherina syndrome (three species), 

Gyromitra syndrome (seven species), Coprinus syndrome 

(one species), Psilocybin syndrome (four species), Paxillus 

syndrome (two species), Gastrointestinal syndrome (32 

species). Furthermore, there are cases of mushrooms that 

may be poisonous if consumed raw (15 species) and 

mushrooms with suspected constituents (five species), as 

well as those reported to be poisonous if consumed with 

alcoholic drinks (three species). Similar poisoning 

accidents have been reported from Northeastern parts of 

North America, but are less common. This study 

demonstrates a comparison of these findings. 

511 - Saprotrophic mycelium mats in the southern rain 

forest of Mexico: effects on fine roots and soil 

arthropod diversity 

R. Guevara * & I. Romero 

Instituto de Ecología, A. C. Departamento de biología de 

Suelos, AP 63 CP 91000 Xalapa, Ver., Mexico. - E-mail: 

roger@ecologia.edu.mx 



Mycelium mats are patches with a high rate of leaf litter 

decomposition and at the same time represent a high 

density of hyphae available for soil fungivores. We 

investigated the spatial and temporal distribution of 

mycelium mats on the soil of a rain forest in Mexico and 

whether fine roots and soil arthropods were influenced by 

mats. The abundance and size of mats varied seasonally. 

There was a low number of mats after the rainy season and 

in general they were small. By contrast in the dry and early 

rains seasons mats were abundant and on average they 

were larger than those found after the rainy season. Also 

there was an effect of mats on the distribution of fine roots. 

In the dry season the amount of observed roots in non-mat 

soil was 4-fold than that observed in mat soil and by early 

rains there was more roots in mat soil than in non-mat soil. 

Also, mats affected the chemical content of fine roots. 

Roots in mat soil had lower concentration of Ca y K over 

the year whereas effects on Na concentration varied among 

seasons. Soil arthropods were also effected by mycelium 

mats. Arthropod diversity was higher in mycelium mat soil 

than in non-mycelium mat soil. This study shows that the 

guild of mycelium mat forming fungi is an important 

component of the tropical rain forest in southern Mexico 

beyond its potential species richness. Mats effect the 

distribution and chemical content fine roots (plants) and 

diversity of soil arthropods two key components of 

ecosystem diversity. 

512 - A database of Italian keratinophilic fungi 

M. Guglielminetti 1* , N. Solari 1 , S. Martellos 2 & G. Del 

Frate 1 

1 Dep. Ecologia del Territorio e degli Ambienti Terrestri - 

Sez. Micologia - Univ. di Pavia, Via S. Epifanio, 14 - 

27100 Pavia, Italy. - 2 Dep. Biologia - Univ. di Trieste, Via 

l. Giorgieri, 9 - 34127 Trieste, Italy. - E-mail: 

mariag@et.unipv.it 

The term 'keratinophilic' is used both for fungi capable of 

decomposing keratin, including those that cause superficial 

mycoses in humans and animals (dermatophyte), and for 

fungi only able to use products of keratin's hydrolysis, or 

material which naturally associates to it (1). Since the 

beginning the 19th century many studies were carried out 

in Italy about this group of fungi, isolated from soil, 

humans or animals. An on-line database is being created 

which could be useful for mycologists, physicians and 

veterinaries needing information on keratinophilic fungi 

isolated in Italy. This database presently includes 118 

infrageneric taxa and will be available on the web before 

the end of 2002. A list of Italian papers in which a species 

is cited is provided as well. This is the first step towards a 

larger nomenclatural database covering all of the Italian 

microfungi. Two other databases are being implemented, 

which will contain morphological and physiological 

information, and will be connected to the first one to 

generate a complex information system. This database is 

part of larger co-operative effort, involving several other 

Italian research centres, the MIUR project 'A network of 

databases on the diversity of terrestrial cryptogams in Italy'. 

References 1. Kirk, P.M., Cannon, P.F., David, J.C. & 

156 


Stalpers, J.A., 2001. Ainsworth & Bisby's Dictionary of the 

fungi. 9th Edition. CAB International, Wallingford UK. 

513 - Biodiversity informatics and the development of 

the GLOPP information system 

G. Hagedorn * , M. Burhenne, M. Gliech, T. Gräfenhan & 

M. Weiss 





The GLOPP project (Global Information System for the 

Biodiversity of Plant Pathogenic Fungi) is creating an 

integrated information system on the biodiversity of plant 

pathogenic fungi and their hosts by gathering descriptive 

and collection data, literature references, resource 

information and taxonomic names. The IT project within 

GLOPP provides data entry applications and develops the 

public access system on the Internet. This task requires the 

development of new concepts and information models as 

well as substantial efforts in application development. To 

simplify analysis and improve collaboration with other 

developers, a modular and component based approach has 

been choosen. This 'DiversityWorkbench' application suite 

ultimately will contain major application components like 

DiversityDescriptions (derived from DeltaAccess version 

1.7), DiversityReferences (with SpecialIndexing for 

organism interactions and distribution), 

DiversityCollection, DiversityTaxonomy and 

DiversityIdentify as well as support modules like 

DiversityUsers, the DiversityGazetteer, 

DiversityResources, or DiversityExsiccatae. Developers 

interested in collaboration are invited to discuss how the 

total development cost and effort can be shared. 

514 - Digitization of important European host-pathogen 

indices for fungi 

G. Hagedorn 1* , G. Deml 1 , W. Brandenburger 2 & M. 

Burhenne 1 

1 Federal Biological Research Center for Agriculture and 



Berlin, Germany. - 2 @THE DATABASE DID NOT ALLOW 

TO LEAVE THIS EMPTY BUT IT SHOULD BE@, 

Ostenallee 48, 59063 Hamm, Germany. - E-mail: 

g.hagedorn@bba.de 

The GLOPP-LIT project focuses on capturing published or 

unpublished scientific information on plant pathogenic 

fungi. In the framework of the collaborative GLOPP 

project (Global Information System for the Biodiversity of 

Plant Pathogenic Fungi) the literature project provides a 

base layer of information for the identification of plant 

pathogenic fungi that is supplemented with more detailed


information from the projects working on specific groups. 

Scientific experts will guarantee the high quality standard 

of the database. Eventually, scientists will not only be able 

to search for specific parasite-host-interactions, also a 

literature reference and geographic relations are given. 

Major achievements of the GLOPP-LIT project are a 

digitization of the major published host-pathogen lists for 

Europe. In addition, highly detailed literature reviews of 

the distribution of Erysiphales, Peronosporales, and 

Uredinales in Germany are currently developed. These will 

lead to an improved understanding of the prevalence and 

distribution of important pathogen groups. It is hoped that 

the tools and processes currently developed for this 

purpose can be used to develop similar data sets in other 

countries as well. 

515 - Identification of basidiomycetes using image 

analysis of pigments and colony morphology 

M.E. Hansen 1* , P.W. Hansen 2 , S. Landvik 3 , M. Sasa 3 , K.F. 

Nielsen 4 & J.M. Carstensen 2 

1 IMM, DTU, Richard Petersens Plads, build 321, DK-2800 

Lyngby, Denmark. - 2 Videometer, Lyngsoe Alle 3, DK-2970 

Hoersholm, Denmark. - 3 NovoZymes A/S, Smoermosevej 

25, DK-2880 Bagsværd, Denmark. - 4 BioCentrum-DTU, 

Soeltofts Plads, build 221, DK-2800 Lyngby, Denmark. - 

E-mail: meh@imm.dtu.dk 

It has been shown that image-analysis can be used for the 

identification of terverticillate species of Penicillium based 

on colour-calibrated images obtained from the 

VideometerLab system. The capability of this newly 

developed method to distinguish between fungal cultures 

that appear identical but are known to be different species 

is very convincing and implies its usefulness in recognizing 

fungal cultures. The establishment of a public database of 

images captured under standardized conditions will allow 

any scientist using the system to compare their data to 

reference information in the database. The species tested so 

far, however, represent only a minor part of the fungal 

diversity. Furthermore, these species often produce 

strongly pigmented cultures. In this study, we have 

challenged the system's ability to recognize and group 

species and strains of two genera of the Basidiomycota; 

three species each of Polyporus (Polyporales) which 

produce whitish/lightly coloured mycelia, and of Pholiota, 

Agaricales, which form mottled cultures. A total of 21 

isolates were cultivated in triplicates on three different 

media in 9 cm Petri dishes, and images were captured of 

each plate on day 8 and 15. The results of the imageanalyses, 

including groupings of strains and their growth 

on the various media were compared to the metabolite 

profile and taxonomy of the groups. The system facilitates 

comparison of subtle visual differences in an accurate and 

reproducible way. 

516 - Multispectral macroscopy for mycology 

P.W. Hansen * & J.M. Carstensen 

Videometer A/S, Lyngsø Allé 3, DK-2970 Hørsholm, 

Denmark. - E-mail: PWH@videometer.com 

Imaging technology has proved very useful for 

classification of fungi which are difficult to separate by 

other means without performing a labor demanding 

chemical analysis. Studies have been carried out using 

traditional trichromatic camera technology, producing three 

images corresponding approximately to the colors red, 

green, and blue, which are sufficient for many purposes 

where information on subtle color differences in the visible 

region is required. A new and innovative technology based 

on light emitting diodes (LEDs) that adds to the advantages 

of the trichromatic technology, is presented. By combining 

LEDs with a black-and-white digital camera, multiple 

advantages are obtained, of which an increased spatial 

resolution (in the megapixel range) and the possibility of 

using wavelengths outside the visible range, such as 

ultraviolet and near-infrared light, are the most notable. At 

present, up to ten relevant wavebands may be combined 

into the same unit producing multispectral images 

incorporating information not visible to the human eye, 

such as information on metabolites or chemical 

composition. In order to reduce the wealth of information 

produced by a multispectral unit to simple properties 

perceivable to the human brain, multivariate statistical 

methods are applied to the image data. By supervised or 

unsupervised learning these methods are used for building 

mathematical models relating image data to properties of 

interest, e.g. species, strain, or clone. 

517 - Host preferences of wood-decaying 

basidiomycetes in a cool-temperate area of Japan 

T. Hattori 

Forestry and For. Prod. Res. Inst., Norin-Kenkyu-Danchi, 

Tsukuba, Ibaraki 305-8687, Japan. - E-mail: 

hattori@ffpri.affrc.go.jp 

I examined host ranges of wood-decaying basidiomycetes 

in a cool-temperate forest in Japan. Fagus spp. and 

Quercus spp. are the main tree species within the forest. I 

marked fallen trees (more than 20 cm diam and 2 m long) 

within a 300 x 200 m plot, then listed polypores, stereoid, 

and hydnoid fungi on each tree. Diameter, tree species 

were also recorded for each tree. In total, 250 trees were 

marked, then 51 species of fungi (44 polypores) were 

recorded. Following species did not show any preference: 

Bjerkandera adusta, Ganoderma applanatum, Rigidoporus 

cinereus, Stereum ostrea, Trametes versicolor, etc. All 

collections of Daedalea dickinsii (26/26) were on Quercus 

spp. (including Castanea crenata). Other species as follows 

were also recurrent on Quercus spp.: Hymenochaete 

rubiginosa (39/41; 2 on undetermined trees), Melanoporia 

castanea (18/18), Piptoporus soloniensis (8/8), Xylobolus 



frustulosus (22/22), etc. The following species were 

frequent on Fagus spp.: Fomes fomentarius L type (7/7), 

Fomes fomentarius S type (19/21; 1 on Styrax obassia, 1 

on an undetermined tree), Protodaedalea hispida (13/14; 1 

on Quercus sp.), etc. Fomitopsis cf. cajanderi was always 

on Prunus spp. (4/4). Melanoporia castanea was usually 

on huge trees with more than 50 cm diam or on branches of 

huge trees. It is suggested that loss of old growth forests 

may reduce diversity of wood-decaying basidiomycetes 

such as those restricted to Fagus spp. and huge Quercus 

spp. 

518 - The discovery of Cryphonectria cubensis on native 

Syzigium spp. from South Africa 

R.N. Heath * , M.V. Gryzenhout, J. Roux & M.J. Wingfield 

University of Pretoria, Department of Microbiology and 

Plant Pathology, Forestry and Agricultural Biotechnology 

Institute, 74 Lunnon road, Hillcrest, Pretoria, 0002, South 

Africa. - E-mail: Ronald.Heath@FABI.up.ac.za 

Cryphonectria cubensis is a pathogen on Eucalyptus 

species and Syzigium aromaticum in tropical and 

subtropical regions worldwide. The pathogen has also been 

reported from Tibouchina species (Melastomataceae) in 

Colombia and South Africa. A previous hypothesis has 

been that C. cubensis was introduced into South Africa 

from South America. However, sequence data derived from 

β-tubulin, histone H3 genes and disease symptoms have 

been used to show that South African isolates of C. 

cubensis are different to those from South America, Central 

Africa and South East Asia. During disease surveys in 

indigenous forests of South Africa, fruiting structures 

resembling the anamorph of C. cubensis were found on 

native S. cordatum and S. guanennsi. Teleomorph 

structures were found on samples from the Northern and 

KwaZulu Natal provinces. The fungus from these native 

Syzigium spp. was broadly identified as C. cubensis based 

on morphology. Comparisons of β-tubulin sequence data 

showed that collections from the native Syzigium spp. 

group together with South African isolates from 

Eucalyptus. Results of this study indicate that the 

Eucalyptus pathogen in South Africa, currently known as 

C. cubensis, occurs on native Syzigium spp. in this country. 

Furthermore, they add convincing evidence to the view that 

C. cubensis in South Africa, is a species different to that 

occurring elsewhere in the world and that it is native to 

South Africa, where it has infected exotic Eucalyptus. 

519 - What does rare wood-associated fungi really 

want? 

J. Heilmann-Clausen 1* & M. Christensen 2 

1 Forest and Landscape Research Institute, Denmark. - 

2 Royal Veterinary and Agricultural University, Denmark. - 

E-mail: jhc@kvl.dk 

158 


Based on studies in Danish deciduous forests, we examine 

the importance of environmental conditions and wood 

quality for macro-fungal diversity (as sporocarps) in 

decaying wood, in order to provide practical conservation 

guidelines for this group. Although we found species 

richness to increase with wood volume, threatened species 

did not appear in general to prefer large logs. Rather, we 

found small logs to host more threatened species per 

volume unit than larger logs. Similarly, branches (diam. 

>10 cm) appeared to be richer in species than logs. Logs in 

intermediate stages of decay were found to be most species 

rich, especially with respect to threatened species. By 

analysing modified data sets we found that even a short gap 

in the supply of dead wood might significantly decrease the 

number of threatened fungi at forest level. Tree species 

diversity was found to be partly reflected in the diversity of 

wood associated fungi. However, some tree species appear 

to have rather similar fungal species assemblages and does 

not independently add to overall species richness at local 

scale. Local variation in microclimate was found to have 

limited importance for fungal diversity. Among localities, 

however, we found huge variation in the frequency of 

threatened species, which seem to be partly reflected by 

variation in microclimatic stress levels, partly by 

differences in forest history. Selecting the right sites for 

forests reserves is crucial for an efficient protection 

strategy. 

520 - Molecular analyses of arbuscular mycorrhizal 

fungal species communities 

I. Hijri 1 , F. Oehl 1 , K. Ineichen 1 , P. Mäder 2 , A. Wiemken 1 & 

D. Redecker 1* 

1 Institute of Botany, University of Basel, Hebelstr.1, 4056 

Basel, Switzerland. - 2 Institute of Organic Agriculture 

(FIBL), Ackerstr., 5070 Frick, Switzerland. - E-mail: 

dirk.redecker@unibas.ch 

Species communities of arbuscular mycorrhizal fungi 

(AMF) were studied using a set of primers specifically 

targeted at the ribosomal RNA genes. Small subunit and 

Internal Transcribed Spacer sequences were amplified from 

colonized roots and analyzed phylogenetically. Soil and 

roots were sampled from field sites under organic or 

conventional agricultural management widely differing in 

input intensity as well as from semi-natural grassland. Trap 

cultures set up from the respective field soils were also 

characterized. All sites are situated in the three-countrycorner 

France/Switzerland/Germany, most of them on 

Loess soils. A wide range of fungi was detected in the trap 

cultures, comprising Glomus, Acaulospora and 

Archaeospora. The occurrence of generalist AMF and 

specialists for specific field sites or types of agricultural 

management was analyzed, as well as the spatial structure 

of AMF communities.


521 - Lobariaceae (Lecanorales, lichenised Ascomycota) 

of Mauritius and La Réunion in the Indian Ocean 

A.C. Holm 1* , R. Gregersen 1 , U. Søchting 1 & E. Timdal 2 

1 Department of Mycology, Botanical Institute, University 

of Copenhagen, O. Farimagsgade 2D, DK-1353 

Copenhagen K, Denmark. - 2 Botanical Museum, University 

of Oslo, Sars gate 1, N-0562 Oslo, Norway. - E-mail: 

annechristine@hotmail.com 

The lichen family Lobariaceae is a prominent part of the 

lichen vegetation in the more humid areas of the tropical 

islands Mauritius and La Reunion situated in the Indian 

Ocean. During several excursions to the islands, the 

authors and also Hildur Krog, Oslo collected about 400 

specimens of Lobaria, Pseudocyphellaria and Sticta from 

numerous sites. The collections include six species of 

Lobaria, five species of Pseudocyphellaria and 10-12 

species of Sticta. Very limited information on the 

Lobariaceae of the islands has been published, and the 

closest comprehensive treatment is the macrolichen flora of 

East Africa by Swinscow and Krog (1988). Lobaria 

holstiana is not previously reported from the islands and 

Sticta cf. cinereoglauca appears to be new to Africa and 

adjacent islands. All of the species are pantropical, except 

Lobaria holstiana and Sticta dichotoma. L. holstiana is 

known from Tanzania, Mauritius and La Réunion, whereas 

S. dichotoma is found in Eastern Africa, the Mascarenes 

and Panama. However, S. dichotoma may be conspecific 

with the pantropical species S. canariensis. The 

photosymbiodemes of Sticta dichotoma and S. 

cinereoglauca are described. 

522 - The merosporagiferous fungi in Taiwan 

H. Hsiao-Man 

Dept. of Natural Sci. Edu.,Natl. Taipei Teachers College., 

134, Sect. 2, Hoping, E. Rd.,Taipei, Taiwan, Taiwan. 

During an investigation of merosporangiferous 

Zygomycetes in Taiwan, eight species of the family 

Piptocephalidaceae (Zoopagales) were isolated. Two of 

these species belong to the genus Piptocephalis. They are 

Piptocephalis cylindrospora and P. sphaerica. Six species 

belong to the genus Syncephalis. They are Syncephalis 

cornu, S. depressa, S. obconica, S. parvula, S. sphaerica 

and S. ventricosa. All species represent new records to 

Taiwan. Previously none of the family Piptocephalidaceae 

was described from Taiwan. These eight species were all 

isolated from soil or dung in Taiwan, mainly mountain 

area. The morphological characters together with their 

hosts are reported. 

523 - Rehabilitation of nutrient-poor grasslands 

(waxcap grasslands) in dunes in the Netherlands: the 

effect of active nature-management on the mycoflora 

L.M. Jalink * & M.M. Nauta 

National Herbarium Netherlands, Universiteit Leiden 

branch, P.O. Box 9514, NL-2300 RA Leiden, The 

Netherlands. - E-mail: l.m.jalink@hccnet.nl 

The effect of active nature-management in grasslands in an 

area in the dunes in the western part of the Netherlands is 

evaluated. The mycoflora in 17 semi natural wet grasslands 

with various management has been followed since 1985, 

two during all years, others irregularly. All 17 grasslands 

are situated in the Amsterdam Waterworks Dunes, a 

protected area of 34 km 2 southwest of Haarlem. They are 

mown, grazed or the topsoil has been removed. The dune 

area has served as a drinking water source for the city of 

Amsterdam which caused desiccation. Therefore 

infiltration started some 30 years ago with unpurified river 

water from the river Rhine. This caused an enormous 

increase of e.g. Calamagrostis, the direct motive to start 

active nature management in the former wet dune slacks. 

Evaluation after 15 years shows positive results for plants 

as well as macrofungi. There is a spectacular increase in 

number of species and fruitbodies of the macrofungi 

representative for Waxcap grasslands (Hygrocybe, 

grassland Entoloma, Geoglossum, Trichoglossum, 

Clavulinopsis, Clavaria). This type of grassland and its 

representative species have been under serious threat in the 

Netherlands since 1900. Of the investigated sites 5 have 

now 10 or more species of Hygrocybe, and two of the 

investigated sites belong now to the best Waxcapgrasslands 

in the Netherlands. There seems to be little 

difference in effect between grazing or mowing. 

524 - Plant pathogenic and saprophytic fungi from 

graminicolous hosts in Thailand 

K. Jaroenthai 1 , L. Manoch 1* , C. Chamswarng 1 & P. 

Suwanarit 2 


Kasetsart University, Bangkok 10900, Thailand. - 

2 Department of Microbiology, Faculty of Science, 

Kasetsart University, Bangkok 10900, Thailand. - E-mail: 


In the present study, corn, rice and grass leaves showing 

leaf blight and leaf spot symptoms were collected from 

various locations in the North, North-East and Central 

Thailand. They were used for isolation of plant pathogenic 

and saprophytic fungi. Tissue transplanting method with 

10% clorox and potato dextrose agar was used. The results 

indicated that dematiaceous hyphomycetes found on Zea 

mays included Bipolaris bicolor, B. maydis, B. zeicola, 

Curvularia akaiiensis, C. eragrostidis. C. lunata, 

Nigrospora sphaerica and Nodulisporium gregarium, 

whereas Alternaria padwickii, Bipolaris hawaiiensis, B. 



oryzae, Curvularia lunata, Drechslera halodes, 

Exserohilum rostratum and Nigrospora oryzae were found 

on rice kernel and diseased leaves. Plant pathogenic and 

saprophytic fungi found on grass leaves included 

Alternaria longipes (on Brachiaria mutica), Beltrania 

rhombica (on Digitaria cialiaris and Pennesitum 

pedicellatum), Curvularia akaii (on D. cialiaris), C. 

borreriae (on Echinochloa crusgalli), C. intermedia (on P. 

polystachyon), C. ovoidea and C. penniseti (on P. 

pedicellatum), C. sorghina (on P. setosum), Drechslera 

spp. (on Rhychelytrum repens, B. ruziziensis, Eleusine 

indica, Penicum maxinum), Exserohilum sp. (on P. 

padicellatum), Nigrospora oryzae (on Imperata cylindrica, 

Dichanthium annulatum and R. repens), N. sphaerica (on 

D. digitaria, R. repens and D. annulatum) and Pyricularia 

grisea (on B. mutica, P. padicellatum, Penicum repens and 

Cenchrus echinatus). 

525 - Diversity of coprophilous fungi from wild and 

domestic animals in Central and North-East Thailand 

O. Jeamjitt 1* , L. Manoch 1 , R. Watling 2 & G.P. Sharples 3 


Kasetsart University, Bangkok, 10900, Thailand. - 2 Royal 

Botanic Garden, Edinburgh, EH3 5LR, Scotland, U.K. - 

3 Institutes of Bioscience, John Moores University, Byrom 

Street, Liverpool L3 3AF, England, U.K. - E-mail: 

onumaj@yahoo.com 

Coprophilous fungi are a large group of saprobic fungi that 

can survive thermal and chemical conditions in herbivore 

digestive tracts. They are adapted to extreme conditions 

and many species show distinctive morphological features. 

This group of fungi have been shown to be a source of 

antibiotics, organic acids, enzymes, biological control 

agents and other secondary metabolites of economic 

importance. In the present studies, 8 dung samples of 

domestic animals and wildlife, including buffalo, cow, 

deer, elephant, goat, rabbit, rat and toad, were collected 

from nine locations in Central and Northeast Thailand. The 

moist chamber, soil plate, alcohol and heat treatment 

methods were used to isolate these fungi. Identification was 

based on morphological characters observed with light and 

scanning electron microscopy.The results revealed that 27 

genera and 14 species were found, including 3 

Zygomycetes: Absidia corymbifera, Cunninghamella and 

Rhizopus; 18 Ascomycetes: Ascobolus, Ascodesmis, 

Cercophora, Chaetomium, C. cupreum, C. globosum, 

Echinopodospora, Eupenicillium osmophilum, Eurotium 

amstelodami, Microascus, Neosartorya fumigata var. 

glager, Podospora curvicolla, Preussia, Saccobolus, 

Sporormiella, Sordaria fimicola, Talaromyces 

bacillisporus and T. rotundus; 13 Hyphomycetes: 

Acremonium, Arthrinium, Arthrobotrys, Aspergillus 

candidus gr., A. fumigatus gr., A. niger gr., Penicillium 

spp., Scopulariopsis, Stachybotrys and Trichoderma 

harzianum and 1 Basidiomycete: Coprinus. 

160 


526 - The habitat ecology and distribution of lichen 

Lobaria pulmonaria in Estonia 

I. Jüriado 

Institute of Botany and Ecology, University of Tartu, 38 Lai 

Str., Tartu, 51005, Estonia. - E-mail: jyriado@ut.ee 

Lobaria pulmonaria grows in biotopes of long ecological 

continuity and favours forests of natural conditions. The 

atmospheric pollution and destructive forest management 

has caused the decline of distribution in many parts of its 

distribution area, especially in Europe. The aim of this 

study was to review the substrate types, habitat demands 

and distribution of L. pulmonaria in Estonia. All the 

available data were accounted. The distribution analysis 

within suitable habitats was performed using Corine 

landcover types database in GIS. L. pulmonaria is totally 

epiphytic lichen in Estonia, it has been found on 15 tree 

species, almost exclusively on deciduous trees. One-third 

of registered records of L. pulmonaria are known on 

Populus tremula; it is common also on Acer platanoides, 

Fraxinus excelsior and Quercus robur. The most of the 

registered specimens (91%) grow in eutrophic borealnemoral, 

eutrophic paludifying and mesotrophic boreal 

forests, 6% are in semi-natural biotopes - wooded 

meadows, and 3% of records are known from parks 

(churchyards, former manor parks). L. pulmonaria is the 

most common in over 100 years old mixed forest, on 

average with 4-5 tree species or spruce dominated forests. 

At present L. pulmonaria is recorded in a few hundred 

localities in Estonia. 

527 - Waxcap grasslands in Slovakia - History, present 

state and perspectives 

I. Kautmanova 

Slovak National Museum - Nature Historical Museum, 

Vajanského nábr.2, 821 05 Bratislava, Slovakia. - E-mail: 

botanika@snm.sk 

Slovakia is rich in seminatural grassland habitats, because a 

small scale farming was the most common way of utilizing 

not very rich nature sources. Small fields, meadows and 

pastures were grazed and/or mowed and sometimes dungfertilized. 

On these grasslands ecosystems very rich in 

biodiversity developed. In the second half of the last 

century many of these small plots were put together and 

managed industrialy, which ment absolute loss of their 

former character. Only at the most inaccessible places 

some of them were preserved, but many of their owners 

lost interest in utilizing them. Research of fungal diversity 

of these ecosystems has just started, but results are 

promising. In 1998 only 28 species of Hygrocybe were 

published and since then 11 new species were recorded and 

also the most endangered species as H. calyptraeformis, H. 

spadicea and H. ovina can be found. The best known 

localities: Landscape protected area (LPA) Biele Karpaty - 

meadows with more than 10 species of Hygrocybe


recorded at single visit (H. reidii, H. marchii, H. 

subpapillata, together with some gasteromycetes and rare 

Clavariaceae. LPA Horná Orava - wet meadows and 

peatbogs with species as H. calyptraeformis, H. 

substrangulata, H. coccineocrenata, H. turunda a.o. 

National Park (NP) Slovenský Raj - seminatural grasslands 

of European importance with very high biodiversity. 

Mycological research has just started, but species as H. 

nitrata, H. marchii, H. perzistens and H. substrangulata 

were recorded. 

528 - Building of the central database of biological 

collections in the museums of the Slovak Republic 

I. Kautmanova * , J. Uhlirova & J. Kautman 

Slovak National Museum - Nature Historical Museum, 

Vajanského nábr.2, 821 05 Bratislava, Slovakia. - E-mail: 

botanika@snm.sk 

In 2001 the project of building national database of 

biological collections, in which 31 Slovak museum take 

part, has started. Guarantee is the Nature Historical 

Museum (NHM) of the Slovak National Museum and the 

project is aimed to the identification and documentation of 

chosen components of the Slovak flora and fauna, with the 

accent on the rare and endangered species. At present the 

structure of database program is prepared and in 2003 first 

data will be filled in. Central node of the database will be at 

the NHM and in 2005 first outputs will be presented on the 

web. Herbary collections preserved in 25 museums contain 

721 405 specimens. Fungi (including lichens) are 157 936 

specimens. Main mycological collections are at: Slovak 

National Museum-Nature Historical Museum, Bratislava 

(BRA) - 146 877, Museum of the Tatra National Park 

(TNP) - 5 348, Forestry and Timber Museum, Zvolen - 1 

910, Museum Nitra (NIM) - 1139, Central-Slovakian 

Museum, Banská Bystrica (SMBB) - 491, Povazie 

Museum, Zilina - 463, Orava Museum of P.O.Hviezdoslav, 

Dolný Kubín - 415, Slovak Museum of Nature Protection 

and Speleology, Liptovský Mikulás (MOP) - 334, Saris 

Museum, Bardejov (SMB) - 200 specimens. 

529 - Role of mobile introns in diversity of mtDNAs of 

imperfect black Aspergilli 

F. Kevei * , Á. Juhász & Zs. Hamari 

Department of Microbiology, Faculty of Sciences, 

Univesity of Szeged, H-6701 SZEGED, P.Ö.Box 533, 

Hungary. - E-mail: kevei@bio.u-szeged.hu 

A high level of intraspecific mtDNA variability was 

reported earlier among black Aspergilli, indicated by 

various RFLP profiles of isolates of Aspergillus niger, A. 

tubingensis, A. carbonarius and A. japonicus. To study the 

reason for the mtDNA polymorphism physical- and 

functional maps of mtDNAs representing different 

RFLPtypes were constructed than the variable regions 

cloned and sequenced. These revealed that apart from 

minor nucleotide changes, the presence of introns at 

different positions and their appearance in various numbers 

in the mtDNAs are responsible for the mitochondrial 

genome diversity and size differences. The reasons of 

variability between two mtDNA types of A. carbonarius 

can be attributed to the presence or absence of a 1.1 kb 

group I intron in their cox2 subunit. Sequence comparison 

of two mtDNA RFLP types of A. japonicus revealed that 

they differed from each other in at least two group I introns 

in cox1 subunit and one group I intron in cob gene. 

Organisation of four A. niger mtDNA types proved to be 

very similar, but one of the three observed variable regions 

included a group I intron in the cox1 subunit was present in 

three different forms. The mobile introns (bearing ORFs) 

play an important role with their homing process 

generating recombinant mtDNAs in the interaction among 

mitochondrial genomes after mitochondrial transmission. 

The homing endonucleases activate a double strand break 

that is finally repaired by a repaire mechanism. 

530 - Arbuscular mycorrhizal status of plant species 

from Western Ghats of Goa, India 

S. Khade * & B. Rodrigues 

Department of Botany, Goa University, India. - E-mail: 

shardakhade@rediffmail.com 

Western Ghats are one of the hot spots of biodiversity in 

the world and the state of Goa lies in the heart of Western 

Ghats. In the present paper, AM fungal status of 62 plant 

species, viz. pteridophytes, herbs, shrubs, trees and plants 

with under storage ground organs from Western Ghats of 

Goa have been discussed. All the plant species selected for 

study, exhibited the presence of AM fungal association. 

The average root colonization ranged from 30% 

(pteridophytes) to 70% (shrubs). Whereas the average 

spore density of AM fungi ranged from 52 spores 100 -1 g 

soil (pteridophytes) to 528 spores 100 -1 g soil (plants with 

under ground storage organs). Species composition of AM 

fungi revealed the presence of five genera viz., 

Acaulospora (9), Glomus (19), Gigiaspora (3), 

Scutellospora (5) and Sclerocystis (5) with species number 

given in parenthesis. A total of 41 species of AM fungi 

were recovered during the study. Scutellospora gregaria, 

Glomus fasiculatum and Sclerocystis taiwanensis were the 

most frequently occurring AM fungal species. The present 

paper is first report on AM fungal association in plant 

species from Western Ghats of Goa and contributes data 

necessary for further studies on AM fungi from this region 

of which very little has been explored so far. 

531 - Field identification of marne yeasts using DNA 

hybridization macroarrays 

T. Kiesling, M. Diaz, A. Statzell-Tallman & J.W. Fell * 

University of Miami, 4600 Rickenbacker Causeway, Key 

Biscayne, Fl, U.S.A. - E-mail: jfell@rsmas.miami.edu 



Yeasts are a natural component of near shore and oceanic 

environments, however the extensive tests required for 

species identifications have hampered studies of their 

ecological roles. Current use of molecular techniques has 

resolved many of these issues. With electronic availability 

of extensive gene sequences in Genbank, species can be 

identified by sequence analysis and by design of speciesspecific 

PCR primers and hybridization probes. In this 

poster, we describe a macroarray hybridization assay, 

which can be used to identify yeasts directly from 

environmental samples or from cultured organisms. The 

technique is in a microtiter format in which speciesspecific 

probes are bound to the wells of the microtiter 

plates; genomic DNA from target species are amplified and 

labeled with biotin labeled universal primers. The probes 

are hybridized with labeled target DNA and a positive 

identification is indicated via a colorimetric assay. The 

results are unambiguous: a color change indicates a 

positive identification. The method is inexpensive, rapid, 

accurate and amenable to shipboard as well as laboratory 

use. 

532 - Fungal diversity in set-aide agricultural soil 

investigated using terminal-restriction fragment length 

polymorphism (T-RFLP) 

M. Klamer * & K. Hedlund 

Lund University, Dept. of Zooecology, Getingevägan 60, S- 


morten.klamer@zooekol.lu.se 

In a newly abandoned agricultural land the fungal 

community response to manipulation of the above ground 

vegetation was investigated. The field site consisted of 20 

plots where the plant diversity was managed by either 

sowing 15 plant species or natural colonization was 

allowed to occur. The plant mixture contained grasses, 

legumes and forbs that all were expected to occur on the 

site. A subset of the plots (5 from each treatment) was 

inoculated with soil cores from a late successional stage. 

Five years after abandonment soil cores were taken from 

the plots and DNA was extracted using Bio101 soil 

extraction kit. The ITS region of the rDNA gene was 

amplified using fluorescently labelled fungal specific 

primers (ITS 1F/ITS 4). The PCR products were digested 

using HinfI and TaqI and analysed on an ABI 377 

sequencer. Results from both restriction enzymes were 

combined and a principal component analysis performed. 

Also the species richness expressed as the number of 

restriction fragments in each sample were analysed. These 

results were compared with field observations of the 

development in plant species composition. 

162 


533 - New and widespread genera of hypocrealean 

Geosmithia on a hidden place 

M. Kolarík 

Faculty of Science, Department of Botany, Benátská 2, 128 

01 Prague, Czech Republic. - E-mail: 

miroslavkolarik@seznam.cz 

The genera of the polyphyletic genus Geosmithia (formerly 

Penicillium) are recently placed to Eurotiales and 

Hypocreales (G. putterilii and G. lavendula). The genetic 

relationships among 90 isolates of Geosmithia spp. 

collected from galleries of phloem boring insects and from 

soil were investigated using RAPD analysis, rDNA, 

microscopy and colony morphology. We found 14 groups 

that were distinguished by both RAPD patterns and 

morphological characters. Based on ITS1-5.8SrDNA-ITS2 

sequences, phylogram relating representatives of RAPD 

groups to the described species was constructed. Two 

highly supported clades were found. First clade consisted 

of type strain of P. pallidum and 3 new groups, second 

clade contained 9 groups, with affinities to G. putterilii and 

G. lavendula. Some groups are very charakteristical by its 

morphology, fingerprint pattern and ITS region and 

represent new genera as well as undescribed 

subpopulations of G. putterilii. The only preference of 

RAPD groups was found for associated insect (S. rugulosus 

× S. intricatus), but not for host tree species or locality. 

This implies that Geosmithia fungi are mostly vectored by 

insect. The critical character of hypocrealean geosmithias 

called 'peg foot' is described by its morphology and 

ontogenesis. The newly found Geosmithia spp. (from G. 

putterillii complex) are very abundant in early fungal 

communities (next to yeast) of feedings of bark beetles. 

534 - Eastern Carpathian lichens: National and 

geomorphologic peculiarities 

S.Ya. Kondratyuk 

M.H. Kholodny Institute of Botany, Tereshchenkivska str. 

2, 01601 Kiev, Ukraine. - E-mail: skondr@botan.keiv.ua 

The first Catalogue of the lichen-forming, lichenicolous 

and allied fungi of Eastern Carpathians Mts. including 

1307 species (Kondratyuk, Lackovicova, Pisut & Popova 

2002) allows to carry out the first attempt of comparison of 

lichen florae of their national portions and geomorphologic 

regions. The highest species diversity of fungi mentioned 

(1095 species) is hitherto known from Ukrainian part of 

Eastern Carpathian Mts., while the biggest Romanian part 

(only 529 lichen species known) is still in urgent need of 

special study. Polish and Slovak portions of Eastern 

Carpathian Mts. are represented consequently by 508 and 

411 species. Among geomorphologic units of the Eastern 

Carpathian Mts. (accepted mainly after Kondraczky 1978), 

Eastern Beskydy region (within Ukrainian part) is the most 

well studied at the moment from lichenological point of 

view. 659 species of lichen-forming and lichenicolous


fungi are hitherto recorded from this region. More than 400 

lichen species (namely 410) is known from Ukrainian 

Volcan Carpathians. From 300 to 400 lichen species is 

recorded from the following areas: Low Bieszczady, High 

Bieszczady, Chornohora, Chyvchyn-Grynyavsky Mts. and 

Marmarosh Mts. More than 200 lichen species (and less of 

300) is known from Bukovsky Vrchy Mts., Gorgany and 

Svidovetz Mts. Similarity and differences of the species 

content of lichens of national and geomorphologic regions 

of the Eastern Carpathian Mts. region are discussed and 

illustrated. 

535 - Epiphytic lichens in Dushanbe 

I. Kudratov 

Tajik State National University, Rudaki str. 17, 252025 

Dushanbe, Tajikistan. - E-mail: botanica@ac.tajik.net 

Dushanbe, the capital of the Tajik Republik, is located in 

the Hissar Valley at a height of 800-900 meters above sea 

level. Dushanbe's climate is sharp continental. The average 

temperature in January is +1 °C and in July +28 °C. 

Epiphytic lichens in Dushanbe and its vicinity were 

investigated in 1997-1998 to learn study their role as air 

pollution indicators. Species composition of the epiphytic 

lichens on Platanus orientalis, Acer negundo, Pinus 

eldaricus was studied and the distribution of certain lichen 

examined. A list of lichens contains 15 species. On the 

ground of received data three zones of air pollution were 

delineated in Dushanbe. Most polluted zone (1) follows the 

distribution of Lecanora hagenii, Phaeophyscia orbicularis 

and Anaptychia ulotricoides etc. This (1) zone includes the 

separate sites along the main motor roads of the city. They 

are the sites with permanent intensive pollution. The zone 

of intermediate pollution (11) was delineated according to 

distribution of Lecanora hagenii, Phaeophyscia 

orbicularis, Anaptychia ulotricoides, Candelariella 

aurella. The third zone is located in the north-eastern 

outskirts of Dushanbe embraces almost the whole hilly 

massif. Therefore the number of epiphytic lichens species 

shows a tendency of decreasing of species number along 

the gradient of air pollution. 

536 - The influence of forestry on wood-inhabiting 

corticioid basidiomycetes in Switzerland 

N. Küffer 1* , D. Job 1 , M. Aragno 1 & B. Senn-Irlet 2 

1 Université de Neuchâtel, Lab. de Microbiologie, Case 

Postale 2, 2007 Neuchâtel, Switzerland. - 2 Swiss Federal 

Research Institute WSL, Zürcherstrasse 111, 8903 

Birmensdorf, Switzerland. - E-mail: 

nicolas.kuffer@unine.ch 

Intensively managed forests show significantly less fruit 

bodies and a lower species richness of corticioid 

basiodiomycetes than forest reserves. Species richness does 

not only seem to be determined by the availability of 

woody debris, since also some strongly managed forests 

host a remarkably high species richness. Even rather rare 

species, e.g. Tubulicrinis sororius or Brevicellicium 

olivascens, could be found in managed forests, whereas 

they were expected to occur more likely in forest reserves. 

Forests, richer in different tree species, harbour a higher 

number of fungal species too, independent of their 

managing history. Nevertheless, the importance of forest 

reserves for the maintenance of a rich species community 

of corticioid basidomycetes is clearly confirmed. In order 

to address the question of diversity, abundance, host age, 

decomposing process, regional factors and the influence of 

modern forestry, 82 plots of 50 m 2 have been selected in 

the main forest types in Switzerland. A total of 3216 

branches with fruit bodies of corticioid basidiomycetes 

have been harvested and thereon 233 species identified. 

Among the most frequent species are Amphinema 

byssoides, an important mycorrhizal partner of conifers and 

Phlebiella vaga or Hyphodontia sambuci, two wooddecomposing 

species. Preliminary results show a few 

species dominating, the majority of the species is found 

only rarily: species rank distribution is best explained with 

a steep power function. 

537 - Nitrogen impact on epiphytic lichens in an ancient 

oak forest in Denmark 

R.S. Larsen * & U. Søchting 

University of Copenhagen, Botanical Institute, Dep. of 

Mycology., Østre Farimagsgade 2D, 1353 Kbh.K, 

Denmark. - E-mail: renesl@bot.ku.dk 

The lichen vegetation was studied in Kås Skov, an ancient 

oak forest in Northern Jutland, Denmark that is under 

protection of EEC habitat directive because of its high 

biodiversity. The very rich lichen vegetation is threatened 

by ammonia and ammonium deposition from animal farms 

in the region. In a floristic survey of epiphytes on tree 

trunks, several old forest indicator lichens previously 

recorded were not refound, however Zamenhofia hibernica 

and Ramonia chrysophaea were found as new to Denmark. 

Lichen vegetation from north and south side of trunks, as 

well as that of north- and southfacing branches of trees 

standing at the forest edge and interior, were compared. 

Multivariate analysis (CCA) showed that the lichen 

occurrence is more dependent on the position on the 

branch, than on where the branch is situated in the forest. 

Indicator Species Analysis (ISA) revealed that species 

generally regarded as nitrophilous, prefer branch parts 10 

to 50 cm from the tip (3-8 years), at the edge of the forest. 

Contrary, thalli of Hypogymnia physodes contain 21% 

more nitrogen in the sheltered part of the forest compared 

to the forest edge. The high nitrogen content in 

Hypogymnia physodes from Kås Skov (2.1%) is compared 

at regional scale and showing that >15 kg N /ha/year is 

deposited. This dose is expected to cause changes in the 

epiphytic lichen flora, and the nearby lichen heathland. 



538 - Wood-inhabiting fungi in primary and secondary 

seasonally dry tropical forest, Costa Rica 

I. Lindblad 

University of Oslo, Kringkollen 13, 0690 Oslo, Norway. - 

E-mail: irene.lindblad@bio.uio.no 

The relation between management regime, quality of 

coarse woody debris, and presence of some groups of 

wood-inhabiting fungi was explored in a tropical dry forest. 

A patch of primary forest and a nearby old secondary forest 

were surveyed in Santa Rosa National Park, Guanacaste, 

Costa Rica. A total of 81 species of poroid and stereoid 

fungi were found. The primary forest had 62 species, while 

54 species were recorded in the secondary forest. Thirtyfive 

species were common between the stands. More 

species were exclusively found in the primary forest than in 

the secondary one (27 vs. 20). The curve for cumulative 

number of species on increasing number of logs was 

slightly steeper for the primary forest than for the 

secondary. At least 84 logs had to be examined before a 

significant difference in species richness between the two 

forests could be detected. The results indicate that species 

richness of poroid and stereoid fungi is returning to the 

level of the primary forest in a regenerating dry forest 

habitat when the woody debris is allowed to stay in the 

forest and there is a patch of original forest in the vicinity. 

Relations between substrate quality and the occurrence of 

fungi in the different forest types were explored and 

reasons for the observed patterns are discussed. 

539 - Growth of mycorrhizal Leucaena leucocephala 

(Lam.) de Wit seedlings in copper contaminated soils 

C.E.L. Lins 1 , G.A. Silva 1* , U.M.T. Cavalcante 2 , A.S. 

Messias 3 , E.V.S.B. Sampaio 4 & L.C. Maia 1 

1 Departamento de Micologia - Universidade Federal de 

Pernambuco, 50670-420 Recife -PE, Brazil. - 

2 Departamento de Biologia - Universidade Federal Rural 

de Pernambuco, 52171-030 Recife - PE, Brazil. - 

3 Departamento de Química - Universidade Católica de 

Pernambuco, 50050-900 Recife - PE, Brazil. - 4 Depto. de 

Energia Nuclear/CTG/Universidade Federal de 

Pernambuco, 50670-420, Brazil. - E-mail: 

gladstonesilva@yahoo.com 

The ability of the mycorrhizal symbiosis in favouring 

nutrient absoption in agricultural systems, specially in arid 

and semiarid areas, as well as the toxicity of heavy metals 

has been well documented. Due to the low infectivy 

potential of the AMF at the study area (Mineradora 

Caraíbas, Bahia State, Brazil), we evaluated the effect of 

the mycorrhizal association in seedlings of Leucaena 

leucocephala in soils with increasing proportions of copper 

contamination. Soils from an 'caatinga' area, not impacted 

(control) and impacted by copper mining (site that receives 

the waste product) were used. Seedlings were inoculated 

with Glomus etunicatum Becker & Gerd. and cultivated in 

164 

substrate constituted by soil from the impacted area diluted 

in soil from the control area, in the following proportions: 

0, 25, 50, 75, and 100%. In general, increasing the 

proportion of copper contaminated soil had a negative 

effect on plant growth. Mycorrhizal plants presented higher 

height, foliar number, and shoot and root biomass than 

those non inoculated, when maintained in soil with up to 

50% contamination. The highest number of AMF spores 

(3.2 g -1 soil) was found in substrate with 25% 

contamination. The arbuscular colonization was higher 

than 40% in the treatments with up to 50% of copper 

contamination. The plant-fungi association promoted 

higher copper tolerance of the seedlings, but the 

mycorrhizal benefit was lost when soil contamination was 

higher than 50%. Financial support: CAPES and CNPq. 

540 - A survey of macrofungi at Pakua mountain range 

in central Taiwan 

P.L. Lu 


Botany Department, National Chung-Hsing University, 

Taichung, Taiwan. - E-mail: lufreda@yahoo.com.tw 

The fungal flora, with special focus on macrofungi of 

Pakua mountain range in the vicinity of Chang-Hua city 

(24 degree 10' N, 120 degree 50' E) was surveyed. The 

elevation is between 20-80 m, the average temperature 

ranges from 18-27 degrees centigrade and the annual 

precipitation is about 1732 mm. Mostly specimens' 

collection was made primarily during rainy days from 

August 2001 to April 2002. Seventy specimens were 

collected and identified. Among the identified collection 60 

specimens belong to basidiomycetes and 4 in ascomycetes. 

Tricholomataceae, Pluteaceae, Cortinariaceae, and 

Amanitaceae are among the dominant families in the flora. 

An estimated 24,000 fungal species inhabit in Taiwan, an 

island located at the northern end of pacific archipelago. 

However, only one-sixth of them have been reported. With 

the increasing pressure of economic development poses an 

urgent need for the investigation of the island's fungal 

resources. Pakua mountain range is among a unique spot 

that has never been systematically investigated. 

541 - Fungi (Basidiomycetes) in the Kielce town 

(Central Poland) 

J. Luszczyñski 

Institute of Biology, Swietokrzyska Academy, Swietokrzyska 

St. 15, PL-25-406 Kielce, Poland. - E-mail: 

jluszcz@pu.kielce.pl 

The Kielce town (112 km 2 , population over 220 000 

inhabitants) has a dense ca 25% forest cover. Comparing to 

the other Polish or European towns, it is characterized by 

the presence of highly differentiated natural vegetation and 

variety of habitats. Macromycetes were mapped using 

carthogram method. The area of town was divided into the


net of 1 km 2 squares. One square was considered as one 

locality. For each species the type of habitat, time of 

fruiting and distribution are given. The studies were 

intensely carried out from 1986 till 1994 and sporadicly 

until now and yielded the collection of over 450 species. 

Among them 6 species were new for Poland (Hypholoma 

capitatum, Phlebia cremeoochracea, Phlebiella 

allantospora, Inocybe cryptocystis, Marasmiellus tremule, 

Pluteus pouzarianus) as well many others rare in Poland 

and in Europe. In Kielce 65 species were listed in the red 

list for Poland (Wojewoda, Lawrynowicz 1992), for 

example: Albatrellus confluens, Artomyces pyxidatus, 

Boletus queletii, Dacrymyces ovisporus, Ganoderma 

carnosum, Geastrum minimum, G. quadrifidum, Gleoporus 

taxicola, Hydropus marginellus, Hygrophorus glyocyclus, 

Phellinus hartigii, Pleurotus calyptratus, Ramaria flava, 

Stropharia albonitens, Tulostoma brumale, Volvariella 

pusilla and 4 species are protected by law: Langermannia 

gigantea, Meripilus giganteus, Phallus impudicus and 

Sparassis crispa. Existing enormous biodiversity of fungi 

within town area is in my consideration a uniqe 

phenomenon. 

542 - Diversity of microfungi from soil and debris at 

termite mounds in Thailand 

L. Manoch * , T. Dethoup & J. Kokaew 

Department of Plant Pathology, Faculty of Agriculture, 

Kasetsart University, Bangkok, 10900, Thailand. - E-mail: 


Termitomyces species, a delicious mushroom, are found in 

the tropics. However, cultivation of this mushroom has not 

been successful for many reasons. Most species of 

microfungi found in termite fungus gardens belong to 

Xylaria and Trichoderma. These fungi may play an 

important role in stimulating the occurrence of 

Termitomyces. It is thus important to study microfungi 

from soil and debris at termite mounds, because some of 

these fungi can produce novel active compounds. For the 

present study, soil samples and debris from termite mounds 

were used to isolate microfungi. Samples were collected 

from various locations in Thailand. The following methods 

were used for isolation: alcohol and heat treatment, soil 

plate and dilution plate. Identification was based on macro- 

and microscopic characters when cultured on different 

media after observation under light - and SEM microscopes 

and making camera lucida drawings. Microfungi found in 

this study included Ascodesmis, Aspergillus clavatus, 

Bartalinia, Beltrania querna, Beltraniella humicola, B. 

odinae, Beltraniopsis esenbeckiae, Bionectria, 

Chaetomium, Eupenicillium parvum, Eupenicillium spp., 

Eurotium spp., Fusarium, Gilmaniella humicola, Hamigera 

avellanea, Humicola fuscoatra, H. grisea, Mammaria, 

Neocosmospora, Neosartorya spp., Nigrospora oryzae, 

Nodulisporium, Paecilomyces spp., Pestalotiopsis, Phoma, 

Rhizoctonia, Scytalidium, Talaromyces spp., Trichoderma 

hamatum, T. harzianum, T. koningii, T virens, T. viride and 

Xylaria. 

543 - Structural characteristics of naturally occurring 

ericoid mycorrhizas on five host species from Eastern 

Canada 

H.B. Massicotte 1 , L.H. Melville 2 & R.L. Peterson 2* 

1 University of Northern British Columbia, College of 

Science and Management, 3333 University Way, Prince 

George, B.C. V2N 4Z9, Canada. - 2 University of Guelph, 

Department of Botany, Guelph, Ontario N1G 2W1, 

Canada. - E-mail: lpeterso@uoguelph.ca 

In spite of numerous published studies, many questions 

remain as to the detailed cellular interactions between the 

symbionts in ericoid mycorrhizas, especially in hosts 

growing in natural habitats. Plant species with ericoid 

mycorrhizas produce many fine roots (termed hair roots) 

that become colonized by few fungal species. This 

colonization is restricted to the enlarged epidermal cells of 

hair roots. A combination of light microscopy (including 

differential interference contrast) and laser scanning 

confocal microscopy was used to document the 

colonization patterns of epidermal cells and to detail the 

hyphal complexes of five native ericaceous hosts 

originating from Ontario and Quebec: Vaccinium 

oxycoccus L. (bog cranberry), Ledum groenlandicum 

Oeder. (Labrador tea), Vaccinium myrtilloides Michx., 

(velvet-leaf blueberry), Kalmia angustifolia L. (sheep 

laurel), Gaultheria procumbens L. (wintergreen). There 

was considerable diversity in colonization patterns among 

hosts, in the morphology of hyphal complexes, and in the 

nature of thick-walled epidermal cells. Of particular 

interest was the frequency of hyphal connections between 

adjacent epidermal cells, indicating that there is not always 

one entry point for each colonized epidermal cell. Further 

observations of other species in the large family Ericaceae 

will help to determine the full range in structural details of 

ericoid mycorrhizas. 

544 - Tatra Mts as the area for mycological 

investigations - presentation of a new project 

A. Miskiewicz 

W. Szafer Institute of Botany, Polish Academy of Sciences, 

Lubicz 46, PL-31-512 Krakow, Poland. - E-mail: 

aniamis@ib-pan.krakow.pl 

Tatra Mts., the highest part of Carpathian range, are 

localized at Polish/Slovak border. Although mountains are 

not very high (Gerlach, 2655 m asl), all vegetation belts are 

well developed in this area, including subalpine and alpine 

zones. Tatra Mts. offer then all kinds of mountain habitats 

and represent excellent model region for complex studies 

on mountain fungi in the Carpathians. Very little is known 

about fungi from this massif. Several papers were 

published (e. g. Kubicka 1963, Frejlak 1973, Fellner, 

Landa 1993), but, in total only ca. 550 and ca. 900 species 

were reported from respectively Polish (Wojewoda 1996) 

and Slovak (Kuthan et al. 1999) part of the massif, and 



systematic observations are completely lacking. The aim of 

this contribution is to present a new project, which may be 

important for considerations on distribution and diversity 

of fungi in European mountains. The systematic research, 

carried out throughout two growing seasons so far, has 

been focused mainly on lower mountain belts, but 

observations in dwarf pine communities and alpine 

meadows have also been done. After only two years of 

study, the number of fungi recorded from this area 

increased considerably and some very interesting and rare 

taxa were found. The most interesting recent findings will 

be presented e.g. Mycena oregonensis, Entoloma 

farinogustus and other. 

545 - Macroscopic basidiomycetes of Sevan National 

Park (Armenia) 

S.G. Nanagulyan * & A.L. Sirunyan 

Yerevan State University, Department of Botany, 

A.Manoogyan str.1, Yerevan, 375025, Armenia. - E-mail: 

snanagulyan@ysu.am 

Sevan National Park was established in 1978 to protect 

largest in Armenia and one of the largest alpine lake in the 

world, lake Sevan and the surrounding areas. Overall 

150.100 ha are protected, including 24.800 ha of dry land. 

It is located between many of mountain ranges, at the 

1897-2100 m above sea level. The aim of this investigation 

is to study the composition of the macroscopic fungies 

communities of the Sevan National Park. Special attention 

is paid to the biodiversity of edible, poisonous and 

medicinal mushrooms. As a material for this investigation 

was the collection of fungi gathered in different part of 

Sevan National Park, as well as critically treated 

collections of the Republic's herbariums and all other 

literature data. As a result of the preliminary investigation 

and special studies of macrofungi's biota in Sevan National 

Park, 186 species and subspecific taxa from 10 orders, 30 

families and 84 genera of Gasteromycetes, Aphyllophoroid 

and Agaricoid mushrooms are identified. The biota is 

characterized by the domination of xylotrophs, humus 

saprotrophs and mycorrhizal fungi. There are also very 

interesting and specific species for this region, some of 

which are rare or endangered and have to be including in 

Armenian Red List. New approaches to the investigation 

and further theoretical study in this field are required. New 

approaches to the investigation and further theoretical 

study in this field are required. 

166 


546 - Mushrooms in Mongolia 

H. Neda 

Forestry and Forest Products Research Institute, 4-11-16, 

Kurokami, Kumamoto, 860-0862, Japan. - E-mail: 

neda@affrc.go.jp 

Mongolia is an nation located between China and Siberia. 

The climate is cold and dry. The steppe and the foreststeppe 

occupy approx. 60% of the land. And the forest 

occupy approx. 10% of the land. One hundred and eighty 

eight mushroom specimens were collected and thirty three 

species were identified. Mushrooms in steppe decompose 

humus, dung and litter. Tricholoma monogolicum Imai is 

the most famous species in Mongolian steppe. This species 

grows on humus and forms fairy ring. Several species of 

Agaricus are common in the steppe. The forest vegetation 

consists Pinus, Larix, Betula, Populus and other trees. 

There are many species of mushroom (Lentinus, Amanita, 

Suillus, Russula, Polyporus, etc.) in the forest. Most of 

them are similar to European and northern Japanese 

species. 

547 - The Douglas-fir epigeous ectomycorrhizal 

basidiomycete community in the western North 

American Northern Spotted Owl zone 

L.L. Norvell 1* & R.L. Exeter 2 

1 Pacific Northwest Mycology Service, 6720 NW Skyline 

Blvd, Portland OR 97229, U.S.A. - 2 USDI-Bureau of Land 

Management, 1717 Fabry Rd SE, Salem OR 97306, U.S.A. 

- E-mail: llnorvell@pnw-ms.com 

A joint private/USA government research team is tracking 

fungal species richness in montane Oregon Douglas-fir 

forests in two different 1998-2003 studies. Target species 

include all 'epi-ecto' basidiomycetes + 40 other non-ecto 

species flagged in the US federal Northwest Forest Plan. A 

chronosequence study incorporates replicate 200 m 2 

permanent strip transects in 25-, 55-, and 150-year old 

stands sampled every 2 weeks during fall & spring fruiting 

seasons. The density management study comprises adjacent 

65-year old stands thinned in 1999 following 5 different 

regimes: untreated (^450 trees/ha), thinned (3 stands with 

^300, ^200, or ^100 residual trees/ha), or regeneration-cut 

(0 residual trees/ha). During the first 4 years, 280 (chrono) 

and 211 (density) target species have been identified from a 

total of 4,100 collections and 515 total (322 target) species. 

Agaricales comprise ^70%, Russulales ^20%, Gomphales 

^5%, Boletales ^3%, and Cantharellales 1.5% of the target 

species total. Cortinariaceae comprise ^84% of the 

Agaricales; Cortinarius, Inocybe, Russula and 

Phaeocollybia represent the most species-rich target 

genera. After thinning, density study post/pre-treatment 

stand species richness ratios were significantly depressed 

only in the two most heavily thinned stands. Absence of 

modern PNW monographs has dictated frequent 

identification re-evaluation and led to development of


temporary identification keys based on individual research 

papers and extralimital monographs. 

548 - Fungal community analysis using environmental 

genomics 

H.E. O'Brien 1 , J.A. Jackson 1 , J.E. Johnson 2 , J.L. Parrent 1 , 

J.-M. Moncalvo 1* & R. Vilgalys 1 

1 Department of Biology, Duke University, Durham NC 

27708, U.S.A. - 2 Department of Biological Sciences, 

Cameron University, Lawson OK 23505, U.S.A. - E-mail: 

jeanmarc@duke.edu 

We applied culture-independent methods developed for 

bacteria to assess fungal community diversity across three 

different forest types within the Duke Forest without 

relying on fruitbody collections. Samples representing 

litter, O, A, and B soil horizons were investigated by direct 

DNA extraction from 50 pooled samples and cloning of 

DNA libraries from PCR fragments for both variable (ITS) 

and conserved (18S) regions within the rDNA genes. Ca. 

1600 ITS sequences were produced and sorted using 

BLAST searches. ITS sequences with > 80% similarity to 

fungal groups for which extensive taxonomic databases are 

already available were further analyzed phylogenetically. 

Two hundred 18S sequences were produced, and a global 

phylogenetic analysis was conducted to place them in a 

broader framework within Eukarya. Results showed that 

species diversity is extremely high, with a large proportion 

of unique sequence types. Fungi, especially 

ectomycorrhizal basidiomycetes, comprise 50-75% of 

eukaryotic diversity in all samples. We are currently in the 

process of developing automated databasing and 

bioinformatic tools to improve our ability to characterize 

fungal communities. Given the relative ease of sequence 

data collection, these tools should enable cross-platform 

comparisons of microbial biodiversity studies, including 

functional ecology and changes in microbial communities. 

549 - Studies in Rocky Mountain alpine Laccaria 

(Basidiomycota, Agaricales, Tricholomataceae) 

T.W. Osmundson * & C.L. Cripps 

Montana State University, Department of Plant Sciences & 

Plant Pathology, 119 Ag-Biosciences Building, Bozeman, 

MT 59717, U.S.A. - E-mail: two@gemini.oscs.montana.edu 

Documenting the diversity of gilled fungi in the North 

American alpine zone is a primary goal of the National 

Science Foundation Biotic Surveys and Inventories - 

sponsored Rocky Mountain Alpine Mycota project. As part 

of this project, a detailed study of the genus Laccaria was 

conducted using material collected at field sites above 

treeline in four mountain ranges located in the central and 

northern Rocky Mountains. Several Laccaria species are 

common components of the alpine mycota, appearing to be 

important ectomycorrhizal symbionts of the alpine dwarf 

willows Salix reticulata and S. arctica and shrub willows S. 

planifolia and S. glauca. Four taxa were identified on the 

basis of macro- and micromorphological characters: L. 

montana, L. pumila, L. cf. bicolor, and L. cf. laccata var. 

pallidifolia. Of these taxa, L. montana is reported only 

from the northern Rocky Mountain alpine sites, L. cf. 

bicolor and L. cf. laccata var. pallidifolia only from the 

central Rocky Mountains, and L. pumila from all four 

mountain ranges. L. proxima, collected at a subalpine site 

close to the northern field sites and in association with 

Salix shrubs, was not found above treeline. Ribosomal ITS 

DNA sequences are being analyzed to study biogeographic 

and phylogenetic patterns. The taxonomy, distributions, 

and ecology of Rocky Mountain alpine Laccaria species 

will be discussed. 

550 - Ribosomal DNA PCR-RFLP analysis of anaerobic 

fungi isolated from rumen and faeces 

E. Ozkose 1* , D.R. Davies 2 , M.S. Ekinci 1 & G.W. Griffith 3 

1 Sutcu Imam University, Animal Science Department, 

Kahramanmaras, Turkey. - 2 Institute of Grassland and 

Environmental Research, Animal Science department, 

Aberystwyth, SY23 3EB, U.K. - 3 University of Wales 

Aberystwyth, Biological Sciences, Aberystwyth, SY23 3BY, 

U.K. - E-mail: eozkose@ksu.edu.tr 

Anaerobic fungi inhabit in the digestive tract of most 

herbivores and they can be isolated from gastro intestinal 

tact and faeces of these animals. In this study we aimed to 

identify anaerobic fungi by PCR-RFLP analysis of ITS 

region. Fresh, frozen (1 month) and fresh rumen digesta of 

3 cows were used as fungal source. Fungal cultures (n=198 

tubes) from highest dilution series of most probable 

number (MPN) technique, which gives rise to axenic 

cultures proliferate from a single cell in the lowest level of 

dilution, were used as DNA source. Primers ITS1F (CTT 

GGT CAT TTA GAG GAA GTA A) and BlastITS4 (TCC 

TCC GCT TAT TAA TAT GC) were used for PCR 

amplification. Successfully amplified samples (133/198) 

produced a single PCR product with ca 960 bp therefore it 

was not possible to identify the isolate at even genus level 

using size of the ITS region alone. Using of restriction 

enzyme DraI, which digested all samples at least once, 

gave rise to 12 different ribotypes. Some ribotypes groups 

were found large (e.g. 45 isolates in one group) whilst few 

ribotypes included only a single isolate. Our results showed 

that PCR-RFLP analysis has a potential use in anaerobic 

fungal discrimination at least to the genera level. 

551 - The role of taxonomists in scientific 

reproducibility 

A. Padmanaban 

Institute of Microbial Technology, Microbial Type Culture 

Collection, Sector 39-A,Chandigarh-160036, India. - Email: 

apnaban@rediffmail.com 



It is generally believed by the scientific community that the 

culture collections and herbaria are managed by trained 

taxonomists who ensure the purity and authenticity of the 

research materials prior to their supply. The quality of 

services provided by such centres are generally of high 

standard vis-à-vis those which lack the expertise. 

Unfortunately, the taxonomists have to prove, time and 

again, that their science is not old-fashioned and 

intellectually dull but is the bed rock of all biological 

research. Nevertheless, based on an appeal by a group of 

taxonomists and with scientific reproducibility gathering 

momentum, the editors of some reputed microbiology 

journals have made it a mandatory requirement of 

providing deposit details of working materials with 

recognized culture collection centres or herbaria before any 

paper is accepted for publication. We propose to discuss a 

few interesting cases encountered with our centre relevant 

to the points mentioned above and the options available 

with the editors of journals for the strict compliance of 

their latest guidelines. 

552 - The spectrum of macrofungi in beech forests 

under different air pollution pressure 

M. Pavlik 

Dept.of forest protection and game management, Technical 

university, Masarykova 24, Zvolen 960 53, Slovakia. - Email: 

mpavlik@vsld.tuzvo.sk 

The base of our investigation is an assumption about 

affection of mycoflora structure by air pollution. The 

occurence of the ectomycorrhizal, saprobic and parazitic 

fungi was observed during three years at three beech 

stands. Conditions for fungi growth were nerly compatible 

on all stands, difference was only in degree of pollution 

pressure. Number of fungal species and fruitbodies, 

together with other characteristics of investigated stands 

(health condition of trees, soil analysis, climatic 

conditions), were analysed using mathematic-statistical 

methods. In this way factors affecting a fungi production in 

beech forest exposing to various pollution pressure was 

choosen. An important result of this work is a possibility to 

use the macromycetes for bioindication of various polluted 

beech forest stands: - The share of mycorrhizal species 

significantly decreases and the share of saprobic ones 

significantly increases at beech stands under pollution 

pressure; - The mycorrhizal species of genus Cortinarius, 

Cantharellus, Hydnum, Ramaria and Tricholoma are 

practically missing at these stands; - At the same time the 

number of saprobic species of genus Clitocybe, Collybia, 

Coprinus and Lepista expressively increases. 

168 


553 - Refuges of heat-demanding polypore fungi near 

geothermal waters in Pribaikalia 

T.A. Penzina 

Siberian Institute of Physiology and Biochemistry Of 

Plants, SD RAS, P/O Box 1243, Irkutsk, Russia. - E-mail: 

tania@webtaiga.com 

Refuges of polypore fungi which are located near 

geothermal waters of Pribaikalia, are distinguished by the 

degree of local heat-provision and the history of site 

evolution. Four types of refuges in the mountainous areas 

of Pribaikalia have been indentified. The fourth type 

includes sites in which all the nemoral complexes have 

have been preserved and concentrated in the warmer steppe 

landscapes. The site near Selenga river which includes 

broad-leaf associations of relics nemoral Japanese Elm 

(Pleshanov, Penzina, 1996). Species composition of the 

first zone is characterized by xerophylies: Gloeophyllum 

sepiarium (Wulf.:Fr.) Karst., G. trabeum (Pers.:Fr.) Murr., 

Trametes ochracea (Pers.) Gilbn.& Ryv. It is a zone of 

direct influence of hot springs. It is necessary to consider 

this biotope as an extreme for wood attaching associations - 

the wood is occupied by fungi for a short period, 

hypothetically, in the winter, when the spring water is 

natually cooler. The biotopes of the Selenga floodplain, 

which belong to the second zone of refuges, are 

distinguished by the maximum size of the fruiting bodies of 

the perennial polypore fungi in Pribaikalia, as well as a 

wealth of species diversity and structure of mycocommunity. 

Striking representatives of the thermophylic 

complex, which have been discovered in relict Elm forest, 

are Trametes conchifer (Schw.:Fr.) Pil. on fallen branches 

of Elm, and Inonotus hispidus (Bul.:Fr.) Karst., living on 

the trunks of live Elm trees. 

554 - New reports of polypores from the Dominican 

Republic, Greater Antilles 

O.P. Perdomo 1* & L. Ryvarden 2 

1 

Dirección Nacional de Vida Silvestre y Biodiversidad, 

Autop. J.F. Kennedy Km 6.5, Jardines del Norte, Santo 

2 

Domingo, Dominican Republic. - Botany Department, 

Biological Institute, University of Oslo, P. O. Box 1045, 

Blindern, N-0316 Oslo, Norway. - E-mail: 

paino@codetel.net.do 

Ciferri (1929a) and Murrill (1915) reported 17 species of 

polypores in 13 genera from the Dominican Republic 

according to modern taxonomy. We surveyed polypores in 

the Dominican Republic beginning in 1996. Decock & 

Ryvarden (2000) described Perenniporia cremeopora and 

Lodge, Ryvarden & Perdomo (2001) described Antrodia 

aurantia from the DR. Minter et al. (2001) listed 80 of our 

new records. In addition to the published records noted 

above, we report here six new species records for the 

Greater Antilles: Antrodiella parasitica, Chaetoporellus 

latitans, Gloeoporus dichrous, Inonotus cf. micantissimus,


Skeletocutis carneola and S. vulgaris. We report here an 

additional 30 species that are new records for the 

Dominican Republic: Amylosporus campbellii, Antrodia 

malicola, A. sinuosa, Antrodiella hydrophila, Aurificaria 

luteo-umbrina, Ceriporia purpurea, Coriolopsis floccosa, 

Cyclomyces iodinus, C. tabacinus, Daedalea aethalodes, 

Fomitopsis spraguei, Gloeophyllum abietinum, G. 

mexicanum, Hapalopilus salmonicolor, Inonotus hispidus, 

Perenniporia cf. tephropora, Phellinus callimorphus, P. 

ferrugineovelutinus, P. linteus, P. merillii, P. orientalis, P. 

pini, P. robustus, Rigidoporus concrescens, Skeletocutis cf. 

carneogrisea, Trametes lactinea, Trichaptum perrottetii, T. 

sector, Tyromyces leucomallus and T. venustus. Including 

the new records from this study, there are 133 species in 46 

genera of polypores known from the DR. Part of each 

collection is deposited at JBSD and O. 

555 - Screening for lichenicolous fungi of Usnea 

D. Persoh 

Lehrstuhl Pflanzensystematik, Universität Bayreuth, 

Universitätsstraße 30 - NW I, D-95440 Bayreuth, 

Germany. - E-mail: derek.persoh@uni-bayreuth.de 

Fungi were isolated from individuals of a species of Usnea 

(U. cf. subfloridana) at three sites in the European Alps, 

one of which was sampled thrice. Under semi-sterile 

conditions small pieces of the lichen were transferred in the 

field to various culture media, torn apart and spread over 

the agar plate. In the laboratory, growing hyphae were 

excised and transferred to fresh media until pure cultures 

were obtained. From these strains DNA was isolated and 

the SSU and ITS regions of the nrDNA were sequenced. 

The SSU sequences were aligned with about 1000 fungal 

sequences from GenBank and a phylogenetic tree, being 

presented, was calculated. A strain was assigned to a 

certain taxonomic group when its sequence clustered as 

ingroup in a clade consisting exclusively of taxa of the 

same taxonomic group according to the current 

classification. As a result, three of the 26 isolated fungi 

were identified as Basidiomycetes, whereas the remaining 

taxa belong to the Pezizomycotina (Ascomycetes). Of the 

latter ones 17 strains could be assigned to the following 

classes: Chaetothyriomycetes, Dothideomycetes, 

Eurotiomycetes, Lecanoromycetes, Leotiomycetes and 

Sordariomycetes and nine to the orders Chaetothyriales, 

Diaporthales, Eurotiales, Lecanorales and Xylariales. Five 

strains were identified at family level (Parmeliaceae, 

Trichocomaceae and Valsaceae). In addition, the nearest - 

yet sequenced - relatives of the isolated fungi were 

specified by an ITS nrDNA blast search in GenBank. 

556 - Endophytic fungi isolated from maize (Zea mays) 

and their possible role for biological control 

I.C. Pimentel 1 , R.A. Campos 2 , W.L. Araujo 3 , N.M. Barros 2 

& J.L. Azevedo 3* 

1 Universidade federal do Paraná, POBox 19001-CEP 

81531-990-Curitiba-PR, Brazil. - 2 Universidade de Caxias 

do Sul, POBox 1352-CEP 95001-970-Caxias do Sul-RS, 

Brazil. - 3 ESALQ-Universidade de São Paulo, POBox 83- 

13400-970-Piracicaba-SP, Brazil. - E-mail: 

jazevedo@esalq.usp.br 

Endophytic fungi were isolated from leaves and stems of 

maize collected from plants growing in greenhouse and 

field conditions. Samples were collected after 20 and 40 

days seed germination. Nine main genera of fungi were 

isolated and the most frequent genera found were 

Colletotrichum, Fusarium and Acremonium. From 1728 

plant fragments quantitative significative differences were 

found being field plants about three times more intensly 

colonized by endophytes when compared to greenhouse 

plants. No significative differences were encoutered from 

number and genera of fungi collected from 40 days old 

plants when compared to 20 days old ones. Endophytes 

from the genera Beauveria and Paecilomyces were isolated 

both from fiels and greenhouse plants. The Beauveria 

isolates were classified as B. bassiana but molecular 

analysis showed that they differ from the main B.bassiana 

strains used in Brazil for insect biological control. B. 

bassiana isolates were tested against maize pest insects and 

against the bovine tick Boophilus microplus. Bioassays 

have shown that endophytic isolates were as effective as 

comercial strains using concentrations varying from 100 to 

100 millions conidia/mL, the last being the most effective 

causing up to 75% mortality in comparison with the 

controls. Our result support the possible biological control 

role of endophytic entomopathogenic fungi as being 

suggested already by other authors. 

557 - New and interesting findings of macrofungi from 

the islands of Andros and Naxos (Cyclades, Greece) 

E. Polemis 1* , G.I. Zervakis 1 & D.M. Dimou 2 

1 National Agricultural Research Foundation, Institute of 

Kalamata, Lakonikis 85, 24100 Kalamata, Greece. - 

2 Agricultural University of Athens, Laboratory of 

Agricultural Microbiology, Iera Odos 75, 11855 Athens, 

Greece. - E-mail: elipolem@otenet.gr 

Reports on the biodiversity of macromycetes in islands and 

coastal areas of southeastern Mediterranean are practically 

non-existent. Investigation of typical biotopes of Andros 

and Naxos (the two largest islands of Cyclades in the 

Aegean archipelago), e.g. xerophytic heaths, sea-shore 

grasslands, dunes and maquis, resulted in significant 

records. The agaricoid taxa Conocybe rickenii, Crepidotus 

epibryus, Lepista rickenii, Macrolepiota phaeodisca, 

Marasmiellus rosellus, Marasmius anomalus var. 



microsporus, M. wynnei f. carpathicus, Melanoleuca 

polioleuca, Mycena zephirus, Panaeolus cintulus, 

Psathyrella microrhiza and Tubaria autochthona were 

recorded for the first time in Greece, while Ceratobasidium 

cornigerum, Mycoaciella bispora, Myxarium laccatum and 

Ramicola haustellaris represent first reports for the 

respective genera in Greece. Of particular interest are 

records of lignicolous species from Mediterranean 

evergreen scrubs and frygana, i.e. Hyphodontia juniperi 

and H. sambuci on Thymus capitatus; Peniophora lycii, P. 

pithya and P. tamaricicola on Pistacia lentiscus; 

Aleurodiscus dextrinoideocerussatus, Hyphodontia pruni, 

Peniophora lycii, Steccherinum fimbriatum and S. 

ochraceum on Genista acanthoclada; Aleurodiscus minor 

and Lentinellus omphalodes on Cistus sp.; Gloeophyllum 

abietinum, Perenniporia rosmarini and Peniophora piceae 

on Juniperus phoenicea; and Dacrymyces lacrymalis and 

Pluteus nanus f. nanus on Nerium oleander. 

558 - BIOTEC Culture Collection (BCC) in Thailand 

W. Potacharoen * , S. Sivichai & M. Tanticharoen 

BIOTEC - Culture Collection, National Center for Genetic 



Khlong 1, Khlong Luang, Pathum Thani12120, Thailand. - 

E-mail: wanchern@biotec.or.th 

BIOTEC Culture Collection is a specialized collection 

formally established in 1996 under the National Science 

and Technology Development Agency (NSTDA). The 

principle objective is to provide safe deposition of cultures 

for researchers; to preserve and maintain microorganisms 

isolated from Thailand for future use; and to manage strain 

data information. Another major role is to supply cultures 

and related information for an in-house screening 

programme for drug discovery from microorganisms. The 

collection comprises 10,000 isolates fungi from various 

sources and habitats: insect fungi (1,618), aquatic 

freshwater and marine habitats (1,324), rice blast fungi 

(1,148), soil including leaf litter and humus (973), wooddecay 

Xylariaceae and Basidiomycota (898), plant seeds 

(389 strains), endophytic fungi (754), lichens (247) and 

taxa from other sources (1,090). The collection also 

includes bacteria and actinomycetes (1,789) and yeasts 

(342). The preservation methods used are freezing at -80 

°C, in a vapor phase of liquid nitrogen at -170 °C; freezedrying 

and under liquid paraffin oil. The essential strain 

data of cultures being maintained are used to create a 

required database and produce the catalogue of cultures. 

On-line database is now available at URL: 

http://bcc.biotec.or.th/. 

170 


559 - Taxonomic verification of basidiomycetes cultures 

in the LE (BIN) collection 

N.V. Psurtseva 

Komarov Botanical Institute RAS, Prof. Popov Str., 2, St. 

Petersburg, 197376, Russia. - E-mail: 

NadyaPsu@NP1512.spb.edu 

The Komarov Botanical Institute Basidiomycetes Culture 

Collection - LE (BIN) maintains over 1000 strains of 428 

species of aphyllophoroid, agaricoid and gasteromycetoid 

macromycetes collected mainly in Russia and former 

USSR. Conservation of Basidiomycetes diversity ex situ 

with a special interest in rare and endangered species as 

well as cultural maintenance of mushroom taxa with 

biological activity, especially those could be used in 

medicine and industry, is one of our primary goals. 

Taxonomic verification of Basidiomycetes cultures 

maintained in the culture collections is of great importance. 

In the process of starting and transferring living cultures 

error can be introduced in two ways. First, cultures can be 

obtained from incorrectly identified basidiomata or the 

identification has been done according to an out of date 

classification. Recent studies on Flammulina and Pleurotus 

species enable us to carry out taxonomic verification of LE 

(BIN) cultures from these genera by mating study 

according to the biological species concept. Second, 

replacement of a Basidiomycetes culture with a a different 

species, mainly from the group of imperfect fungi, can 

occur during the strain isolation or subculturing. This often 

happens in culturing of slowly growing and 

ectomycorrhizal fungi. To veryfy a cultural strain's identity 

and purity detailed studies of cultural characters, fruiting of 

strains in culture and molecular analysis are being used in 

the LE(BIN) Collection. 

560 - Contribution of hyphal length in predicting P 

influx in maize and groundnut under field condition 

A. Rakshit 1* , P.B.S. Bhadoria 1 & N. Claassen 2 

1 Agri and Food Engg Dept., IIT, Kharagpur-721302, W.B, 

India. - 2 Agricultural Chemistry, University of Gottingen, 

University of Gottingen,Germany, Germany. - E-mail: 

amitabha_1975@yahoo.co.in 

When P is deficient in soil solution, the critical root 

parameter controlling its uptake is its surface area. Hyphae 

of mycorrhizal fungi have the potential to greatly increase 

the absorbing surface area of the root. Modified gridline 

intersect method was employed for extracting hyphae from 

the soil and quantifying length under field conditions in a 

Benomyl treated and non treated soil with the following 

three P levels: P 0,P 50 and P 400mgkg-1 soil as single 

super phosphate. To asses the importance of mycorrhizal 

hypha nutrient uptake model was used. Wettable powder of 

the fungicide Benomyl was mixed thoroughly to 15 cm soil 

depth at 500 kg ha-1. Treated plots were covered by paddy 

straw for two weeks to avoid excessive heating. There was


significant interaction between benomyl treatment and 

phosphorus application on hyphal length. P 0 recorded 

maximum hyphal length and was significantly higher as 

compared to other P application levels for both the crops. 

Maximum hyphal length was observed for maize crop than 

groundnut. When hyphal length was included in model 

calculations it gave better prediction of P influx in to the 

roots of maize and groundnut. When hyphal length was not 

included in model calculation, under prediction of P influx 

was more affected at lower P levels. It indicates that P 

solubilization by root exudates might have played more 

important role than AM in P uptake by maize and 

groundnut. 

561 - Rare taxa in Estonian lichen flora - are they really 

rare? 

T. Randlane * & A. Saag 

University of Tartu, Lai 38, 51005 Tartu, Estonia. - E-mail: 

randlane@ut.ee 

The recent checklist of lichenized and lichenicolous fungi 

of Estonia (Randlane & Saag 1999), which is mainly based 

on herbarium material, contains 851 species from 200 

genera. Latest data available 

(http://www.ut.ee/lichens/fce.html) indicate the presence of 

909 species in Estonia. In the checklist, the distribution of 

taxa in five regions of Estonia and their frequency 

according to six frequency classes has been presented. Rare 

species with 10 or less localities form 64% of the entire 

Estonian lichen flora. The analogous figure for vascular 

plants in Estonia is 36% and for bryophytes - 45%. The 

unusually large share of rare lichen taxa is partly due to the 

great diversity of different habitats and substrata suitable 

for lichenized fungi. Still, these data might be misleading 

because of the unsufficient recording of different species. 

Several new localities have been reported for 'rare' species 

during the last three years; six lichens that were considered 

to be extinct have been re-found. The analysis of worldwide 

distributional patterns of taxa would help us sort out 

these species which are locally really rare (and therefore 

also endangered) because they occur at the periphery of 

their range. In Estonia, e.g. Arctoparmelia incurva, 

Caloplaca jungermanniae, Flavocetraria cucullata occur 

at their southern limits while Cladonia convoluta, 

Flavoparmelia caperata, Phaeophyscia chloantha are 

recorded at the northern limits of their distribution. 

562 - Conservation and valorisation of Pleurotus spp. 

germ plams 

P. Rapanà * & F. Di Mario 

CNR IBAF - IBESM, Area della Ricerca di Roma, via 

Salaria km 29,300 Monterotondo sc 00016 (Roma), Italy. - 

E-mail: pompilio.rapana@mlib.cnr.it 

Germ plasm collection must assure the conservation of 

genetic diversity also in view of their possible 

technological application. To be exchanged and used, germ 

plasm must be characterized by an univocal fingerprint 

able to assure its identity and stability. This is an important 

condition for comparing the results obtained in different 

studies. Pleurotus spp. are widespread populations among 

them, Pleurotus eryngii, P. e. ferulae and P. e. nebrodensis 

are typical Mediterranean populations. The International 

Bank of Edible Saprophytic Mushrooms of Italian National 

Council of Research owns 36 isolates of P. ostreatus, 313 

of P. eryngii, 48 of P. e. ferulae and 22 of P. e. 

nebrodensis. These isolates were collected in view of their 

applications in biotechnologies as agents able to transform 

agricultural wastes into both an edible biomass and starting 

material for high value products extraction. 

Polysaccharides in particular 3 β glucans have been 

purified. The isolates, supplied by Research Institute 

donors, have been genetically characterised by RAPDs 

fingerprint. Moreover genetic approaches aimed at 

evaluating population biodiversity and genetic relationship 

among species are running with molecular markers as 

microsatellites, AFLP, and RFLP of amplified ITS and IGS 

rDNA region. A good level of variability in P. eryngii and 

in the correlated population/species P. e. ferulae and P. e. 

nebrodensis have been evaluated. A similar behaviour has 

been made evident also in P. ostreatus. 

563 - Comparative diversity of vesicular arbuscular 

mycorrhizas in New Zealand vineyards under organic 

and conventional management regimes 

H.J. Ridgway * & A. Stewart 

Lincoln University, PO Box 84, Canterbury, New Zealand. 

- E-mail: ridgwh@lincoln.ac.nz 

Many economically important plant species form complex 

symbioses with vesicular arbuscular mycorrhizal (VAM) 

fungi. Considerable research data indicates that these fungi 

impact on productivity, nutrient acquisition, pathogen 

defence and ecosystem stability. In natural systems, 

grapevines are normally infected with VAM and these 

associations have been shown to improve grape growth. In 

this study, soil samples were removed from the base of 

three common grapevine rootstock varieties at two South 

Island vineyards, one under conventional management, and 

the other under organic production (certified in 2000). 

Spores were collected by wet sieving, the genera identified 

by microscopic examination and expressed on a per gram 

basis. The individual VAM species present were 

determined using a combination of morphological criteria 

and sequencing of the ribosomal DNA. Root fragments 

were also cleared, stained and assessed for VAM 

colonisation. Implications of VAM diversity in New 

Zealand vineyards and the effect of management systems 

on population structures are discussed. 



564 - Diversity and conservation of fungi on Danube 

flood-plain forests 

S. Ripková 1 & P. Lizon 2* 

1 Dept. of Botany, Komensky University, Révová 39, SK-811 

02 Bratislava, Slovakia. - 2 Dept. of Cryptogams, Institute 

of Botany, Dúbravská 14, SK-842 23 Bratislava, Slovakia. 

- E-mail: botupali@savba.sk 

River floodplain forests become extremely endangered in 

last century and there are only scarce data on fungi of those 

habitats. Field research on biodiversity of macrofungi in 

two Danube islands, Sihot' and Sedláckov ostrov (city of 

Bratislava) resulted in 211 species of Ascomycota and 

Basidiomycota. Following taxa recorded during our 

research are, according to the 3 rd edition of the Red List of 

Slovak fungi, endangered: Ceriporia purpurea, Crepidotus 

crocophyllus, Hypoxylon ticinense, Hyphodontia latitans, 

Marasmius minutus, Ossicaulis lignatilis, Oxyporus 

latemarginatus, Oxyporus philadelphi, Phlebia ryvardenii, 

Rhodotus palmatus, Scutellinia legaliae, Scutellinia 

sinensis, Spongipellis fractipes, Trichophaea variornata, 

Trichophaeopsis bicuspis. 

565 - Different approaches on rice blast disease in Italy: 

an in vitro assay for assessing pathogenicity of the 

Italian Pyricularia grisea collection 

D. Rodino 1* , G. Forlani 2 , A.M. Picco 1 , A. Rozkowicz 2 , E. 

Lorenzi 1 & M. Rodolfi 1 

1 Dip.to Ecologia del Territorio e degli Ambienti Terrestri, 

Sezione di Micologia, Università degli Studi di Pavia, Via 

S. Epifanio, 14 - 27100 Pavia, Italy. - 2 Dip.to di Biologia, 

Sezione di Biologia Evolutiva, Università degli Studi di 

Ferrara, Via L. Borsari, 46 - 44100 Ferrara, Italy. - Email: 

dorianarodino@hotmail.com 

Blast disease, caused by Pyricularia grisea (teleomorph: 

Magnaporthe grisea), represents the main threat to rice 

crop. Blast pathogen population is divided into races, or 

lineages, defined according to virulence spectrum on 

differential rice cultivars; unfortunately, pathogenicity of 

races identified in a country cannot be compared with that 

of races in other countries, and in vivo screening 

procedures are both complex and time-consuming. Italian 

lineages have been only partially investigated, and a lot of 

work is still needed as many (125) rice cultivars are grown 

in Italy. A survey on a collection of P. grisea isolates, 

currently in progress in our laboratories, is the first step to 

characterise the Italian population. To date 103 strains have 

been isolated, and are available for further analysis. In 

order to characterise fungal isolates different methods 

(greenhouse and molecular screening, in vitro assays) 

could be used. An in vitro test was set up using partially 

purified elicitors, instead of viable pathogen, and rice calli. 

Suspension cultures of two rice cultivars (Gigante Vercelli 

and Maratelli, resistant and susceptible to blast, 

respectively) were used. In all cases a sharp rise of 

172 


hydrogen peroxide production was found soon after elicitor 

addition. Interestingly, both control and elicited rates were 

3 to 5-fold higher in the tolerant cultivar than in the 

susceptible one, thus strengthening the possibility to use 

this model to simulate in vivo infection. 

566 - Wild mushrooms: valorisation and conservation 

A.P. Rodrigues 1* & S.P.B. Neves 2 

1 Parque Natural Montesinho, Apartado 90, 5301-901 

Bragança, Portugal. - 2 Área Paisagem Protegida Serra do 

Açor, Mata Margaraça, Casa Grande, 3305 Benfeita, 

Portugal. - E-mail: pnm.rodriguesap@icn.pt 

In recent years, in Portugal, the harvest of wild edible 

mushrooms has intensified greatly motivated by 

commercial valorisation of this resource in the international 

markets. The reduction of the mycological patrimony, and 

biodiversity in general, is mentioned as the main 

consequence of this process. The absence of regulation that 

discipline the harvest and commercialisation of wild edible 

mushrooms, as well as the inexistence of a global strategy 

that promotes the conservation, management and 

sustainable use of the fungus species and its habitat, are 

considered as the main factors that contribute to this 

situation. In this area, representatives of several entities 

under the coordination of the Instituto da Conservação da 

Natureza (Nature Conservation Institute) elaborated a 

proposal of intervention that created guidelines and 

indispensable measures to conserve and regulate the 

mycological resources. Among the several measures, 

suggested are highlighted: urgent regulation of the harvest, 

reception, concentration and commercialisation of wild 

edible mushrooms; creation of a good practices code; 

implementation of measures to promote the knowledge on 

portuguese fungal diversity and monotorising of particular 

interesting species; promotion of training courses, 

sensibilisation and environmental education which 

emphasize the ecological and economical importance of 

fungi. The present work intends, in a summary form, to 

promote the main aspects that resulted from this initiative. 

567 - Measuring macrofungal diversity in four 

Portuguese protected areas 

A.P. Rodrigues 1 , P. Gonçalves 2 , R. Neiva 3 , S.M. Branco 1 , 

S. Baptista 3 , S. Mota 2 & S.P.B. Neves 4* 

1 Parque Natural de Montesinho, Apartado 90, 5301-901 

Bragança, Portugal. - 2 Reserva Natural Serra Malcata, 

Largo de S. Tiago, 6320-325 Sabugal, Portugal. - 3 Parque 

Natural Serra Estrela, Rua 1° Maio, 2, 6260 Manteigas, 

Portugal. - 4 Área Paisagem Protegida Serra do Açor, Mata 

Margaraça, Casa Grande, 3305 Benfeita, Portugal. - Email: 

spbneves@hotmail.com 

The struggle to bring mycological patrimony to the list of 

issues involved in biological research and conservation


prioritiy, and as an important item to the management of 

protected areas, is recent in Portugal. The increasing 

perception that the harvest of wild mushrooms has greatly 

intensified in recent years and the negative influences both 

on general biodiversity and on mycological diversity that it 

might have, and also the general lack of knowledge on 

Portuguese fungal patrimony, were the main reasons to 

initiate a project involving four protected areas in Central 

and Northern Portugal coordinated by the Portuguese 

Institute for the Conservation of Nature. This project 

involves the measurement of macrofungal diversity in 

several priority habitats listed in the Habitats Directive 

using a common sampling method. The macrofungal data 

will be complemented with environmental and vegetation 

parameters necessary for the assessment of the micota's 

ecological context. 

568 - Foliicolous lichens of La Réunion 

S. Rønhede, S. Johansson * & U. Søchting 

Department of Mycology, Bothanical Institute, University 


Copenhagen K, Denmark. - E-mail: 

s.johansson@get2net.dk 

The foliicolous lichen flora of the French island La 

Réunion was investigated based on 113 leaf samples 

collected at seven localities. They represent different types 

of rain forest at altitudes from 160 to 1700 m. 27 species 

were identified, of which 16 are new to La Réunion. 

Tricharia amazonum is reported for the first time outside 

the neotropics, and Porina atriceps and Badimia pallidula 

are new to the African continent with adjacent islands. 

569 - Vertical distribution of ectomycorrhizal fungi in a 

podzol profile 

A. Rosling 1* , R. Landeweert 2 , B.D. Lindahl 1 , K.-J. 

Larsson 3 & T.W. Kuyper 2 

1 Department of Forest Mycology and Pathology, SLU, Box 

7026, 750 07 Uppsala, Sweden. - 2 Sub-dept of Soil Quality, 

Wageningen University, Box 8005, NL-6700 EC, 

Wageningen, The Netherlands. - 3 Systematic Botany / 

Herbarium GB, Botanical Institute, Göteborg University, 

Box 461, SE-405 30 Göteborg, Sweden. - E-mail: 

Anna.Rosling@mykopat.slu.se 

Ectomycorrhizal (ECM) fungi form symbiotic associations 

with roots of woody plants and are important in nutrient 

uptake in boreal forests. Podzols are highly stratified soils. 

Most studies of ECM species composition focus on the fine 

roots in the organic soil. The variation in ECM community 

structure between different soil horizons is thus poorly 

understood. We studied the vertical distribution of ECM 

roots in a podzol soil in the north of Sweden. In three 

continuous soil columns, seven horizons were 

distinguished and sampled, from the top organic layer to 

the bottom parental material at 50 cm depth. Root tips from 

each horizon were sorted into morphotypic groups. These 

were verified and identified by sequencing of the rDNA 

ITS region. Root tip density decreased from the O horizon 

down to the E2 horizon but increased again in the B 

horizon before declining to low values in the C horizon. 

Half of the root tips were found in the mineral soil. The 

degree of ECM colonization varied from 60 to 98%, but 

did not follow a clear pattern with respect to depth. We 

distinguished 22 ECM taxa, half of these were found only 

in the mineral horizons. The abundance of taxa in different 

horizons was analysed by correspondence analysis. We 

found a significant relationship between ECM taxon and 

soil horizon. The different horizons in a podzol profile 

represent different environments and the specific 

distribution of certain fungi could be explained by their 

functional role in the system. 

570 - Leprose lichens in Estonia 

L. Saag 

University of Tartu, Lai street 38, Tartu, Estonia. - E-mail: 

lsag@ut.ee 

Over 60 usually sterile sorediate crustaceous lichen species 

have been registered from Estonia. Such species occur in 

many genera which ordinarily produce ascocarps. Majority 

of the constantly asexual taxa are gathered into g. Lepraria 

and g. Leproloma (Lichenes Imperfecti), some of them 

have tentatively been assigned to other genera (Fuscidea 

pusilla Tonsberg, Lecanora norvegica Tonsberg et al.). 

The first checklist of Estonian lichens published in 1970 

included 8 Lepraria species, only one of them - L. incana - 

belongs to that genus nowadays. Primitive morphology and 

often complex secondary chemistry of representatives of 

the leprose growth form makes their identification 

complicated and chemical analysis (TLC) essential. 

Currently 11 Lepraria and 1 Leproloma species are 

reported from Estonia, occurrence of lately described 

Lepraria atlantica Orange is in question. 

571 - Experimental study for increasing productivity of 

Boletus edulis s.l. in Italy 

E. Salerni * & C. Perini 

University of Siena Dep. Environmental Sciences, Via P.A. 

Mattioli - 4 - 53100 Siena, Italy. - E-mail: salerni@unisi.it 

Species grouped together as Boletus edulis s.l. are the 

mushrooms in highest demand and best known in Italy: 

they are a typical minor forest product of the hill and 

mountain belt of the penisula, of major economic 

importance in some cases surpassing the commercial value 

of timber. More than 140 ectomycorrizhal fungi are 

regarded as edible but only a small percentage have been 

successfully cultivated. For the other species, including the 

highly prized penny bun mushroom, no technique of stable 



inoculation on a regular basis in the host plant has jet been 

found. This means that the entire demand for this product, 

which amounts to more than 70,000 tons/year in Italy is 

met by mushrooms gathered in the wild. The present study 

proposes to clarify the ecological requirements of these 

species through manipulation of certain environmental 

parameters, in order to provide indications for 

environmental management aimed at conserving the ideal 

habitat of this culinary delicacy. The study area consists of 

four reforestations in two age classes (30 and 60 years) of 

Abies alba Miller on the Mt. Amiata (Tuscany, Italy). This 

area is historically famous for its mushrooms, but has 

recorded a decline in production in recent years. Two years 

of preliminary studies have shown that certain conditions 

are necessary for increasing productivity of B. edulis s.l. in 

these woods: forest age must not exceed 50 years, forest 

density should be intermediate and litter should be 

maintained. 

572 - An update on the biogeographical distribution 

area of the genus Anthracoidea (Ustilaginomycetidae) in 

light of new species 

V. Salo 

University of Helsinki, Department of Applied 

Biology/Botany, P.O.Box 27 (Latokartanonkaari 5), FIN- 

00014 University of Helsinki, Finland. - E-mail: 

vanamo.salo@helsinki.fi 

Fungal samples loaned from herbaria were examined with 

light and scanning electron microscopy. The material 

consisted of various plant parasitic fungi on inflorescences 

of Carex spp. from all over the world. Most of the fungi 

were identified as belonging to the smut genus 

Anthracoidea. After spore morphological comparisons, the 

conclusion of the study was that several Anthracoidea 

species new to science can be described. The smut flora of 

Europe is well known and hence, new species from 

European material could not be found. All other areas are 

poorly studied despite of some published local smut 

surveys. New species could be found from Asia, North and 

South America. The distribution areas of host plants and 

their biodiversity are very likely to have a profound effect 

on their parasitic fungi. Compared to Europe many parts of 

Asia and America have a much richer sedge flora but still 

fairly few species of Anthracoidea have been found. 

Reasons for this are certainly manifold. Numerous species 

occurring on related sedge plants are reported to be 

common to e.g. Europe and Asia or Europe and North 

America. However, Anthracoidea flora of Central Asia is 

seemingly very unique and differs from Northern or 

Eastern Asian flora. On the other hand, some sedges may 

bear different smut species but they occur in different 

geographical areas. Some interesting examples will be 

presented with area maps. 

174 


573 - Mycorrhizal interaction in the wild relatives of 

grain legumes in Palney Hills of Western Ghats of 

South India 

P. Saravana Kumar 1* & S. Ignacimuthu 2 

1 University of Peradeniya, Senior Lecturer,Department of 

Botany, Faculty of Science, University of Peradeniya, 

Peradeniya, Sri Lanka. - 2 Bharathiyar University, Vice 

Chancellor, Bharathiyar University,Coimbatore, Tamil 

Nadu, India. - E-mail: psaravan@pdn.ac.lk 

Pulse crops have certain uniform features which together 

make them indespensible if not difficult to replace. In 

India, as many as 28 wild legumes are known to be 

consumed as pulses by different tribal communities. The 

importance of legumes with arbuscular mycorrhizae are 

well defined. But available information on wild legumes is 

highly limited, especially in Indian scenario. The diversity 

of mycorrhizal fungi associated directly with wild legumes 

is not well understood. Estimation of mycorrhizal fungi in 

rhizosphere has little relationship with colonization of roots 

by individual species. Our presentiation will discuss a 

complete analysis of three interactions including growth 

and biochemical variations with reference to natural 

landscape conditions. 

574 - Entolomatoid fungi from Korup National Park, 

Cameroon 

E.M. Saunders * & P.J. Roberts 

The Mycology Section, Royal Botanic Gardens, Kew, 

Richmond, Surrey, TW9 3AE, U.K. - E-mail: 

E.Saunders@rbgkew.org.uk 

The Korup National Park consists of ancient, mainly 

lowland, rain forest in the South West Province of 

Cameroon and is acknowledged to be one of the most 

species-diverse areas in equatorial Africa. Macrofungi were 

collected in April - May 1996 and March - April 1997, at 

the beginning of the rainy season. As a result of these 

visits, an initial three papers have been published on the 

clavarioid fungi, heterobasidiomycetes, and corticioid fungi 

of the National Park. Several collections were made of 

entolomatoid agarics, representing at least 20 taxa. These 

are now being examined at the Royal Botanic Gardens, 

Kew, and provisional information, including descriptions 

and photographs, is presented here. It is hoped this work 

will result in the identification of some previously 

undescribed taxa and will provide essential baseline data 

for the better understanding of fungal biodiversity in the 

African wet tropics.


575 - Micromycetes isolated from air and soil in an 

Italian carsic cave 

E. Savino 1* , C. Buratti 1 , G. Caretta 1 , A. Fiorina 2 & E. 

Salerni 3 

1 Dep. Ecologia Territorio & Ambienti Terrestri, Via S. 

Epifanio, 14 - 27100 Pavia, Italy. - 2 Servizio Pneumologico 

A.S.L. 2 Savonese, Via Adelasia, 57 - 17021 Alassio (SV), 

Italy. - 3 Dep. Environmental Sciences, Via P. A. Mattioli, 4 

- 53100 Siena, Italy. - E-mail: esavino@et.unipv.it 

Caves are an hostile habitat to biota: do not support plant 

life and few animal species are adapted to live inside. Very 

little investigation has been devoted to fungal ecology of 

caverns and it is believed that no species is peculiar of this 

habitat. Micromycetes present in an Italian carsic 

showcave, situated at Toirano (SV), have been studied. 

Sixteen sites were sampled in a distance of about 400 

metres from the entrance. Air sampling was carried out by 

settle plate technique and by surface air sampler (ParTrap 

Fa 52); soil analysis by hair baiting technique. Statistical 

analysis of the results, involving chi-square, has been 

performed. A total of 76 species, belonging to 47 genera, 

was recorded. Alternaria alternata, Aureobasidium 

pullulans var. pullulans, Cladosporium cladosporioides, C. 

herbarum, Penicillium brevicompactum, Trichophyton 

terrestre and Simplicillium lamellicola (abundant because 

it is involved in the process of stalactite growth), were 

dominants. Among the sporadic species, should be noticed 

Conidiobolus coronatus, mostly saprotrophic but also 

insect parasite and causal agent of nasal granuloma in man. 

Comparison between the number of species and the 

different methodological techniques used results 

statistically significant. In conclusion, mycoflora in a cave 

can be rather diversified; that can be mainly due to the 

atmospheric circulation and the presence of animals or 

their debris. 

576 - Chemotaxonomic groups in species of Phyllosticta 

based on the cell wall polysaccharides 

R. Senthil Kumaran * & J. Muthumary 

Centre for Advanced Studies in Botany, University of 

Madras, Guindy Campus, Chennai - 600 025, India, India. 

- E-mail: sendil75@yahoo.com 

Phyllosticta is a Coelomycetous fungus producing 

pycnidial fruit bodies and causing leaf spot diseases in 

plants. It is a dimorph fungus having both perfect and 

imperfect states in their life cycle. Studies on the various 

species of Phyllosticta were collected from South India has 

been taken with a view to build up a genetic resources 

collection in our centre. For this purpose, fungal collections 

were made from various places in and around Tamilnadu, 

South India. So far more than 50 different species of 

Phyllosticta were collected and 30 of them were 

successfully grown in culture. In the case of Phyllosticta, 

identification of species is merely based upon the host 

specificity. But it is worthwhile to study the composition of 

cell wall polysaccharides along with the combination of 

morphological characters make an advantage in identifying 

the higher taxa at species level. In the present study cell 

wall Polysaccharides have been proved as 

chemotaxonomic markers used in the determination of 

natural groupings in species of Phyllosticta. In order to find 

out the chemical composition and structures of 

polysaccharides (alkali extractable & water-soluble) of cell 

wall and culture filtrate of the fungi were carried out with 

the above isolates. The inter specific relationship of 

different species of Phyllosticta was compared for the 

species studied based on the polysaccharide type of the cell 

wall. The results and significance of the findings are 

discussed in detail. 

577 - Mycorrhizal and ethanopharmacological value of 

some threatened orchid species of Shevaroy's, S. India 

S. Senthilkumar 

Dept. of Botany, St. Joseph's College (Autonomous), 

Tiruchirappalli-620 002, India. - E-mail: 

sen_kumar66@yahoo.co.uk 

Orchid mycorrhizas are symbiotic organisms, which 

establish an obligate symbiotic association with the roots of 

orchids. The orchid mycorrhizas represent the most 

widespread and the large majority of orchids must have 

evolved with compatibility system towards the fungal 

symbionts. In India, more than 2000 species of orchids 

have so far been reported in which mostly belong to 

epiphytic form received poor considerations than terrestrial 

form. All the threatened species of orchid under study 

penetrated an intracellular fungus called pelotons. The 

fungal hyphae may traverse various layers of velamen 

before entering into passage cells. From velamen itself the 

hyphae forming pelotons followed by exodermal cells and 

finally it reaches the interior cortex. Different 

histochemical, histoenzymological and fluorescent 

techniques were applied on peloton of the species viz. 

Bulbophyllum kaintense (Weight) Reichb.F., Coelogyne 

nervosa A.Rich and Gastrochillus acaulis (Lindley) Kuntz. 

However, antibacterial activity of organic and aquous 

extracts (leaf, root) of the former was carried out against 

different bacterial strains. From this study we can conclude 

that these threatened orchid species showed high 

antibacterial activity against the tested bacteria. These 

findings strongly imply that the involved orchid species is a 

major source for different ailments. Supported by: 

Department of Science and Technology, Government of 

India, No:SR/FTP/LS-217/2000. 



578 - Micromycetes in book depositories 

L.E. Sergeeva 

National Library of Russia, 18, Sadovaya st., 191069, 

St.Petersburg, Russia. - E-mail: kasht@aport.ru 

The micromycetes inhabiting air and the surface of the 

books were studied in 7 large book depositories of the 

National Library of Russia (Saint-Petersburg). The study 

was performed during 1995-2001. Micromycetes were 

isolated from numerous characteristic points and 

investigated in vitro. Total concentration of airborne fungal 

spores were examined. Furthermore physical 

characterististics of studied lodgings measured too. Total of 

100 fungal species were recorded. The list included species 

belonging to 33 genera, mainly from Deuteromycetes. 

Some of them had a wide occurance, whereas for some 

species it was the rather limited. The dominant and the rare 

species were indicated. Most likely the distinction of the 

fungal microcommunities in the studied book depositories 

usually were resulting from presence of accidental species. 

Season changes on fungal composition of revealed 

microcommunities were also recorded. Considerable 

fluctuations of concentration of airborne fungi depended 

upon ecological factors variation. Between them were 

tempaterature, humidity and chemical properties of paper 

type. There were determined the conditions increasing the 

content of viable and conversely, factors completely 

inhibiting the concentration of micromycetes in book 

depositories. 

579 - Biodiversity of keratinophyles in Heritage Taj 

City- Agra (India) 

J.N. Shrivastava, G.P. Satsangi, N. Govil * , M. Mathur & P. 

Saxena 


of Botany, Faculty of Science, Dayalbagh Educational 



Abstract In present investigation 360 soil samples from 

urban and rural areas of Agra, were analysed. Out of which 

296 were recorded positive for urban area and 238 for rural 

area. The percentage prevalence of dermatophytic and 

keratinophilic fungi were calculated 82.2% and 66.7% for 

urban and rural areas respectively. Total 21 species were 

recorded and isolated from baits amended soil collected 

different sites, out of which 9 sps were common in both 

sites. However, their prevalence varies with respect to sites. 

The major fungal species isolated from urban sites includes 

Chrysosporium (11 sps), Trichophyton (5 sps), while from 

rural area includes Chrysosporium (5 sps), Trichophyton (4 

sps), Myceliophthora (2 sps), Epidermophyton (1 sps) and 

Microsporum (1 sps). The maximum percentage prevalence 

of C. carmichaelii was observed in urban are (22.2%). 

While in rural areas M. vellerea was recorded. Thus, it is 

concluded that maximum prevalence of dermatophytes 

176 


were recorded in rural area due to prevailing unhygienic 

conditions as compared to urban areas. 

580 - Biodiversity of arbuscular mycorrhizal fungi in 

Indonesia 

R.D.M Simanungkalit 1* , K. Kramadibrata 2 & A.G. 

Gunawan 3 

1 Research Institute for Agricultural Biotechnology and 

Genetic Resources, Jl. Tentara Pelajar 3A,, Bogor, 

Indonesia. - 2 Research Centre for Biology, Jl. Ir. H. 

Juanda, Bogor, Indonesia. - 3 Faculty of Mathematics and 

Natural Sciences, Bogor Agricultural University, Jl. 

Padjadjaran, Bogor, Indonesia. - E-mail: 

borif@indo.net.id 

This paper reviews the results of several studies on the 

biodiversity of arbuscular mycorrhizal (AM) fungi in a 

wide range of ecosystems in Indonesia, including natural 

forest, agricultural crops, plantation crops, botanic garden 

and peat swamp forest. Glomus and Acaulospora are the 

most common genera found in those ecosystems. At least 

33 spesies of AM fungi in the 39 locations surveyed were 

described. Glomus with 17 species comprises G. 

aggregatum, G. albidum, G. citricolum, G. clavisporum, G. 

etunicatum, G. fasciculatum, G. fuegianum, G. geosporum, 

G. lacteum, G. macrocarpum, G. manihotis, G. 

microaggregatum, G. microcarpum, G. multicaulis, G. 

rubiforme, G. sinuosum and G. versiforme. Acaulospora 

with 10 species comprise A. bireticulata, A. delicata, A. 

foveata, A. longula, A. mellea, A. rehmii, A. scrobiculata, 

A. spinosa, A. tuberculata, and A. walkeri. Scutellospora 

with 4 species comprise S. calospora, S. heterogama, S. 

pellucida, and S. projectura. The genera Entrophospora 

and Gigaspora are represented with one species each, 

Entrophospora infrequens and Gigaspora gigantea. 

Acaulospora walkeri and Scutellospora projectura are the 

two new species reported from Indonesia. The number of 

AMF species varies from ecosystem to ecosystem, 

suggesting that their biodiversity can be influenced by soil 

conditions, cultural practices and vegetation types. 

581 - Diversity and host preference of leaf endophytic 

fungi in the Iwokrama Forest Reserve, Guyana 

C.M. Simmons 1 & P.F. Cannon 2* 

1 Iwokrama International Centre for Rainforest 

Conservation & Development, 67 Bel Air, Georgetown, 

Guyana. - 2 CABI Bioscience, Bakeham Lane, Egham, 

Surrey TW20 9TY, U.K. - E-mail: pcannon@cabi.org 

Endophytic fungi were isolated from living symptomless 

leaves of 12 tree species from two locations in the 

Iwokrama Forest Reserve, Guyana. 64 fungal morphotaxa 

were characterized from 2492 cultures, which were derived 

from a total of 2520 sample units. Species of 

Colletotrichum, Nodulisporium, Pestalotiopsis and


Phomopsis were most frequently isolated. Colonization 

was greater in samples from the midrib than in those from 

laminar tissue, and slightly greater at the tip of the lamina 

compared with the base of the leaf. In contrast to studies in 

temperate ecosystems, no distinct fungal communities were 

identified for individual plant species, suggesting that the 

degree of host preference is low. The implications for 

estimation of fungal diversity in tropical systems are 

explored. 

582 - Conservation status of the Agaricales, Boletales, 

Cantharellales, Lycoperdales, Phallales and Russulales 

of South Australia 

J.A. Simpson 1* & C.A. Grgurinovic 2 

1 

State Forests of NSW, Research Division, P.O. Box 100, 

2 

Beecroft 2119, Australia. - Australian Biological 

Resources Study, GPO Box 787, Canberra 2601, Australia. 

- E-mail: jacks@sf.nsw.gov.au 

The conservation status of 443 species of 

Holobasidiomycetidae known from the State of South 

Australia in southern Australia has been evaluated. The 

World Conservation Union Red List criteria endorsed in 

1994 were found to be inappropriate as the decision rules 

are based on quantitative threshholds of population size, 

distributional ranges, rates of decline and extinction risk. 

For most of the past 70 years there has been no taxonomic 

mycologist working on the Agaricales, Boletales, 

Cantharellales, Lycoperdales, Phallales or Russulales of 

South Australia. Therefore few quantitative data were 

available. We adopted a modified version of the system 

used to prepare the list of Rare or Threatened Australian 

Plants (ROTAP). In the ROTAP system threat is 

qualitatively assessed according to available expert 

knowledge. Species known only from the holotype, 

lectotype or neotype collection constitute 22% of the taxa 

studied. Ectomycorrhizal fungi comprise 35% of the 

mycoflora studied. Fifty-six species (13%) have been 

introduced from overseas. Of these 19 taxa are 

ectomycorrhizal with exotic trees. More than half of the 

taxa (57%) was described from a collection from South 

Australia. However, only 34% of the taxa are known from 

conservation reserves in South Australia. A remarkable 

26% of the total taxa studied are known from one small 

national park of 840 ha close to Adelaide. The importance 

of specimen based collections rather than observational 

surveys cannot be over emphasised. 

583 - Host determinants of mycorrhizal colonization 

and functioning in maize (Zea maize L.) 

C. Singh 1* , A.K Sharma 2 & B.N Johri 3 

1 M.B.G.E., G.B.P.U.A&T, Pantnagar, 263145 Uttranchal, 

India. - 2 Directorate of Experiment station, G.B.P.U.A&T, 

Pantnagar, 263145 Uttranchal, India. - 3 Microbiology, 

G.B.P.U.A&T, Pantnagar, 263145 Uttranchal, India. - Email: 

charusing2@rediffmail.com 

Enzymes related to metabolism of AM fungi i.e. alkaline 

phosphatase, acid phosphatase and malate dehydrogenase 

and host defence mechanism i.e. peroxidase were measured 

in mycorrhizal and non-mycorrhizal roots of four maize 

genotypes namely HIM 29, VL 90, VL16 and CM129 

differing in mycorrhizal responsiveness. There was 

remarkable enhancement in Enzymes related to metabolism 

of AM fungi i.e. alkaline phosphatase, acid phosphatase 

and malate dehydrogenase and host defence mechanism i.e. 

peroxidase were measured in mycorrhizal and nonmycorrhizal 

roots of four maize genotypes namely HIM 29, 

VL 90, VL16 and CM129 differing in mycorrhizal 

responsiveness. There was remarkable enhancement in the 

level of enzymes related to mycorrhizal functioning in 

genotype VL 90 and HIM 129 while VL 16 and CM 129 

did not respond as well in spite of considerable mycorrhizal 

colonization. The levels of mycorrhizal colonization as 

judged by arbuscular, vesicular to arbuscular senescence. 

584 - Biodiversity of arbuscular mycorrhizal fungi 

(AMF) in different agroclimatic regions of India 

R. Singh * & A. Adholeya 

Centre for Mycorrhizal Research,Tata Energy Research 

Institute, Habitat Centre, Lodi Road, New Delhi-110003, 

India. - E-mail: reenas@teri.res.in 

Arbuscular mycorrhizal fungi (the symbiotic soil fungi 

forming association with plant roots) are an important 

component of biodiversity as about 80% of the world's 

plants are associated with AM fungi. India is a land of 

many climates and varieties of soils affording scope for 

much diversity. The diversity of AMF was studied in 10 

different wheat growing regions of India as wheat is the 

second important food crop of India. This study would then 

help in exploiting the potential of these fungi in sustainable 

agriculture, particularly in wheat. The rhizosphere soil 

samples were collected from wheat fields. The mycorrhizal 

and soil parameters were analyzed and the trap cultures 

were raised from the collected samples for their further 

multiplication. The mycorrhizal parameters analysed 

include: total AMF spore count, species diversity of AMF, 

dominant AMF species, percent AMF colonization inside 

the roots and the infectivity potential of the AMF isolate. 

The soil parameters include analysis of pH, electrical 

conductivity, % total nitrogen, available phosphorus and 

available potassium. The spore count, root colonization, 

species diversity, dominant species and inoculum potential 

of AM fungi were found to vary with the agroclimatic 

region as well as the soil nutrient conditions. The number 

of species in a given region ranged from 1-20. 



585 - Long term decomposition studies of wooden test 

blocks at Khao Yai National Park, Thailand 

S. Sivichai * , N. Boonyeun, N.L. Hywel-Jones & E.B.G. 

Jones 




Khlong 1, Khlong Luang, Pathum Thani12120, Thailand. 

In a survey of wood decay, 4 woods (Dipterocarpus alatus, 

Xylia dolabriformis, Alstonia scholaris and Anisoptera 

oblonga) were immersed in two streams: Tad Tha Phu 

flows year-round; km29 is ephemeral. These were 

recovered each month over 1 year. 236 spp. of fungi were 

found. Importantly, after 1 year decomposition had barely 

begun. Using the same sites the work was extended to 

examine other woods and to allow for recovery of up to 10 

yr. Work after 1 yr allows direct comparison with the 

preliminary study. 5 spp. were used: Buchanania latifolia, 

Mangifera caloneura, Sindora siamensis, Stereospermum 

neuranthum and Tectona grandis. Buchanania and Tectona 

supported the highest diversity (20+ spp.) while Mangifera 

and Sindora had


alpine lichen heath, moist and dry screes. The dominant 

species occurring in most soils were: Pseudogymnoascus 

roseus and its anamorph, Mortierella parvispora, 

Cladosporium cladosporioides, C. herbarum, Mucor 

hiemalis, Tolypocladium inflatum, Penicillium 

simplicissimum, and Fusarium sambucinum. 

Correspondence analysis showed that alpine fens and 

Rhododendron bushes had the most distinctive microfungal 

communities. The latter communities somewhat resembled 

those of podzolic soils under boreal forests. Some 

taxonomically interesting isolates were obtained. A taxon 

that was rather frequent in alpine fen soils was 

provisionally identified as Leptodontidium sp. Two groups 

of isolates differed slightly morphologically from one 

another. A detailed study including sequencing of ITS and 

18S regions revealed that these fungi are rather unrelated to 

the type of the genus Leptodontidium but close to the 

teleomorph genus Pseudeurotium. Observation of 

conidiogenous structures of Pseudeurotium confirmed this 

affinity. A similar strain, CBS 326.81, had been isolated 

from potato meal factory sewage water in the Netherlands. 

Partial sexual reaction with infertile cleistothecia was 

observed between one Caucasian strain and CBS 326.81. 

589 - A preliminary investigation on coprophilous fungi 

in Northeastern Thailand 

S. Somrithipol & N.L. Hywel-Jones * 




Khlong1, Khlong Luang, Pathum Thani 12120, Thailand. - 


Coprophilous fungi are associated with herbivore dung, 

playing an important role in recycling of nutrients in 

animal faeces. This group is a source of biocontrol agents, 

enzymes, antibiotics, etc. Surveys were made in Khao Yai 

National Park and areas in northeastern Thailand between 

May, 2001 and April, 2002. Over 150 isolates were made 

from 85 dung samples of sambar deer (Cervus unicolor), 

barking deer (Muntiacus muntjak), Asian elephant (Elephas 

maximus) and cattle. Of these, 52% were Ascomycetes and 

10% their anamorphs. Basidiomycetes made up a second 

group with 25% of isolates while Zygomycetes accounted 

for only 12%. 14 genera were recognized with 

Ascomycetes accounting for 10 of these. The common 

genera were Podospora, Coprinus and Pilobolus which 

accounted for 54% of collections: most were successfully 

isolated. A succession was demonstrated. Pilobolus 

appeared after 3 days incubation, followed by the 

apothecial-forming ascomycetes such as Ascobolus and 

Saccobolus from 4-7 days. Perithecia of pyrenomycetes 

e.g. Delitschia, Podospora and Sporormia appeared from 

7-14 days and were often associated with basidiocarps of 

Coprinus. These genera continued to fruit for a further 3 

wks. Mitosporic fungi appeared late in the succession. 

Notable species were: Coprinus, Delitschia pachylospora, 

Pilobolus, Podospora communis, Poronia gigantia, 

Saccobolus citrinus, S. thaxteri, and Zygospermella. 

590 - Detection of fossil fungi in Cretaceous amber from 

Álava Northern Spain by different microscopy 

techniques 

M. Speranza 1 , M.J. Martínez 1 , A.T. Martínez 1 , J. 

Wierzchos 2 , J. Alonso 3* & C. Ascaso 4 

1 Centro de Investigaciones Biológicas, CSIC, Velázquez 

144, E-28006 Madrid, Spain. - 2 Servicio Micrópia 

Electònica, Universita de LLeida, E-25198, Spain. - 

3 Museo de Ciencias Naturales, Siervas de Jesús 24 E- 

01001 Vitoria, Alava, Spain. - 4 Centro de Ciencias 

Medioambientales, CSIC, c/ Serrano 115 dpdo., 28006 

Madrid, Spain. 

Amber deposits dated from the Lower Cretaceous period 

were recently found at Álava, Northern Spain, being one of 

the more highly fossiliferous resin deposits. Although some 

reports on the characterization of fossil fungi exists, very 

few is known about their relationships with other fossil 

microorganisms. In this work we reported the use of 

different microscopy techniques to obtain simultaneously 

structural, ultrastructural and analytical information about 

the entire amber microscenosis and their individual 

components. A very well preserved microcenosis, which 

corresponds to a semiaquatic habitat, including different 

types of protozoas, microalgae and fungi was present in 

this amber. By scanning electron microscopy using 

secondary electron (SEM-SE) and backscattered secondary 

electron detectors (SEM-BSE), we found an abundant 

mycelium of hyaline and coenocytic hyphae in close 

association with numerous zooflagellates (probably fungal 

feeders). The elemental distribution spectroscopy 

microanalysis during SEM-BSE revealed Fe in the hyphae 

suggesting the nature of mineralization process. Using light 

and confocal laser scanning microscopy a basidiomycete 

mycelium with numerous clamp connections, crystals and 

encrusted cystidia like structures was detected in the amber 

outside section. Other hyphae, gametangia or sporangia 

like fungal structures and spores were abundant in this 

amber sample. Current work attempts to compare these 

fungi with extant and modern genera. 

591 - A preliminary survey of lichen fungi from 

Thailand 

V. Sri-indrasutdhi, S. Sivichai * & N.L. Hywel-Jones 





E-mail: sivichai@biotec.or.th 

The first published records of fungi from Thailand were 

lichens from Ko Chang Island at the turn of the last 

Century. They were neglected for the next 90 yr. However, 

the last 10 yr has seen increasing interest in Thai lichen 

fungi. Importantly, these are now being isolated and made 

available for further research. A lichen collection/isolation 

programme has been in place at BIOTEC for 2 yr. Natural 

forest was surveyed every 2 months. Forest was selected to 



provide a range of micro-habitats and plant communities. 

Since October 2000, lichens were collected from 13 

National Parks in Thailand. These sites were from the far 

south (Hala Bala at sea level) to the far north (Doi Inthanon 

at 2,565 metres above sea level). These collections yielded 

1116 samples classified into crustose 80%, foliose 14%, 

fruticose 1%, others 2% and unidentified 3%. These have 

been identified to 2 orders including 14 families, 32 genera 

and over 150 species. Common genera of crustose lichens 

were Graphis, Haematomma, Laurera and Lecanora. 

Foliose genera included Heterodermia, Parmotrema and 

Phyllopsora. Cladonia and Usnea were common examples 

of fruticose genera. Canoparmelia and Hypotrachyna were 

notable rarities of foliose lichen. Isolations were made from 

ascospores by direct streaking of spores and spore 

discharge. 

592 - Lignin degrading enzymes of Xylariacious fungi in 

Thailand 

P. Srikitikulchai * & E.B.G. Jones 

BIOTEC-Fermentation Laboratory, National Center for 



RD., Khlong 1, Khlong Luang, Pathum Thani 12120, 

Thailand. - E-mail: prasert@biotec.or.th 

Thailand supports a rich diversity of xylariaceous 

ascomycetes, with most occurring on wood, although a 

number have been found on leaves, elephant dung, fruits 

and seeds, while others have been shown to be endophytes 

of forest trees (Thienhirun, 1997; Whalley et al. 1995). 

Nineteen genera in the Xylariaceae have been documented 

for Thailand with a total of 91 species. However, some 15 

Hypoxylon and 30 Xylaria species await description 

(Thienhirun, 1997). Members of the Xylariales are active 

degraders of timber in nature and have been shown to 

cause soft rot and white rot attack of wood. However, little 

is known about the enzymes they produce. We have 

screened 51 strains of xylariaceous fungi over the past 2 

years for their lignin degrading enzymes. For this study we 

have used Poly R-478, ABTS and azure-B medium to 

screen for general lignin degrading enzymes, laccase and 

peroxidase activity, respectively. Ninety four percent of the 

isolates gave positive results for laccase, 21% positive for 

peroxidases, while 43% decolourized the dye Poly-R 478. 

593 - Fungal diversity on woody substrates in 

Scandinavia 

J.N. Stokland 1* & A. Dahlberg 2 

1 Norwegian institute of land inventory, P.O.Box 115, 1430 

Ås, Norway. - 2 Swedish Species Information Centre, 

P.O.Box 7007, SE-750 07 Uppsala, Sweden. - E-mail: 

jns@nijos.no 

180 


Species diversity patterns have been derived from speciesspecific 

information of more than 1500 species of 

Ascomycetes, Heterobasidiomycetes and Basidiomycetes. 

Several functional associations to woody debris were 

defined, including primary parasite, primary saprotroph, 

secondary myco-parasite, secondary myco-saprotroph, and 

mycorrhiza species (using wood for sporocarp production). 

Most species are probably primary saprotrophs, but all 

functional associations are represented. Most species are 

associated to either coniferous (40%) or hardwood hosts 

(40%), but many are host generalists (20%). Most 

taxonomic groups exhibit this pattern, but at least the 

Ascomycetes show an elevated species richness on 

hardwood hosts. The degree of decomposition of the wood 

is important and four patterns were found: early decay 

preference (less than 10%), medium decay preference 

(50%), late decay preference (15%) and no decay 

preference (25%). These proportions varied across 

taxonomic groups. The dimension of woody substrates is a 

third factor that many fungi respond to. One group prefer 

small dimensions (twigs and thin stems) and another is 

strongly associated to trunks of large dimensions, but the 

majority of the species (60%) show no strong dimension 

preference. There are additional factors that woodinhabiting 

fungi respond to like sun exposure/shade, part of 

the tree and standing/lying. For these factors there are 

major knowledge gaps that prevent compilation of diversity 

patterns. 

594 - Lichenicolous fungi in Estonia 

A. Suija 

Institute of Botany and Ecology, University of Tartu, 38 Lai 

Str., Tartu, 51005, Estonia. - E-mail: avesuija@ut.ee 

The history of floristical investigation of lichen-habiting 

fungi in Estonia is short. Until the 1990s, only two 

lichenicolous fungi (Athelia arachnoidea and Rimularia 

insularis) were known from Estonia. During the last five 

years, 69 species from 47 genera of these fungi have been 

registered from Estonia. Most of the lichenicolous species 

are ascomycetes (45 species); fewer are deuteromycetes 

(23) and basidiomycetes (7). The most species-rich genera 

are Abrothallus (5 species), Licheniconium (4), Muellerella 

(3), Stigmidium (4) and Tremella (4). Most species are 

categorized as very rare i.e. known from one or two 

localities in Estonia. Still, there are some exceptions. The 

distribution area of the basidiomycete Athelia arachnoidea, 

an aggressive parasite of different protococcoid algae and 

lichens, has extended since the 1940s. Since the 1990s, this 

species is especially widespread in towns, roadsides etc. 

The deuteromycete Vouauxiella lichenicola, a parasite of 

Lecanora chlarotera and allied taxa, was reported as very 

rare in 1999. However, the species is now found in about 

30 places and its distribution partially coincides with the 

distribution of Lecanora chlarotera.


595 - Interesting ascomycetes on Juncaceae and 

Apiaceae collected in the Czech Republic 

M. Suková 

National Museum, Mycological Department, Václavské 

nám. 68, 11579 Praha 1, Czech Republic. - E-mail: 

marketa.sukova@nm.cz 

Saprotrophic ascomycetes and mitosporic fungi on 

Juncaceae and Apiaceae in the Czech Republic were 

collected and studied during the years 1997-2001. Eightyfive 

species belonging to 56 genera were found. Fifteen of 

these species are reported from the Czech Republic for the 

first time. They include representatives of Helotiales 

(Hysteropezizella diminuens, Mollisia fusco-paraphysata, 

Naeviopsis carneola, Scutomollisia stenospora), 

Pleosporales (Lophiotrema grandispora, Nodulosphaeria 

mathieui, N. spectabilis), Mycosphaerellales 

(Mycosphaerella hypostomatica), and mitosporic fungi 

(Arthrinium cuspidatum, Chalara urceolata, Coremiella 

cubispora, Dendryphiella vinosa, Phragmocephala 

prolifera, Pyricularia juncicola, Stachybotrys 

cylindrospora). The most interesting species are discussed 

and illustrated. 

596 - NITE Biological Resource Center (NBRC), a 

newly established biological resource center in Japan 

K. Suzuki & S. Iwamoto * 

NITE Biological Resource Center, National Institute of 

Technology and Evaluation, 2-5-8, Kazusakamatari, 

Kisarazu, Chiba 292-0812, Japan. - E-mail: iwamotosusumu@meti.go.jp 

According to the report of OECD, biological resource 

centers (BRCs) are an essential part of the infrastructure 

underpinning life sciences and biotechnology. NITE, a 

non-profit institute under Ministry of Economy, Trade and 

Industry, has established a new biological resource center, 

NBRC with the following concept: NBRC is to preserve 

and supply the cultures of reference microorganisms 

including fungi, yeasts, bacteria and archaea as an essential 

function of a culture collection. NBRC isolates, 

characterizes and develops the original biological resources 

for its own collection. In addition, NBRC also provides 

DNA and clones based on the genomic sciences. To access 

the appropriate resources, NBRC develops advanced 

databases containing application data and taxonomic data. 

As the so-called intelligent administration, NBRC have to 

take care of the ownership of the biological resources to 

encourage the deposit of valuable resources. Convention 

for Biological Diversity should be considered when 

biological resources are transferred between countries. 

Based on these, NBRC intensively make international 

cooperative studies with the resource countries especially 

in the South East Asia to produce benefits for both sides. 

597 - Taxonomical investigations of Hungarian 

protected fungi 

Gy. Szedlay 1* & I. Siller 2 

1 Eötvös University, Dept. of Plant Anatomy, 1117 

Budapest, Pázmány P. s. 1/c, Hungary. - 2 Szent István 

University, Dept. of Botany, 1074 Budapest, Rottenbiller u. 

50., Hungary. - E-mail: szedlay@ludens.elte.hu 

In the last decades fungal taxonomy has undergone 

enormous change. The traditionally used morphological 

description of species has been completed with molecular 

and interfertility results. We examined phylogenetic 

relationships of the holotypes of fungi described in 

Hungary using morphological and molecular methods 

(DNA sequencing). Species of Agaricus, Leucoagaricus, 

Macrolepiota, Lepista and Cortinarius genera were 

investigated. In some cases our results confirmed, in others 

questioned the isolation of the examined species from their 

near relatives. 

598 - Diversity of fungi on wild fruits in Hong Kong 

A.M.C. Tang 

The University of Hong Kong, 2N-16 Kadoorie Biological 

Science Building, The University of Hong Kong, Pokfulam 

Road, Hong Kong, China. - E-mail: alvtang@hkem.com 

The postharvest diseases of cultivated fruits have been well 

studied, however, there have been few surveys of the 

diseases of wild fruits. Small fleshy fruits are of special 

interest due to the potentially conflicting relationship with 

frugivorous birds and microbes. The fruits need to attract 

dispersers after ripening but repel microbes. In the current 

survey, 15 species of wild fruits were examined from the 

mature stage to the senescent stage for visible symptoms 

and signs of diseases in the field and after incubation. Half 

of the fruit samples were also surface-sterilized to 

eliminate surface fungi. A total of 104 fungal taxa have 

been isolated so far, including 21 ascomycetes, 57 

coelomycetes and 24 hyphomycete. The most common 

ascomycete genera were Gaeumannomyces, Guignardia 

and Massarina spp. and the most common mitosporic 

genera were Cladosporium, Colletotrichum, Penicillium, 

Pestalotiopsis, Phoma, and Phomopsis spp. 



599 - Species of Cordyceps attacking Hymenoptera in 

Thailand 

K. Tasanatai & N.L. Hywel-Jones * 






In Thailand 88 Cordyceps spp. are known: Coleoptera (15 

spp.) and Lepidoptera (28 spp.) hosts dominate with 9 spp. 

from Hymenoptera. 3 spp. (C. sphecocephala, C. humberti 

and Cordyceps sp.) are off wasps and 6 off ants. Of these 6, 

4 are named - C. irangiensis, C. myrmecophila, C. 

pseudolloydii and C. unilateralis. 5 spp. have Hymenostilbe 

anamorphs; 4 Hirsutella. In contrast to coleopteran and 

lepidopteran Cordyceps, those off Hymenoptera are hard to 

isolate. Usually, Hirsutella are isolated easily while 

Hymenostilbe is considered problematic. With 

hymenopteran Cordyceps the converse holds. The 

anamorph of C. unilateralis is H. formicarum: the most 

common Hirsutella in natural forest in Thailand, it was 

isolated once during a 10 yr period. A programme to isolate 

this species has produced 3 more isolates. Conditions 

needed to induce a vegetative colony from ascospores are 

under investigation. However, for suborder 

Ophiocordyceps evidence suggests that ascospores act as 

carrier vehicles - the role being to produce spores that 

germinate to form vegetative colonies. Hymenopteran 

Cordyceps with Hymenostilbe anamorphs are assigned to 

suborder Neocordyceps. Molecular work (with limited 

species) shows evidence of long-branch attractions. 

Circumstantial evidence suggests a link between the 

hymenopteran Cordyceps with Ophiocordyceps appearing 

ancestral to Neocordyceps. 

600 - A preliminary study of the prevalence of 

endophytes in leaves and seed pods of Colophospermum 

mopane (Leguminosae) in Botswana 

J.E. Taylor 1* , B.W. Bojosi 1 & A. Jordaan 2 

1 Department of Biological Sciences, University of 

Botswana, UB 0022, Gaborone, Botswana. - 2 Electron 

Microscope Unit, Department of Physics, University of 

Botswana, UB 0022, Gaborone, Botswana. - E-mail: 

taylor@mopipi.ub.bw 

Colophospermum mopane (Mopane) is a deciduous tree or 

multi-stemmed shrub dominating the vegetative structure 

in parts of Southern Africa. It is browsed by wildlife and is 

utilised by local people for domestic animal fodder and 

firewood. Mopane is the primary host plant of the 'mopane 

worm', the caterpillar state of Imbrasia belina which is 

harvested by local people and forms an important source of 

protein in their diet and provides income. No previous 

studies have been carried out on the fungi associated with 

this regionally important tree. Young (soft green), mature 

182 


(leathery green), senescent (leathery yellow) and dead 

(brittle yellow-brown) leaves, and mature seed pods, were 

collected directly from selected trees. Tissue samples were 

dissected from the leaves and pods after surface 

sterilisation. The endophyte assemblage was dominated by 

species of Alternaria and Phoma, and to a lesser extent 

Phomopsis. The members of the assemblage remained in 

similar proportions in different aged leaves, but increased 

in frequency with increasing leaf age. Alternaria spp. were 

proportionally less common in petiole than lamina tissue. 

TEM studies on seed pods, which yielded endophytes, 

were carried out to investigate the role of the endophytes in 

degradation of lignified parts of the pericarp. 

Lignocellulolytic enzyme production by the endophytes 

was assessed with qualitative assays. 

601 - A preliminary report on a moss pathogenic 

heterothallic Pythium sp. from Spitsbergen Island, 

Svalbard 

M. Tojo 1* , H. Fujii 1 , T. Hoshino 2 , H. Kanda 3 , M.L. 

Herrero 4 , S.S. Klemsdal 4 , A.M. Tronsmo 4 & S.T. Ohki 1 

1 Graduate School of Agriculture and Biological Sciences, 

Osaka Prefecture University, Gakuen-cho, Sakai, Osaka 

599-8531, Japan. - 2 National Institute of Advanced 

Industrial Science and Technology (AIST), Toyohira-ku, 

Sapporo 062-8517, Japan. - 3 National Institute of Polar 

Research, Itabashi-ku, Kaga, Tokyo 173-8515, Japan. - 

4 Norwegian Crop Research Institute, Fellesbygget, N-1432 

Aas, Norway. - E-mail: tojo@plant.osakafu-u.ac.jp 

In the course of studies on moss pathogenic fungi in arctic 

zone, compatible isolates of heterothallic Pythium sp. were 

frequently isolated from dying moss (Sanionia uncinata) in 

Ny-Ålesund (79°N, 12°E), Spitsbergen Island, Svalbard. 

The fungus caused discoloration of the moss after artificial 

inoculation at 0.5 and 10 °C. Morphological analysis of the 

isolates showed the following features. The main hyphae 

were up to 6 µm wide. Oogonia were mostly terminal, 

globose, smooth-walled, 18-30 µm in diameter. Antheridia 

were terminal, 1 to 5 per oogonium, sac-like and diclinous. 

Oospores were single, aplerotic, rarely plerotic, globose, 

16-26 µm in diameter. The thickness of the oospore wall 

was up to 1 µm. Sporangia were globose, mostly terminal, 

occasionally intercalary. Diameter of encysted zoospores 

ranged from 10-11 µm. Optimal growth of mycelia 

occurred at 25 °C, the minimum temperature for the growth 

was 0.5 °C, and maximum was 28 °C. The rDNA internal 

transcribed spacer sequences of the isolates were different 

to those of Pythium spp. in the GenBank database. The 

results suggested that a new heterothallic Pythium sp. 

caused the discoloration of moss occurred on Spitsbergen 

Island, Svalbard.


602 - Distribution of macrofungi in the forest in Taiwan 

J.S.M. Tschen, I.C. Ho, H.S. Hsu & E.F.T. Tschen * 

Department of Botany, National Chung Hsing University, 

250 Kuokang Road, 402 Taichung, Taiwan. - E-mail: 

smtschen@mail.nchu.edu.tw 

The distribution of macrofungi at the long-term ecological 

research site in the Guandaushi forest in the middle Taiwan 

was ecologically studied. Several experimental plots were 

set up for the study. The important macrofungi genera were 

Mycena, Marasmius, Xylobolus, Trametes, Russula, 

Coriolus, Lactarius, Amanita and Stereum. Most of these 

macrofungi occurred in July during the summer period. A 

lesser amount of macrofungi grew in March during the 

winter period. Mycena and Marasmius were the common 

macrofungi that occurred both in summer and winter. The 

Russula, Coriolus, Lactarius, Amanita, Armillaria, 

Fomitopsis, Ganoderma, Microsporus and Rhodophyllus 

species were present in the summer period. Xylobolus, 

Trametes, Stereum and Peniophora fungi were found in the 

winter. The macrofungi distribution in the experimental 

plots varied. The occurrence and diversity of the 

macrofungi depended upon the type of predominant 

vegetation. The plants favorable to macrofungi were 

Blastus cochinchinesis, Cinnamomum randaiense, Helicia 

rengetiensis, Syzygium buxifolium, Engelhardtia 

roxburghiana, Smilax china, Cunninghamia lanceolata, 

Nephrolepis ariculata and Plagiogyria dunnii. The 

unfavorable plants to fungi were Illicium arborescens and 

Diplopterygium chinensis. 

603 - South African National Collection of Fungi: 

collections and services 

E.J. van der Linde, O.M. O'Brien, I.H. Rong * & C. Roux 

ARC-Plant Protection Research Institute, Private Bag 

X134, Pretoria 0001, South Africa. 

The National Collection of Fungi of South Africa is 

internationally accredited and affiliated at the World 

Federation of Culture Collections as PPRI (for the living 

culture collection) and PREM (for dried reference 

material). Both collections contain important type material 

and researchers are encouraged to deposit reference 

material in these collections to ensure the expansion of this 

invaluable national asset to the benefit of the scientific 

community. The types contained in PREM are integrated 

onto a database. The rest of the almost 57 000 specimens 

will shortly be incorporated as well. The list of types is 

available electronically. The specimens are available on 

loan for a period of six months to researchers nationally 

and internationally. About 4500 specimens are contained in 

the PPRI collection. This list is also available 

electronically. Both collections are managed by the staff of 

the Mycology unit of the Plant Protection Research 

Institute (ARC-PPRI), an institute of the Agricultural 

Research Council. Apart from managing the above 

mentioned collections, the unit also provides the following 

services: processing of infected material, isolation and 

identification of fungi, fungal analyses including testing of 

efficacy of fungicides, provision of biological information, 

as well as presentation of mycological courses such as 

basic laboratory techniques and taxonomy of major groups. 

604 - Biodiversity of fungi on reed (Phragmites 

australis) 

G. Van Ryckegem * & A. Verbeken 

Ghent University, Dpt. Biology, Group Mycology, K.L. 

Ledeganckstraat 35, B-9000 Gent, Belgium. - E-mail: 

Gunther.vanryckegem@rug.ac.be 

Reed is a dominant emergent macrophyte in many wetland 

ecosystems all over the world. Fungal communities 

associated with this high productive grass are demonstrated 

to be taxonomically diverse. A field survey in Belgium 

(mainly Schelde estuary) of Fungi occurring on reed during 

growth and decomposition results in about 230 taxa of 

which 40% Ascomycetes, 10% Basidiomycetes, 30% 

Coelomycetes and 20% Hyphomycetes. 1 Basidiomycete 

represents a new species, about 13 species (14%) of the 

Ascomycetes are undescribed; in the Mitosporic Fungi this 

proportion is probably even higher. A compilation of our 

research and world literature available on phragmiticole 

Fungi leads to the hypothesis that the total number of fungi 

occurring on reed will exceed 650 species. This poster 

presents results which will be published in a paper on 

biodiversity of fungi on reed. This biodiversity study is part 

of a broader ecological study concerning succession of 

fungal species and a monitoring of ergosterolconcentration 

(fungal biomass measure) during decomposition. 

605 - Towards a Mediterranean check-list of fungi: 

macromycetes from beech woods of Sicily and Greece 

G. Venturella 1* , D.M. Dimou 2 , A. Saitta 1 , E. Polemis 3 & 

G.I. Zervakis 3 

1 Department of Botany, University of Palermo, Via 

Archirafi 38, I-90123 Palermo, Italy. - 2 Agricultural 

University of Athens, Laboratory of gricultural 

Microbiology, Iera Odos 75, 11855 Athens, Greece. - 

3 National Agricultural Research Foundation, Institute of 

Kalamata, Lakonikis 85, 24100 Kalamata, Greece. - Email: 

gvent@unipa.it 

The importance to increase the knowledge on 

macromycetes diversity in southern Europe has been 

repeatedly stressed by the OPTIMA (Organization for the 

Phyto-Taxonomic Investigation in the Mediterranean Area) 

Commission on Fungi, which considers the check-list of 

Mediterranean fungi as its main goal for the near future. In 

recent times, in Sicily (South Italy) and Greece many 

investigations on fungal diversity and distribution were 

carried out. According to recent literature data, 1248 and 



1300 species of macrofungi have been recorded in Sicily 

and Greece respectively. As regards beech woods in 

particular, 402 taxa are reported from Sicily (which 

account to 35% of the total number of fungi recorded from 

Italian beech woods), whereas an equivalent number of 

fungi (i.e. 405 taxa) were found to occur in beech woods of 

Greece. In addition, the two investigated areas presented 

137 taxa in common. Among the most representative 

genera, Amanita, Boletus, Russula and Tricholoma are 

prevalent in Sicily, whereas Cortinarius and the corticioid 

fungi are quantitatively more represented in Greece. 

Noteworthy is the occurrence of species strictly linked to 

the thermophilous beech woods such as B. fragrans and B. 

satanas (only in Sicily), and B. aereus, B. radicans (in both 

areas examined). 

606 - Biodiversity and distribution of aeroaquatic fungi 

H. Voglmayr 

Institute of Botany, Rennweg 14, A-1030 Vienna, Austria. - 

E-mail: hermann.voglmayr@univie.ac.at 

Aeroaquatic fungi are an ecological group of fungi 

inhabiting shallow stagnant to slow-flowing freshwater 

bodies. They grow on submerged plant litter and often 

show pronounced ability to survive micro-aerobic 

conditions. However, their dispersal units are only 

produced if the substrate is exposed to the air; they consist 

of complicated, usually multicellular structures of an 

astonishing morphological diversity, which entrap air and 

are therefore well buoyant. Buoyancy is often enhanced by 

warts and hydrophobic encrustations. Dispersal takes place 

on the water surface. Up to data, about 90 aero-aquatic 

species are known world-wide. Most species have been 

recorded exclusively from temperate climates (more than 

75%), comparatively few from the tropics (less than 20%), 

and very few were found both in tropical and temperate 

habitats (about 5%). Many species have a wide geographic 

distribution. Recent investigations revealed many new taxa 

even in comparatively well-studied Europe, indicating that 

its biodiversity is very imperfectly known world-wide. 

Systematically, the aero-aquatic fungi comprise members 

of the oomycetes, basidiomycetes and ascomycetes; 

however, for the majority of species teleomorphs are 

unknown. Concluded from known teleomorph stages and 

DNA sequence data, the largest aeroaquatic genera 

Helicoon and Helicodendron are polyphyletic, which may 

be also true of other genera. This clearly demonstrates that 

the aeroaquatic lifestyle evolved many times 

independently. 

184 


607 - Utilisation of the population patterns and 

pathogenicity of soilborne and root-colonising fungi as 

bioindicators of soil health in agricultural soils 

N.W. Waipara 

HortResearch, Canterbury Research Centre, P.O.Box 51, 

Lincoln, New Zealand. - E-mail: 

nwaipara@hortresearch.co.nz 

Biodiversity and population dynamics of microbes change 

due to anthropogenic soil disturbances such as different 

agricultural land management practices. Mycological 

surveys, using several isolation methods, were undertaken 

to determine species diversity of soil and root-colonising 

fungi in pastoral, cropping, native grasslands and forest 

soils in New Zealand. Soilborne plant pathogenic species, 

such as Cylindrocarpon spp. and Fusarium spp., were 

frequently isolated during the surveys. Pathogenicity of 

these fungi was determined using axenically grown 

seedlings to provide an index whereby the relative 

pathogenicity of species could be compared between soil 

and land management types. Results indicated that more 

pathogenic species and strains were present in highly 

disturbed agricultural soils compared to relatively 

undisturbed native grassland forest and soils. These species 

and methods may have potential to be used as bioindicators 

of soil health and disease suppressiveness. 

608 - Ecology of rare species of Hericium in Britain 

P.M. Wald * & L. Boddy 

Cardiff Scool of Biosciences, Park Place, PO Box 915, 

Cardiff CF10 3TL, U.K. - E-mail: paul_wald@hotmail.com 

The wood-decomposing species of Hericium are not 

common in Britain. Hericium erinaceus is protected under 

the Wildlife and Countryside Act (1981), reported as being 

'vulnerable' in the British Red Data List, and is a UK 

Biodiversity Action Plan priority species. H. coralloides, 

H. cirrhatum and H. alpestre are all listed in the Red Data 

List as being 'vulnerable'. With the exception of H. 

alpestre, which is found decaying coniferous wood, 

Hericium spp. occupy the heart wood of deciduous trees 

and all species cause white rot. The ecology of these 

species of Hericium is little understood, thus we have 

examined their establishment following inoculation into 

felled logs and standing trees, how fungal community 

structure develops once these fungi have become 

established, and the outcome of interactions with other 

wood-rotting fungi. The work has revealed that it is 

relatively easy to establish these fungi by inoculation of 

colonised dowels, they are quite combative against other 

basidiomycetes, and they fruit readily. Preliminary 

evidence, however, suggests that spore germination may be 

poor. Thus, mode of colonisation may be an important 

factor contributing to their apparent rarity. These studies 

should provide the information required to conserve extant


Hericium populations and make recommendations for their 

artificial reintroduction, if necessary. 

609 - Encapsulation cryopreservation of Serpula 

lacrymans and the development of methods to identify 

molecular damage 

N.A. White 1* , A. Ariza 1 , E.E. Benson 1 , M.J. Ryan 2 & A. 

Buddie 2 

1 SIMBIOS, University of Abertay, Kydd Building, Bell St., 

Dundee, DD1 1HG, Scotland, U.K. - 2 CABI Bioscience, 

Bakeham Lane, Egham, Surrey, TW20 9TY, England, U.K. 

- E-mail: mltnaw@aberaty.ac.uk 

We present a generic encapsulation-dehydration method 

for the successful cryopreservation of building and 

Himalayan woodland isolates of Serpula lacrymans and S. 

himantioides. Mycelial cores from actively growing 

cultures were encapsulated in alginate beads and pretreated 

for 72 h in a 0.75 M sucrose solution. Three 

desiccation methods were tested on the pre-treated beads: 4 

h in a flow of sterile air while cooling the beads on ice (c. 

19-26 °C), 4 h in a flow of sterile air without cooling the 

beads (c. 28-40 °C) and 18 h in a specially designed 

desiccator with silica gel (c. 22 °C). Desiccated beads were 

stored in liquid nitrogen for 30 min, then left to recover and 

the effect of treatment on the survival, lag phase, radial 

colony extension rate and morphology measured. A 

comparison of the different experimental methodologies 

employed clearly highlights the sensitivity of this approach 

to the operational parameters, in particular temperature. 

Results are described and compared for individuals of 

different origin and in view of the known temperature 

sensitivities of Serpula species. The development of 

molecular techniques such as ISSR-PCR to identify genetic 

damage accumulated during different storage and 

preservation treatments applied to S. lacrymans are 

described. 

610 - The biomarkers and fossils of cryptoendolithic 

Antarctic fungi and lichens 

J. Wierzchos 1 & C. Ascaso 2* 

1 Universitat de Lleida, Servei de Microscòpia Electrònica, 

c/ Rovira Roure 44, 25198 Lleida, Spain. - 2 Centro de 

Ciencias Medioambientales, CSIC, c/ Serrano 115 dpdo., 

28006 Madrid, Spain. - E-mail: ascaso@ccma.csic.es 

Lithobiontic microbial communities living within are an 

example of survival in an extremely cold and dry 

environment. Any unfavourable change in external 

conditions can result in the death and disappearance of 

microbes, and this may be followed by the appearance of 

trace biomarkers and/or their microbial fossils. The results 

reveal the presence of inorganic biomarkers in the form of 

physicochemical bioweathering mineral patterns. The 

presence of Fe-hydroxide nanocrystals and biogenic clays 

around endolithic hyphae cells have been also observed. 

Others biomarkers, including deposits of calcium oxalates 

and silica accumulations, are clear signs of endolithic 

hyphae activity. The interior of the sandstone rocks reveal 

the presence of microbial fossils of cryptoendolithic 

lichens. These microbial fossils, detected for the first time 

within Antarctic rocks, contain well preserved relics of 

ultrastructural cytoplasm elements. Recognising internal 

cell structures within fossilised cells is proposed as a new 

criterion for defining the biogenicity of biomorphs. We 

suggest the use of SEM-BSE and EDS in situ 

microscopical and microanalytical investigation strategies, 

which make it possible to clearly determine inorganic 

traces of Antarctic endoliths. The implications of our 

findings are potentially significant for astrobiology because 

Antarctic cold and dry ecosystem are considered a 

terrestrial analogue for possible life on early Mars. 

611 - Study on the biodiversity of inoperculate 

discomycetes in Yamingshan, Taiwan 

M.L. Wu 

Dept. of Natural Science Education, Taipei Municipal 

Teachers' College, No. 1, Ai-Kuo West Rd. Taipei, Taiwan. 

- E-mail: wuml@mail1.tmtc.edu.tw 

Thirty sampling areas with 1 square meter had been 

constructed at the Silvergrass Grasslands and Arrow 

Bamboo Bushes near Hsiaoyukeng as well as Broadleaved 

Forest near Lujankenghsi in Yangmingshan National Park, 

Taiwan. One hundred and twelve samples including 43 

from decayed twigs, 10 from arrow bamboo and 59 from 

silvergrass and other straws had been collected from the 

above sampling areas at the elevations of 510-830 m since 

August, 2000 to July, 2001. Eighteen species of 

Dermateaceae, 4 species of Leotiaceae, 3 species of 

Orbiliaceae, 4 species of Sclerotiniaceae, 18 species of 

Hyaloscyphaceae had been identified. The dominant 

species of the above three different areas of Lujankenghsi, 

Silvergrass Grasslands and Arrow Bamboo Bushes are 

respectively Lachnum sclerotii,Lachnum cf. apala and the 

species of Mollisia. 

612 - Fungi, rotted wood and beetles: Implications for 

native forest management in Tasmania 

M. Yee 1 , Z.Q. Yuan 2 & C. Mohammed 3* 

1 Co-operative Research Centre for Sustainable Production 

Forestry, GPO Box 252-12, Hobart, Tasmania, 7001, 

Australia. - 2 School of Agricultural Science, University of 

Tasmania, GPO Box 252-54, Hobart, Tasmania, 7001, 

Australia. - 3 CSIRO Forestry and Forest Products, GPO 

Box 252-12, Hobart, Tasmania, 7001, Australia. - E-mail: 

caroline.mohammed@csiro.au 

Shorter rotation lengths of around 80 years in commercial 

native forests in Tasmania will eventually lead to the 



elimination of large decaying logs on the forest floor. This 

poster describes an investigation of whether small trees, 

which could be left as a future decaying log habitat at the 

end of the rotation, are likely to follow the same 

successional pathways as larger logs, providing the full 

range of decay types and hence fungal and invertebrate 

biodiversity. The four objectives of the project were, in 

regard to Eucalyptus obliqua coarse woody debris (CWD) 

in Tasmanian wet sclerophyll forests: 1) To better 

understand the ecology of decomposing CWD, in terms of 

the associated wood decay fungi and saproxylic insects; 2) 

To investigate differences in fungal and insect biodiversity 

between small and large diameter logs; 3) To provide a 

checklist of the fungal and insect species dependent on 

Eucalyptus obliqua CWD in wet sclerophyll forests; 4) To 

look for direct associations between decayed wood, wood 

decay fungi and saproxylic insects; The ecological 

information from this research will contribute to assessing 

whether current forest practices are ecologically 

sustainable, developing appropriate indicators for 

monitoring forest diversity, and proposing prescriptions 

that ensure the conservation of these diverse wood 

decomposing communities. 

613 - Preliminary biodiversity inventory of dictyostelid 

slime molds in Taiwan 

Z.-Y. Yeh 

National Taiwan Normal University, 88, Tingchou Rd., 

Sec. 4, 117, Taipei, Taiwan. - E-mail: 

biozyy@scc.ntnu.edu.tw 

Dictyostelid cellular slime molds are widely distributed in 

the world and there are most numerous and of the greatest 

variety in forest soils. Taiwan is an island located in the 

subtropics with many high mountains. Due to the vertical 

geographical and climatic complexity of this island, plant 

vegetation is luxuriant and the fungal biodiversity is 

abundant. The dictyostelid taxa were obtained from soils of 

various vegetations such as forest, grassland or cultivated 

field of Taiwan. Up to date, three genera and seventeen 

species have been recorded, namely: Acytostelium 

leptosomum Raper, Dictyostelium aureo-stipes Cavender, 

Raper et Norberg, D. brefeldianum Hagiwara, D. clavatum 

Hagiwara, D. coeruleo-stipes Raper and Fennell, D. 

delicatum Hagiwara, D. giganteum Singh, D. lavandulum 

Raper and Fennell, D. macrocephalum Hagiwara, Yeh et 

Chien, D. minutum Raper, D. monochasioides Hagiwara, 

D. polycephalum Raper, D. purpureum Olive, D. 

rhizopodium Raper et Fennell, Polysphondylium pallidum 

Olive, P. pseudo-candidum Hagiwara and P. violaceum 

Brefeld. Currently, D. exiguum Hagiwara was newly 

isolated from forest soils in Nantou County of Taiwan. The 

species was examined and illustrated in this text. 

Concerning ecological distribution and keys for the above 

eighteen species were also discussed and provided. 

186 


614 - Preliminary assessment of diversity and 

productivity of edible mushrooms in savanna 

woodlands of Western Africa 

S.N. Yorou 1* , A. De Kesel 2 & B. Sinsin 1 

1 Laboratoire d'écologie Appliquée, Faculté des Sciences 

Agronomiques, Université d'Abomey Calavi au Bénin., 01 

BP 526, Cotonou, Bénin. - 2 National Botanic Garden of 

Belgium, Domein van Bouchout, B-1860 Meise, Belgium. - 

E-mail: nourou@avu.org 

A simplfied method for preliminary assessment of the 

diversity and productivity of wild edible mushrooms in 

different types of savanna woodlands is presented. The 

results of a case study in Bénin (West-Africa) are given. 

Thirty edible macromycetes were found in 18 fixed plots 

from six Soudano-guinean vegetation types. The 

mushroom productions were assessed per species and per 

plot, using the number of produced carpophores and fresh 

weight (kg/ha). Edible macromycetes from the studied 

areas are for 86.6% ectomycorrhizal, mainly belonging to 

Lactarius, Russula and Amanita. The total edible 

macromycetes fresh weight production varies from 0.2 to 

225 kg/ha per season, depending on the forest type. Open 

forests, i.e. savanna and woodland savanna, dominated by 

Caesalpiniaceae, yield qualitatively and quantitatively 

more edible taxa than the denser and rivulet associated 

forest types. Lactarius gymnocarpoides is the most 

common and abundant edible species in the savannas and 

woodland savannas. Specific productions of edible taxa are 

affected by host tree compositions reflected in the forest 

type. The study illustrates the importance of local 

indigenous knowledge, and indicates that the edible 

macromycetes from West-African savanna woodlands have 

substantial importance as a renewable, non cultivatable, 

non timber forest product. 

615 - Arbuscular mycorrhizas from the rhizosphere of 

Macaranga denticulata and their effects on the host 

plant at different levels of fertilizers 

S. Youpensuk 1* , B. Rerkasem 2 , B. Dell 3 , S. Lumyong 1 & P. 

Lumyong 4 

1 Biology Department, Science Faculty, Chiang Mai 

University, Chiang Mai 50200, Thailand. - 2 Agronomy 

Department, Agriculture Faculty, Chiang Mai University, 

Chiang Mai 50200, Thailand. - 3 School of Biological 

Sciences and Biotechnology, Murdoch University, Perth, 

Australia. - 4 Plant Pathology Department, Agriculture 

Faculty, Chiang Mai University, Chiang Mai 50200, 

Thailand. - E-mail: scboi027@chiangmai.ac.th 

The hill tribe (Karen) farmers in Mae Hong Son province 

of northern Thailand believe in beneficial effects of 

Macaranga denticulata Müll. Arg. on upland rice yield. 

The farmers manage the M. denticulata canopy in fallow 

plots of their rotational shifting cultivation in an acid soil 

(pH 4.9). Diversity of arbuscular mycorrhizal (AM) fungi


was found 30 species from the indigenous rhizosphere of 

M. denticulata. The effects of indigenous AM fungi on 

growth of M. denticulata were assessed by growing the 

plants in pots containing soil (sandy loam and equal 

available P as indigenous soil, 3.5 mg/kg) that was set up to 

have pH 4.9 by mixing with soluble aluminium. AM 

colonization in roots of M. denticulata was highest (87.8%) 

in the treatment of no added N and P that was similar to 

AM colonization in the natural rhizosphere in wet season 

and lowest in the treatment of added N and P 50 mg/kg soil 

(51.0%). Spore numbers of AM fungi ranged from 13 to 37 

spores/g fresh soil. In treatments with inoculated AM fungi 

the growth of M. denticulata was higher than in noninoculated 

AM fungi. The highest growth of M. denticulata 

was obserbed in inoculated soil with 50 mg/kg soil of N 

and no added P (AM colonization, 70.4%). 

616 - Lichens on littoral of the White and Barents seas 

(Northwest Russia) 

A.A. Zavarzin * & D.E. Himelbrant 

St.Petersburg State University, Universitetskaya emb., 7/9, 

St.Petersburg 199034, Russia. - E-mail: 

zavarzin@yahoo.com 

Littoral lichen communities composed of species adapted 

to both periodic over wetting and high salinity are among 

the least studied by lichenologists. Investigations of littoral 

lichens along the extensive coastal strip of Russia are 

lacking. Lichen diversity of the White (Kandalaksha bay) 

and Barents (Dalnie Zelentsy) seas' coasts were studied 

during the field trips, supported by Federal Program 

Integratsia. Coinciding with data on British coasts three 

groups of lichensit were revealed: (1) coastal, (2) 

supralittoral, (3) littoral, with the latter being the least 

diverse but most specific. The littoral group is restricted to 

lower supralittoral and whole littoral zones. Barents coasts 

are dominated by lichen communities formed by 

Verrucaria mucosa in middle and lower littoral (were it is 

competing with red alga Hildenbrantia sp.), substituted by 

Verrucaria maura and satellite Verrucaria species in upper 

littoral. Both species form clear belts with coverage up to 

98%. On the White sea coasts the same species are present, 

though with less developed coverage and V. mucosa does 

not extend into lower littoral. Additionally filamentous 

Lichina confinis is common in midlittoral on granite 

outcrops. In contrast with the former sea coast Verrucaria 

species here prefer littorals that are not exposed to direct 

waving. All littoral lichens are growing on basic rocks, and 

only Pyrenocollema sublitorale inhabits calcareous shells 

of Semibalanus balanoides. 

617 - Mycocoenological studies in beech-woods in 

Western Liguria (Italy) 

M. Zotti 1* , S. Zappatore 2 & G. Barberis 1 

1 DIP.TE.RIS - University of Genoa, Corso Dogali 1M - I- 

16136 Genoa, Italy. - 2 DIST - Univeristy of Genoa, Via 

Opera Pia 13 - I 16145 Genoa, Italy. - E-mail: 

milla@klaatu.com.dist.unige.it 

The work presents a study carried out in selected plots of 

several beech woods in Western Liguria (Italy) through 

five years. The zone investigated were the subject of some 

previous researches, devoted to study the mycological 

flora. From a phytosociological point of view and despite 

the limited diversity of the flora, the woods belong to 

Trochiscantho-Fagetum association (Geranio-Fagion 

alliance), quite common in Northern and Central 

Apennines of Italy. The macrofungi collected and 

identified amount to three hundred and fifty seven species. 

These species were attributed to the following five 

ecological groups: 173 ectomycorrizal fungi, 75 

saprotrophic on dead wood, 60 saprotrophic on soil or 

humus, 47 saprotrophic in the litter, 2 parasitic on leaving 

trees. Gathered data point out that the mycological flora is 

rich in species which can be deemed strictly related to the 

beech, but they have a rather wide synecological relevance, 

occuring in all kinds of beech woods. Finally, some 

preliminary results are also reported, exploiting statistical 

analysis techniques based on both myco-floristic and 

enviromental data. 

618 - Filamentous fungi associated with the bivalve 

mollusk Corbicula japonica Prime 

L.V. Zvereva * & A.M. Vysotskaya 

Institute of Marine Biology, Far East Branch, Russian 

Academy of Sciences, Palchevskogo Street, 17, 

Vladivostok-41, 690041, Russia. - E-mail: 

inmarbio@mail.primorye.ru 

The bivalve mollusk Corbicula japonica Prime 

(Corbiculidae) is a commercially valuable species and has 

unique pharmacological (hepatoprotective) properties. 

Mollusks were collected in the brackish water Ainskoye 

Lake (Sakhalin) on September 25, 2000. Adult individuals 

with shells 25 - 35 cm wide were selected. To remove sand 

and mud from the internal organs, animals were maintained 

for a day in flowing water. Then the mollusks were frozen 

and kept in a refrigerator at -18°C until the mycological 

investigation. Internal organs of the mollusks - mantle, 

muscles, gills, kidneys, digestive glands, male and female 

gonads - were prepared for the experimental study. Before 

inoculation to the liquid nutrient media, the organs of the 

mollusks were kept for two hours in sterilized distillated 

water with antibiotics to inhibit the bacterial flora. Several 

nutrient media were used: glucoso-yeast extract medium, 

Tubaki's medium, and others. We isolated and identified 

ten species of the filamentous fungi associated with the 

mollusk Corbicula japonica. They are Mortierella 

longicollis, M. vinacea, M. sp., Mucor circinelloides, 

Gymnoascus sp., Trichoderma aureoviride, T. hamatum, 

Penicillium atramentosum, Aspergillus ochraceum, and 

Cladosporium sphaerospermum. Growth of the yeast fungi 

was observed. Fungi were found in the digestive gland, in 

the female gonads, in the male gonads, in the mantle, and 

in the kidneys. Fungi were not found in the gills and 

muscles. 


IMC7 Main Congress Theme II: SYSTEMATICS, PHYLOGENY AND EVOLUTION Posters 

619 - Coevolution between fungus-growing termites and 

Termitomyces fungi 

D.K. Aanen 1* , P. Eggleton 2 , C. Rouland 3 , T. Guldberg- 

Froslev 4 , S. Rosendahl 4 & J.J. Boomsma 1 

1 Zoological Institute, University of Copenhagen, 

universitetsparken 15, 2100 Copenhagen, Denmark. - 2 The 

Natural History Museum, Cromwell Road, London, U.K. - 

3 Laboratoire décophysiology des invertebrates, University 

of Paris XII, Val de Marne 61, avenue du General Charles 

de Gaulle, 94010 Creteil Cedex, France. - 4 Botanical 

Institute, University of Copenhagen, Oster Frarimagsgade 

2D, DK-1353 Copenhagen, Denmark. - E-mail: 

dkaanen@zi.ku.dk 

Termites of the exclusively Old World subfamily 

Macrotermitinae live in an obligate symbiosis with fungi of 

the genus Termitomyces (Basidiomycotina). Here we 

present phylogenies of both partners in this symbiosis 

(estimated using Bayesian analyses of DNA sequences). 

Our sample consists of 43 colonies of termites and their 

fungi (belonging to 32 termite species, covering 9 of the 11 

genera) from three African and three Asian localities. 

Fungal sequences were obtained using comb material, 

basidiocarps and termite gut contents (using Termitomyces 

specific primers). For information on the taxonomic 

affiliation of the Termitomyces symbionts we also obtained 

DNA sequences from herbarium specimens of seven 

relatively well defined species. We show that the symbiosis 

has a single African origin and that secondary 

domestications of other fungi or reversals to a free-living 

state have not occurred. Host switching at low taxonomic 

levels is common and single termite species can have 

different symbionts. This is consistent with fungal 

reproduction, independent of termite reproduction 

(horizontal transmission), which is inferred to be the 

ancestral transmission mode. Specificity increases towards 

the higher taxonomic levels and the four main clades of 

fungus-growing termites are generally associated with 

specific clades of fungi. The inferred patterns of coevolution 

challenge the hypothesis that vertical 

transmission is a prerequisite for maintaining advanced 

mutualistic symbiosis. 

620 - Species delimitation in the Puccinia striiformis 

complex 

M. Abbasi 1 , S.B. Goodwin 2* , M.S. Scholler 3 & Gh.A. 

Hedjaroude 4 

1 Plant Pests & Diseases Research Institute, Botany 

Department, Tehran, Iran. - 2 USDA-ARS / Purdue 

University, Department of Botany and Plant Pathology, 

1155 Lilly Hall, West Lafayette, IN 47907-1155, U.S.A. - 

3 Purdue University, Arthur and Kriebel Herbaria, 1155 

Lilly Hall, West Lafayette, IN 47907-1155, U.S.A. - 

4 Tehran University, Plant Protection Department, College 

of Agriculture, Karaj, Iran. - E-mail: 

sgoodwin@purdue.edu 

188 


Puccinia striiformis (Uredinales), the cause of yellow rust 

or stripe rust, is a species forming uredinia and telia on 

various species of Poaceae. It is assumed to be 

heteroecious, but the aecial host is not known. Currently, 

two varieties, P. striiformis var. striiformis and P. 

striiformis var. dactylidis are accepted by most authors. We 

sequenced the variable internal transcribed spacer (ITS 

1+2) region and the more conservative 5.8S gene of the 

ribosomal DNA and found considerable differences among 

specimens on Triticum (Triticae), Dactylis glomerata 

(Poae) and Poa pratensis (Poae). These results correspond 

with a previous isozyme phenotype study and also with 

analyses of morphological features. Based on these 

differences we suggest a three-species system including P. 

striiformis on numerous hosts (mainly on Triticeae), P. 

pseudostriiformis sp. nov. on Poa pratensis (Poae), and P. 

striiformioides nom. et stat. nov. on Dactylis glomerata 

(Poae). 

621 - Nomenclatural and taxonomic status of some taxa 

in Russula sect. Xerampelinae 

S. Adamcík & P. Lizon * 

Institute of Botany, Dúbravská 14, SK-842 23 Bratislava, 

Slovakia. - E-mail: botupali@savba.sk 

Numerous validly published names in agarics have not 

been accepted by current authors. Due to short and 

insuficient original descriptions and missing type 

specimens their identity is doubtful. We have studied 

members of Russula sect. Xerampelinae with greenish 

pileipes those taxonomic delimitation and nomenclature are 

confusing. Only three taxa in that group are clearly 

delimited: Russula clavipes Velen., R. cicatricata Romagn. 

ex Bon and R. schaefferi Kärcher. R. schaefferi differs in 

having narrow terminal cels of generative hyphae in 

pileipes and spores with long spines. R. clavipes and R. 

cicatricata have terminal cells of generative hyphae in the 

center of the pileipes inflated. Illustrations are available for 

lectotypification (even Latin diagnoses are missing) of 

Russula elaeodes and R. fuscoochracea R. Schulz. Later 

name is superfluous (homonym of R. fuscoochracea 

Velen.) but R. elaeodes seems to represent a distict taxon. 

No type specimen was designated for R. barlae, R. fusca, 

R. ochracea and R. olivascens and original descriptions 

lack characters typical for the section. Another names, such 

as R. citrinocincta, R. cookeiana and R. duportii need also 

clarification. Complexe knowledge of the members of 

Russula sect. Xerampelinae and consequent rejection of 

several names are the only way how to stabilize the 

nomenclature and the taxonomy of this section.


622 - Impact of rhizomorph structure on systematics of 

Hymenomycetes (Basidiomycota) 

R. Agerer 

University of München, Department Biology I, Biodiversity 

Research: Systematic Mycology, Menzinger Str. 67, D- 

80638 München, Germany. - E-mail: 

myrrhmyk@botanik.biologie.uni-muenchen.de 

Rhizomorphs can be used to confirm or to clarify 

relationships within Hymenomycetes (Agerer 1999, 2002, 

Hahn et al. 2001). Recent studies have shown that the final 

structure in combination with the ontogeny of rhizomorphs 

are important means to discern fungal relationships. 

Virtually all families of Boletales s. l. (Gomphidiaceae, 

Tapinellaceae, and Truncocolumellaceae excluded) have an 

identical type of rhizomorphs, the boletoid type (Agerer 

1999). The recently by DNA-analyses repeatedly 

confirmed relationship between Gomphales, Geastrales and 

Gautieriales is characterized by ramarioid rhizomorphs 

(Agerer 1999, Hahn et al. 2001), which are unknown in any 

other fungal group. Agaricoid rhizomorphs (Agerer 1999, 

2002) are typical of Agaricaceae and Lycoperdales. The 

unique rhizomorph structures are shown and their impact 

on Hymenomycetes systematics discussed. References: 

Agerer R (1999) Never change a functionally successful 

principle: the evolution of Boletales s. l. (Hymenomycetes, 

Basidiomycota) as seen from below-ground features. 

Sendtnera 6: 5-91. Agerer R (2002) Rhizomorph structures 

confirm the relationship between Lycoperdales and 

Agaricaceae (Hymenomycetes, Basidiomycota). Nova 

Hedwigia (subm.) Hahn C, Agerer R, Wanner G (2001) 

Anatomische und ultrastrukturelle Analyse von 

Ramaricium ochraceoalbum, einer seltenen Art der 

Gomphales und seine verwandtschaftliche Beziehung zu 

Geastrum und Gautieria. Hoppea 61: 115-125. 

623 - Preliminary results in assessing phylogenetic and 

biogeographic relationships among species of Russula 

subgenus Ingratula 

R. Aldana-Gomez 1* , G.M. Mueller 1 & B. Buyck 2 

1 Department of Botany. The Field Museum of Natural 

History, 1400 S. Lake Shore Dr. Chicago, IL 60605-2496, 

U.S.A. - 2 Laboratoire de Cryptogamie. Museum National 

d'Histoire Naturelle, 12 rue Buffon, 75005 Paris, France. - 

E-mail: raldanagomez@fmnh.org 

Phylogenetic and biogeographic relationships among 

species of Russula subgenus Ingratula are being studied 

using partial nuclear Ribosomal DNA Large Subunit 

(rnDNA LSU) and the complete Internal Transcribed 

Spacer region (ITS1, ITS2 and 5.8S rDNA gene). Species 

of Russula subgenus Ingratula are macromorphologically 

characterized by the tuberculate-striate margin of the 

pileus, the brownish-yellow extracellular pigments and the 

often disagreable or pronounced smell. 

Micromorphological characters vary broadly between 

groups of species. Based on these morphological features, 

species in this subgenus have been grouped into several 

sections and series, but their phylogenetic relationships had 

not been evaluated. Phylogenetic analyses of 35 LSU and 

ITS sequences, respectively, are being performed using 

Maximum Parsimony, Maximum Likelihood and Bayesian 

methods. The distinction between sections and between 

series is not supported in preliminary analyses. Species in 

the subsection Foetentinae are scattered in the cladogram, 

while species in section Subvelatae fall within two separate 

and unrelated clades. Sister taxon relationships for some 

taxa are suggested, e.g. neotropical R. arcyospora and 

cosmopolitan R. laurocerasi, American R. pulverulenta and 

European R. insignis. Test of Congruence for the 

combinability of the two data set together with sequencing 

of additional genes and additional taxon sampling are 

ongoing. 

624 - The 'Torula' infections on Cetraria spp. 

V. Alstrup 

Botanical Museum, University of Copenhagen, 

Gothersgade 130, DK 1123 Copenhagen K, Denmark. - Email: 

vagna@bot.ku.dk 

The so-called sorediate forms of the lichen genus Cetraria 

was revised and found to be pathogenic conditions caused 

by parasitic hyphomycetes of which only one species had 

been formally described. The new genus Erichseniomyces 

with the type species E. sorediella comb. et stat. nov. and 

E. islandica sp. nov. are described as are the new species 

Trimmatostroma ahlneri and T. danica. 

625 - Locating the position of the Micareaceae within 

the Lecanorales (lichenized Ascomycota) 

H.L. Andersen 

Department of Botany, University of Bergen, Allégaten 41, 

5007 Bergen, Norway. - E-mail: 

heidi.andersen@bot.uib.no 

The phylogeny of the Micareaceae and the genus Micarea 

Fr. was studied using 39 nuclear small subunit ribosomal 

DNA sequences, of which four were new. Phylogenetic 

analyses using maximum parsimony, maximum likelihood, 

and Bayesian inference were carried out. These resulted in 

18 most parsimonious trees, of which one was 

corresponding to the 50% majority rule consensus tree 

from the Bayesian inference. The maximum likelihood tree 

was identical except for one collapsed branch. In all trees, 

Micarea adnata and Byssoloma leucoblepharum formed a 

strongly supported group. This study supports the 

placement of the Micareaceae together with the 

Pilocarpaceae inside the Lecanorales, as a sistergroup to 

the Bacidiaceae. Several null hypotheses concerning 

monophyly were tested: the monophyly of the family 

Micareaceae in the sense of Eriksson & Hawksworth, and 



the genus Micarea was rejected. Scoliciosporum A. Massal 

has sometimes been considered close to Micarea, but null 

hypotheses assuming monophyly together with the 

Micareaceae or the Bacidiaceae plus Micareaceae were 

rejected. 

626 - A Monograph of tribus Myceneae (Mycena 

excluded) in Europe 

V. Antonín 1* & M.E. Noordeloos 2 

1 Moravian Museum, Botanical Department, Zelny trh. 6 

Brno, Czech Republic. - 2 National Herbarium of the 

Netherlands, Leiden University Branch, P.O.Box 9514 

NL2300 Leiden, The Netherlands. - E-mail: 

vantonin@mzm.cz 

Tribus Myceneae, in the tradition of the Flora agaricina 

neerlandica, vol. 2 (Bas et al. 1995) is characterized by 

fruitbodies with an omphalioid, mycenoid or 

tricholomatoid habit, a pileipellis in form of a cutis or 

ixocutis, sometimes with transitions to a trichoderm, with 

smooth, coralloid, diverticulate elements, amyloid or 

inamyloid spores, and white spore print. Revisions of all 

genera, except Mycena, are given, based on a thorough 

examination of material from all parts of Europe, including 

type-specimens and additional information. Particularly the 

genus Hemimycena appeared to be in need of a critical 

revision, because no serious monograph had been 

published after the Mycena monograph of Kühner (1938). 

Revision is hampered by the lack of type material, 

particularly of the species described from the (sub)alpine 

belt in Haute Savoie, France by Valla (1957). The present 

authors now accept 32 species, including 3 new ones. Type 

revisions result in a few name changes. In the difficult 

complex of H. mauretanica, a wide species concept is 

proposed. The genus Xeromphalina now is represented 

with eight species in Europe, including the North American 

X. brunneola and X. campanelloides. The genus Rickenella 

embraces after revision also the genus Jacobia, and is 

represented with 7 species. 

627 - Methods for the interpretation and the evaluation 

of spores in the genus Cortinarius 

D. Antonini 1* , M. Antonini 1 & G. Consiglio 2 

1 c/o Orto Botanico Forestale Abetone, Via F. Ferrucci 626 

- I 51036 Larciano (PT), Italy. - 2 c/o Associazione 

Micologica Bresadola, Via C. Ronzani 61 - 40033 

Casalecchio di Reno (BO), Italy. - E-mail: dantonini@tin.it 

In the genus Cortinarius sl. spores play a fundamental role 

in differentiating taxa, as to dimensions, form and 

ornamentations. In order to carry out an objective 

sporographic analysis, for each collection n spores (at least 

32) have been measured with regard to length (L), width (l) 

and quotient (Q). Then, the analytic equation of the 

isoprobability ellipse at 68,26% level and the equations of 

190 


the tangents to this ellipse have been calculated so as to 

obtain the length-width measures of six singular 

equiprobable spores, that is, the longest, the widest, the 

shortest, the narrowest and those which show the highest 

and the lowest Q, respectively. Starting from the basic 

spore, with given form and ornamentations, is thus possible 

to construct a sporogram with a highly descriptive 

effectiveness. The typology of ornamentations has been 

evaluated by observations made with both the optical and 

the SEM microscope. The results obtained have allowed us 

to find out some ranges based on the following features: 

ornamentation height [≤ 1 µm (smooth or hardly visible by 

the optical microscope); from 1 to 2 µm ; ≥ 2 µm]; 

development of the ornamentations on the spore surface 

(more concentrated towards the apex or in the upper half or 

more or less spread on the whole surface); warts form 

(rounded, pyramidal or spiny); presence or lack of hilar 

plage. 

628 - The taxonomy of the Lecanora allophana group 

using DNA data 

U. Arup * & P. Frödén 

Botanical Museum, Östra Vallgatan 18, SE-223 61 Lund, 

Sweden. - E-mail: ulf.arup@sysbot.lu.se 

Lecanora allophana belongs to the large genus Lecanora 

of lichenized ascomycetes. Within the genus there are 

several groups of species and L. allophana belongs to the 

L. subfusca group, that is spread all over the world. L. 

allophana is also the type species of the genus. There are a 

number of species related to L. allophana that are more or 

less difficult to seperate from L. allophana using 

morphological characters. Therefore, there has been much 

doubt in some of the species of the group. Molecular 

studies, using nuclear ribosomal ITS data, show that most 

species in the group are well separated and that there are 

even more species than previously understood. However, 

within L. allophana and L. horiza the molecular data are 

not homogenous and indicates that there may be so called 

cryptic species within these taxa. Further studies of more 

genes will have to be carried out to settle this matter. 

629 - Application of pectic zymogram in the 

identification and genetic variation of Fusarium species 

G.R. Balali * & M. Iranpour 

Dept of biology, University of Isfahn, Isfahan 81744, Iran. 

- E-mail: rbalali@sci.ui.ac.ir 

The genus Fusarium includes several species which 

classified based on morphological characters. Different 

media are required to study the morphological characters of 

different species. Regarding molecular and biochemical 

techniques for identifying fungi, pectic zymograms have 

been used to characterized different fungi. Zymogram 

technique was used to identify species and form species of


Fusarium isolates. 318 isolates identified as the genus 

Fusarium were collected from different areas and hosts. 

Species of the isolates were identified based on 

morphological characters. The pectic enzyme solution was 

prepared for each isolate using liquid media containing 

citrus pectin as a sole carbon source. Electrophoresis was 

performed using pectin acrylamide gel. Several zymogram 

phenotypes were obtained for polygalacturonase and pectin 

esterase. In total, 12 zymogram patterns were determined 

for 318 isolates tested. The results showed that there is a 

considerable intraspecific variation for Fusarium species. 

There were 3, 5 and 2 zymogram electrophoretic patterns 

for Fusarium oxysporum, F. solani and F. culmorum 

respectively. However, there were only one zymogram 

pattern for F. subglutinans and also one for F. equiseti. 

Although the intraspecific variation based on zymogram 

was not correlated to the form species of Fusarium, the 

species of Fusarium were distinguished using this 

technique and there was no common zymogram pattern 

among species. 

630 - The lichen genus Caloplaca on Svalbard 

L. Balschmidt * & U. Søchting 

Botanical Institute, Department of Mycology, University of 

Copenhagen, Øster Farimagsgade 2D, 1353 Copenhagen 

K, Denmark. - E-mail: lineb@bot.ku.dk 

The Svalbard Archipelago hosts a highly diverse flora of 

high Arctic Caloplaca species with more than 40 species 

growing primarily on rock, soil, lignum and bone. A 

comprehensive study of the Caloplaca flora adds to the 

understanding of Arctic species distribution and clarifies 

the species concept of several formerly poorly understood 

species, e.g. Caloplaca invadens. 

631 - Molecular phylogeny of North American species 

of Laetiporus and related genera 

M.T. Banik 1 , D.L.L. Czederpiltz 2* & J.A. Micales 1 

1 Center for Forest Mycology Research, USDA Forest 

Service, Forest Products Laboratory, One Gifford Pinchot 

Drive, Madison, WI 53705-2398, U.S.A. - 2 University of 

Wisconsin-Madison, Dept. of Plant Pathology, 1630 

Linden Dr., Madison, WI 53706, U.S.A. - E-mail: 

dlindner@facstaff.wisc.edu 

Relationships among species in the genera Laetiporus, 

Phaeolus, Pycnoporellus, Wolfiporia, and Leptoporus were 

investigated using nuclear 25S and ITS rDNA and 

mitochondrial 18S rDNA sequences. Members of these 

genera have poroid hymenophores and simple septate 

hyphae, and cause brown rots in a variety of substrates. 

Results based on parsimony and maximum likelihood 

analyses indicate that the genus Laetiporus as currently 

defined is not monophyletic. The ITS regions of all 

Laetiporus species examined were easily aligned, with the 

exception of L. persicinus. Analyses suggest that L. 

persicinus is no more closely related to other Laetiporus 

species than to species of Phaeolus, Leptoporus or 

Pycnoporellus. The six other Laetiporus species examined 

(L. cincinnatus, L. conifericola, L. gilbertsonii, L. 

huroniensis, L. sulphureus, and an undescribed species 

from the Caribbean) appear to make up a well-supported, 

monophyletic group. Results confirmed that L. conifericola 

and L. huroniensis, two species that occur on conifers, are 

very closely related but distinct species. The existence of a 

previously undescribed yellow-pored species from the 

Caribbean was also confirmed. This species may be more 

closely related to L. gilbertsonii than to any other species 

of Laetiporus. Two varieties of L. gilbertsonii, which are 

compatible in mating tests and differ only in pore color, 

were indistinguishable in these analyses. 

632 - Using parsimony networks, migration estimates 

and coalescence approaches to resolve the 

phylogeography of Mycosphaerella graminicola 

S. Banke * & B.A. McDonald 

Institute of Plant Sciences, Phytopathology, ETH, 

Universitaetstr. 2 / LFW-B28, CH-8092 Zurich, 

Switzerland. - E-mail: soren.banke@ipw.agrl.ethz.ch 

DNA sequences of nuclear-encoded loci can be used to 

determine the phylogeographical history of fungal 

populations. Four nuclear loci were sequenced in the fungal 

wheat pathogen M. graminicola. Two neutral non-coding 

RFLP loci and two nuclear single copy genes were 

selected. The coding genes were β-tubulin and 3isopropylmalate 

dehydrogenase (LeuC). A total of 300 M. 

graminicola isolates originating from 14 wheat fields 

around the world were sequenced. All sequence loci 

showed useful diversity. The RFLP locus STS2 had 29 

polymorphic sites in a 490 bp region, while STL10 had 31 

polymorphic sites in a 1265 bp region. For the β-tubulin 

locus, 24 polymorphic sites were found among 366 bp 

sequenced. The leuC gene contained 65 polymorphic sites 

within the 870 bp region sequenced. Gene genealogies 

were generated for all sequenced loci following clonecorrection 

for each locus/population. Trees were rooted 

using Septoria passerinii, the closest known relative of M. 

graminicola. Parsimonious networks were constructed 

using a nested cladistic approach, and migration rates 

between populations were estimated. A coalescence 

approach was used to determine the temporal scale of 

historical migration events among populations. All loci 

were also tested for neutrality and intergenic 

recombination. Our results suggest that the centre of origin 

for this pathogen is most likely in the Fertile Crescent, and 

that migration occurred first into the European continent 

and later into the New World. 



633 - A new winter-fruiting hyphomycete on wood of 

black locust (Robinia pseudoacacia) in the eastern 

United States 

G.F. Bills 

Centro de Investigación Básica, Merck Sharp & Dohme de 

España, S.A., Josefa Valcárcel 38, E-28027 Madrid, Spain. 

- E-mail: gerald_bills@merck.com 

A new hyphomycete, tentatively assigned to the genus 

Cadophora, is described based on studies of field 

collections and culture morphology, and analyses of the 

ITS region of ribosomal DNA. The fungus is found 

consistently on the undersides of decorticated logs, 

branches, and stumps of Robinia pseudoacacia in the 

eastern United States. Collections have been observed from 

Connecticut southward to Virginia and westward into 

Ohio. The fungus forms membranous, sporodochial 

conidiomata on decorticated wood of R. pseudoacacia. The 

conidiomata develop early in the winter and persist until 

spring. The fungus can be isolated from soil in the 

proximity of R. pseudoacacia and was once recovered from 

river silt in Maryland. Ribosomal DNA ITS sequences 

exhibit high homology with the ITS sequences of 

Phialocephala fortinii, various ericoid mycorrhizal fungi, 

and Mollisia species. Phylogenetic placement near Mollisia 

is consistent with the habitat on decorticated wood and 

darkly pigmented, phialidic conidiogenous cells that 

produce simple ameroconidia. 

634 - Expansion of the genus Trichoderma: addition of 

seven new species from Eurasia 

J. Bissett 1 , I. Druzhinina 2 , C.M. Kullnig-Gradinger 2 , G. 

Szakacs 3 & C.P. Kubicek 2* 

1 Agriculture and Agri-Food Canada, Eastern Cereal and 

Oilseed Research Center, Central Experimental Farm, 

Ottawa, Ontario, Canada K1A 0C6, Canada. - 2 Institute 

for Chemical Engineering, Microbial Biochemistry and 

Gene Technology Group, Technical University of Vienna, 

Getreidemarkt 9/166, A-1060 Vienna, Austria. - 

3 Department of Agricultural Chemical Technology, 

Technical University of Budapest, 1111 Budapest, Gellert 

ter 4, Hungary. - E-mail: 

ckubicek@mail.zserv.tuwien.ac.at 

Species of the deuteromyceteous genus Trichoderma are 

cosmopolitan and typically soil-borne or wood-decaying 

fungi, some of them being economically important because 

of their production of industrial enzymes (cellulases and 

hemicellulases), antibiotics, and their action as biocontrol 

agents. In order to fully exploit this potential, we started a 

study of the global biodiversity of the genus. Thereby a 

number of unique strains were obtained which were 

culturally and genetically dissimilar to all of the about 40 

known taxa of Trichoderma. Here we characterize seven of 

these strains as new species, all originating from Eurasia, 

and assign them to the established sections of the genus: T. 

192 


taiwanense and T. effusum to section Longibrachiatum; T. 

helicum, T. rossicum, T. velutinum, and T. cerinum to 

section Pachybasium; and T. erinaceum to section 

Trichoderma. Parsimony analysis, using ITS1 and 2 and 

tef1 gene sequences, strongly confirm these placements and 

also clearly distinguish these new species from 

morphologically similar species. 

635 - Phylogenetic significance of adaptive structures of 

macromycete sporophors 

M.A. Bondartseva 

V.L. Komarov Botanical Institute of Russian Academy of 

Sciences, 2 Prof. Popov Str. 197376 St. Petersburg, Russia. 

- E-mail: mbond@iz6284.spb.edu 

During last two centuries the approaches in study of fungi 

changed from the recognition of macromorphological 

characters to the microscopic and molecular ones 

nowadays. The last ones are available to the limited 

number of taxa with extrapolation of the morphologically 

similar species to the same group. New knowledge 

stimulates the diversity in interpretation of the data and 

reflects in multiplicity of the systems. New approach in 

regarding of fungi relationships consists in dividing 

characters to the groups of ecologically active, that 

connected directly with fungus adaptation to the 

environment (morphology of fruit body etc.) and 

ecologically inert (basidia and spores features, clamps etc.). 

As genetic relationships reflect in morphological similarity, 

and adaptation to the environment comes to be on the base 

of definite hereditary, modus variability being historically 

limited, it is possible to differentiate phylogenetic 

relationships and convergent likeness, as well as progress 

with the rise of evolutionary level and adaptive allogenetic 

evolution on the same evolutionary level. There are many 

examples to confirm this view. When evaluation of 

taxonomic characters in dependence on its adaptation to the 

environment agree with the molecular data it means the 

selected criteria are right, if not - attentive study to find the 

reason must be made and conclusions have to be more 

cautious. 

636 - Taxonomic monograph of Fusicladium, Pollaccia 

and Spilocaea (Hyphomycetes, Venturia anamorphs 

p.p.) 

U. Braun 1* , A. Ritschel 2 & K. Schubert 1 

1 

Martin-Luther-Universität, Institut für Geobotanik und 

Botanischer Garten, Neuwerk 21, D-06099 Halle, 

Germany. - 

2 

Eberhard-Karls-Universität, Botanisches 

Institut, Spezille Botanik/Mykologie, Auf der Morgenstelle 

1, D-72076 Tübingen, Germany. - E-mail: 

braun@botanik.uni-halle.de 

The genera Fusicladium Bonord., Pollaccia E. Bald. & Cif. 

and Spilocaea Fr. (hyphomycetes) have recently been


monographed (Ritschel 2001, Schubert 2001), based on 

light and scanning electron microscopical as well as 

molecular (rDNA ITS sequences) examinations, and 

includes comprehensive descriptions and illustrations of 40 

Fusicladium, 7 Pollaccia and 5 Spilocaea species. Venturia 

and its anamorphs proved to be monophyletic. Based on a 

combination of morphological and molecular examinations 

and data, the generic delimitation and the species concept 

of Venturia anamorphs have been re-evaluated. Features 

suitable for the delimitation of species and other features 

unsuitable for this purpose have been pointed out. The 

structure of the conidiogenous loci (scars) and conidial 

hila, which is uniform within Venturia anamorphs, proved 

to be the basic characteristic for the delimitation of genera. 

Morphological and molecular examinations showed that 

the three anamorphic genera Fusicladium, Pollaccia and 

Spilocaea are not tenable. It is suggested to merge these 

genera under the well-known name Fusicladium, which is 

proposed to be conserved over the older name Spilocaea. 

637 - Cultural and microscopic studies of Schizophyllum 

commune and Grifola frondosda (higher 

basidiomycetes) strains isolated in Ukraine 

A.S. Buchalo 1* , S.P. Wasser 2 & V.M. Linovitska 3 

1 

N. G. Kholodny Institute of Botany, National Academy of 

Science of Ulraine, 2 Tereshchynkivska, Kiev, 01601, 

Ukraine. - 2 International Center for Cryptogamic Plants 

and Fungi, Institute of Evolution, University of Haifa, 

Institute of Evolution, University of Haifa, Mt. Carmel, 

Haifa 31905, Israel. - 

3 NTUU 'KPI', Faculty of 

Biotechnology and Biotechnique, 37 Peremogy Ave., Kiev, 

03057, Ukraine. - E-mail: asbuchalo@kiev.botan.ua 

The increasing interest in the cultivation of Schizophyllum 

commune and Grifola frondosa depends on their 

extraordinary medicinal properties. Biotechnological 

application of S. commune and G. frondosa mycelial 

cultures requires isolation of new strains from nature and 

more detailed studies of their morphological 

characteristics. Pure cultures of G. frondosa and S. 

commune were isolated from fruiting bodies collected on 

the territory of the Ukraine. Cultures were screened on 

different agar media at the temperature interval +4 - 38 °C. 

The growth rate of strains and enzyme tests on oxidases in 

cultures were determined. Fast growing strains, which have 

strong enzyme reactions were selected for the following 

investigation. Scanning electron microscopic studies 

showed that on hyphae of G. frondosa numerous clamp 

connections of medallion type were present. In old parts of 

colony often sprouting clamps and anastomoses are typical. 

In the younger parts of a colony thin (≤ 1 mm wide) 

branched hyphae (dichohyphidia) are forming. Apical and 

intercallar chlamydospores are usual. In S. commune clamp 

connections of a classic shape are registered. Some hypha 

are densely covered with projections described by some 

authors as pseudoconidia or secretory conidia. Ellipsoid 

chlamydospores are often present. Teleomorph is typically 

formed during 5-6 weeks. Described characteristics are 

significant. For instance, clamps can be used for the control 

of purity and physiologic state under cultivation. 

638 - Two new hyphomycetes from Spain 

M. Calduch * , J. Gené, A.M. Stchigel & J. Guarro 

Rovira i Virgili University, C/ Sant Llorenç 21, 43201- 

Reus, Spain. - E-mail: umb@fmcs.urv.es 

Spanish mycobiota is considered highly diverse due to the 

regional variety of climate and vegetation. However, the 

hyphomycete check-list is still very reduced. Over recent 

years, we have conducted an extensive survey of those 

fungi from different habitats. Recently, we have found two 

undescribed specimens belonging to Oidiodendron Robak 

and Phialophora Medlar, isolated from litter and soil, 

respectively. Plant debris samples were placed into moist 

chambers, and soil samples were plated onto sterile Petri 

dishes which were moistened thoroughly with sterile 

distilled water and covered with small pieces of sterile 

wood (soil bait technique). Oidiodendron myxotrichoides 

sp. nov. is characterized by the presence of dark brown 

reticulate sporodochia, which resemble gymnothecia 

(ascomata) of Myxotrichum, and by its globose or 

subglobose arthroconidia. The presence of sporodochia 

constitutes a new character in Oidiodendron. Phialophora 

vesiculosa sp. nov. is characterized by possessing erect and 

branched conidiophores with branches ending in sterile 

vesicles, and by its mono- or polyphialidic, terminal or 

lateral conidiogenous cells with flared collarettes, from 

which spherical, brown conidia with a protuberant basal 

hilum are produced. 

639 - Evolutionary relationships between aquatic 

anamorphs and teleomorphs: Lemonniera 

J. Campbell 1* , C.A. Shearer 1 & L. Marvanová 2 

1 UIUC, Dept of Plant Biology, 265 Morrill Hall, 505 S. 

Goodwin Ave, Urbana, IL, U.S.A. - 2 Masaryk University 

Brno, Faculty of Science, Tvrdeho 14, 602 00 Brno, Czech 

Republic. - E-mail: jcampbe2@life.uiuc.edu 

Mitosporic fungi have traditionally been classified based 

on their method of conidiogenesis, conidial secession, and 

conidium and conidiophore morphology. We wanted to 

determine whether these were important phylogenetic 

characters for a group of morphologically and 

developmentally similar aquatic hyphomycetes, or if this 

was due to convergent evolution. Cladistic analyses were 

performed on the 28S rDNA of eight species of 

Lemonniera and one species of Margaritispora. These two 

genera both have phialidic conidiogenesis and schizolytic 

conidial secession. Lemonniera has tetraradiate conidia and 

Margaritispora has unbranched conidia. Molecular 

analyses demonstrate that Lemonniera species are placed in 

two distinct clades: one within the Leotiomycetes; the other 

within the Pleosporales, Dothideomycetes. Margaritispora 

is placed with Lemonniera species within the 

Leotiomycetes. This demonstrates that for this group of 

species conidiogenesis, conidial secession and conidium 



shape are not phylogenetically informative, but are the 

result of convergent evolution. 

640 - Determination of biological species and analysis of 

genetic variability by RAPD of strains of Pleurotus 

subgenus Coremiopleurotus 

M. Capelari 1* & M.H.P. Fungaro 2 

1 Instituto de Botânica, Caixa Postal 4005, São Paulo, SP, 

CEP 01061-970, Brazil. - 2 Universidade Estadual de 

Londrina, Londrina, PR, Brazil. - E-mail: 

mcapelariibot@yahoo.com 

Pleurotus cystidiosus and P. smithii strains of different 

geographic origins were studied in terms of growth rate in 

culture, mono-dikaryotic matings and genetic variability 

determined by RADP in order to prove the occurrence of P. 

cystidiosus in South America. The present results show that 

the criteria used to separate the two species are 

unsatisfactory and that P. smithii should be considered a 

synonym of P. cystidiosus, thus extending the distribution 

of this species to Central and South America. 

641 - Phylogenetic relationships among the bunt fungi 

L.A. Castlebury 1* & L.M. Carris 2 

1 USDA ARS Systematic Botany and Mycology Laboratory, 

10300 Baltimore Ave., Beltsville, MD 20705, U.S.A. - 

2 Dept. of Plant Pathology, Washington State University, 

Pullman, WA 99164-6430, U.S.A. - E-mail: lisa@nt.arsgrin.gov 

There are approximately 120 species of bunt fungi in 

Tilletia and related genera that parasitize members of the 

Poaceae. Most species are distinguished by teliospore 

ornamentation and host. A number of non-reticulate-spored 

species in Tilletia have been placed in other genera in the 

Tilletiales including Neovossia, Conidiosporomyces, 

Ingoldiomyces, and Oberwinkleria. Two economically 

important species, T. indica (Karnal bunt) and T. horrida 

(rice kernel smut) have been placed in Neovossia on the 

basis of tuberculate teliospore ornamentation, floret 

infection and the production of large number of nonconjugating 

basidiospores. Phylogenetic analysis of large 

subunit nuclear rDNA sequences shows that reticulatespored 

taxa with hosts in the Pooideae, including T. tritici, 

T. laevis and T. controversa, form a well-supported 

monophyletic group (bootstrap >95%). Tilletia indica and 

T. walkeri, also on members of the Pooideae, form a wellsupported 

group (100%) and group with reticulate-spored 

taxa and Ingoldiomyces hyalospora on hosts in the 

Pooideae (bootstrap >70%). A few reticulate-spored taxa 

and other tuberculate-spored taxa on non-Pooideae hosts, 

including T. horrida, fall in a separate, unsupported group 

although a number of well-supported lineages exist within 

this group. Conidiosporomyces ayresii groups strongly 

with T. vittata. Results suggest that host may be more 

194 


phylogenetically informative than morphological 

characters. 

642 - Taxonomic revision of the lichen genus 

Heterodermia (Physciaceae) from China's mainland 

J.B. Chen 

Institute of Microbiology, Academia Sinica, P.O.Box 2714, 

Beijing 100080, China. - E-mail: chenjbin@yahoo.com 

Thirty-one species of Heterodermia are recognized from 

China's mainland, based on examination of more than 1000 

specimens kept in HAMS. Of which, H. orientalis and H. 

sinocomosa were described as new (Chen, 2001), and H. 

galactophylla, H. pseudosquamulosa and H. rubescens are 

added to Chinese lichen flora. The previous records of H. 

erinacea and H. granulifera are proved to be mistaken 

reports. They are actually H. pellucida and H. isidiophora 

respectively and should be excluded from Chinese lichen 

flora. The previous records of H. albicans, H. barbifera 

and H. dactilyza from China are doubtful. 

Biogeographically, H. boryi, H. comosa, H. diademata, H. 

flabellata, H. isidiophora, H. leucomela, H. lutescens, H. 

obscurata, H. podocarpa, H. propagulifera, H. 

pseudospeciosa and H. speciosa are tropical and/or 

temperate. H. angustiloba, H. dendritica, H. firmula, H. 

fragilissima, H. microphylla, H. orientalis, H. pellucida, H. 

pseudosquamulosa, H. rubescens, H. sinocomosa, H. 

subascendens and H. togashii are eastern Asian or southern 

Asian. Of which H. orientalis, H. pseudosquamulosa and 

H. sinocomosa are endemic to China. H. dissecta and H. 

hypoleuca are disjunctive in eastern Asia and eastern North 

America. H. pacifica occurs in Pacific region. The 

geographical affinities of H. galactophylla, H. hypochraea, 

H. hypocaesia and H. japonica are still uncertain. H. 

corallophora, H. incana, H. pandurata and H. spinula 

recorded in Taiwan of China have not been found in 

China's mainland. 

643 - Some merosporangiferous Mucorales indigenous 

to dungs of house mice in Taiwan 

C.-Y. Chien 

Dept. of Biology, National Taiwan Normal University, 88 

Tingchou Rd., Sec. 4, Taipei, 117, Taiwan. - E-mail: 

cychien@cc.ntnu.edu.tw 

During studies of coprophilous fungi indigenous to dung of 

house mice, some merosporangiferous Mucorales were 

isolated. By the use of corn meal agar (CMA) and malt 

extract agar (MEA) media as well as technique favor the 

isolation of merosporangiferous fungi in Taiwan. Dung of 

samples of mice in houses or garages was collected directly 

into glass vials for transport to laboratory. The materials 

then were dispended into aseptic damp chambers prepared 

by moistening of filter paper discs in Petri dishes having 

depths of two centimeters or they were put directly onto


CMA plates. The author selected Mucor rouxianus 

(Calmette) Wehmer as the standard host for culturing 

member of Dimargaris cristalligena van Tieghem and 

Circinella umbellata van Tieghem & Le Monnier for 

Syncephalis nodosa van Tieghem, Dispira cornuta van 

Tieghem and D. simplex Benjamin. Samples of house mice 

dung were assayed for species of representatives of the 

merosporangiferous Mucorales and identified as following 

species, namely, Syncephalis nodosa van Tieghem, 

Dimargaris cristalligena van Tieghem, Dispira cornuta 

van Tieghem, D. simplex van Tieghem, Coemansia 

breviramosa Linder and C. interrupta Linder. All of above 

listed six species were illustrated and described as new 

record to Taiwan. 

644 - A comparison of Peronospora farinosa isolates 

from Chenopodium album and Spinacia oleracea using 

morphological and molecular characteristics 

Y.J. Choi 1 , S.B Hong 2 & H.D. Shin 3* 

1 

Korea University, Department of Agricultural Biology, 

2 

Graduate School, Korea. - National Institute of 

Agricultural Biotechnology, Korean Agricultural Culture 

Collection, Korea. - 3 Korea University, Division of 

Environmental Science and Ecological Engineering, 

Korea. - E-mail: hdshin@korea.ac.kr 

Downy mildews of any chenopodiaceous host are currently 

treated as a single species, Peronospora farinosa. 

Microscopic examination of the Korean isolates from 

Chenopodium album (CAI) and Spinacia oleracea (SOI) 

revealed that they are morphologically different. The 

conidia are minutely papillate in CAI, but non-papillate in 

SOI. The ultimate branchlets of CAI are mostly longer than 

those of SOI. Sequence analyses of the internal transcribed 

spacer region 1 of ribosomal DNA showed that they are 

clustered into two different groups. SOI shared only ca. 

91% nucleotide sequence homology with CAI, but showed 

ca. 94% homology with Peronospora corydalis isolates. 

On the other hand, CAI had higher sequence homology 

with Peronospora destructor isolate than with SOI. 

Consequently, we believe that downy mildews from C. 

album and S. oleracea could be considered as separate 

species. 

645 - Taxonomic revision of Uromyces species on 

cultivated legumes in Japan 

W.H. Chung 1 , T. Tsukiboshi 2 & M. Kakishima 1* 

1 Institute of Agriculture and Forestry, University of 

Tsukuba, Tsukuba 305-8572, Japan. - 2 National Institute of 

Agro-Environmental Sciences, Tsukuba 305-0856, Japan. - 

E-mail: kaki@sakura.cc.tsukuba.ac.jp 

Cultivated legumes are important crops throughout the 

world. Three Uromyces species; U. appendiculatus, U. 

vignae, and U. viciae-fabae, including varieties on 

cultivated legumes, have been reported in Japan. Although 

these species and varieties were classified mainly by 

morphology of the teliospores and urediniospores, their 

morphological delimitation is obscure. Therefore, the 

morphological characteristics of teliospores and 

urediniospores were clarified and molecular phylogenetic 

analyses based on the sequences of ribosomal DNA regions 

were carried out. More than 300 specimens collected from 

various areas of Japan were used for morphological 

observations. We examined the morphological 

characteristics of teliospores and urediniospores by light 

and scanning electron microscopy. Based on these 

observations, specimens were classified into three groups. 

About 80 specimens were selected from the specimens 

used for morphological observations and sequences of LSU 

rDNA (D1/D2) and ITS regions were analyzed. Specimens 

were separated into two clades in phylogenetic trees by the 

D1/D2 regions. However, they were separated into three 

clades in phylogenetic trees by the ITS regions. 

Correlations between morphological groups and 

phylogenetic groups based on these results suggest a 

revision of these species and varieties. 

646 - Identification of Armillaria isolates from Bhutan 

based on ITS and IGS-1 sequences 

M.P.A. Coetzee 1* , B.D. Wingfield 1 , T. Kirisits 2 , D.B. 

Chhetri 3 & M.J. Wingfield 1 



Pretoria, South Africa. - 2 Institute of Forest Entomology, 

Forest Pathology and Forest Protection, Universität für 

Bodenkultur, Hasenauerstrasse 38, A-1190 Vienna, 

Austria. - 3 Renewable Natural Resources Research Centre 

(RNR-RC), Yusipang, P.O Box 212, Thimphu, Bhutan. - Email: 

martin.coetzee@fabi.up.ac.za 

Armillaria root rot is known to occur in conifer forests in 

Bhutan. The species causing this disease have, however, 

not been identified. The aim of this study was to determine 

the identity of field isolates collected from Pinus 

wallichiana and Abies densa at two localities in western 

Bhutan. DNA sequence data were obtained for the ITS and 

IGS-1 regions of the rRNA operon and compared with 

those of Armillaria spp. available on GenBank and our 

own database. IGS-1 sequences for isolates from P. 

wallichiana had the greatest level of similarity (98%) with 

sequences from the same DNA region of A. mellea subsp. 

nipponica from Japan. Although ITS sequence data are not 

available for A. mellea subsp. nipponica, sequences from 

this DNA region were most similar to the closely related A. 

mellea s.s. from Japan. Based on these findings, we believe 

that the isolates from P. wallichiana included in this study, 

represent A. mellea subsp. nipponica. ITS sequences for 

isolates from A. densa were most similar (99%) to A. 

cepistipes on GenBank. Parsimony analysis placed these 

isolates in a sister group to A. cepistipes, but also indicated 

that they are very closely related to A. gallica and A. 

sinapina. We believe that the isolates from A. densa 

represent A. cepistipes, but sexual compatibility tests are 

required to confirm this view, especially because A. 

cepistipes is closely related to A. gallica and A. sinapina. 



647 - Molecular phylogeny of ascomycetes from the 

Helotiales 

J. Collado 1* , V. Rubio 2 , R. Galán 3 , G. Platas 1 , F. Arenal 2 , 

V. González 2 , J. Sánches-Ballesteros 2 , M. Villareal 2 , H.-O. 

Baral 4 & F. Peláez 1 

1 Centro de Investigación Básica, Merck Research 

Laboratories, Merck Sharp & Dohme de España S.A., 

Josefa Valcárcel 38, Madrid 28027, Spain. - 

2 Departamento de Biotecnología Microbiana, Centro 

Nacional de Biotecnología (CNB-CSIC), Campus 

Cantoblanco, Universidad Autónoma de Madrid, Madrid 

28049, Spain. - 3 Departamento de Biología Vegetal, 

Facultad de Biología, Universidad de Alcalá, Alcalá de 

Henares (Madrid) 28871, Spain. - 4 Blaihofstr. 42, D- 

72074 Tübingen, Germany. - E-mail: 

javier_colladomartinez@merck.com 

This work presents a preliminary assessment of the 

phylogenetic relationships among species in the Helotiales 

based on rDNA sequence analysis. Except for the 

Sclerotiniaceae, the Helotiales have not been subjected to 

extensive molecular phylogenetic studies. We have 

analyzed the sequences of the ITS1-5.8s-ITS2 region from 

more than 100 strains corresponding at least to 42 species 

in the Hyaloscyphaceae and 32 in the Helotiaceae, with a 

special emphasis on the genera Lachnum and 

Hymenoscyphus. In addition, a subset of the species has 

been subjected to sequence analysis of the D1-D2 region of 

the 28s rRNA gene. The result of the cladistic analysis 

confirmed the heterogeneity of the taxa defined within the 

two families. The topology did not support the current 

morphology-based classification schemes of some of the 

taxa examined. In particular, a significant degree of 

incongruity was evident for several species in the genus 

Hymenoscyphus and less notably for Lachnum spp. The 

results are discussed in comparison with the morphological 

characters used to define taxa within the two families 

studied. The suitability of the ITS region for phylogenetic 

analysis of this heterogeneous group of fungi is also 

discussed. 

648 - Lumping Tricholoma sulphureum and T. 

bufonium? 

O. Comandini 1* , I. Haug 2 , A.C. Rinaldi 3 & T.W. Kuyper 4 

1 

Dipartimento di Scienze Ambientali, Università 

dell'Aquila, via Vetoio, Loc. Coppito, I-67100 L'Aquila, 

2 

Italy. - Spezielle Botanik, Mykologie, Universität 

Tübingen, auf der Morgenstelle 1, D-72076 Tübingen, 

Germany. - 3 Cattedra di Chimica Biologica, Dipartimento 

di Scienze Mediche Internistiche, Università di Cagliari, I- 

09042 Monserrato, Cagliari, Italy. - 4 Department of 

Environmental Sciences, Section Soil Quality, Wageningen 

University, P.O. Box 8005, NL-6700 EC Wageningen, The 

Netherlands. - E-mail: comandin@univaq.it 

196 


The taxonomical status of Tricholoma bufonium, generally 

considered a rare species, has been a matter of discussion 

for a long time, some authors questioning the validity itself 

of the taxon and claiming that it is actually a mere variant 

of the closely related T. sulphureum. Others accept T. 

bufonium as a valid species on the basis of the constancy of 

the features distinguishing it from T. sulphureum, although, 

admittedly, intermediary forms exist. The two 'species' are 

clearly depicted in colour by some authors. We report the 

complete morpho-anatomical and molecular 

characterisation of the naturally occurring ECM type 

formed on silver fir by Tricholoma bufonium, as classified 

by sporocarp features. Molecular information has also been 

obtained for T. bufonium sporocarps collected on silver fir 

and oak and for fruitbodies classified as T. sulphureum 

collected on spruce and chestnut. Our data show that both 

ITS and LSU sequences were virtually identical in 

sporocarps referred to T. sulphureum and T. bufonium on 

the basis of colouration. However, strains from conifers 

and broad-leaved trees are to some extent different in 

sequence data. The taxonomic and ecological implications 

of our findings, together with already published 

information, notably the anatomical characteristics of T. 

sulphureum ECM on spruce, will be discussed. 

649 - Cryptic speciation, evolution and morphological 

plasticity in metalliferous Acarospora (lichenised 

Ascomycota) 

A.T. Crewe 1* , O.W. Purvis 2 & M. Wedin 1 

1 Dept. of Ecology and Environmental Science, Umeå 

University, SE-901 87 Umeå, Sweden. - 2 Department of 

Botany, The Natural History Museum, Cromwell Road, 

London SW7 5BD, U.K. - E-mail: anna.crewe@eg.umu.se 

The Acarospora smaragdula (Wahlenb.) A. Massal. and A. 

rugulosa Körb. species complexes are excellent models for 

studying the pattern of evolution and adaptation in a group 

where traditional taxonomic interpretation is complicated, 

and where molecular phylogeny can be utilised to 

investigate and test hypotheses of the evolution of 

morphological / anatomical traits. We are investigating the 

molecular phylogeny of populations of the Acarospora 

smaragdula and A. rugulosa complexes, to enable 

description of the pattern of evolution of metalliferous 

lichens in these groups, and to test the hypothesis that 

morphologically ± well-characterised taxa occurring on 

different metalliferous substrates are phylogenetic species. 

Here, we will present preliminary results from a pilot study 

focussing on A. smaragdula s. lat. and A. sinopica 

(Wahlenb.) Körb., utilizing well-known loci where fungalspecific 

PCR-primers are readily available (the nuclear ITS 

rDNA and mitochondrial SSU rDNA). Future directions, 

including extended phylogenies with additional markers, 

studies of the localisation and accumulation of metals in 

the lichens utilizing light microscopy, confocal scanning 

microscopy, SEM, mineralogical techniques and electron 

probe microanalysis, are briefly discussed.


650 - The mating relationships of the S and F groups of 

Heterobasidion annosum in Northern Hemisphere 

Y.C. Dai 1* , T. Niemelä 2 & K. Korhonen 3 

1 Institute of Applied Ecology, Academia Sinica, Shenyang 

110016, Wenhua Road 72, China. - 2 Botanical Museum, 

University of Helsinki, P.O. Box 47, FIN-00017, Finland. - 

3 Finnish Forest Research Institute, P.O. Box 18, FIN- 

01301 Vantaa, Finland. - E-mail: yuchengd@yahoo.com 

Pairings made among homokaryons of Heterobasidion 

parviporum Niemelä & Korhonen (S group of the H. 

annosum complex) from Europe, Northeast China and 

Northwest North America, and the European H. abietinum 

Niemelä & Korhonen (F group) showed that the Northeast 

Chinese S has the strongest mating capacity among all the 

testers, being closely related with the European S and F, 

and also the North American S. The North American S is 

more readily compatible with the European F than with the 

European S, and it seems that the so-called North 

American S group should be referred to the European F 

group rather than the European S. Samples collected from 

North Yunnan (eastern foothills of the Himalayas, China) 

were paired with testers of the S group from NE China, 

with P, S and F groups from Europe, and S group from NW 

North America; the Yunnan stocks mated with all the 

testers by frequency over 90%. This suggests that the 

Yunnanese stocks have the largest effective population 

size. 

651 - Systematics and evolution of Gomphales 

(Basidiomycetes) 

P.P. Daniëls * , M.P. Martín & M.T. Tellería 

Real Jardín Botánico, C.S.I.C., Plaza de Murillo, 2, 28014 

Madrid, Spain. - E-mail: daniels@ma-rjb.csic.es 

The order Gomphales enclose eight genera in the world and 

Ramaria is the main one in relation to species number. 

Previous systematic arrangement in subgenus Ramaria was 

proposed by Frachi & Marchetti (2001, Fungi non 

delineati. 16). This subgenus was divided into ten sections 

only based upon the presence/absence of clamped hyphae 

and the basidiome colour. As part of a monograph of the 

Iberian Gomphales a phylogenetic study in Gomphales 

focused on Ramaria has been undertaken. Thirty-four 

species of Ramaria as well as other genera of Gomphales 

were included in the phylogenetic analysis based on ITS 

and 5.8S rDNA sequences. Moreover, phylogenetic 

analysis of the morphological characters were done on 

clavarioid basidiomes. The very variable ITS regions gave 

low bootstrap values but it can be deduced some 

considerations about the systematics and evolution of the 

Gomphales: 1) Subgenus Echinoramaria is monophyletic. 

2) Ramaria is a genus related with several taxa among the 

Gomphales, Gautieria morchellaeformis is related with 

Section Ramariae and Gomphus clavatus with Section 

Fennicae. 3) Ramaria bataillei and R. pumila seems to be a 

bridge between the ectomycorrhizal and the saprobic 

Ramarias. 4) Lentaria and Hydnocristella are closely 

related with subgenus Lentoramaria. 5) The gelatinose 

texture of the fruitbodies seems to be a synapomorphy. 6) 

Clamp connections and fruitbody colour are very 

homoplasic characters and do not serve to divide sections 

among Ramaria. 

652 - Another canker-causing aerial Phytophthora from 

forest trees in California and Oregon 

J.M. Davidson 1 , M.M. Garbelotto 2 , E.M. Hansen 3* , P. 

Reeser 3 & D.M. Rizzo 1 

1 Plant Pathology, University of California, Davis CA 

95616, U.S.A. - 2 Environmental Science Policy and 

Management, U. C. Berkeley, Berkeley CA 94720, U.S.A. - 

3 Botany and Plant Pathology, Oregon State University, 

Corvallis OR 97331, U.S.A. - E-mail: 

hansene@bcc.orst.edu 

Two distinct Phytophthora species cause similar symptoms 

on several western forest trees. P. ramorum causes lethal 

cankers (sudden oak death) on tanoak and coast live oak 

trees and foliar and dieback symptoms on other tree and 

shrub species in some western forests. A second 

Phytophthora species, previously undescribed, is 

occasionally isolated from lethal cankers on tanoak and 

coast live oak, and from foliar lesions on Umbellularia and 

other hosts, in areas where P. ramorum is also active. ITS 

DNA sequence indicates close relationship to P. ilicis (a 

foliar pathogen of holly) and P. psychrophila (newly 

described from European oak forest soils). It is homothallic 

with amphigynous antheridia, and has deciduous sporangia. 

It grows more slowly, with a lower temperature optimum, 

than P. ramorum. In log inoculation tests it is nearly as 

pathogenic to tanoak as P. ramorum. It does not infect 

holly leaves in leaf inoculation tests. In the forest it is 

usually associated with single killed trees, in contrast to the 

expanding patches of mortality caused by P. ramorum. 

653 - Endophytes from leafy liverworts: a molecular 

phylogenetic perspective 

C. Davis 

Duke University, Biological Sciences, Box 90338, Durham, 

NC, U.S.A. - E-mail: ecf5@duke.edu 

It has long been known that leafy liverworts possess 

endophytic fungi, but these fungi have not previously been 

identified with precision. Here, I present results of 

molecular phylogeny as the means to identify these 

endosymbionts from selected leafy liverworts. I address 

possible explanations for the nature of such symbioses. I 

discuss hypotheses regarding liverwort structural 

adaptations for endosymbiosis, and explore methodology to 

test these hypotheses. 



654 - A molecular taxonomic revision of the genus 

Pythium 

A.W.A.M. de Cock 1* , C.A. Lévesque 2 , J.E.J. Bedard 3 , 

A.M. Schurko 3 & G.R. Klassen 3 

1 Centraalbureau voor Schimmelcultures, P.O.Box 85167, 

NL-3508 AD Utrecht, The Netherlands. - 2 Agriculture & 

Agri-Food Canada, ECORC, Summerland, B.C., V0H 1Z0, 

Canada. - 3 University of Manitoba, Dept. Microbiology, 

Winnipeg, Manitiba, R3T 2N2, Canada. - E-mail: 

decock@cbs.knaw.nl 

The genus Pythium mainly consists of plant pathogens. 

Identification is based on morphology and is notoriously 

difficult due to variability and overlapping of characacters. 

Therefore a molecular approach was chosen to evaluate the 

existing taxonomy, and to find more reliable methods for 

identification and detection. All available species (approx. 

100), represented by more than 500 isolates were included. 

One representative isolate of each species was used for 

sequencing of ITS and D1-D3 regions of the lsrDNA: all 

ex-type, neo-type, authentic or otherwise well defined 

representative strains. Sequences were used for both 

reconstruction of phylogeny and development of speciesspecific 

oligonucleotides. The latter were used in DNAarrays 

for simultaneous detection of multiple pathogens in 

soil samples. All strains were used in studies of RFLPs of 

PCR amplified ribosomal IGS and ITS and RAPD patterns, 

to reveal intraspecific variation and to establish species 

boundaries. RAPD patterns were most variable; IGS 

patterns proved to be useful for identification. Studies were 

performed simultaneously. Phylogenetic trees showed 

which species were to be compared side by side on RFLP 

and RAPD gels; the latter patterns showed which 

additional strains should be sequenced in case of 

intraspecific variation. Results will be used to redescribe 

species based on molecular groups, to provide a database 

of molecular characters for identification and to develop 

specific probes for detection. 

655 - Elucidating the 'enigma' of Aenigmatomyces 

ampullisporus which is now found as a Zygomycetous 

fungus parasitizes a spermatophore of collembolan 

insect 

Y. Degawa 

Kanagawa Prefectural Museum of Natural History, Iryuda 

499, Odawara, Kanagawa 250-0031, Japan. - E-mail: 

degawa@pat-net.ne.jp 

Aenigmatomyces ampullisporus Castañeda and Kendrick 

was described from Canada in 1993. Although the authors 

assigned it to the kingdom Fungi, they could not decide to 

which phylum it belonged. They gave it a name 'enigmatic 

fungus' and left its appropriate taxonomic position as a 

mystery to be resolved in future. From 1999 to 2002, I 

collected the fungus repeatedly from Japan, which was 

identified as A. ampullisporus in reference with the 

198 


holotype specimen. As a result of observations, the 

'enigma' was resolved as follows; 1) The 'host hyphae' as 

explained in the original description can no longer be a 

fungal structure, but are stalks of spermatophores of 

Collembola. 2) Spores infecting the head of 

spermatophores swelled to become hyphal bodies. While 

one of the hyphal arms elongates downward functioning as 

an anchor, the other arm elongates upwards bearing 

abundant sporogenous structures. In maturation, 

spermatozoon in a spermatophore gradually degrades and 

finally disappears entirely. 3) Spherical liquid-drops were 

stained in blue at the apices of spores when mounted in 

Lacto-aniline blue. 4) 'Oospores of the host' are directly 

connected with the hyphal bodies. Their pigmentation and 

ornamentation suggests that they are zygospores. 

Consequently, A. ampullisporus is not an Oomycetesparasite 

but a zygomycetous fungus parasitizing the 

spermatophore of Collembola, and is probably a relative of 

holocarpoic amoeba-parasites of Cochlonemataceae, 

Zoopagales. 

656 - Inter- and infraspecific length variability in 

minisatellites in the ITS region of Leccinum (Boletales, 

Boletaceae) 

H.C den Bakker 1* , B. Gravendeel 1 , T.W. Kuyper 2 & M.E. 

Noordeloos 1 

1 Nationaal Herbarium Nederland, Leiden university 

Branch, P.O.box 9514, 2300 RA, Leiden, The Netherlands. 

- 2 Wageningen Agricultural University, Department of 

Environmental Sciences, P.O. box 8005, 6700 EC, 

Wageningen, The Netherlands. - E-mail: 

bakker@nhn.leidenuniv.nl 

Phylogenetic relationships of the European species of 

Leccinum (Boletaceae) were investigated by maximum 

parsimony, likelihood analyses and Bayesian analyses of 

nrITS1-5.8S-ITS2 and 28S sequences. The separate gene 

trees inferred were largely concordant, and their combined 

analysis indicates that several traditional sectional and 

species-level taxonomic schemes are artificial. In 

Leccinum, the nrITS region ranges in size from 694 to 1480 

bp. This extreme length heterogeneity is localized to a part 

of the ITS1 spacer that contains a minisatellite with 

tandemly repeated copies including the six-basepair 

GAAAAG motif. The number of tandem repeats varied 

from 2 to 6, but there were many repeat variants. 

Secondary structure analysis revealed that the ITS1 spacer 

in Leccinum appears to be still functional despite the 

presence of the minisatellite. Intraindividual sequence 

variation of the minisatellite was always smaller than 

between different species, indicating that concerted 

evolution proceeds rapidly enough to retain phylogenetic 

signal at the infraspecific level. In contrast, the 

evolutionary pattern exhibited by the major ITS1 repeat 

types found was homoplastic when mapped onto the 

species lineages inferred from the combined 5.8S-ITS2 

sequences. The minisatellite therefore appears not to be 

useful for phylogeny reconstruction at the species level.


657 - Notes on the tribes Leucocoprineae and Lepioteae 

(higher basidiomycetes) of Israel mycobiota 

M.Ya. Didukh 

M. G. Kholodny Institute of Botany, National Academy of 

Sciences of Ukraine, 2 Tereshchenkivska Str., Kyiv, MSP, 

252601 Ukraine, Ukraine. - E-mail: manuuzz@yahoo.com 

Agaricaceae (Fr.) Cohn has been of particular interest to 

scientists all over the globe for years due to peculiarities of 

its representatives ecology, biochemistry, biotechnological 

application. However, for some areas - Israel in particular - 

even the first step of study of this family - revision of 

species diversity - has not been done yet. Out of the four 

tribes of the family only Agariceae Pat. has been revised 

(Wasser & Binyamini, 1992; Wasser, 1996, 1998, 2002). 

The Leucocoprineae Sing. and Lepioteae Fay. in Israel 

tribes are in need of critical study and inventory of the 

species content. Analysis of existing fragmentary data in a 

number of articles (Reichart & Avizohar-Hershenzon 1955, 

1959; Avizohar-Hershenzon, 1967; Binyamini, 1975; 1976; 

1984) showed that up to now 15 species (9 of genus 

Lepiota, 1 of Leucocoprinus, 1 of Chlorophyllum, 1 of 

Leucoagaricus and 3 of Macrolepiota) are registered. We 

report of seven new for the Israel territory species and 

intraspecies taxa found during 2001-2002 growing seasons: 

Chamaemyces fracidus (Fr.) Donk.; Chamaemyces fracidus 

var. pseudocastaneus Bon et Boiff.; Lepiota 

brunneolilacea Bon et Boiff.; Leucoagaricus littoralis 

(Meiner) Bon et Boiff.; Leucoagaricus wichanskyi (Pil.) 

Bon et Boiff.; Leucoagaricus carneifolius (Gill.) S. 

Wasser; Macrolepiota fuligineosquarrosa Malenc. 

658 - Out of Gondwana? - Relationship between 

European and African Lactarius sp. of the subgenera 

Lactifluus and Lactariospsis 

U. Eberhardt 1* & A. Verbeken 2 

1 SLU, Dept. of Forest Mycology and Pathology, Box 7026, 

SE-75007 Uppsala, Sweden. - 2 Gent University, Dept. 

Biology, Group Mycology, K.L.Ledeganckstraat 35, B- 

9000 Gent, Belgium. - E-mail: 

ursula.eberhardt@mykopat.slu.se 

The forests of tropical Africa are remarkably rich in 

Russulales species, particularly in species displaying what 

are considered to be conservative characters. Therefore, the 

hypothesis was put forward that the Russulales evolved on 

the Gondwana continent and spread from there to the 

northern hemisphere. The subgenera Lactifluus and 

Lactariopsis of the genus Lactarius belong to these 

supposedly primitive groups with a strong tropical African 

foothold, in terms of species numbers, infra-subgeneric 

diversity, and distribution. Both subgenera are also 

relatively well-represented in tropical lowlands of Central- 

and South-America. In contrast, only few and 

morphologically outstanding European Lactarius species 

were assigned to these subgenera. In molecular 

phylogenetic analysis, these species were indeed placed on 

very long branches, separated from the majority of 

European species that are comparatively closely related. 

Molecular phylogenetic analysis methods were applied to 

investigate the relationship between selected European and 

tropical African species of the subgenera Lactifluus and 

Lactariopsis and to test the provisional infra-subgeneric 

classification of African Lactarius species that was entirely 

based on morphological characters. 

659 - The phylogenetic position of the genera Lepraria 

and Leproloma 

S. Ekman 1* & T. Tønsberg 2 

1 Department of Botany, University of Bergen, Allégaten 41, 

N-5007 Bergen, Norway. - 2 Museum of Botany, University 

of Bergen, Allégaten 41, N-5007 Bergen, Norway. - Email: 

stefan.ekman@bot.uib.no 

The phylogenetic position of the entirely asexually 

reproducing genera Lepraria and Leproloma was 

investigated using sequence data from the ITS1-5.8S-ITS2 

and small subunit (SSU) nuclear ribosomal DNA. 

Phylogenetic reconstructions were carried out using a 

Bayesian Markov chain Monte Carlo (MCMC) tree 

sampling technique and an alignment-free maximum 

likelihood distance method. The results indicate that most 

species currently referred to the genera Lepraria and 

Leproloma form a single, monophyletic group. This 

monophyletic group is sister to a well-known group of 

lichens in the Stereocaulaceae (Lecanorales, 

Lecanoromycetes, Ascomycota), namely Stereocaulon and 

Muhria. Leproloma is polyphyletic and nested within 

Lepraria s. str. A few species, however, do not belong in 

Lepraria s. str., viz. Lepraria flavescens, which belongs in 

Lecanora, and Lepraria lesdainii and L. obtusatica, the 

positions of which are currently unknown. 

660 - Toninia species in Hungary 

E. Farkas 1* & L. Lökös 2 

1 Institute of Ecology and Botany, Hungarian Academy of 

Sciences, H-2163 Vácrátót, Hungary. - 2 Department of 

Botany, Hungarian Natural History Museum, H-1476 

Budapest, Pf. 222., Hungary. - E-mail: 

efarkas@botanika.hu 

Species of Toninia are dominant in the sandy steppe and 

rocky grassland areas in Hungary. In the Hungarian lichen 

flora of Verseghy (1994) nine species were listed under 

Toninia: T. candida, T. caradocensis, T. cinereovirens, T. 

coeruleonigricans, T. lobulata, T. toniniana, T. tristis, T. 

tumidula, T. zsakii. A revision of the c. 300 Hungarian 

Toninia specimens was highly needed, since the Toninia 

monograph of Timdal (1991) was not considered by 

Verseghy. Three of the above species are now belong to 

other genera: Hypocenomyce caradocensis (syn. T. 



caradocensis), Mycobilimbia lobulata (syn. T. lobulata), 

Micarea melaenida (syn. T. zsakii). Toninia sedifolia with 

c. 200 specimens is the most frequent species of Toninia in 

Hungary. One specimen of T. alutacea (under T. 

intermedia from 1933) and two specimens of T. aromatica 

(from 1917 and 1955) kept in BP were regarded as 

uncertain data by Verseghy. No recent collections could 

confirm their occurrence in Hungary. T. physaroides and T. 

verrucarioides were indicated with Hungarian distribution 

by Timdal (1991). T. physaroides must have been 

overlooked because of its similarity with T. sedifolia. 

Parasitic and lichenicolous lichens are generally 

undercollected in Hungary. Most of the recent specimens 

also belong to T. sedifolia. Nevertheless T. diffracta was 

identified from a recent collection as new to Hungary. 

[Supported by the Hungarian Research Fund (OTKA 

T030209) and 'Research Grant Bolyai János'.] 

661 - To morphological and anatomical variability of 

lichen-forming fungus Xanthoria patietina s. lat. 

(Teloschistaceae, Ascomycotina) 

N.M. Fedorenko * & S.Ya. Kondratyuk 


2, 01601 Kiev, Ukraine. - E-mail: skondr@botan.kiev.ua 

Xanthoria parietina (L.) Th. Fr. is characterized by large 

thallus with plane and wide lobes. More than 40 

infraspecific taxa of various status are hitherto described 

for this very polymorph species. However there are only a 

few morphological characters, which have being hitherto 

used for taxonomy of this group, namely presence of 

soredia or isidia, morphology of thallus or single lobes and 

ecological characters of habitats. The aim of this study is to 

select a number of correlated characters, which may be 

applied for taxonomy of the whole X. parietina group. 

Three morphological types of Xanthoria parietina were 

selected on the basis of results obtained. The first one, 

thallus is very small (up to 2 cm diam.) with very dissected 

narrow lobes which never overlapping by marginal zone 

(lobes of the second level are well developed). The second 

one, with large entire thallus (4-16 cm diam.), only slightly 

dissected at marginal zone, lobe margins not overlapping 

(lobes of the second level are absent). The third one, thallus 

is very large (10 cm and more in diam.), margin almost 

entire, almost undissected, lobe margins (if developed) 

overlapping. Involving additional material from other 

regions of Eurasia and the further comparative study of 

groups selected with type material of X. parietina and X. 

microspora B. de Lesd. will show taxonomical status of 

these groups mentioned. 

200 


662 - Identification and intra-specific characterization 

of penicillia and aspergilli by Fourier-Transform 

Infrared Spectroscopy (FT-IR) 

G. Fischer * , S. Braun & W. Dott 

Institute of Hygiene and Environmental Health, University 

Hospital Aachen, Pauwelsstr. 30, 52224 Aachen, Germany. 

- E-mail: Guido.Fischer@post.rwth-aachen.de 

Airborne microfungi are a matter of discussion in public 

and occupational hygiene, they can be allergenic, toxigenic 

or some may cause infections. To determine health 

hazards, reliable species identification and physiological 

characterization of isolates is needed. Conidial suspensions 

of airborne fungi from the genera Aspergillus and 

Penicillium were characterized using Fourier-Transform 

Infrared Spectroscopy (FT-IR). A new method was 

developed for the preparation of conidia. A database was 

established containing reference strains from culture 

collections as well as fresh isolates. The results obtained by 

cluster analysis were compared to data from 

chemotaxonomic investigations, where the production of 

secondary metabolites and the presence of mycotoxins in 

conidia has been investigated. The taxonomic relationship 

was reflected in the dendrograms resulting from the cluster 

analysis. Identification of newly isolated strains was 

possible by comparison to the database established. In 

some species, intra-specific variation concerning 

mycotoxin production occurred, which was equally 

reflected in the discrimination of strains. The new method 

seems to be particularly suited to be applied in different 

areas of applied and systematic mycology: microbial 

inventories, rapid identification of pathogens, 

biotechnology, quality control in culture collections. 

Moreover, the method proved to be a powerful tool in 

taxonomic studies. 

663 - Phylogeny of Termitomyces and related taxa based 

on nuclaer (nLSU) and mitochondrial (mtSSU) 

ribosomal DNA 

T. G. Frøslev 1* , D.K. Aanen 2 , T. Læssøe 1 & S. Rosendahl 1 

1 Copenhagen University, Botanical Institute, Mycological 

Department, Østre Farimagsgade 2D, 1353 Kbh K., 

Denmark. - 2 Copenhagen University, Zoological Institute, 

Department of Population Ecology, Universitetsparken 15, 

2100 Copenhagen, Denmark. - E-mail: 

TobiasGF@bot.ku.ku 

A phylogeny of the symbiotic fungi of the fungus-growing 

higher termites (subfamily Macrotermitinae) and related 

taxa is estimated with Bayesian methods. Our sample 

consists of 130 sequences of nLSU rDNA obtained from 

basidiocarps, termite nests and termite gut material and 

represents extensive geographic and taxonomic sampling. 

Results of the Bayesian analysis are in agreement with 

those of traditional maximum parsimony and maximum 

likelihood methods. Inclusion of mtSSU rDNA sequence


data from several samples agrees with the overall 

topography of the phylogeny. The results confirm that 

Termitomyces constitutes a monophyletic group within 

Lyophylleae. Podabrella (T. microcarpus) is derived 

within Termitomyces. The proposed related genus 

Sinotermitomyces belongs to Termitomyces but does not 

constitute a monophyletic group. Gerhardtia 

brunneoincarnata, proposed to have affinities to 

Termitomyces due to absence of clamps, belongs to 

Lyophylleae, but does not belong to Termitomyces. There 

seems to be a geographical differentiation of clades as well 

as a differentiation in relation to host genera. 

664 - Characterisation of yeast communities in aquatic 

environmental samples by Temperature Gradient Gel 

Electrophoresis (TGGE) 

M. Gadanho 1* , R. Tenreiro 2 & J.P. Sampaio 1 

1 Centro de Recursos Microbiológicos, Secção Autónoma 

de Biotecnologia, Faculdade de Ciências e Tecnologia, 

Universidade Nova de Lisboa, 2829-516, Portugal. - 

2 Centro de Genética e Biologia Molecular, Departamento 

de Biologia Vegetal, Faculdade de Ciências, Universidade 

de Lisboa, Portugal. - E-mail: majg@mail.fct.unl.pt 

Several difficulties affect the characterisation of microbial 

communities in environmental samples. Traditionally, 

culture-dependent methods were used for studies of 

microbial diversity. However, this approach is too 

laborious to be used intensively. To overcome the 

disadvantages of the isolation step and the selective effect 

induced by culture media, several culture-independent 

approaches have been proposed. One of the most popular is 

PCR-DGGE/TGGE. This method is based on the PCR 

amplification of DNA fragments from the pool of total 

DNA extracted from a natural sample. The separation of 

the amplicons is done in a denaturing electrophoresis. In 

this study we optimised this approach for the detection and 

characterisation of yeasts in aquatic samples. Water 

volumes of 5 L were filtered and the DNA of the organisms 

present in the filter was extracted. Then, a set of primers 

specific for fungi and designed for the 26S rDNA, was 

used. Finally, the amplicons were separated using a TGGE 

apparatus. It was observed that yeasts were present in 

relatively low numbers, which difficulted their detection. 

Simultaneously, liquid medium enrichments were 

performed during periods of 1-12 days. After incubation, 

DNA was extracted and analysed by PCR-TGGE. The 

electrophoretic profiles obtained from enriched and nonenriched 

samples from different aquatic environments were 

compared and the composition of the yeast community was 

analysed by direct DNA sequencing of the TGGE bands. 

665 - Nexomyces cubensis gen. et sp. nov. from Cuban 

soil 

D. García * , A.M. Stchigel, J. Cano & J. Guarro 

Rovira i Virgili University, c/ Sant Llorenç 21 43201, Reus, 

Spain. - E-mail: umb@fmcs.urv.es 

During the course of a study of soil ascomycetes from 

Cuba, an undescribed ascomycete was isolated from 

Ciénaga de Zapata (Matanzas province). Due to endemic, 

rare and endangered organisms, which can be found in this 

region, it has been declared a Biosphere Reserve by 

UNESCO. A bait technique was used for recovering the 

fungi. Petri dishes were half-filled with the soil samples 

and moistened with sterile distilled water. Several pieces of 

sterile wood (approx. 1x2 cm) were placed on the soil 

surface or partially buried in it. The new ascomycete 

Nexomyces cubensis gen. et sp. nov. is characterized by 

ostiolate and non-ostiolate setose ascomata and ellipsoidal, 

thick- and smooth- walled, brown to dark brown, opaque 

ascospores, which are umbonated at the apex with a 

protuberant apical germ pore. Due to its morphological 

features, this taxon was easily recognized as belonging to 

the Sordariales, although its exact taxonomic placement 

was difficult to ascertain. In order to establish its 

phylogenetic relationships, the ITS rDNA region sequence 

was compared with those of other genera of the 

Sordariales. Analyses of the sequences suggest that this 

taxon is close to Sordariaceae. 

666 - Phylogenetic boundaries in Ramariopsis 

R. García-Sandoval 1* , J. Cifuentes 1 , E. de Luna 2 , A. 

Estrada-Torres 3 & M. Villegas 1 

1 FCME Herbaria, UNAM, Po.Box 70-399, CP 04510, 

Coyoacán DF, Mexico. - 2 Instituto de Ecología AC., 

Po.Box 63, CP 9100, Xalapa, Veracruz, Mexico. - 3 Centro 

de Investigación en Ciencias Biológicas, UATX, 

Km10.5SnMartín Texmelucan-Tlaxcala, 90120, Tlaxcala, 

Mexico. - E-mail: gsr@minervaux.fciencias.unam.mx 

The genus Ramariopsis was first described by Corner 

(1950) who separated from the genus Clavaria those 

species having branched fruit bodies, whitish colors and 

spiny hyaline spores. Petersen (1966, 1969 & 1978) 

proposed an emendation in order to include species with 

smooth spores and simple fruit bodies. The present work 

comprehends a phylogenetic analyses based on 

morphology, whitch pretends to clarify the phylogenetical 

boundaries of Ramariopsis. Thirty-six morphological 

character were considered for four species of Ramariopsis 

sensu Corner (1970) and six species of Ramariopsis sensu 

Petersen, eleven close outgroups from genus Clavaria, 

Multiclavula, Clavariadelphus, and Scytinopogon, included 

to test phylogenetical boundaries; three additional far 

outgroups were included for cladogram orientation 

purposes, a total of twenty-four species were included. 

Outgroups were selected by performing a phylogenetical 



prospection of the Clavariaceae family. Ramariopsis sensu 

Corner was found as a monophyletic group, with 

Scytinopogon as its sister group, which agree with Corner 

(1970) proposals. Ramariopsis sensu Petersen was rejected 

as a monophyletic natural genus. 

667 - Molecular study of the Caloplaca saxicola group 

on the background of morphological taxa 

E. Gaya 1* , P. Navarro-Rosinés 1 , F. Lutzoni 2 & N. Hladun 1 

1 Dept.Biologia Vegetal.Facultat de Biologia.Universitat de 

Barcelona, Av.Diagonal 645. 08028 Barcelona, Spain. - 

2 Dept. of Biology. Duke University, Box 90338. Durham, 

North Carolina, U.S.A. - E-mail: gaya@porthos.bio.ub.es 

Caloplaca (Teloschistaceae, Ascomycota) with more than 

1000 species can be considered one of the most complex 

and diversified genus among crustose lichens. Its 

delimitation has been always problematic due mostly to the 

strong similarity between lobed species of Caloplaca and 

species of other genera within this family, such as 

Xanthoria. Among all lobed Caloplaca species, the 

Saxicola group has been the most controversial 

taxonomically. Here we report the results from 

phylogenetic analyses of the nuclear rDNA internal 

trancribed spacers (ITS) sequence data for 60 specimens 

belonging to the Saxicola group. Collections from Europe, 

North America and Asia are included. We have used these 

molecular data to compare the current phenotypic 

delimitation of certain species. Hence, a morphological 

comprehensive study of the group is provided with an 

emphasis on the enigmatic C. arnoldii s. auct., the 

phenotypically variable C. saxicola s. str. and the concept 

of C. biatorina in the different continents. A proposal for 

achieving a new and stable classification of this group will 

be presented. 

668 - Phylogeny of the genus Arachnomyces and its 

anamorphs 

C. F. Gibas 1* , L. Sigler 1 , R.C. Summerbell 2 & R.S. Currah 3 

1 University of Alberta Microfungus Collection and 

Herbarium, Edmonton, Alberta, T6G 2E1, Canada. - 

2 Centraalbureau voor Schimmelcultures, Baarn, The 

Netherlands. - 

3 Department of Biological Sciences, 

University of Alberta, Edmonton, Alberta, T6G 2E9, 

Canada. - E-mail: cgibas@ualberta.ca 

Arachnomyces is a genus of cleistothecial ascomycetes that 

has morphological similarities to the Onygenaceae and the 

Gymnoascaceae but is not accommodated well in either 

taxon. The phylogeny of the genus Arachnomyces and 

related anamorphs was studied using nuclear SSU rDNA 

gene sequences. Partial sequences were determined from 

ex-type cultures representing A. minimus, A. nodosetosus 

(anamorph Onychocola canadensis), A. kanei (anamorph 

O. kanei) and A. gracilis (anamorph Malbranchea sp.) and 

202 


aligned together with corresponding sequences of forty 

other ascomycetes. Phylogenetic analysis using maximum 

parsimony showed the genus Arachnomyces is 

monophyletic, includes the hyphomycete Malbranchea 

sclerotica, and forms a separate lineage within the 

Eurotiomycetes. All known anamorphs in this lineage are 

arthroconidial and have been placed either in Onychocola 

(A. nodosetosus, A. kanei) or in Malbranchea (A. gracilis). 

Onychocola is chosen as the most appropriate taxon for the 

arthroconidial states of Arachnomyces and thus 

Malbranchea sclerotica and the anamorph of A. gracilis 

are treated as species within Onychocola. 

669 - Analysis of 18S ribosomal RNA gene sequences 

reveals the phylogenetic relationships of the genus 

Mycosphaerella 

S.B. Goodwin * & J.R. Cavaletto 

USDA-ARS / Purdue University, Department of Botany and 

Plant Pathology, 1155 Lilly Hall, West Lafayette, IN 

47907-1155, U.S.A. - E-mail: sgoodwin@purdue.edu 

Mycosphaerella is one of the largest groups of plant 

pathogenic fungi with more than 1800 species names. 

Anamorphs associated with Mycosphaerella number more 

than 40, many of which also are economically important 

with large numbers of species. For example, Cercospora 

and Septoria contain more than 3000 and 1000 species 

names, respectively, most of which probably were derived 

from species with a Mycosphaerella teleomorph. Despite 

their common occurrence and high economic importance, 

members of this genus have been under-represented in 

phylogenetic analyses and the taxonomic position of 

Mycosphaerella is uncertain. Most taxonomists place the 

genus in the order Dothideales. However, this conclusion 

has not been supported by limited analyses of 18S 

ribosomal RNA gene sequences. To test whether the genus 

Mycosphaerella belongs in the order Dothideales, 18S 

rDNA sequences were obtained from five species 

representing the major groups within the genus identified 

in previous analyses of ITS sequences. All five species of 

Mycosphaerella clustered together but separate from 

species of Dothidea and formed a basal group to the other 

loculoascomycetes in a neighbor-joining analysis of 109 

18S sequences. Species of Dothidea clustered with those of 

Botryosphaeria, another genus of uncertain taxonomic 

position. Inclusion of Mycosphaerella within the 

Dothideales was not supported by this analysis and a new 

order may be needed to accommodate species of 

Mycosphaerella and related genera. 

670 - Biogeography and species concepts - Lentinus 

lindquistii is Lentinus tigrinus, a circum-global species 

E.A. Grand * , R.H. Petersen & K.W. Hughes 

The University of Tennessee, 437 Hesler, Knoxville, 

Tennessee 37996-1100, U.S.A.


Lentinus lindquistii (Sing.) Lechner & Alberto (=Pleurotus 

lindquistii Singer) is known only from its type locality in 

Argentina. Lentinus lindquistii has been separated from L. 

tigrinus s. s. based on non-inflated generative hyphae and 

other slight morphological differences (i.e. spore size). We 

employed tester strains of L. lindquistii and singlebasidiospore 

isolates (SBIs) of L. tigrinus (Bull.: Fr.) from 

10 widely scattered geographic locations to ascertain 

congruence of morphological, biological, and genetical 

species concepts. SBIs of L. tigrinus were obtained from 

fresh material collected in the field and basidiomata 

produced in vitro. SBIs from each collection were paired 

with those of all other collections (n = 4 or 8) to ascertain 

sexual intercompatibility. Results showed complete 

compatibility among collections, indicating that all 

collections represented the same biological species. Our 

data demonstrate the morphological variability of two 'taxa' 

in the L. tigrinus complex. Sequence analysis of the ITS1- 

5.8S-ITS2 nrDNA suggests that widely scattered 

geographical populations of the L. tigrinus complex are 

genetically divergent. Although these populations have 

been separated long enough to allow sequence evolution, 

they are still capable of compatible mating. 

671 - Ascocarp impalement: a novel mechanism of 

dispersal in cleistothecial ascomycetes 

M.D. Greif * & R.S. Currah 

Department of Biological Sciences, University of Alberta, 

CW-405, Edmonton T6G 2E9, Canada. - E-mail: 

mattgreif@hotmail.com 

Ascomycetes have evolved a variety of mechanisms to 

ensure the dispersal of their meiospores by animals. Some 

of these include the production of sticky spores (e.g. 

Ophiostomatales) or even the development of the ascocarp 

directly on the body of an arthropod (e.g. Laboulbeniales), 

both ensuring that propagules are transferred economically 

from one suitable habitat to another. Here, we report a 

novel mechanism that has developed independently in two 

ascomycete lineages, the Leotiomycetes and the 

Eurotiomycetes. In representatives of these classes, namely 

Myxotrichum and Auxarthron respectively, the ascocarp is 

a cage-like cleistothecium with an internal mass of minute, 

pale coloured ascospores borne within a lattice of rigid 

thick walled hyphae. By exposing mature ascocarps of 

these two fungi to active Dipterans (Sarcophagidae and 

Calliphoridae) for 24 hours we demonstrated that this type 

of ascocarp is picked up through an impalement event in 

which the arthropod hairs pass between the spaces of the 

cleistothecial lattice, glancing or piercing the ascospore 

mass and affixing the entire ascocarp to the body of the 

vector. We suggest that this impalement mechanism is the 

driving force behind the convergence in ascocarp form in 

these cleistothecial fungi. 

672 - Cryphonectria cubensis resides in a genus outside 

Cryphonectria 

M.V. Gryzenhout 1* , H. Myburg 2 , M.J. Wingfield 1 & B.D. 

Wingfield 2 

1 Department of Microbiology and Plant Pathology, 

Forestry & Agricultural Biotechnology Institute (FABI), 

University of Pretoria, Pretoria, 0002, South Africa. - 

2 Department of Genetics, Forestry & Agricultural 


Pretoria, 0002, South Africa. - E-mail: 

Marieka.Gryzenhout@fabi.up.ac.za 

Cryphonectria cubensis is a serious pathogen of Eucalyptus 

spp., clove (Syzigium aromaticum) and Tibouchina spp. in 

the tropics and sub-tropics. In previous studies, sequence 

data from the ITS1/ITS2 regions of the ribosomal DNA 

operon, and LSU and SSU DNA, suggested that C. 

cubensis does not group together with other Cryphonectria 

spp. The aim of the present study was to pursue this 

question further and to determine an appropriate generic 

placement for C. cubensis. An expanded collection of 

Cryphonectria and Endothia spp. was included and we 

produced additional sequences for the ITS1/ITS2 regions 

and the beta--tubulin genes. Herbarium specimens, 

including type collections, of C. cubensis and other species 

of Cryphonectria were also examined in detail. DNA 

sequences revealed that a world-wide collection of C. 

cubensis isolates, grouped separately from Cryphonectria 

and Endothia species in a strongly resolved clade. The 

anamorph of C. cubensis is blackened, pycnidium-like and 

superficial, and the teleomorph has black perithecial necks 

extending beyond the stromatal surface. This is in contrast 

to Cryphonectria that has orange stromata with semiimmersed 

anamorphs and orange perithecial necks. 

Morphological characteristics support DNA-based 

analyses, showing that C. cubensis should reside in a genus 

separate from Cryphonectria. In contrast to previous views, 

anamorph morphology and colour appear to be more 

taxonomically informative than teleomorph structure and 

ascospores. 

673 - Reconsideration of the conspecificity of Endothia 

eugeniae and Cryphonectria cubensis 

M.V. Gryzenhout 1* , H. Myburg 2 , M.J. Wingfield 1 & B.D. 

Wingfield 2 


Forestry & Agricultural Biotechnology Institute (FABI), 


2 Department of Genetics, Forestry & Agricultural 



Marieka.Gryzenhout@fabi.up.ac.za 

Endothia eugeniae, a canker pathogen of clove (Syzigium 

aromaticum), was synonymised with the Eucalyptus canker 

pathogen Cryphonectria cubensis. This synonymy emerged 



from morphological studies, cross-inoculations and 

isozyme analyses. However, for stromatal morphology, the 

type description of E. eugeniae does not concur with that of 

C. cubensis. New collections of Cryphonectria have 

become available to us and the aim of this study was to 

consider the validity of the synonymy of these two fungi. 

Isolates of C. cubensis and E. eugeniae were compared 

based on DNA sequences from the ITS1/ITS2 region and 

beta-tubulin genes. Specimens of clove in Zanzibar and 

South East Asia, were also compared morphologically with 

specimens of C. cubensis from Eucalyptus. Phylogenetic 

analyses showed that the majority of isolates from clove 

grouped together with C. cubensis isolates. A smaller 

number of isolates from Indonesia, however, grouped 

separately from the Cryphonectria and C. cubensis clades. 

Two fungi with different tissue types were observed on 

clove bark. One of these resembled C. cubensis. The other 

fungus is a species morphologically similar to 

Cryphonectria. The C. cubensis anamorph was 

morphologically variable on clove, which might explain 

anomalies regarding the description of E. eugeniae. Our 

results suggest that the synonymy of C. cubensis and E. 

eugeniae is valid. However, there is another fungus, closely 

related to Cryphonectria, also present on clove. 

674 - The genus Urocystis (Ustilaginomycetes) in China 

L. Guo 

Institute of Microbiology, Academia Sinica, Beijing 

100080, China. - E-mail: guol@sun.im.ac.cn 

For the genus Urocystis in China, Ling(1953) described 4 

species and Wang (1963) published 11 species. In 1979 Tai 

Fanlan in his 'Sylloge Fungorum Sinicorum' recorded 13 

species. So far a total of 36 species of the genus Urocystis 

are reported from China, distributed among 33 genera of 9 

families of the host plants. These smut species are arranged 

alphabetically as follows: Urocystis achnatheri, U. 

agropyri, U. agropyri-campestris, U. agrostidis, U. 

antipolitana, U. beijingensis, U. bolivari, U. brassicae, U. 

bromi, U. calamagrostidis, U. clintoniae, U. delphinii, U. 

dioscoreae, U. fischeri, U. granulosa, U. helanensis, U. 

hierochloae, U. irregularis, U. japonica, U. leersiae, U. 

magica, U. melicae, U. nevodovskyi, U. occulta, U. paridis, 

U. poae, U. primulicola, U. pulsatillae, U. qinghaiensis, U. 

ranunculi, U. rodgersiae, U. sorosporioides, U. stipae, U. 

tritici, U. ulei and U. yunnanensis. Among them, U. 

achnatheri, U. beijingensis, U. helanensis, U. qinghaiensis 

and U. yunnanensis were recently discovered as new 

species, known only from type locality and possibly 

endemic. 

204 


675 - Traditional infrageneric classifications of 

Gymnopilus (Agaricales, Cortinariaceae) are not 

supported by ribosomal DNA sequence data 

L. Guzmán-Dávalos 1* , G.M. Mueller 2 , J. Cifuentes 3 , A.N. 

Miller 2 & A. Santerre 4 

1 Departamento de Botánica y Zoología, Universidad de 

Guadalajara, Apdo. postal 1-139, Zapopan, Jal., 45101, 

Mexico. - 2 The Field Museum of Natural History, 1400 

South Lake Shore Drive, Chicago, Ill 60605-2496, U.S.A. - 

3 Facultad de Ciencias, UNAM, Circuito Exterior, Cd. 

UNiversitaria, México, D.F., 04510, Mexico. - 

4 Departamento de Biología Celular y Molecular, 

Universidad de Guadalajara, Zapopan, Jal., Mexico. - Email: 

lguzman@cucba.udg.mx 

Fulvidula (Gymnopilus P. Karst.) was divided by 

Romagnesi (1942) into Annulatae and Cortinatae (= 

section Gymnopilus Singer). The first includes species with 

a persistent, membranous annulus and the second species 

with an arachnoid veil. Hesler (1969) elevated Singer's 

sections to subgenera and divided Gymnopilus into two 

sections: Microspori and Gymnopilus. Guzmán-Dávalos 

(1995) proposed a third section, Macrospori, within 

Gymnopilus. This classification has been accepted by most 

authors; even though the assignment of a species is 

oftentimes ambiguous. Here we test the monophyly of 

Gymnopilus and the appropriateness of the current 

classification system using ribosomal nuclear ITS1-5.8S- 

ITS2 sequence data. Phylogenetic analyses of 59 sequences 

(53 ingroup and 6 outgroup) were performed using 

maximum parsimony. Initial analyses support the 

monophyly of Gymnopilus. Its division into the two 

traditional subgenera was not supported, neither was the 

recognition of the three sections in subgenus Gymnopilus. 

Four well-supported clades were identified in the genus: 1) 

the spectabilis-imperialis group; 2) the penetrans-sapineus 

complex; 3) a clade formed by G. underwoodii, G. 

validipes and G. cf. flavidellus; and 4) the dilepis-medius 

group. Thus, while the presence of a membranous veil and 

the size of the basidiospores traditionally have been used to 

support infrageneric groups, these characters have been 

shown to be highly homoplastic, and of little use at this 

taxonomic level. 

676 - Species level identification of the barley pathogens 

in Pyrenophora: Morphological versus molecular 

evidence 

S. Hambleton * , R.A. Shoemaker, K.A. Seifert & H. Boulay 

Eastern Cereal and Oilseed Research Centre, Agriculture 

and Agri-Food Canada, 960 Carling Ave, Ottawa, Ontario 

K1A 0C6, Canada. - E-mail: hambletons@em.agr.ca 

Pyrenophora teres and P. graminea cause two important 

foliar diseases of barley, net blotch and leaf stripe, 

respectively. P. teres comprises two pathovars (P. teres f. 

teres and P. teres f. maculata) differentiated by lesion type


(net blotch and spotted net blotch). Molecular phylogenetic 

analyses were used to examine species concepts within 

Pyrenophora. The initial ITS sequence analysis revealed 

species-specific sequence motifs in the ITS1 region for 

most species sampled but failed to definitively differentiate 

among barley isolates at the species level. This result 

supported prior mating experiments suggesting that P. teres 

and P. graminea represent one biological species, despite 

morphological differences, and differing mode of infection 

and disease symptoms on a single host. Further parsimony 

analyses were performed using sequence data from three 

genetic loci for multiple strains. Several isolates were 

included whose P. teres pathovar identification had been 

confirmed in inoculation trials on the host and several 

strains of P. graminea newly isolated from distinctive leaf 

stripe lesions. Two distinct clusters corresponded to P. 

teres f. teres and P. graminea, while numerous strains 

variously identified but including those confirmed as P. 

teres f. maculata, were intermediate. 

677 - Molecular and morphological characterization of 

a cosmopolitan root endophyte allied to the 

Leotiomycetes 

S. Hambleton 1* , L. Sigler 2 & R.S. Currah 3 


and Agri-Food Canada, 960 Carling Ave., Ottawa, Ontario 

K1A 0C6, Canada. - 2 University of Alberta Micorfungus 

Collection and Herbarium, University of Alberta, 

Edmonton, Alberta T6G 2E1, Canada. - 3 Department of 

Biological Sciences, University of Alberta, Edmonton, 

Alberta T6G 2E9, Canada. - E-mail: 

hambletons@em.agr.ca 

Sterile fungi isolated from surface-sterilized roots of the 

Ericaceae were hypothesized to be conspecific based 

primarily on restriction fragment length polymorphisms 

and provisionally named as Variable White Taxon (VWT). 

In preliminary re-synthesis trials with Vaccinium 

myrtilloides and V. vitis-idaea, two isolates tested did not 

form ericoid mycorrhizas. Several isolates of culturally 

similar fungi were obtained from roots of the Orchidaceae 

and Pinaceae and suspected of being VWT based on colony 

morphology and characters of the mycelium. To assess 

conspecificity and infer their phylogenetic affinities, partial 

nuclear ribosomal DNA sequences were determined. 

Parsimony analyses supported a species level distinction 

for VWT isolates and indicated that the taxon is closely 

related to, but distinct from, Hymenoscyphus ericae 

(Leotiomycetes). A new anamorph genus and species is 

proposed to accommodate this endophyte, which so far has 

remained sterile in culture, to facilitate information 

retrieval and discussion and provide a sound foundation for 

additional experimental work. Strains are identified using a 

suite of morphological characters in conjunction with ITS 

sequence data. 

678 - Knufia cryptophialidica, close phylogenetic relative 

of Coniosporium spp. (Ascomycota, incertae sedis) 

S. Hambleton 1* , A. Tsuneda 2 & K.A. Seifert 1 


and Agri-Food Canada, 960 Carling Ave, Ottawa, Ontario 

K1A 0C6, Canada. - 2 Northern Forestry Centre, Canadian 

Forest Service, Edmonton, Alberta T6H 3S5, Canada. - Email: 

hambletons@em.agr.ca 

When first described, the taxonomic position of Knufia 

cryptophialidica (monotypic genus, Hyphomycetes), 

isolated from galls on stems and branches of Populus 

tremuloides, was hypothesized to be related to the 

metacapnodiaceous sooty molds. Similar characteristics of 

the group include slow-growing, black, cerebriform growth 

on agar, the production of undifferentiated conidiogenous 

cells on moniliform hyphae, and phialidic conidiogenesis 

in some species. In a study focused on the identification 

and phylogenetic placement of a sterile black cerebriform 

fungus from Sphagnum leaves also suspected of being 

related to the Metacapnodiaceae, DNA sequences from the 

nuclear ribosomal RNA gene were determined for 4 strains 

of Knufia cryptophialidica for comparative purposes. 

Parsimony analyses indicated that K. cryptophialidica was 

distant from the sooty molds in the Capnodiales but closely 

related to Coniosporium apollinis and C. perforans, two 

species shown previously to form a strongly supported 

sister clade to the Chaetothyriomycetes. The Coniosporium 

spp. were isolated from stone and are superficially similar 

in morphology to K. cryptophialidica, but develop catenate 

muriform conidia from meristematic conidiophores. 

679 - Hawaiian puffballs, earthstars and stinkhorns 

D.E. Hemmes 1* & D.E. Desjardin 2 

1 University of Hawaii at Hilo, Biology Department, 200 W. 

Kawili St., Hilo, Hawaii 96720, U.S.A. - 2 San Francisco 

State University, Department of Biology, 1600 Holloway 

Ave., San Francisco, California 94132, U.S.A. - E-mail: 

hemmes@hawaii.edu 

An annotated list of gasteromycetes found in the Hawaiian 

Islands including members of the Lycoperdaceae, 

Geastraceae, Nidulariaceae, Tulostomataceae, Phallaceae, 

Clathraceae, and Sclerodermataceae will be presented from 

collections taken over the past ten years on all the major 

islands. Alien habitats such as coastal Casuarina groves, 

coastal Prosopsis groves, cypress plantings, and hillside 

Leucaena thickets harbor a number of earthstars and 

stalked puffballs including Tulostoma and Battarreoides. 

The native mamane-naio vegetation zones on the leeward 

sides of the high volcanic mountains are also excellent 

habitats for xerophytic earthstars, Battarrea, and 

Tulostoma. Members of the Phallaceae including 

Dictyophora, Mutinus, Pseudocolus, and Phallus are found 

in agricultural and horticultural settings whereas Aseroë 

rubra, along with various species of Scleroderma are 



associated with Eucalyptus. Vascellum, Bovista, Calvatia, 

and Dictyophora are common inhabitants of grass on 

lawns, golf courses, and pastures. A pictorial summary of 

the individual species will be presented. 

680 - Population structure of Septobasidium curtisii, a 

fungal symbiont of scale insects 

D.A. Henk * & R. Vilgalys 

Duke University, Box 90338 Durham, NC 27708, U.S.A. - 

E-mail: dah@duke.edu 

Septobasidium curtisii lives symbiotically with scale 

insects on trees and is common in the southeastern United 

States. The fungus forms thin black mats of hyphae over 

colonies of scale insects infecting some but not all. Species 

of Septobasidium are thought to disperse only as spores 

infecting young scale insects, and subsequently, hyphae 

from infected insects fuse to form a single fungal colony. If 

this is true for S. curtisii then fungal colonies may be 

genetic mosaics, and populations may be differentiated on 

small spatial scales within a stand of trees or on a single 

tree in accordance with host insect population structure. To 

test these hypotheses we use DNA sequence data from 

multiple loci to determine patterns of genetic diversity in 

single colonies and across geographic and host ranges in S. 

curtisii. Sequence data from five loci from single spore 

isolates show that colonies are composed of a single 

heterokaryotic individual that produces recombinant 

spores. Preliminary data from collections across the 

southeastern United States indicate that population 

structure of S. curtisii and host insects are correlated but at 

a larger spatial scale than expected. 

681 - Puccinia hemerocallidis, cause of daylily rust, a 

newly introduced disease in the Americas 

J.R. Hernandez 1* , M.E. Palm Hernandez 2 & L.A. 

Castlebury 1 

1 USDA, Agricultural Research Service, Systematic Botany 

and Mycology Laboratory (ARS, SBML), Rm. 304, B-011A, 

BARC-West, Beltsville, MD 20705-2350, U.S.A. - 

2 USDA/APHIS, Systematic Botany and Mycology 

Laboratory, Rm. 329, B-011A, BARC-West, Beltsville, MD 

20705-2350, U.S.A. - E-mail: jose@nt.ars-grin.gov 

Daylilies (Hemerocallis spp.) are the most important 

perennial flowering plant grown in the U.S. and are traded 

extensively by hobbyists. A rust of daylilies was introduced 

recently into North and Central America. Puccinia 

hemerocallidis is known on daylilies in Asia (stages II, III) 

with Patrinia (Valerianaceae) as the alternate host (stages 

0, I), also native to Asia. Specimens from Costa Rica, 

Japan, Russia and the United States as well as previously 

collected specimens from its native eastern Asia were 

examined morphologically to confirm that this rust was P. 

hemerocallidis. In addition, the ITS region of the ribosomal 

206 


DNA was sequenced from six representative fresh 

specimens from the Americas and Asia. The range of 

variability of certain morphological characteristics, 

including the ratio of non-septate to 1-septate teliospores, 

was greater than indicated in the current literature. 

Teliospore and urediniospore sizes varied among 

specimens but statistical analysis indicated that there was 

no significant difference in size between the Asian and the 

American spores. Three synapomorphies (including one 

indel) were present in the ITS region that distinguished the 

two Asian specimens from the four American specimens. 

However the ITS variation within the American or Asian 

specimens was approximately equal to variation between 

specimens from the two broad geographic areas. We 

conclude that the rust introduced into the Americas is 

Puccinia hemerocallidis. 

682 - Exceptional morphological diversification in 

Cladia and phylogenetic related lineages in Australasia 

N. Hoffmann * & P.T. DePriest 

Smithsonian Institution, Department of Botany, National 

museum of Natural History, P.O. Box 37012 Washington, 

DC 20013-7012, U.S.A. - E-mail: 

nikolaus.hoffmann@kfunigraz.ac.at 

Cladia is a genus of lichenized fungi with most of its 

species endemic to the Australasian region. Of the 14 

described species in the genus only four occur outside this 

region. Three other lichen genera endemic to this region, 

Heterodea, Ramalea and Thysanothecium, share many 

anatomical features with Cladia, and its family 

Cladoniaceae in general, but have very distinct vegetative 

morphologies. In our phylogenetic analysis of three regions 

of the nuclear and mitochondrial rDNA, the species of the 

genus Cladia were divided into two clades. The first clade, 

containing C. aggregata, the type species of the genus, 

consisted exclusively of members of the C. aggregata 

complex. This complex of very similar, hard to distinguish 

species is characterized by rather irregular perforations and 

a dark green-brown to black color. The second clade, in 

some analyses a grade, includes regularly perforated 

species of Cladia, such as C. ferdinandii and C. sullivanii. 

The latter species are paraphyletic relative to Heterodea, 

Ramalea and Thysanothecium. On the basis of this result 

the name Cladia should be restricted to species of the 

Cladia aggregata-complex; the correct generic names for 

the remaining species requires further investigation. Our 

phylogenetic hypothesis is consistent with the recent 

diversification of these endemic 'genera' in Australasia. 

Their previous treatment as separated genera reflects the 

striking morphological evolution that has accompanied 

their diversification.


683 - The genus Gymnopilus (Basidiomycetes, 

Agaricales) in the Czech Republic 

J. Holec 

National Museum, Mycological Department, Vaclavske 

nam. 68, 115 79 Praha 1, Czech Republic. - E-mail: 

jan.holec@nm.cz 

A project dealing with the taxonomy and distribution of the 

genus Gymnopilus in the Czech Republic was started in 

2001. Up to now, 7 species were recognised. Gymnopilus 

picreus is common in mountains but scarce to rare in 

highlands and lowlands. Gymnopilus bellulus is found 

almost exclusively in natural to virgin montane forests. 

Gymnopilus josserandii (= G. subsphaerosporus) is very 

rare and grows in the same habitats like G. bellulus. 

Gymnopilus flavus is rarely found in forest meadows of 

Moravia. Gymnopilus fulgens is also rare. It was collected 

on peaty soil in a forest near Prague. Gymnopilus 

spectabilis is scarce in lowlands and hilly country. 

Concerning the group of G. sapineus and G. penetrans, my 

preliminary results show that there are no differences in 

microcharacters between collections with smooth and scaly 

pileus. For the present I agree with Høiland that there is 

only one species, namely Gymnopilus sapineus s.l. It is the 

most abundant species of Gymnopilus in the Czech 

Republic. The species growing on burnt places (G. odini, 

G. decipiens) are extremely rare or even absent in the 

Czech Republic as no collections are kept in Czech 

herbaria and no fresh fruitbodies were found in the year 

2001. The taxonomic value of some interesting collections 

(e.g a whitish form of G. spectabilis or a species with the 

vinaceous red pileus growing on soil among mosses) will 

be evaluated in the future. 

684 - Delimitation of the genus Ciboria (Sclerotiniaceae, 

Helotiales, Ascomycota) based on rDNA sequence, 

morphology, ultrastructure and life history data 

A. Holst-Jensen 1* , T. Oftedal 2 & T. Schumacher 3 

1 National Veterinary Institute, Section of Feed and Food 

Microbiology, Ullevålsveien 68, P.O.Box 8156 Dep., 0033 

Oslo, Norway. - 2 Mycoteam AS, Forskningsvn. 3B, 

P.O.Box 5, Blindern, 0313 Oslo, Norway. - 3 University of 

Oslo, Department of Biology, Ascomycete Research group 

Oslo (ARON), P.O.Box 1045, Blindern, 0316 Oslo, 

Norway. - E-mail: arne.holst-jensen@vetinst.no 

The current delimitation of the genus Ciboria includes taxa 

with cup-shaped to plane to recurved, brownish, stipitate 

apothecia with an outer excipulum of globose cells, 

producing ellipsoid, hyaline, unicellular meiospores and 

substratal stromata, and lacking a macroconidial mitosporic 

state. Species of Ciboria infect a wide range of host plants 

and tissues, and is morphologically heterogeneous. Recent 

studies based on rDNA sequence data and ultrastructural 

data have indicated that the genus is polyphyletic. Here we 

present a more comprehensive study including new as well 

as previously published rDNA sequence data and 

phylogenetic evidence, and discuss the resulting phylogeny 

in comparison with available data on morphology, 

ultrastructure and life history. We conclude that the genus 

should be restricted to comprise only a group of taxa 

infecting and mummifying catkins of trees in the plant 

families Betulaceae, Corylaceae and Salicaceae. Another 

distinct group currently classified in Ciboria but 

characterised by production of verrucose ascospores, 

infects seeds of host in Betulaceae, develops very dense 

and hard stromata and appears to be obligate biotrophs. We 

conclude that this group deserves rank as a separate genus. 

Several taxa currently classified as Ciboria should be 

excluded from the genus, but their taxonomic status remain 

uncertain until additional phylogenetic evidence is made 

available. 

685 - Taxonomy of Rhytismatales in China 

C.-L. Hou 1* , M. Piepenbring 2 & F. Oberwinkler 1 

1 Lehrstuhl Spezielle Botanik/Mykologie, Botanisches 

Institut, Universität Tübingen, Auf der Morgenstelle 1, 

72076 Tuebingen, Germany. - 2 Botanisches Institut, 

Universität Frankfurt, Senckenberganlage 31-33, 60054 

Frankfurt/M, Germany. - E-mail: Cheng-lin.heu@unituebingen.de 

The Rhytismatales are a well-known order of ascomycetes, 

which is widely distributed in temperate and tropical 

regions of the world. Some members of this order can 

cause considerable economic damage on trees. 

Lophodermium seditiosum which causes serious needle 

cast of Pinus sylvestris and Ploioderma pini-armandii 

causing withering of needles of Pinus armandi, for 

example, are known from Western China.The abundance of 

members of Rhytismatales has been documented for 

Europe, North America, and Australia, but in the past only 

few species were reported from China. Since 1990, 

however, many new members were described, especially 

from tropical and subtropical regions of China. A total of 

17 genera and 117 species were documented by the end of 

2000. As a continuation of this effort the authors made an 

expedition to the Yunnan Province located in Southwestern 

China and to the Anhui province in Southeastern China in 

2001. A microscopic analysis of more than 120 collections 

revealed 12 genera, Bifusella, Coccomyces, Colpoma, 

Davisomycella, Duplicaria, Hypoderma, Lirula, 

Lophodermium, Naemacyclus, Neococcomyces, 

Ploioderma, and Soleella - represented by 38 species. 

Among these 16 are species new for science. Three genera 

and five species are recorded for China for the first time. 



686 - A re-evaluation of Lojkania Rehm, with an 

addition of two new species 

W.H. Hsieh & C.Y. Chen * 

Department of Plant Pathology, National Chung Hsing 

University, Taichung, Taiwan. - E-mail: 

chiyu86@yahoo.com.tw 

The circumscription of Lojkania is amended to merely 

include species with germ slits on ascospores. Additional to 

the germ slit, there are characters found to be remarkably 

correlated and consistent, which coincides with the new 

definition of this genus. The ascomatal wall is composed of 

characteristically thin sclerotial cells. Smooth ascospores 

are 1-septate and uniseriate-arranged in ascus. Asci exhibit 

different extent of development in an ascoma, and alike 

most bitunicate genera, have no ocular chamber at the tip. 

Accordingly, the segregations with its similar genera are, 

therefore, readily distinguishable. Two new species are 

hitherto erected in Lojkania. They are L. incrassata and L. 

wushensis. 

687 - Phylogenetic relationships between the genus 

Saprolegnia and related genera inferred from ITS 

sequences 

S. Inaba * & S. Tokumasu 

Sugadaira Montane Research Center, University of 

Tsukuba, 1278-294 Osa, Sanada-machi, Chiisagata-gun, 

Nagano Prefecture, Japan. - E-mail: 

s975116@icho.ipe.tsukuba.ac.jp 

Phylogenetic relationships among the genera of Isoachlya, 

Pythiopsis and Saprolegnia (Saprolegniales, Oomycetes) 

were studied based on the sequence data of internal 

transcribed spacers including 5.8S ribosomal DNA regions. 

The genera share the saprolegnoid type of zoospore 

discharge, but they are different in some points. Thus, the 

zoospores of Pythiopsis show monoplanetic behavior, 

while those of the other genera are diplanetic. Isoachlya is 

distinguishable from Saprolegnia by its manner of 

zoosporangial renewal. Several species selected from three 

genera were sequenced. The data were analyzed with 

supplement data of Achlya spp. obtained from GenBank. 

The results of the neighbor-joining and the maximum 

parsimony analysis showed sequenced species separated 

into three clusters. Cluster I consisted of several species of 

Isoachlya and Saprolegnia. Cluster II included two species 

of Pythiopsis (P. cymosa and P. terrestris) and S. 

megasperma. It was discovered that the mucilage outer 

layer surrounding young oogonia is a common 

characteristic to all of them. Cluster III was composed of 

three species with eccentric oospore, i. e. I. eccentrica, P. 

humphreyana and S. anisospora. These results suggest that 

the orthodox generic classification reflects only the part of 

phylogenetic relationships among these organisms. We 

reexamine the phylogenetic value of the various 

208 


taxonomical criteria that have been used in the orthodox 

classification of the family Saprolegniaceae. 

688 - Towards a monograph of the Parmulariaceae 

C.A. Inacio * , P.F. Cannon & D.W. Minter 

CABI Bioscience, Bakeham Lane, Egham, Surrey TW20 

9TY, U.K. - E-mail: c.inacio@cabi.org 

The family Parmulariaceae is widespread in the tropics and 

contains fungi with a large variety of morphological 

characters and host species. 31 genera and more than 100 

species are included. Type species of the genera Aldonata, 

Aldona, Aulacostroma, Coccodothis, Cocconia, 

Cycloschizon, Cyclostomella, Cocconia, Dictyocyclus, 

Dothidasteroma, Englerodothis, Ferrarisia, 

Hysterostomella, Inocyclus, Kentingia, Kiehlia, 

Pachypatella, Palawaniella, Parmularia, 

Parmulariopsella, Parmulariopsis, Parmulina, 

Perischizon, Polycyclina, Polycyclus, Protothyrium, 

Pseudolembosia, Rhagadolobium, Rhipidocarpon, 

Symphaeophyma, Thallomyces and their synonyms have 

been studied. Illustrations with drawings and 

microphotographs have been prepared, and all genera redescribed. 

Some characters, including ascospores in a 

mucous coat, asci with rostrate dehiscence, centrum 

immersed in a gelatinous layer which sometimes becomes 

blue in iodine and superficial mycelia with hyphopodiumlike 

structures are useful as diagnostic features. Useful 

characters for generic delimitation include ascomata 

(position in the substrate, shape, outer wall, connection to 

the host), superficial mycelium (presence or absence, 

position, presence of hyphopodium-like structures) and 

ascospores (colour, shape, septa). Comparison will be 

made with the possibly related family Asterinaceae and a 

new concept for the Parmulariaceae introduced. 

689 - Molecular systematics of phylloplane yeasts: 

Pigmented species belonging to the Erythrobasidium 

lineage of the Urediniomycetes (Basidiomycota) 

J. Inácio * , A. Fonseca & I. Spencer-Martins 

CREM, SABT, Universidade Nova de Lisboa, Quinta da 

Torre, 2829-516 CAPARICA, Portugal. - E-mail: 

jjis@mail.fct.unl.pt 

A screening of the mycobiota on leaves from selected plant 

species (Acer monspessulanum, Quercus faginea, Cistus 

albidus, Pistacia lentiscus and Osyris quadripartita), 

collected at the 'Arrábida Natural Park', an ecosystem of 

Mediterranean characteristics in Portugal, yielded about 

830 yeast isolates. Two isolation methods were used: the 

conventional method, based on plating of leaf washings 

onto solid medium, and the spore-fall method. About 30% 

of the isolates, a significant fraction of which producing 

ballistoconidia, presented the following characteristics: 

production of orange to red pigments, inability to produce


starch-like compounds, assimilation of D-glucuronic and/or 

protocatechuic acids but not inositol. These yeasts were 

assigned to the Erythrobasidium lineage of the 

Urediniomycetes using a combination of conventional 

phenotypic identification methods with molecular 

techniques: PCR fingerprinting (MSP-PCR) with primer 

m13 and rDNA sequence analysis (26S rDNA, D1/D1 

domains). The results showed that some of the isolates 

corresponded, or were similar, to recognised species, viz. 

Sporobolomyces coprosmae, Sp. salicinus, Sp. gracilis, 

Rhodotorula aurantiaca and Erythrobasidium 

hasegawianum, whereas others seemed to represent 

undescribed species. 

690 - Evolution of mating type gene arrangement in the 

genus Stemphylium (Ascomycetes) 

P. Inderbitzin * , J. Harkness & M.L. Berbee 

University of British Columbia, Department of Botany, 

#3529-6270 University Blvd., Vancouver, BC V6T 1Z4, 

Canada. - E-mail: bhpatrik@mail.botany.ubc.ca 

In this study we are investigating distribution and 

arrangement of the mating type genes MAT-1 and MAT-2 

in 96 isolates of the genus Stemphylium. The mating types 

were assessed with primer sets amplifying the diagnostic 

alpha and HMG boxes of the MAT-1 and MAT-2 genes 

respectively. For most of the isolates, both MAT-1 and 

MAT-2 genes could be demonstrated. In the genus 

Cochliobolus, which is closely related to Stemphylium, the 

occurrence of both mating type genes in one isolate is 

indicative of homothallism. In homothallic species of 

Cochliobolus, MAT-1 and MAT-2 are arranged in different 

ways. In Stemphylium, several species are known 

homothallics, including S. vesicarium, and the anamorph of 

the type species of Pleospora, S. herbarum. MAT-1 and 

MAT-2 were present in both S. vesicarium and S. 

herbarum. The remaining 94 isolates included in this study 

belonged to 17 other species, as well as a number of yet 

undescribed species. Some of these isolates contained only 

one mating type gene. The distribution of mating type 

genes and their arrangement was mapped into phylogenetic 

trees inferred from ITS, partial GPD, and EF-1a DNA 

sequences. It is generally thought that homothallism 

derived from heterothallism. To test this hypothesis, we 

used phylogenetic analyses to investigate whether 

heterothallism or homothallism was the ancestral state in 

Stemphylium, and how many times a switch from 

homothallism to heterothallism or vice versa occurred. 

691 - A preliminary catologue of the Saprolegniaceae of 

the Iberican peninsula 

E.P. Iniesta 1* , M.T. Tellería 1 & M. Dueñas 2 

1 Real Jardín Botánico de Madrid, CSIC, Plaza de Murillo 

2, Madrid 28014, Spain. - 2 Universidad Autónoma de 

Madrid., Ctr.a Colmenar viejo, km 15, Madrid 28049, 

Spain. - E-mail: epiniesta@ma-rjb.csic.es 

The Saprolegniaceae are one of the lest known and 

investigated group of aquatic fungi in the Iberican 

Peninsula. According to the bibliographic data it was only 

known two genera over twenty of the orden. In view of the 

interest concerning to the data in this area, we have been 

undertaken a biosystematic study of the Saprolegniaceae in 

the Iberican Peninsula, as a Phd included in the project 

'Flora Micológica Iberica'. Up to now samples has been 

colleted from different areas of West, North-East and South 

of Iberican Peninsula and species from five genera of these 

orden have been identificated: Saprolegnia, Achlya, 

Dyctiuchus, Aphanomyces and Thraustotheca. 

692 - Molecular and morphological comparisons of 

Fusarium species accommodated in the Fusarium 

subglutinans sensu lato complex 

A. Jacobs 1 , E.T. Steenkamp 1 , T.A. Coutinho 1* , G. 

Ramotshodi 2 , B.D. Wingfield 2 & M.J. Wingfield 1 

1 Dept of Microbiology and Plant Pathology, Forestry and 

Agricultural Biotechnology Institute (FABI), University of 

Pretoria, Pretoria, 0002, South Africa. - 2 Dept of Genetics, 


University of Pretoria, Pretoria, 0002, South Africa. - Email: 

teresa.coutinho@fabi.up.ac.za 

A total of 13 species formed part of the Fusarium 

subglutinans sensu lato species complex at the time of this 

study. These species are pathogens of many hosts including 

pine, mango and pineapple but are morphologically similar. 

They are all characterised by typical F. subglutinans 

morphology including false heads, absence of 

chlamydospores and microconidia produced in 

polyphialides. The aim of the study was to distinguish 

between these species based on morphology and DNA 

sequences. Phylogenetic relationships were determined 

based on sequences from the histone, elongation 1α, and βtubulin 

genes. Furthermore, sequences of the EF-1α gene 

were subjected to restriction analysis using four different 

restriction enzymes. Based on their recognition sites, 

unique EF-1α restriction patterns were generated for the 

species. The morphological characteristics that were 

informative included the origin of conidiophores on the 

aerial mycelium, conidiophore branching, the number of 

conidiogenous openings on the polyphialides, 

macroconidial septation and the presence or absence of 

sterile coiled hyphae. All 13 of the species in the Fusarium 

subglutinans sensu lato could thus be distinguished based 

on morphological and molecular characteristics. 

693 - Ophiostoma kryptum, prov. nom. from larch and 

spruce in Europe, similar to Ophiostoma minus 

K. Jacobs 1* & T. Kirisits 2 


and Agri-Food Canada, 960 Carling Avenue, Ottawa, 

Ontario, K1A 0C6, Canada. - 2 Institute of Forest 

Entomology, Forest Pathology and Forest Protection, 



Universität für Bodenkultur Wien, Hasenauerstrasse 38, A- 


karin.jacobs@fabi.up.ac.za 

Tetropium species (Coleoptera: Cerambycidae) are 

secondary, though important pests of their host trees in 

Europe. Tetropium fuscum and T. castaneum principally 

infest Norway spruce (Picea abies), while Tetropium 

gabrieli attacks primarily European larch (Larix decidua). 

Recently, the fungal associates of Tetropium spp. in Europe 

and Canada came under scrutiny after the invasion of T. 

fuscum into Atlantic Canada. Outside its native habitat in 

Europe, T. fuscum proved to be a potentially devastating 

pest and with it came Ophiostoma tetropii, a fungus that 

has previously not been recorded in Canada. An unknown 

species of Ophiostoma was isolated from Norway spruce, 

infested by Tetropium sp. on European larch, infested by T. 

gabrieli in Europe. Although similar to O. minus, this 

fungus could be distinguished from this species and is 

described here as Ophiostoma kryptum prov. nom. 

Ophiostoma kryptum is characterized by perithecia with 

short necks and a Hyalorhinocladiella anamorph as well as 

a Leptographium-like anamorph. Although O. kryptum and 

O. minus are morphologically very similar, molecular data, 

significant differences in their colony morphologies on OA 

and MEA and differences in their habitat and insect vectors 

distinguish them. Ophiostoma minus and O. kryptum thus 

represent another example of morphologically similar, yet 

genetically and ecologically separated species in the 

ophiostomatoid fungi. 

694 - Taxonomic re-evaluation of three related species 

of Graphium, based on phylogeny, morphology and 

ecology 

K. Jacobs 1* , T. Kirisits 2 & M.J. Wingfield 3 


and Agrifood Canada, 960 Carling Avenue, Ottawa, K1A 

0C6, Canada. - 2 Institute of Forest Entomology, Forest 

Pathology and Forest Protection, Universität für 

Bodenkultur Wien, Hasenauerstrasse 38, A-1190 Vienna, 

Austria. - 3 Forestry and Agricultural Biotechnology 

Institute (FABI), University of Pretoria, Pretoria, 0002, 

South Africa. - E-mail: karin.jacobs@fabi.up.ac.za 

Graphium pseudormiticum was described in 1994 from the 

galleries of the bark beetle, Orthotomicus erosus, 

introduced into South Africa from Europe. This fungus is 

characterized by conidia with conspicious basal frills that 

are produced in false chains, a trait not observed in other 

species of Graphium. Rhexographium fimbriasporum was 

described in 1995 from Ips typographus on Picea abies and 

was distinguished from Graphium based on its apparently 

unique conidiogenesis. The conidia of R. fimbriasporum 

are produced in false chains and have conspicious basal 

frills. In this study we compared G. pseudormiticum with 

R. fimbriasporum based on DNA sequences, morphology 

and ecology. We included a large collection of isolates, 

tentatively identified as Graphium spp., collected from I. 

typographus on Picea abies, Ips cembrae on Larix decidua 

and Tomicus minor on Pinus sylvestris. Graphium 

pseudormiticum and R. fimbriasporum were 

210 


morphologically similar and based on rDNA sequence (ITS 

and 18S), phylogenetically closely related but different, 

grouping with species of Graphium. Rhexographium 

fimbriasporum will thus be transferred to Graphium, but 

remains distinct from G. pseudormiticum. Another fungus 

with conidia and conidium development similar to G. 

pseudormiticum, is one of the dominant associates of I. 

cembrae on L. decidua in Europe. This fungus is 

distinguished from G. pseudormiticum and G. 

fimbriasporum based on morphology and phylogeny and is 

described as G. laricis prov. nom. 

695 - Taxonomy of the lichen family Pertusariaceae in 

Thailand 

S. Jariangprasert 1* , A.W. Archer 2 , J.A. Elix 3 & V. 

Anusarnsunthorn 4 

1 Maejo University, Biology Department, Faculty of 

Science, Chiang Mai 50290, Thailand. - 2 National 

Herbarium of New South Wales, 14 Romford, Epping, New 

South Wales 2121, Australia. - 3 Australian National 

University, Department of Chemistry, The Faculties, 

Australian National University, GPO Box 4, Canberra, 

ACT, 2601, Australia. - 4 Chiang Mai University, Biology 

Department, Faculty of Science, Chiang Mai 50200, 

Thailand. - E-mail: s_vipoosunti@yahoo.com 

The methods to identify the crustose lichen family 

Pertusariaceae are composed with morphological and 

anatomical studies, chemical test by spot test, UV test, 

recrystallization, thin layer chromatography (TLC) and 

high performance liquid chromatography (HPLC). That can 

identify Pertusariaceae into two genera Pertusaria and 

Ochrolechia. The different distinctly characters of 

Ochrolechia from Pertusaria are well-defined prominent 

exciple, vivid colour disc and the chemical, gyrophoric 

acid with a little amount of others. While as Pertusaria has 

two forms of apothecia, disciform and verruciform without 

superfacial thallus and more chemicals, xanthones, orcinol 

depsides, depsones and depsidones. Species of Pertusaria 

are P. albescens, P. amara, P. asiana, P. cicatricosa, P. 

cinchonae, P. coccopoda, P. commutata, P. leiocarpella, P. 

melaleucoides, P. patellifera, P. pertusa, P. scaberula, P. 

scutellifera, P. thwaitesii, P. tetrathalamia, P. tropica, P. 

velata, P. wattiana, P. xantholeuca and P. xylophyes. Most 

of them are new records in Thailand. Many specimens 

could be new species. Species of Ochrolechia are during 

identification. 

696 - Phylogenetic relationships of Phellinus and allied 

taxa based on ITS1-5.8S-ITS2 sequences 

W. Jeong, Y. Lim, J. Lee * & H. Jung 

School of Biological Sciences, Seoul National University, 

56-1 Shillim-dong, Kwanak-gu, Seoul 151-742, Korea. - Email: 

xylaria@empal.com


To elucidate phylogenetic relationships of the 

Hymenochaetales, sequences of the nuclear ribosomal 

ITS1-5.8S-ITS2 region from 96 strains were analyzed. The 

results showed that Trichaptum and Schizopora are closely 

related to the Hymenochaetales. But Asterostroma, 

Phaeolus, and Vararia are placed outside the 

Hymenochaetales. Total twelve groups were recognized as 

separate clades. Eight groups were same as those 

recognized by early mycologists: Phellinus robustus 

complex (= Fomitiporia Murrill 1907), P. pini complex (= 

Porodaedalea Murrill 1905), P. igniarius complex (= 

Phellinus Quélet 1886), P. rimosus complex (= Fulvifomes 

Murrill 1914), P. ferruginosus complex (= Fuscoporia 

Murrill 1907), Inonotus radiatus complex (= Mensularia 

Lazaro 1916), I. hispidus complex (= Inonotus P. Karst. 

1880) and the eighth group (= Inocutis Fiasson & Niemelä 

1984). Four groups were newly recognized in this study. 

The first group was temporarily named as P. linteus 

complex. Hymenochaete formed two separate groups that 

did not include any species of Phellinus and Inonotus. The 

last group, consisted of three Phellinus species and two 

Inonotus species, occupied a basal position of the main 

clade and clustered with Schizopora and Trichaptum. The 

present study confirmed that Phellinus is polyphyletic and, 

furthermore, rejected the widely acknowledged monophyly 

concept of Inonotus. The species of Inonotus were so 

intermingled with Phellinus species that the genus became 

topologically polyphyletic. 

697 - Secondary metabolite syndromes in the lichen 

genus Letrouitia (Ascomycota, Letrouitiaceae) 

S. Johansson * & U. Søchting 

Department of Mycology, Bothanical Institute, University 


Copenhagen K., Denmark. - E-mail: 

s.johansson@get2net.dk 

Secondary metabolites from the lichen genus Letrouitia 

Haf. & Bell. were analysed with HPLC. Most of the 

species contained the anthraquinones parietin and fragilin, 

and a few also 7-chloroemodin. In addition, a number of 

additional secondary compounds were detected, presenting 

chemosyndromes that in most cases are characteristic for 

each species. 

698 - Biology and systematics of the Saprolegniaceae: A 

treatise 

T.W. Johnson 1 , R.L. Seymour 2 & D.E. Padgett 3* 

1 Duke Univ. (retired), Durham, NC, U.S.A. - 2 The Ohio 

State Univ. (retired), Columbus, Ohio, U.S.A. - 3 The Univ. 

of NC at Wilmington, Wilmington, NC, U.S.A. - E-mail: 

Padgett@uncwil.edu 

We are pleased to announce the impending publication of a 

comprehensive treatise on the biology and systematics of 

the watermolds (Mastigomycota: Saprolegniaceae). This 

work critically analyzes the world literature on this family 

covering the period from 1750 through 1987 - citing >2800 

references - with a brief excursion to 2000 in the 

systematic section. It includes 30 chapters (with 49 tables 

and 58 figures) dealing with important aspects of 

watermold biology and an additional 20 in which we 

present a comprehensive revision of the systematics. Keys 

to the family and genera are presented together with 

extensive illustrations of included taxa. This work will be 

available to the world scientific community at no charge on 

the worldwide web probably by August 2002. It can be 

accessed using the URL http://www.ilumina-dlib.org/ by 

selecting "advanced search". In the advanced search 

window, type "Padgett" in the "contributor search 

window", tic "book" in the "learning resource type" 

window directly below, then click on the "submit" button. 

We hope that our effort will stimulate renewed interest in 

this fascinating group of organisms. 

699 - Systematics of Australian Dermocybe 

R.H. Jones 1 , T.W. May 2* & F. Udovicic 2 

1 School of Botany, The University of Melbourne, Victoria 

3010, Australia. - 2 Royal Botanic Gardens Melbourne, 

Private Bag 2000, South Yarra, Victoria 3141, Australia. - 

E-mail: tom.may@rbg.vic.gov.au 

The ectomycorrhizal basidiomycete genus Dermocybe is 

well represented in a diverse range of ecosystems on most 

continents. There has been a paucity of reliable taxonomic 

information for Australian species, and many collections of 

Dermocybe from Australia have previously been 

incorrectly assigned to Northern Hemisphere species. 

Several Australian species have considerable differences in 

pigment chemistry to their Northern counterparts, including 

a number of unique chemical compounds. Morphology and 

pigment chemistry (thin-layer chromatography) were used 

in phenetic analyses of 110 Australian Dermocybe 

collections. Nineteen taxonomic groups were identified at 

species level, of which 11 are undescribed. Representatives 

from 12 of these groups were combined with 25 other 

members of Cortinarius and Dermocybe from the Northern 

and Southern Hemispheres in parsimony analyses using 

ITS rDNA sequence data. Trees show Dermocybe as 

monophyletic, but Cortinarius as paraphyletic. Dermocybe 

aff. sanguinea from the Southern Hemisphere is sister 

group to the clade of Northern Hemisphere dermocybes. 

Northern Hemisphere members of Dermocybe, with the 

exception of D. olivaceopicta, form a separate clade to the 

Southern Hemisphere taxa. Southern Hemisphere 

Dermocybe are consistently placed basal to Northern 

Hemisphere members. 



700 - Anatomical and molecular characterization of 

Tomentella ectomycorrhizae 

E. Jukucs 1* , G. Kovács 2 & C. Vágvölgyi 3 

1 Department of Plant Anatomy, Eötvös Loránd University, 

H-1117 Budapest Pázmány Péter sétány 1/c, Hungary. - 

2 Veterinary Medical Research Institute, Hungarian 

Academy of Sciences, H-1581 Budapest, P.O. Box 18, 

Hungary. - 3 Institute of Microbiology, University Szeged, 

H-6701 Szeged, P.O. Box 533, Hungary. - E-mail: 

jakucse@ludens.elte.hu 

The members of the genus Tomentella (Thelephoraceae) 

are among the most widespread ectomycorrhizal (ECM) 

partners of trees in needle and deciduous forests all over 

the world. Although being abundant in the 

mycorrhizosphere, fruitbodies appear rarely and are usually 

overlooked. That is the reason why identification of 

mycorrhizae of this group is mainly carried out using 

molecular tools (e.g. DNS sequence analysis). Investigation 

of morphological and anatomical characteristics using the 

methods introduced by Agerer (PhC, Nomarski 

microscopy, SEM and histochemical tests) is also useful in 

determining ECMs but more precise results can be got by 

combining morphological and molecular methods. Up to 

now only few tomentelloid ECMs have been described 

comprehensively. From the continental deciduous forests 

of the Hungarian Plain several different morphotypes of 

tomentelloid mycorrhizae have been isolated and 

characterized by Agerer's methods. The majority of them 

have been detected in this territory for the first time. 

Molecular methods (PCR and sequence analysis of the 

rDNA ITS region) were successful in identification, 

comparing ITS-sequences of mycorrhizae with those 

included in DNA sequence databases of fruitbodies. 

Results of molecular identification and morphologicalanatomical 

characterization of tomentelloid ECM 

morphotypes collected in drought-adapted oak and poplar 

forests, forming symbioses with Quercus robur, Q. cerris 

and Populus alba trees, have been demonstrated. 

701 - Phylogeny of rDNA its region in smut fungi 

J.T. Juuti 1 & V. Salo 2* 

1 University of Helsinki, Department of Biosciences, 

Division of Plant Physiology, P.O.Box 56 (Viikinkaari 9), 

FIN-00014 University of Helsinki, Finland. - 2 University of 

Helsinki, Department of Applied Biology/Botany, P.O.Box 

27 (Latokartanonkaari 5), FIN-00014 University of 

Helsinki, Finland. - E-mail: vanamo.salo@helsinki.fi 

Nuclear ribosomal ITS regions were amplified from spore 

samples of several Anthracoidea Brefeld species collected 

from nature. PCR products identified as anthracoideal 

showed unexpectedly large length variation among the 

species. Two species viz. A. aspera (Liro) Kukkonen and 

A. fischeri (P. Karst.) Kukkonen with a considerable size 

difference were taken for sequence analysis. ITS products 

212 


were cloned into pGEM-T vector and sequenced. For full 

reliable bidirectional sequencing of the longer ITS 

fragment additional internal primers had to be designed. By 

sequence alignment it was found that 358 bp length 

difference of the sequences was caused by single gap in the 

ITS1 region. To evaluate taxonomic position of the genus 

Anthracoidea among the Ustilaginales multiple alignment 

was generated with other smut ITS sequences downloaded 

from GenBank. It became soon evident that comparison 

had to be expanded to more distantly related groups. The 

sampling of fungal sequences was biassed to cover much of 

the biodiversity of plant parasites, especially species which 

had been formerly or are presently recognised as smuts. 

Alignment revealed that the highly conservative and slowly 

evolving 5.8S rRNA distinguishes smuts on grasses and 

sedges of the present subclass Ustilaginomycetidae from all 

other fungi. Phylogenetic analyses will be presented and 

taxonomic status of the genus Anthracoidea will be 

discussed. 

702 - Phylogenetic studies of some Terfezia and 

Choiromyces species 

V. Kagan Zur 1* , S. Aviram 2 , Y. Ferdman 2 , Y. Li 1 & N. 

Roth-Bejerano 2 

1 Ben-Gurion Univ. Inst. Appl. ResR, POB 653 Beer-Sheva 

84105, Israel. - 2 Ben-Gurion Univ. Life SCi. Dept.D, POB 

653 Beer-Sheva 84105, Israel. - E-mail: 

zur@bgumail.bgu.ac.il 

A phylogenetic study of the genus Terfezia was undertaken 

based on analysis of the ITS and the 5 prime end of the 25S 

rDNA regions. Phylogenetic analysis of Terfezia and 

Choiromyces spp. (Terfeziaceae) revealed that all but one 

of the Terfeziaspecies form a single clade, while all but one 

of the Choiromyces species group, as expected, with Tuber 

melanosporum (Tuberaceae). Several discrepancies 

detected between the two trees indicate a separate 

evolutionary path for the two regions studied. C. 

echinulatus and Terfezia pfeilii, both from the Kalahari 

Desert, are closer to each other than to any other species, a 

point which will be discussed further. Closer analysis of a 

group of sequences obtained for T. boudieri suggests that 

this group comprises at least two distinct though 

morphologically indistinguishable species. Further analysis 

of a mycelial isolate from T. boudieri fruit-bodies 

exhibiting a double ITS profile belonging to one of the two 

proposed species provides tentative evidence for long-term 

in vitro subsistence of an ascomycetous dikaryon. 

703 - New systematics in xanthorioid lichens 

I. Kärnefelt 1 , U. Søchting 2 & S.Ya. Kondratyuk 3* 

1 Botanical Museum, Ö Vallg. 18, S-223 61 Lund, Sweden. - 

2 Botanical Institute, Ø Farimagsgade 2D, DK 1353 

Copenhagen, Denmark. - 

3 Institute of botany, 

Terescchenkivska 2, 2526o1 Kiev-4, Ukraine. - E-mail: 

skondr@botan.kiev.ua


The genus Xanthoria has been delimited on its foliose 

thallus morphology and the presence of upper and lower 

cortical layers. The morphology and anatomy of the genus 

have recently been described in several papers, and 

molecular phylogenetic studies have shown it to be 

polyphyletic. One of these lineages is the X. fallax (or 

ulophyllodes) group. The Xanthoria fallax group is 

characterised primarily by having bacilliform spermatia 

and well-developed rhizinae. Furthermore, many of the 

species produce soredia. At present the group includes the 

seventeen species. Molecular studies based on nuclear 

ribosomal DNA suggest that the X. fallax group is a 

separate lineage diverging at the very base of the genus 

Xanthoria, and HPLC-studies of the anthraquinones in 

Xanthoria also pointed at particular characteristics of the X. 

fallax group. The strong indications that the X. fallax group 

was circumscribed by a number of different and unrelated 

character sets prompted us to perform a closer study of the 

morphology, anatomy, secondary chemistry, and molecular 

phylogeny of the group. The monotypic, recently described 

genus Xanthomendoza, was included in the study as it was 

known from molecular studies also to have a basal position 

relative to Xanthoria. Apart from the new generic 

scircumscription of the xanthorioid lichens many new 

species, especieally from the southern hemisphere are 

being been described. 

704 - Phylogeography and molecular variation in 

Serpula lacrymans and S. himantioides 

H. Kauserud 1* , A. Holst-Jensen 2 & T. Schumacher 1 

1 University of Oslo, Department of Biology, P.O.Box 1045 

Blindern, 0316 Oslo, Norway. - 2 National Veterinary 

Institute, Department of Feed and Food Microbiology, 

P.O.Box 8156 Dep, 0033 Oslo, Norway. - E-mail: 

haavarka@bio.uio.no 

The dry rot fungus Serpula lacrymans, predominantly 

found in houses, and its sister taxon S. himantioides, 

mostly occurring outdoors, have global distributions in 

temperate regions. The phylogeography of both species 

were investigated on a global scale using DNA-sequences 

from three target loci; the nrDNA internal transcribed 

spacer (ITS) region, and partial sequences of the bettatubulin 

(btub) and translation elongation factor 1 alpha 

(efa) genes. Preliminary data suggest the existence of two 

varieties of S. lacrymans, one predominantly occurring 

indoor, and one growing in the wild. Little genetic 

variation was found in worldwide S. lacrymans ITS, btub 

and efa sequences of the domesticated variety; obtained 

ITS sequences from Canada, China, Europe, Mexico and 

New Zealand were identical to ITS GeneBank sequences 

from Himalaya and Australia. In S. himantioides, on the 

other hand, extensive sequence variation was found in all 

three target loci. The preliminary data suggest that the 

domesticated S. lacrymans variety has experienced a 

bottleneck event coupled to a recent worldwide spread-out. 

705 - Genotypes of Lobaria pulmonaria along an East - 

West transect in Eurasia 

C. Keller 1* , C. Cornejo 1 , J.-C. Walser 1 , I. Mikhailova 2 , T. 

Pystina 3 , S. Chabanenko 4 , S.Ya. Kondratyuk 5 & C. 

Scheidegger 1 

1 WSL Swiss Federal Research Institute, CH-8903 

Birmensdorf, Switzerland. - 2 Institute of Plant and Animal 

Ecology, 8 Marta Str. 202, 620141 Ekaterinburg, Russia. - 

3 Institute of Biology, Komi Science Centre, 

Kommunisticheskaya str. 28, 167610 Syktyvkar, Komi 

Republic, Russia. - 4 Sakhalin Botanical Garden, Gorky 

street 25, 693023 Yuzhno-Sakhalinsk, Russia. - 5 Kholodny 

Institute of Botany, Tereshchenkivska str. 2, Kiew MSP-1, 

Ukraine. - E-mail: christine.keller@wsl.ch 

Lobaria is a conspicuous foliose lichen species. The 

species was common in pre-industrial periods in humid 

temperate and boreal regions of the Northern Hemisphere, 

and cooler parts of the Tropics. Unfortunately this species 

suffered a tremendous decline during the past decades and 

the species is now considered endangered in most parts of 

Central Europe and other industrialised countries. North- 

Eastern Asia is considered a diversity hotspot of L. 

pulmonaria and related species. In Switzerland, recent 

studies based on ITS I revealed genetically diverse and 

impoverished populations. A geographic differentiation 

among the local populations was not detected within this 

small geographic area. In this paper we will test the 

hypothesis that the genetic diversity of L. pulmonaria is 

higher in the Far East of Russia compared to more western 

populations. Specimens were collected in Sakhalin, 

Northern Urals, Komi Republic (Russia), Ukrainian 

Carpathians (Ukraine), Finland, Norway, Switzerland, 

France and Portugal. We analysed a fragment, which 

included the internal transcribed spacer ITS1, the 5.8S 

ribosomal RNA gene, and parts of ITS2. The fungusspecific 

primer ITS1-F and the recently developed L. 

pulmonaria specific primer nu-ITSII-137-3' were used to 

amplify the fragment. 

706 - Survey of ophiostomatoid fungi in Austria, 

Central Europe 

T. Kirisits 1* , H. Konrad 1 , K. Jacobs 2 & M.J. Wingfield 3 

1 Institute of Forest Entomology, Forest Pathology and 


Wien, Hasenauerstrasse 38, A-1190 Vienna, Austria. - 


and Agri-Food Canada, Ottawa, Ontario, K1A 0C6, 

Canada. - 3 Forestry and Agricultural Biotechnology 

Institute (FABI), Faculty of Biological and Agricultural 

Sciences, University of Pretoria, Pretoria 0002, South 

Africa. - E-mail: kirisits@edv1.boku.ac.at 

Fungi belonging to the ascomycetous genera Ceratocystis, 

Ceratocystiopsis and Ophiostoma and related anamorph 

genera such as Leptographium, Pesotum and Graphium are 



also known as the ophiostomatoid fungi. Ophiostomatoid 

fungi are dispersed by bark beetles (Coleoptera: 

Scolytidae) and other phloem feeding and wood boring 

beetles or by air-borne and rain-splash inoculum. Since 

1992 the assemblages of ophiostomatoid fungi associated 

with bark beetles on Norway spruce, Picea abies (Ips 

typographus, Ips amitinus, Pityogenes chalcographus, 

Hylurgops palliatus, Hylurgops glabratus, Dryocoetes 

autographus), European larch, Larix decidua (Ips 

cembrae), Swiss stone pine, Pinus cembra (Ips amitinus), 

Scots pine, Pinus sylvestris and Austrian pine, Pinus nigra 

(Tomicus piniperda, Tomicus minor, Ips sexdentatus), elm, 

Ulmus spp. (Scolytus spp.) and European beech, Fagus 

sylvatica (Taphrorychus bicolor) have been studied in 

Austria. The mycobiota of Tetropium spp. (Coleoptera: 

Cerambycidae) on spruce and larch was also investigated. 

In addition, a small number of ophiostomatoid fungi were 

isolated from conifers and hardwoods without signs of 

insect infestation. In total, 40 species of ophiostomatoid 

fungi were isolated. These included 3 Ceratocystis spp., 3 

Ceratocystiopsis spp., 22 species of Ophiostoma, 5 

Leptographium spp., 6 Graphium spp. and 1 Pesotum sp. 

This ongoing study has greatly improved our knowledge of 

the occurrence, hosts and the vectors of ophiostomatoid 

fungi in Austria. 

707 - Phylogenetic analyses of four taxa of Fusarium, 

based on partial sequences of the translation elongation 

factor-1 alpha gene 

A.K. Knutsen * , M. Torp & A. Holst-Jensen 

National Veterinary Institute, Section of Feed and Food 

Microbiology, Ullevaalsveien 68, P.O. Box 8156 Dep, 

0033 Oslo, Norway. - E-mail: annkristin.knutsen@vetinst.no 

Phylogenetic relationships between four Fusarium species 

were studied using parts of the nuclear EF-1α-gene as a 

phylogenetic marker. Sequences from 12 isolates of F. 

poae, 10 isolates of F. sporotrichioides and 12 isolates of 

F. langsethiae Torp & Nirenberg ined. yielded 4, 5 and 5 

genotypes respectively. In addition we included one isolate 

of F. kyushuense. The aligned sequences were subjected to 

neighbor-joining, maximum parsimony and maximum 

likelihood analyses. The results from the different analyses 

were highly concordant. The EF-1α-based phylogenies 

support the classification of F. langsethiae as a separate 

taxon in the section Sporotrichiella of Fusarium, as the 

closest sister taxon to F. sporotrichioides while F. 

kyushuense is the sister taxon to F. poae, corresponding 

well with the ability of the former taxa to produce T-2 and 

HT-2 toxins. In contrast morphological characters indicate 

a closer relationship between F. langsethiae and F. poae on 

the one hand, and between F. sporotrichioides and F. 

kyushuense on the other hand. 

214 


708 - Chlamydospore formation of Entoloma clypeatum 

f. hybridum on mycorrhizas and rhizomorphs associated 

with Rosa multiflora 

H. Kobayashi 1* & A. Yamada 2 

1 Ibaraki Prefectural Forestry Research Institute, Nakamachi, 

Naka-gun, Ibaraki 311-0122, Japan. - 2 Faculty of 

Agriculture, Shinshu University, Minami-minowa-mura, 

kamiina-gun, Nagano 399-4588, Japan. - E-mail: 

hisakoba@deneb.freemail.ne.jp 

Chlamydospores of Entoloma clypeatum f. hybridum were 

described on the mycorrhizas and rhizomorphs associated 

with Rosa multiflora. Pinkish mycelia were observed 

around rhizomorphs and mycorrhizas of E. clypeatum f. 

hybridum associated with R. multiflora. Rhizomorphal 

connections with fruiting bodies were traced to identify the 

colored mycelia. They were thick walled with roughened 

surface, ellipsoid with marginal segments, 12-16 x 5-7 

Âµm (including segments), and hyaline to pinkish color. 

Hyaline, roughened-surface and swollen cells were 

terminally observed in vegetative hyphae with clamp 

connections. Surface view was the same both in the 

swollen cells and the spores. Two spores arranged in a 

chain were also observed. Fragmented clamp connections 

were observed on several hyphal tips. Developmental 

pattern of chlamydospore seems to be the Nyctalis type. 

This is the first report on chlamydospore formation on the 

mycorrhizas in entolomatoid fungi. 

709 - A putative hybrid or introgressant between 

Ophiostoma ulmi and Ophiostoma novo-ulmi from 

Austria, Central Europe 

H. Konrad * & T. Kirisits 

Institute of Forest Entomology, Forest Pathology and 


Wien, Hasenauerstrasse 38, A-1190 Vienna, Austria. - Email: 

hkonrad@edv1.boku.ac.at 

The ascomycete fungi Ophiostoma ulmi and Ophiostoma 

novo-ulmi have been responsible for the two destructive 

epidemics of Dutch elm disease since the early 20. century. 

Although a strong reproductive barrier operates between 

these two species, natural hybridization between them has 

been reported (Brasier et al., 1998, Mycol. Res. 102, 45- 

57). During recent surveys of the Dutch elm disease 

pathogens in Austria an unusual Ophiostoma isolate was 

obtained from a twig sample of a diseased elm tree. This 

isolate has an unique colony morphology neither 

resembling that of O. ulmi nor that of O. novo-ulmi, but 

similar to that of certain O. ulmi x O. novo-ulmi laboratory 

generated hybrids (Kirisits et al., 2001, Forstwiss. Cbl. 120, 

231-241). In laboratory crosses with authenticated isolates 

of the Dutch elm disease pathogens this strain proved to be 

sterile as recipient (female), while it behaved like O. novoulmi 

ssp. americana as donor (male) in crosses with both 

subspecies of O. novo-ulmi as recipient. The DNA


sequences of the cerato-ulmin gene and the colony type 

gene of this unusual isolate were also identical to those of 

O. novo-ulmi ssp. americana. The unusual colony 

morphology of this Ophiostoma isolate and its female 

sterility gives suspicion that it could have been arisen from 

hybridization between O. ulmi and O. novo-ulmi or from 

introgression of O. ulmi into O. novo-ulmi. Examination of 

this isolate is continuing in order to prove its possible 

interspecies hybrid origin. 

710 - Hybridization between the subspecies of 

Ophiostoma novo-ulmi in Austria, Central Europe 

H. Konrad * , C. Stauffer, E. Halmschlager & T. Kirisits 

Institute of Forest Entomology, Forest Pathology and 


Wien, Hasenauerstrasse 38, A-1190 Vienna, Austria. - Email: 

hkonrad@ento.boku.ac.at 

The second epidemic of Dutch elm disease has been caused 

by the ascomycete fungus Ophiostoma novo-ulmi. O. novoulmi 

is separated into the subspecies novo-ulmi and 

americana, both of which occur in Europe. We have 

recently presented evidence for recombination between two 

genes in isolates of O. novo-ulmi from Central Europe, 

indicating hybridization between O. novo-ulmi ssp. novoulmi 

and O. novo-ulmi ssp. americana (Konrad et al., Plant 

Pathology 51, 78-84). In the present study a geographically 

diverse population of 99 isolates of O. novo-ulmi from 

Austria was investigated. Isolates were examined for their 

fertility response as donor (male) to authenticated strains of 

O. novo-ulmi spp. novo-ulmi and ssp. americana. They 

were also studied by PCR-RFLP of the cerato-ulmin (cu) 

gene and the colony type (col1) gene. In 71 isolates fertility 

tests, as well as PCR-RFLP of cu and col1 gave consistent 

results. 49 and 22 isolates were identified as ssp. novo-ulmi 

and ssp. americana, respectively. 23 isolates showed 

recombination between cu and col1 and were thus 

identified as hybrids between ssp. novo-ulmi and ssp. 

americana. In further 5 isolates the fertility tests and the 

molecular analyses gave conflicting results and these 

isolates are suggested to be subspecies hybrids as well. 

This study has shown that a large portion of isolates of O. 

novo-ulmi from Austria consists of hybrids between ssp. 

novo-ulmi and ssp. americana. 

711 - Analysis of pathogenicity and molecular markers 

in sexual hybrids of Phytophthora infestans and P. 

mirabilis 

L.P.N.M. Kroon * & W.G. Flier 

Plant Research International, P.O. Box 16 6700AA 


l.p.n.m.kroon@plant.wag-ur.nl 

Isolates of Phytophthora infestans and P. mirabilis, 

originating from the presumed centre of origin of P. 

infestans (Toluca valley, Mexico), were crossed in vitro. 

Both species are heterothallic, and can act both as male and 

female partners in the cross. Viable offspring was obtained 

with an increased in vitro growth rate and abundant 

sporulation. DNA-fingerprinting (AFLP) showed that the 

F1 progeny consisted of true sexual hybrids. The offspring 

was (mildly) pathogenic on potato and Mirabilis jalapa, the 

respective hosts of P. infestans and P. mirabilis, and on 

tomato. F1 isolates were successfully crossed to obtain F2 

and back-cross offspring. In F2 and BC populations, 

pathogenicity on tomato and potato (and possibly on M. 

jalapa) was a segregating trait. Mitochondrial DNA 

haplotype was checked for all offspring and almost all 

isolates contained the P. mirabilis type mitochondrial 

DNA, indicating that P. mirabilis acts preferably as the 

maternal partner. By analysing the molecular make-up and 

host specificity of P. infestans/P. mirabilis F1, F2 and BC 

hybrids, the possible role of hybridisation in the speciation 

process of Phytophthora species in Toluca valley could be 

assessed. 

712 - Lewia teleomorph of Alternaria triticicola from oat 

grain in Norway is now described 

H. Kwasna 1* & B. Kosiak 2 

1 Department of Forest Pathology, Agricultural University, 

ul. Wojska Polskiegio 71 c, Poland. - 2 National Veterinary 

Institute, Department of Feed and Food Hygiene, P. O. Box 

8156 Dept., N-OO33 Oslo, Norway. - E-mail: 

barbara.kosiak@vetinst.no 

Lewia triticicola Kosiak & Kwaúna ined., the teleomorph 

of Alternaria triticicola isolated from oat grain in Norway, 

has recently been described. During a survey of Fusarium 

and Alternaria species in Norwegian grains of reduced 

quality, a fungus with characteristics of Alternaria 

triticicola Vasant Rao isolated from surface disinfected oat 

(Avena sativa L.) kernels has been identified. The oat 

sample was collected from Volden in Trøndelag, Norway 

in September 1998. Stored on slant agar at 4 °C in darkness 

for a year, the fungus produced ascomata. The ascomata 

were single or often gathered in groups, 2-3 mm in 

diameter, submerged mainly in the surface layer of the 

medium. Single-ascospore cultures produced anamorphic 

and teleomorphic states by the same isolate. This is the first 

report on the production of a teleomorph by A. triticicola. 

The teleomorph of A. triticicola was named Lewia 

triticicola Kosiak & Kwaúna ined. Although the fungus 

originated from Avena and not Triticum, the specific name 

triticicola is retained to conform with that of the anamorph, 

as is conventional for Lewia / Alternaria relationships. 

Lewia Barr & Simmons species are known as teleomorphs 

of Alternaria Nees: Fries. Including this new species, five 

species of Lewia connected with Alternaria anamorphs 

have been described so far. Apart from L. infectoria, which 

is a common inhabitant of Gramineae, no other Lewia 

species have been described from cereals. 



713 - A comparison of Metarhizium isolates from 

natural forest with those from agricultural ecosystems: 

A preliminary study 

1* 1 1 

T.F.N. Kwong , R.Y.C. Kong , L.L.P. 

Vrijmoed , E.B.G. 

2 2 

Jones & N. L. Hywel-Jones 

1 Department of Biology and Chemistry, City University of 

Hong Kong, 83 Tat Chee Avenue, Kowloon, Hong Kong 

S.A.R., China. - 2 BIOTEC-Mycology, National Center for 



RD., Khlong 1, Khlong Luang, Pathum Thani 12120, 

Thailand. - E-mail: 97472973@student.cityu.edu.hk 

Metarhizium is one of the most studied of insect fungi. 

However, most isolates are derived from agro-ecosystems. 

We posit that the rate and pattern of evolution of 

Metarhizium spp. from natural forest may prove different 

from those of agricultural origin. To test this hypothesis a 

molecular approach was adopted using selected strains 

from the natural forests in Thailand compared with 

sequences of key isolates from agricultural strains 

downloaded from the Genbank. A region ITS-5.8S rDNA 

was sequenced for isolates of Metarhizium from forests in 

Thailand. With the inclusion of strains from nature, the 

sequence data further confirms the distinct separation of 

the Metarhizium isolates into two clades which are well 

supported by high bootstrap values: the M. anisopliae and 

M. flavoviride clades. The anisopliae clade formed a sister 

group to a species from forest off Coleoptera which is 

provisionally identified as a Metarhizium anamorph of 

Cordyceps taii. No Cordyceps state has been linked with 

the flavoviride clade. Long considered an Homopteran 

clade, it was notable that two isolates from nature, but off 

cockroaches, grouped within flavoviride sensu lato. 

Significantly, forest flavoviride from Homoptera (including 

leafhoppers and adult cicadas) formed the base of this 

clade. Current evidence presented here suggests the 

flavoviride clade has host-fidelity while the anisopliae 

clade demonstrates host-promiscuity. 

714 - The plant tree, roots and clades: mycorrhizae and 

plant phylogeny 

F.C. Landis, A. Gargas * & T.J. Givnish 

University of Wisconsin, Madison, 132 Birge Hall, 430 

Lincoln Dr, Madison, WI 53706, U.S.A. - E-mail: 

agargas@facstaff.wisc.edu 

Most plants obtain at least some nutrients through hyphae 

of mycorrhizal fungi, and these symbioses appear to date 

back to the early days of plants' land colonization. Since 

then, several mycorrhizal types have evolved, each type 

allowing exploitation of different nutrient pools and 

habitats, yet limiting the symbionts' ranges to areas of 

mutual occurrence. As mycorrhizae play important roles in 

community and ecosystem ecology, understanding the 

evolution of different mycorrhizal states allows researchers 

216 


to connect plant evolution, community and ecosystem 

ecology. Recent substantial advances in our understanding 

of plant phylogeny and mycorrhizae in general prompted a 

re-evaluation of previously published reports. We mapped 

available mycorrhizal data onto current phylogenetic 

models of the plant kingdom. The resulting trees show 

some interesting patterns, such as a concentration of 

ectomycorrhizae in the Eurosids, and they also reveal 

significant gaps in our knowledge. These trees, overlays, 

and evolutionary commentary are available on the web 

(http://www.wisc.edu/botit/MPP/Index.htm or 

http://www.botany.wisc.edu/) to provide a practical tool for 

teaching about mycorrhizae to systematists and ecologists 

of all levels. These comparative data also provide 

suggestions for future research leading to elucidation of the 

biological mechanisms underlying evolution of these 

symbioses. 

715 - Incipient sympatric speciation of a fungal 

endophyte after host shift 

A. Leuchtmann * & D. Brem 

Geobotanisches Institut, ETH Zürich, Zollikerstrasse 107, 

8008 Zürich, Switzerland. - E-mail: 

leuchtmann@geobot.umnw.ethz.ch 

Host shifts which promote adaptational changes are 

considered to be important in processes of sympatric 

speciation. Here, we report on sympatric host races of the 

fungal parasite Epichloë bromicola (Ascomycota, 

Clavicipitaceae) which presumably emerged after the shift 

from the grass Bromus erectus to other Bromus hosts. The 

life cycle of E. bromicola infecting B. erectus is 

characterized by a conspicuous sexual state (stromata) that 

enables the fungus to transmit contagiously, and which 

prevents seed production of the host. Asexual strains 

infecting the two woodland species, B. benekenii and B. 

ramosus, are asymptomatic and transmit vertically by seed. 

Analyses of AFLP fingerprinting and of sequences of the 

tub2 and tef1 genes of isolates from the three Bromus hosts 

collected in Switzerland and nearby France revealed that 

isolates are genetically differentiated according to their 

host and that the seed-transmitted isolates are derived from 

ancestral, sexual strains on B. erectus. These sexual strains 

appeared to be compatible with all three hosts, while seedtransmitted 

isolates from B. benekenii and B. ramosus were 

host specific on these two hosts. However, experimental 

matings indicated still unrestricted interfertility among 

isolates of all three hosts. We suggest that isolates infecting 

B. benekenii and B. ramosus represent long-standing host 

races or incipient species, which emerged after host shifts 

and which are evolving through host-mediated, disruptive 

selection.


716 - Molecular characterization of red yeasts from 

aquatic environments in Patagonia, Argentina 

D. Libkind 1 , S. Brizzio 1 , A. Ruffini 1 , M. Gadanho 2 , M. van 

Broock 1 & J.P. Sampaio 2* 

1 Laboratorio de Microbiología Aplicada y Biotecnología, 

Universidad Nacional del Comahue - Consejo Nacional de 

Investigaciones Científicas y Tecnológicas (CONICET), 

Centro Regional Universitario Bariloche, Quintral 1250, 

Bariloche, Río Negro CP(8400), Argentina. - 2 Centro de 

Recursos Microbiológicos, Secção Autónoma de 

Biotecnologia, Faculdade de Ciências e Tecnologia, 

Universidade Nova de Lisboa, 2829-516, Portugal. - Email: 

jss@mail.fct.unl.pt 

Main features of aquatic environments of glacial origin in 

Patagonia (Argentina) are low pollution level and ultraoligotrophy. 

Eight of these aquatic reservoirs were 

surveyed for the occurrence of red yeasts. Subsurface water 

samples were filtered and used for colony counting and 

yeast isolation. A set of 64 isolates was characterised based 

on the mini/micro satellite-primed technique (MSP-PCR). 

DNA fingerprinting with M13 and (GTG)5 primers 

allowed the grouping of 34% and 56% of all isolates, 

respectively. Comparison of representative fingerprints of 

each group with those of type strains of various pigmented 

yeast, made possible the quick identification of an 87% of 

isolates. The 26S rDNA (D1/D2 domain) sequence analysis 

was performed for one representative of the remaining 

unidentified groups. Rhodotorula mucilaginosa was found 

in almost all samples and represented ca. 50% of the 

isolates studied. Less frequent species corresponded to 6 

genera and 16 species including five new yeast species of 

the genera Rhodotorula (2), Cryptococcus (1) and 

Sporobolomyces (2). Total yeast counting usually ranged 1- 

150 CFU l-1 and rarely exceeded 200 CFU l-1. In contrast 

with other studies on yeast biodiversity in freshwater, red 

yeasts did not often predominated in our water samples. 

This study represents the first report on red yeast 

occurrence and biodiversity in North-western Patagonia. 

717 - Polyphasic taxonomy of the genus Rhizopus 

G.-Y. Liou * , S.-R. Chen, H.-M. Fu, F.-L. Lee & G.-F. 

Yuan 

Food Industry Research & Development Institute, P. O. 

Box 246, Hsinchu 300, Taiwan. - E-mail: gyl@firdi.org.tw 

Rhizopus Ehrenberg, established by Ehrenberg in 1820, 

was monographed by Schipper & Stalpers, who divided the 

genus into three groups, the R. stolonifer-, R. oryzae-, and 

R. microsporus-groups, according to characteristics of the 

sporangial apparatus and growth temperatures. Thereafter, 

the characteristics of morphology, growth temperature and 

the data from mating experiments became the basic criteria 

for species delimitation of Rhizopus. In the zymogram, 

three distinct groups, the same as those obtained by 

traditional methods are recognizable. On the other hand, a 

number of specific taxonomic problems were addressed. 

Then, sequencing of 28S rDNA D1 & D2 regions, G+C 

content, and DNA-DNA hybridization were proceeded. 

The analyses of 28S rDNA sequences resolved the 

Rhizopus strains into three divergent phylogenetic clades 

consistent with the morphological characteristics and 

zymogram. The results of G+C content and DNA-DNA 

hybridization support the recognition of R. azygosporus 

and R. reflexus. Taxonomy of the genus Rhizopus was 

reevaluated. 

718 - Phylogenetic analysis of selected Aschersonia 

(Hypocrella) species and their allies 

M. Liu * & K.T. Hodge 

Cornell University, 334 Plant Science Bldg., Ithaca, NY 

14850, U.S.A. - E-mail: ml276@cornell.edu 

Despite its potential in biocontrol of scale insects and 

whiteflies, Aschersonia (teleomorph: Hypocrella) has 

seldom been included in phylogenetic studies. 

Nevertheless, it sits in an important position in the family 

of Clavicipitaceae. In this study, we investigated the 

phylogenetic relationships among Aschersonia 

(Hypocrella) isolates and species from related genera based 

on several genes. Parsimony analysis based on molecular 

data shows that Aschersonia (Hypocrella) isolates form a 

monophyletic clade, which is sister to an insect pathogen 

clade including Cordyceps and allies. A third clade 

represents grass endophytes. Within the Aschersonia clade, 

A. aleyrodis and A. goldiana form a clade; while another 

isolate referred to A. goldiana exists in another clade that 

includes A. andropogonis. Some authors have considered 

A. aleyrodis and A. goldiana to be conspecific, while others 

treat them as separate species based on morphological 

characters, especially pigmentation. We intend to clarify 

the relationships between these two species using 

molecular data and detailed morphological study. 

719 - A revision of the terverticillate Penicillia 

F. Lund 1* , J.A.M.P. Houbraken 2 , A.F.A Kuijpers 2 , R.A. 

Samson 2 & J.C. Frisvad 1 

1 Mycology Group, BioCentrum-DTU, Building 221, 


Denmark. - 2 Centraalbureau voor Schimmelcultures, P.O. 

Box 85167, NL-3508 AD Utrecht, The Netherlands. - Email: 

fl@biocentrum.dtu.dk 

Some of the most common fungi in stored foods are the 

terverticillate Penicillia. Correct identification of these 

Penicillia is important in order to predict and avoid fungal 

spoilage and mycotoxin contamination of foods, feeds and 

in indoor environments. A revision of these species is 

overdue. We have chosen a polyphasic approach for 

classifying and cladifying these Penicillia (approximately 

50 species), using a series of morphological, physiological 



and secondary metabolite data for classifying and betatubulin 

sequencing for cladifying them. Effective 

identification keys can then be developed after this 

important step. In general classification and cladification 

pointed to the same species and species series (natural 

series). For example the series Olsonii is unique both 

concerning morphology, physiology and profiles of 

secondary metabolites and this is in agreement with 

cladistic analysis of beta-tubulin sequence data. Image 

analysis of colonies and chromatographic data or other 

rapid methods combined with chemometrics can also be 

used at a later stage once the classification is clear. 

Examples of the use of the different methods will be given. 

720 - Tuberculina: relatives of rusts parasitize rusts 

M. Lutz 1* , R. Bauer 1 & D. Triebel 2 

1 Universität Tübingen, Spezielle Botanik/Mykologie, Auf 

der Morgenstelle 1, 72072 Tübingen, Germany. - 

2 Botanische Staatssammlung München, Menzinger Strasse 

67, 80638 München, Germany. - E-mail: 

matthias.lutz@uni-tuebingen.de 

The morphology, ultrastructure, molecular phylogeny, and 

infection biology of some species of the hyphomycetous 

genus Tuberculina Sacc. were examined. The presented 

results indicate that (1) Tuberculina belongs to the 

Basidiomycetes and is closely related to the Uredinales, (2) 

Tuberculina species are parasites on the haplontic stage of 

their rust hosts and show a distinct mode of interaction, and 

that (3) the commonly used species concept within the 

genus does not meet all aspects of the phylogeny inferred 

from molecular data. 

721 - Identification of a basidiomycetous fungus with 

high ability to degrade dioxins 

N. Maekawa 1* , H. Suhara 2 , R. Kondo 2 & K. Sakai 2 

1 The Tottori Mycological Institute, 211, Kokoge, Tottori 

689-1125, Japan. - 2 Faculty of Agriculture, Kyushu 

University, 6-10-1, Hakozaki, Higashi-ku, Fukuoka 812- 

8581, Japan. - E-mail: kin-maek@infosakyu.ne.jp 

During screening for efficient biodegraders of various 

recalcitrant organopollutants, including dioxins, an 

interesting fungus, which possesses a high ability to 

degrade toxic dioxins, was isolated from white rot of a 

broad-leaved tree from Kyushu Island, Japan. An isolate of 

this fungus was inoculated on a sterilized log of Quercus 

acutissima. After four months of incubation, it produced a 

resupinate and poroid basidioma on the inoculated log. The 

basidioma is microscopically characterized by having 

monomitic hyphal system, clampless hyphae and clavate 

basidia, and by lacking cystidia. These morphological 

characteristics and the partial sequencing data of 18S 

rDNA indicate that this fungus belongs to the genus 

Ceriporia (Basidiomycetes). Among the described species 

218 


of the genus, the fungus is microscopically similar to C. 

alachuana and C. ferruginicincta. However, its 

basidiospores are broader than those of C. alachuana and 

narrower than C. ferruginicincta. Further, the specimens of 

C. alachuana and C. ferruginicincta have distinctly poroid 

hymenial surface with small angular pores (3-8 and 6-8 per 

mm, respectively), whereas hymenial surface of the present 

fungus is variable in shape, i.e., poroid with larger pores 

(2-6 per mm), lacerate to dentate. These results strongly 

indicate that the present fungus is an undescribed species 

belonging to the genus Ceriporia. 

722 - Phenotypic and molecular variability amongst 

isolates of Ceratocystis fimbriata, causing coffee tree 

canker in Colombia 

M. Marin 1* , B. Castro 2 , A. Gaitan 2 , O. Preisig 1 , B.D. 

Wingfield 3 & M.J. Wingfield 1 




2 Centro Nacional de Investigaciones de Café 

(CENICAFE), Chinchiná, A.A. 2427, Colombia. - 




mauricio.marin@fabi.up.ac.za 

One of the most important diseases that reduces the 

profitability of coffee plantations in Colombia is coffee tree 

canker. This disease, caused by Ceratocystis fimbriata, has 

increased in severity and incidence in recent years, 

resulting in substantial losses to resource-poor farmers. The 

aim of this investigation was to consider the relatedness of 

C. fimbriata isolates from different Colombian coffeegrowing 

regions, and thus to provide a foundation for 

future development of resistant coffee varieties. Fifty 

isolates from 11 provinces were characterized based on 

morphology and pathogenicity. In addition, isolates were 

compared using RAPD and microsatellite markers. DNA 

sequence comparsions for the ITS region, Mat-2 HMG Box 

and partial β-tubulin gene, were also undertaken. Results of 

all comparisions clearly showed that Colombian isolates of 

C. fimbriata are highly variable. All molecular parameters 

used for these comparisons also showed that C. fimbriata 

isolates from Colombia, reside in two major phylogenetic 

lineages. We believe that these probably represent distinct 

taxa, which we are in the process of characterizing further. 

Isolates had different levels of pathogenicity on coffee, 

with some causing death of more that 90% and others 

killing less than 5% of inoculated plants. There was also a 

clear pattern to the distribution of the most pathogenic 

strains within coffee growing areas. Our results represent 

the first intensive study of this plant pathogen in Colombia.


723 - Phylogenetic relationships within the Boletales 

(Basidiomycotina) 

M.P. Martín * & M.T. Tellería 

Real Jardin Botanico, CSIC, Plaza de Murillo 2, 28014 

Madrid, Spain. - E-mail: maripaz@ma-rjb.csic.es 

The Boletales is a morphologically diverse order of 

Basidiomycetes that includes poroid, gilled, resupinate and 

epigeous gasteroid fungi. The work presented here is 

included in the Flora Micologica Iberica project. The main 

purpose was to establish the relationships, based on 

morphological, molecular and chemical data, between the 

Boletales. Although, the main collections belong to the 

Iberian Peninsula, material from all over the world were 

included to compare with the Iberian ones. The 

phylogenetic analyses, based on sequences of the ITS 

regions including the 5.8S rDNA, allow to arise the 

following conclusions: a) The boletoid, agaricoid, secotioid 

and gasteroid basidiomes appear in different groups. The 

genera with gills (Chroogomphus (Singer) O.K. Mill, 

Gomphidius Fr., Hygrophoropsis (J. Schröt) Maire ex 

Martin-Sanz, Neopaxillus Singer, Omphalotus Fayod, and 

Paxillus F.) included in three families according to 

Hawksworth et al. (Dictionary of the fungi, 1995) appear in 

five clades. Only the family Gomphidiaceae is 

monophyletic. b) The family Rhizopogonaceae is not 

monophyletic. Alpova C.W.Dodge is not included between 

the species of the genus Rhizopogon Fr. & Nordholm but 

within the clade of the Boletus species. On the other hand, 

Truncocolumella Zeller is closely related to Suillus species. 

c) The family Suillaceae should include not only Suillus 

and Gastrosuillus, but also Truncocolumella and 

Rhizopogon. 

724 - Morchella conica fruited in several waves a season 

in one site in Northern Israel 

S. Masaphy * , L. Zabari & D. Goldberg 

Migal, Galilee Technological Center, Kiryat Shmona., 

P.O.Box 90000 Rosh Pina 12100, Israel. - E-mail: 

segula@migal.org.il 

Morels (Morchella spp.) are known to grow in a wide 

range of habitats, in various places around the world, 

usually for one extended wave of three weeks per season in 

one site. However, in Northern Israel we have located 

morel fruitbodies (M. conica), which appeared in several 

waves from December until the end of April in one site. 

The morels emerged on roadside, nearby various types of 

trees. We have observed and recorded the changes in the 

mushroom population size and the distribution of fruitbody 

size, as well as changes in weather conditions. Over 150 

initials, up to 1.5 cm long were counted in the beginning of 

the first wave, but only less then half survived 2 days later, 

some continue to develop up to 10 cm long. A correlation 

was found between cold rainy days followed by warm dry 

days and between fruitbody initiation and development. 

These observations suggest that Morchella species are 

adjusted to their geographical location, and can fruit for 

several waves if weather conditions permit. 

725 - Leptographium species isolated from bark beetles 

infesting Japanese red pine in Japan 

H. Masuya 1* , Y. Yamaoka 2 & S. Kaneko 3 

1 JST Domestic Research Fellow, Tohoku Research Center 

of Forestry and Forest Products Research Institute, 

Nabeyashiki 92-25, Shimo-Kuriyagawa, Morioka, Iwate 

020-0123, Japan. - 2 Laboratory of Plant Pathology and 

Mycology, Institute of Agriculture and Forestry, Univ. 

Tsukuba, enoudai 1-1-1, Tsukuba, Ibaraki 305-8572, 

Japan. - 3 ansai Research Center of Forestry and Forest 

Products Research Institute, omoyama, Fushimi, Kyoto 

612-0855, Japan. - E-mail: mass@amy.hi-ho.ne.jp 

A survey was conducted to accumulate knowledge on the 

biogeography of Leptographium and their association with 

beetles in Japan. Seven species of Leptographium were 

isolated from bark beetles infesting Japanese red pine in 

Japan. They were L. pini-densiflorae, L. lundbergii, L. 

serpens, L. wingfieldii, L. yunnanense, Leptographium sp.1 

and Leptographium sp.2. Leptographium sp.1 was similar 

to L. neomexicanum but different in the respect of having 

more ellipsoidal conidia. Leptographium sp.2 resembled L. 

procerum but was distinguishable with well-developed 

base of stipe. These unidentified species are considered to 

be undescribed. L. serpens and L. yunnanense are firstly 

recorded in Japan. Others are already-known species in 

Japan. Leptographium sp.1 was isolated from Hylastes 

pulmbeus together with L. serpens. L. lundbergii and 

Leptographium sp.2 was isolated from five beetle species, 

Tomicus piniperda, T. minor, Hylurgops interstitialis, 

Hylastes paralleus, and H. plumbeus. L. pini-densiflorae 

was isolated from T. piniperda, T. minor, Cryphalus fulvus 

and Orthotomicus angulatus. Leptographium wingfieldii 

and L. yunnanense were isolated exclusively from T. 

piniperda. 

726 - The Genera Gymnopus and Rhodocollybia in the 

southern oak forests of Costa Rica 

J.L. Mata * , K.W. Hughes & R.H. Petersen 

The University of Tennessee, 437 Hesler Biology Bldg., 

Knoxville, TN 37996-1100, U.S.A. - E-mail: 

jmata@utk.edu 

Floristic and monographic studies in the genera Gymnopus 

and Rhodocollybia in Europe and in northeastern United 

States have been compiled recently (Halling, 1983; 

Antonín & Noordeloos, 1997). Such studies have been 

scarce, however, for these collybioid species collected in 

tropical locations. Morphological studies on specimens 

collected in the oak forests in the Talamanca Mountains of 

southern Costa Rica have revealed eight species in 



Rhodocollybia and 18 in Gymnopus. Most represent 

previously described tropical - subtropical species like G. 

neotropicus, G. lodgeae, G. fibrosipes, and R. turpis 

reported from South America and the Caribbean, but some 

are considered to be of temperate origin, such as G. 

confluens, G. dryophilus, and R. prolixa var. distorta. 

Presence of such diverse fungal taxa from the north and 

south, especially those associated with oak forests, put 

perceivable importance to the biotic exchange that took 

place over the Central American Isthmus in recent geologic 

times. Sequences from the ITS1-5.8S-ITS2 nuclear 

ribosomal DNA region were obtained for most of these 

tropical specimens and have been used to generate a 

phylogenetic tree. Costa Rican species, including newly 

proposed species, are highlighted to show their relationship 

to those from temperate locations. Current taxonomic 

placement of these taxa based on morphological 

characteristics is also analyzed. 

727 - Ascomycetous mitosis in basidiomycetous yeasts: 

its evolutionary implications 

D.J. McLaughlin 1* , R.W. Hanson, Jr. 1 , E.M. Frieders 2 , E.C. 

Swann 3 & L.J. Szabo 4 

1 University of Minnesota, Dept. of Plant Biology, St. Paul, 

MN, U.S.A. - 2 University of Wisconsin, Dept. of Biology, 

Platteville, WI, U.S.A. - 3 EMSL Analytical Inc., Plymouth, 

MN, U.S.A. - 4 USDA Agricultural Research Service, Cereal 

Disease Laboratory, St. Paul, MN, U.S.A. - E-mail: 

davem@tc.umn.edu 

In budding cells of ascomycetous yeasts, mitosis occurs in 

the parent, while in basidiomyceteous yeasts it occurs in 

the bud. However, in the basidiomycete Agaricostilbum 

pulcherrimum mitosis occurs in the parent and parent-bud 

junction. To test whether A. pulcherrimum has a novel 

mitotic pattern, investigations of additional yeasts in the 

Agaricostilbomycetidae (Urediniomycetes) utilized 

immunofluorescence localization of freeze-substituted 

material to visualize mitosis. In Bensingtonia yuccicola 

mitosis occurred in the bud, as in all other basidiomycetous 

yeasts studied to date, but in Stilbum vulgare it occurred in 

the parent. Stilbum vulgare also exhibited predominantly 

binucleate yeast cells. Nuclear rDNA sequence data 

showed that A. pulcherrimum and S. vulgare are more 

closely related to each other than to B. yuccicola in the 

Agaricostilbomycetidae. Evolutionary implications of these 

mitotic patterns will be considered. 

728 - Uredo sp. nov., a first rust species on the 

Penaeaceae 

M. Mennicken * & F. Oberwinkler 

University of Tuebingen, Botanical Institute, Systematic 

Botany and Mycology, Auf der Morgenstelle 1, D-72076 

Tuebingen, Germany. - E-mail: 

mechthilde.mennicken@gmx.de 

220 


In the context of our surveying and mapping from rust 

fungi within the BIOTA-project Southern Africa, we have 

found a new rust fungus in the BIOTA-observatory near 

Olifantsbos in the Cape Peninsula National Park. The host 

is Saltera sarcocolla (L.) Bullock. Till this day there is no 

other rust fungus known neither from the genus Saltera 

Bullock, which is endemic in the SW Western Cape, nor 

from the family Penaeaceae, which is endemic in the S and 

SW Western Cape and S Eastern Cape in South Africa (1, 

2). 1. Crous, P. W., Phillips, A. J. L., Baxter, A. P. 2000. 

Phytopathogenic fungi from South Africa, 1-358 2. 

Bredenkamp, C. L. 2000. Penaeaceae. In O. A. Leistner 

(ed.), Seed plants of southern Africa: families and genera. 

Strelitzia 10: 440-442. 

729 - Ongoing speciation within Cryptococcus 

neoformans revealed by global patterns of genetic 

variation 

W. Meyer 1* , S. Kidd 1 , A. Castaneda 2 , S. Jackson 1 & G.N. 

Latouche 1 

1 University of Sydney at Westmead Hospital, Molecular 

Mycology Laboratory, ICPMR, Level 3, Room 3114A, 

Westemead Hospital, Westmead, NSW 2145, Australia. - 

2 Grupo de Microbiologia, Instituto Nacional de Salud, 

Bogota, Colombia. - E-mail: meyer@angis.usyd.edu.au 

Taxonomy within C. neoformans is an open question, with 

3 varieties and 5 serotypes currently recognised. Variety 

grubii (serotype A), var. neoformans (serotype D) and the 

hybrid serotype AD) correspond all to the teleomorph 

Filobasidiella neoformans var. neoformans, and var. gattii 

(serotypes B, C) corresponds to the teleomorph F. n. var. 

bacillispora. A global epidemiological study using PCRfingerprinting 

with a minisatellite specific primer (M13) 

and RFLP analysis of the orotidine monophosphate 

pyrophosphorylase (URA5) and phospholipase (PLB1) 

genes revealed ongoing speciation within C. neoformans. 

All 3 typing techniques divided over 1000 clinical, 

veterinary and environmental isolates from around the 

world into 8 molecular types (VNI serotype A, var. grubii; 

VNII serotype A, var. grubii; VNIII serotype AD hybrid; 

VNIV serotype D (var. neoformans); VGI, VGII, VGIII 

and VGIV serotypes B and C, var. gattii. VNIII, AD hybrid 

isolates revealed two RFLP patterns one corresponding to 

VNI, VNII and VNIV and the other to VNII and VNIV 

suggesting different recombination events between var. 

grubii and var. neoformans leading to diploid or triploid 

strains, having two or three different copies of the 

respective genes. The variation found in the PCRfingerprinting 

profiles and RFLP patterns lies within a 

range comparable to that between established species in 

other fungal genera, indicating that evolution and 

speciation within the cryptococcal complex is an ongoing 

process. 

730 - New approach to an old problem - resolving the 

Peltigera canina species complex (Peltigeraceae, 

lichenized Ascomycota)


J. Miadlikowska * & F. Lutzoni 

Department of Biology, Duke University, Durham, NC 

27707, U.S.A. - E-mail: jolantam@duke.edu 

The Peltigera canina species complex represents the most 

recently derived section within the genus Peltigera. 

Morphology and secondary compounds were the only 

taxonomic evidences used to circumscribe the species 

forming this complex of highly polymorphic group of 

foliose lichens. To evaluate the putative morphospecies 

within the canina complex, maximum parsimony (MP) and 

maximum likelihood (ML) phylogenetic analyses were 

conducted on separate and combined ITS and LSU nrDNA 

sequences for 17 recognized species and eight potential 

taxa, represented by 65 individuals. Patterns of variation in 

length and structure found within a hypervariable region of 

ITS1 were used as supplementary data for delimiting 

genetic boundaries among closely related species. In 

addition to 20 coded characters derived from ambiguously 

aligned portions of alignments (INAASE), 24 coded 

discrete characters were provided by the hypervariable 

ITS1 marker. Based on optimal topologies derived from 

MP and ML searches we found complete concordance 

between phylogenetic and morphological species 

circumscription for 13 Peltigera species from this complex. 

Phylogenetic distinctness between North American and 

European populations of the morphologically uniform 

species P. degenii was detected and highly supported. Our 

results confirm recognition of three newly proposed 

undescribed species. No evidence for recombination was 

found in the ITS and LSU regions within the P. canina 

complex. 

731 - PCR-RELP analysis of intergenic spacer(IGS) of 

ribosomal DNA among Fusarium oxysporum formae 

speciales 

B.R. Min 1* & Y.K. Choi 2 

1 Dept. of Biology, Sangmyung Univ., 7 Hongji-Dong, 

Chongno-Gu, Seoul, Korea. - 2 Dept. of Life science, 

Hanyang Univ., 17 Haengdang-Dong, Seongdong-Gu, 

Seoul, Korea. - E-mail: genebang@sangmyung.ac.kr 

Nuclear rDNA provides useful inter- and intra-specific 

polymorphism eukaryotic organism. The intergenic spacer 

(IGS) or non-transcribed spacer (NTS) lies between the 

large subunit and small subunit encoding regions of 

consecutive cistrons. Closely related species may show 

considerable diversity in IGS of few reflecting both length 

and sequence variation. Fusarium oxysporum has one of 

the broadest host ranges of many plant pathogenic fungi 

and over go different formae speciales of F. oxysporum are 

recognized. Variation within the intergenic spacer (IGS) of 

the ribosomal DNA gene for twnety-two strains of F. 

oxysporum and its formae speciales was examined by PCR, 

compled with RELP analysis. The length of the amplified 

IGS region was about 2.6 Kb in all strains except F. 

oxysporum f. sp. cucumerium from Korea and F. 

oxysporum f. sp. niveum from Japan. Those two strains 

were 2.5 Kb long. Restriction digestion of IGS-RELP 

regions by EcoRI, NruI, HincII, SalI, SmaI, BgeII, HindIII, 

XhoI, and KpnI gave rise to nine IGS haplotypes among all 

strains. Cluster analysis based on the prosence or absence 

of comingrating restriction fragments show the two groups 

based on 44% genetic similarity. These results 

demonstrated that analysis of IGS showed some difference 

within and between F. oxysporum formae speciales. 

732 - Variability analysis of the Fumonisin producing 

species F. verticillioides using the IGS region 

S. Mirete 1* , B. Patiño 2 , C. Vázquez 2 & M.T. González- 

Jaén 1 

1 Dp. Genetics, University Complutense of Madrid, Fac. 

Biology, Ciudad Universitaria, 28040-Madrid, Spain. - 

2 Dp. Microbiology III, University Complutense of Madrid, 

Fac. Biology, Ciudad Universitaria, 28040-Madrid, Spain. 

- E-mail: smirete@bio.ucm.es 

F. verticillioides, included in the so-called Gibberella 

fujikuroi species complex, is one of the most important 

source of fumonisins contaminating food products because 

of the widespread occurrence of this fungus on maize, 

among other crops. Detection methods rely on the previous 

knowledge about the degree of intraspecific variability. In 

this work we have analysed the partial sequence of the 

intergenic spacer of rDNA (IGS) of a wide sample of F. 

verticillioides strains from different geographic location, 

hosts, mating type and fumonisin production. considering 

the incidence of asexual reproduction. The analysis 

revealed the existence of two groups of strains: fumonisin 

producing and fumonisin non producing strains which 

would indicate that some divergence is taking place within 

F. verticillioides. Within the group of fumonisin producing 

strains no clustering of isolates regarding geographic origin 

nor host could be detected. The variability observed was 

compared with data from other Fusarium species taking 

into account their mode of reproduction. 

733 - Contribution to the knowledge of Termitomyces 

(tropical basidiomycetes) from Cameroon: Ecology and 

systematics 

D.C. Mossebo 

University of Yaoundé 1, Mycological Laboratory, B.P. 

1456 Yaoundé, Cameroon. - E-mail: 

mossebo@uycdc.uninet.cm 

Species of the genus Termitomyces (Agaricales, 

Pluteaceae) are good edible tropical mushrooms 

characterized by a small to very large pileus generally 

bearing a more or less prominent umbo called 

perforatorium and a central and solid stipe producing a 

more or less long underground pseudorrhiza. Their other 

main characteristic is that they are exclusively symbiotic 

species living in symbiosis with termites on or around 

termitaries built by the latter. They arise from funguscombs 

within these termitaries. The first description of 



species growing in Cameroon was done by Heim in 1952 

and the author reported just 8 species in his account. A 

field study recently carried out enabled us to describe 14 

species of Termitomyces from Cameroon among which 4 

new and 5 new forms of Termitomyces striatus (3) and 

Termitomyces subclypeatus sp. nov. (2). The new species 

and forms described are Termitomyces grisumbo, T. 

mboudaeïna, T. subumkowaani, T. subclypeatus forma 

bisporus, T. subclypeatus forma tetrasporus, T. striatus 

forma grisumbo, T. striatus forma subperforiata and T. 

striatus forma tricystidiata. Also as shown in this colour 

poster with species described mostly in their natural 

habitat, the description of 5 other species (Termitomyces 

mammiformis Heim, T. aurantiacus Heim, T. striatus 

forma annulatus (Beeli) Heim, T. schimperi (Pat.) Heim, T. 

letestui (Pat.) Heim and T. microcarpus (Berk. & Br.) 

Heim previously described by Heim in 1952 is revised and 

completed. 

734 - Lichenicolous lichens: Independent lines of 

evolution? - evidence from Acarospora stafiana 

T.H. Nash III 1* , D. Persoh 2 , E. Barreno 3 , M. 

Wojciechowski 1 & G. Rambold 2 

1 Dept. of Plant Biology, Arizona State University, Box 

871601, Tempe, AZ 85287-1601, U.S.A. - 2 Lehrstuhl 

Pflanzensystematik, Universitaet Bayreuth, 

Universitaetsstrasse 30 - NWI, D-95440 Bayreuth, 

Germany. - 3 Plant Biology, Botany, Universitat de 

Valencia, C/Dr. Moliner 50, E-46100 Burjasot, Valencia, 

Spain. - E-mail: tom.nash@asu.edu 

Several common lichenized fungal genera, such as 

Acarospora, Buellia, Caloplaca, Lecidea and Rhizocarpon, 

comprise lichenicolous as well as free-living species. An 

implicit assumption of recognizing lichenicolous species is 

that they represent independent lines of evolution. 

Molecular approaches offer a powerful tool for testing the 

phylogenetic relationships among lichenicolous and 

independent taxa. This is a preliminary investigation of 

Acarospora, and focuses on A. stafiana, a relatively 

common lichenicolous species on Caloplaca trachyphylla 

in western North America. We analysed nrDNA ITS 

sequences of A. stafiana, its host, other free-living 

Acarosporae, other Acarospora hosts, a lichenicolous 

fungus (Stigmidium epixanthum) occurring on A. stapfiana, 

and a Candelariella sp. that appears at least facultatively 

lichenicolous on the same Caloplaca. The results support 

the inference that A. stapfiana is an obligate, lichenicolous 

species, in so far as the adjacent, free-living Acarosporae 

were distinct from the lichenicolous one. But one 

surprising result was that the ITS sequence of A. stafiana 

was identical with a free-living specimen of Acarospora 

hilaris collected from Spain. If this finding is supported by 

additional investigations, then the assumption of 

independent evolution of the lichenicolous growth-form for 

this species is not supported and it may be that Acarospora 

stapfiana and A. hilaris should be considered conspecific. 

222 


735 - Phylogenetic studies in the asexual lichen genus 

Lepraria 

M.P. Nelsen * & A. Gargas 

University of Wisconsin - Madison, Department of Botany, 

430 Lincoln Drive, Madison, WI 53706-1381, U.S.A. - Email: 

mpnelsen@students.wisc.edu 

Lepraria, a genus of lichenized fungi, is known to 

reproduce solely by asexual propagules. Widespread in 

distribution, members of Lepraria are often found growing 

on moist, shaded rocks and trees, frequently among 

mosses. Morphologically, Lepraria appears quite simple 

and bears little similarity to other genera, thus preventing 

its classification at a rank higher than genus. We 

investigate the phylogenetic position of selected Lepraria 

species in relation to other lichenized fungi using rDNA 

SSU and ITS sequence data. We also present preliminary 

data testing the hypothesis that in the absence of sexual 

reproduction, co-cladogenesis between symbionts occurs 

using rDNA ITS sequence data from both fungi and algae. 

Future studies will include testing for cryptic 

recombination within the species L. lobificans. 

736 - Molecular studies of the Hyphoderma setigerum 

complex 

R.H. Nilsson 1* , N. Hallenberg 1 , B. Nordén 1 & N. 

Maekawa 2 

1 Göteborg University, Botanical Institute, Box 461 ; 405 30 

Göteborg, Sweden. - 2 Tottori Mycological Institute, 211 

Kokoge ; Tottori ; 689-1125, Japan. - E-mail: 

henrik.nilsson@systbot.gu.se 

Hyphoderma setigerum (Basidiomycotina, 

Homobasidiomycetes) is a corticioid fungus associated 

with white rot and is primarily found on dead wood of 

deciduous trees. Morphological variation and intersterility 

tests within the taxonomical unit formerly known as H. 

setigerum have lead to the recognition of a species 

complex. The present study uses DNA sequence 

information from the ITS region of 50 specimens from 

Europe, N America, and Asia to address this variation in a 

phylogenetic context. A considerable amount of variation 

was found in the DNA sequences; based on DNA sequence 

analysis and interfertility tests, several hitherto undescribed 

species are shown to exist inside the species complex. The 

relation of H. setigerum to H. subsetigerum and H. 

nudicephalum is discussed.


737 - On the presence of ectomycorrhizae in the genus 

Sistotrema 

R.H. Nilsson 1* , K.-H. Larsson 1 , E. Larsson 1 & U. Kõljalg 2 

1 Göteborg University, Botanical Institute, Box 461 ; 40530 

Göteborg, Sweden. - 2 University of Tartu, Estonia, Riia 

181, EE51014 Tartu, Estonia. - E-mail: 

henrik.nilsson@systbot.gu.se 

The genus Sistotrema makes up a large part of the 

cantharelloid clade of Homobasidiomycete evolution. 

While ectomycorrhiza is in strong evidence for other 

genera of the cantharelloid clade (like Cantharellus, 

Hydnum, and Clavulina), it has hitherto not been recorded 

in Sistotrema. Using a combination of fruit-body / 

mycelium co-occurrence and molecular analysis (ITS, 

nLSU), the present study proves Sistotrema to contain at 

least two species capable of engaging in ectomycorrhizal 

relationships. Since the cantharelloid clade is a basal 

evolutionary lineage in the Homobasidiomycetes, the 

presence of ectomycorrhiza forming species in the clade 

may provide important clues to the nature of evolution in 

the Homobasidiomycetes. Some possible implications to 

phylogenetic analysis in the Homobasidiomycetes are 

discussed. 

738 - The Flora agaricina neerlandica project: the 

importance of morphological studies 

M.E. Noordeloos * & M.M. Nauta 

National Herbarium Netherlands, Universiteit Leiden 

branch, P.O.Box 9514, NL-2300 RA Leiden, The 

Netherlands. - E-mail: noordeloos@nhn.leidenuniv.nl 

The Flora agaricina neerlandica is a critical flora, covering 

the agarics and boleti occurring in the Netherlands and 

adjacent regions. It is composed of contributions from 

various authors, edited by Noordeloos, Kuyper and 

Vellinga. It provides keys, extensive descriptions and 

illustrations of all taxa, and data on ecology and 

distribution. The flora is essentially based on own 

observations of the authors on fresh material and herbarium 

specimens collected by Dutch mycologists over more than 

50 years. The research for the Flora is carried out according 

to standard methods and it uses in principal a 

morphological species concept, though the results of 

molecular and other studies are incorporated if available. 

As such the flora is a standard work and can serve as a base 

for further research. The importance of thorough, 

standardized morphological studies is nowadays often 

underestimated by molecular mycologists. The project has 

been accelerated by substantial financial support from the 

Kits van Waveren Foundation. Five volumes have been 

published; the sixth, containing revisions of the 

Bolbitiaceae and Coprinus, is currently in print. Volumes 

7-11 will follow with an interval of 2-3 years. Current 

projects: M. Noordeloos is revising the Boletes; M. Nauta 

concentrates on Psathyrella, Cystoderma and Lyophyllum; 

M. Noordeloos, A. Verbeken and J. Wisman work on 

Russulaceae; various Cortinariaceae genera are under 

revision by E. Arnolds, N. Dam and Th. Kuyper. 

739 - Anamorph form in the Xylariaceae 

L.K. Nugent * , M.A. Whalley, G.P. Sharples & A.J.S. 

Whalley 

School of Biomolecular Sciences, Liverpool John Moores 

University, Byrom street, Liverpool, L3 3AF, U.K. - Email: 

beslnuge@livjm.ac.uk 

Modern classification of the Xylariaceae relies more 

heavily on anamorph form than normally applied in 

traditional systematics. The form genera involved can be 

highly teleomorph genus specific e.g. Camillea and 

Xyloclodium, Astrocystis and Acanthodochium, Poronia, 

Podosordaria and Lindquistia. In other genera the 

anamorph is not genus specific and, for example 

Nodulisporium occurs in Hypoxylon, Daldinia, Entonaema, 

Thamnomyces and other genera. Similarily 

Geniculosporium is associated with Nemania, Euepixylon, 

Anthostomella, Rosellinia and several other genera. The 

distinction between Nodulisporium, Periconiella and even 

Geniculosporium is not always clear although the 

occurrence of these form genera is seen as important in 

genus recognition. The relationship between anamorph and 

teleomorph and differences between a member of the form 

genera are presented based on light and Scanning Electron 

Microscope studies. The reliability of anamorph form is 

also discussed. 

740 - Morphological and molecular study of Lactarius 

section Deliciosi in Europe 

J. Nuytinck * & A. Verbeken 

Ghent University, Dpt. Biology, Group mycology, K.L. 

Ledeganckstraat 35, B-9000 Gent, Belgium. - E-mail: 

jorinde.nuytinck@rug.ac.be 

Representatives of the section Deliciosi are easily 

distinguished from other members of the genus Lactarius 

by their orange, red or blue latex and the remarkable colour 

changes of the context. There is however no consensus on 

the species concept. Microscopic features are rather similar 

in almost all taxa and the taxonomic value of the striking 

field characters is still discussed. Carefully observed 

macro- and micromorphological characters are combined 

here with molecular data. ITS rDNA sequences are 

provided for 68 specimens collected all over Europe. The 

resulting cladogram is shown and the monophyly of the 

section and species relationships are discussed. A try-out of 

AFLP has shown that this technique is suitable to generate 

very detailed fingerprints. Eight European species are 

clearly supported from these data. 



741 - Characterization of Lactarius tesquorum 

ectomycorrhizae on Cistus sp. and molecular phylogeny 

of related European Lactarius taxa 

J. Nuytinck 1 , A. Verbeken 1 , A.C. Rinaldi 2 & O. 

Comandini 3* 

1 Ghent University, Dpt. Biology, Group Mycology, K.L. 

Ledeganckstraat 35, B-9000 Gent, Belgium. - 2 Cattedra di 

Chimica Biologica, Università di Cagliari, I-09042 

Monserrato Cagliari, Italy. - 3 Dipartimento di Scienze 

Ambientali, Università de LAquila, Via Vetoio Loc. 

Coppito, I-67100 LAquila, Italy. - E-mail: 

comandin@univaq.it 

Lactarius is one of the larger genera of ectomycorrhizal 

Basidiomycota, with about 400 species recognized 

worldwide. Members of the genus have been reported in 

association with numerous trees and shrubs, and their 

important ecological role as late-stage root colonizers in a 

range of ectotrophic plant communities is largely 

appreciated [1]. We have started a study aimed at 

extending our knowledge of the distribution, phylogeny, 

and ectomycorrhizal biology of Lactarius species occurring 

in selected ecosystems [2]. In the frame of this research, we 

have identified the ectomycorrhizae formed by Lactarius 

tesquorum on Cistus sp., one of the most common and 

ecologically relevant shrubs present in the semi-arid 

regions in the Mediterranean basin, and described them in 

terms of morphological, anatomical, and molecular 

features. In addition, an ITS rDNA sequence-based 

phylogenetic analysis of the related European Lactarius 

taxa currently classified together with L. tesquorum, 

subgenus Piperites section Piperites, was carried out in 

order to clarify species-level relationships. [1] Hutchison, 

L.J. (1999) Lactarius. In: Ectomycorrhizal Fungi: Key 

Genera in Profile. Cairney, J.W.G. and Chambers, S.M. 

(eds.), 269-285. Springer-Verlag, Berlin Heidelberg [2] 

Eberhardt, U. et al. (2000) Mycologia 92, 860-873. 

742 - A molecular phylogeny of coprophilous fungi of 

the Delitschiaceae and Sporormiaceae sensu Barr 

(Pleosporales, Ascomycota) 

Å. Nyberg * & M. Wedin 



asa.nyberg@eg.umu.se 

Preliminary results are presented from an ongoing study of 

the phylogeny of Pleosporales utilising nLSU-sequences, 

with special focus on the families Delitschiaceae and 

Sporormiaceae sensu Barr. The main aims of the study are 

to test the circumscription of these families, and to trace the 

evolution of coprophilous taxa. 

224 


743 - Karyological and molecular phylogenetic 

evaluation on Wallemia sebi 

G. Okada * & M. Takashima 

Japan Collection of Microorganisms, RIKEN (The Institute 

of Physical and Chemical Research), 2-1 Hirosawa, Wako, 

Saitama 351-0198, Japan. - E-mail: 

okada@jcm.riken.go.jp 

Wallemia sebi (Fr.) Arx is a unique osmophilic fungus with 

some mysterious aspects. Its phylogenetic relationship and 

accurate life cycle have not yet been revealed. It can be 

treated as an anamorphic ascomycete 

(http://www.ncbi.nlm.nih.gov/; Kirk et al., 2001) or a 

teleomorphic basidiomycete (Moore, 1986). Pioneer 

researches on nuclear behavior in sporogenesis and septal 

pore structure were carried out by Hashmi and Morgan- 

Jones (1973) and Terracina (1974), respectively. In this 

study, we tried to evaluate W. sebi with the focus on 

molecular phylogeny based on nuclear rDNA sequences 

and karyology especially in the very unique sporogenesis 

using some authentic strains (e.g., JCM 8329, 11230, 

11231; http://www.jcm.riken.go.jp/). Based on 

nSSU/nLSU rDNA sequence analyses (this study) and 

dolipore-like septum (Terracina, 1974; Moore, 1980), it 

became clear that W. sebi has basidiomycetous affinities. 

Our molecular phylogenetic researches showed that this 

fungus presumably locates at the base in the 

Ustilaginomycetes as a new clade. Moreover, our 

karyological research focusing on the number and size of 

nuclei in sporogenesis of W. sebi by using fluorescence 

microscopy and nuclear staining method agreed fairly well 

with the hypothesis proposed by Moore (1986). As a 

conclusion of the moment, we believe that W. sebi is very 

possibly a teleomorphic basidiomycete. 

744 - A revision of Gymnopilus in New Zealand 

A.-M.B. Oliver & D.A. Orlovich * 

Department of Botany, University of Otago, PO Box 56, 

Dunedin, New Zealand. - E-mail: 

david.orlovich@botany.otago.ac.nz 

Gymnopilus P. Karsten is a genus of wood-rotting 

basidiomycetes with over 150 species distributed 

worldwide. Gymnopilus produces a rusty-brown spore print 

and has rough spore ornamentation, commonly without a 

plage. The taxonomy of Gymnopilus in New Zealand is 

revised using numerical analysis. Ten species are recorded 

and described, including one new species, six new records 

for New Zealand (G. ferruginosus, G. allantopus, G. 

tyallus, G. austropicreus, G. norfolkensis and G. 

mullaunius) and three that were previously known (G. 

junonius, G. crociphyllus and G. mesosporus). This 

revision indicates a close relationship between New 

Zealand and Australian Gymnopilus and it is suggested that 

studies of related genera may reveal similar close 

associations of species.


745 - Characterization of Laetiporus sulphureus 

complex in Japan using phylogenetic analyses, 

morphological study and incompatibility tests 

Y. Ota * & T. Hattori 

Forestry and Forest Products Research Institute, Tsukuba, 

305-8687, Japan. - E-mail: yuota@ffpri.affrc.go.jp 

Phylogenetic analyses, morphological studies and 

incompatibility tests were conducted to define intra-generic 

taxa of Japanese Laetiporus sulphureus complex. 

Phylogenetic analyses based on the nucleotide sequences of 

the ITS regions of the ribosomal DNA and beta-tubulin 

gene suggest that Japanese Laetiporus spp. are divided into 

four distinct groups (groups A-D). European population of 

L. sulphureus is clearly distinct from all Japanese groups. 

Each group shares similar morphological characteristics 

and host range (conifers vs. hardwoods). Group A has a 

white pore layer, pinkish orange pileus surface, and 

imbricated pilei. This group is distributed in the cooltemperate 

area and associated with hardwoods. Group B 

has lemon yellow pore layer, pinkish orange pileus surface, 

and imbricated pilei. This group is associated with conifers. 

Group C has a lemon yellow pore layer, yellow pileus 

surface and non-imbricated pilei. Sometimes it produces 

semi-globose basidiocarps with abundant chlamydospores 

in the context. Group C is distributed in the warmtemperate 

area of central parts of Japan. Group D has a 

lemon yellow pore layer and pinkish orange to yellow 

pileus surface. This group also produces semi-globose 

form. This group is distributed in the warm-temperate to 

subtropical areas of southern part of Japan. Single spore 

isolates from groups A, B and C were incompatible each 

other, which revealed that L. sulphureus complex in Japan 

is composed of at least three species. 

746 - Penicillium tulipae and P. carotae, two new species 

belonging to the series Corymbifera 

D.P. Overy * & J.C. Frisvad 

Mycology Group, Bio-Centrum, DTU, Building 221, 

Danish Technical University, DK-2800, Kgs. Lyngby, 

Denmark. - E-mail: do@biocentrum.dtu.dk 

Taxa of the Penicillium series Corymbifera are known for 

their synnemata and association with the rhizosphere of 

vegetables, most commonly with bulbs. The known species 

P. allii, causing a storage rot of garlic, P. hordei, associated 

with cereals, and P. hirsutum associated with a variety of 

vegetables can easily be separated based on colony 

morphology. The concepts of P. albocoremium and P. 

venetum, reported to cause a severe bulb rot on a variety of 

commercially grown flower and Allium species, later 

evolved from P. hirustum senso lato as a response to 

apparent differences in secondary metabolism. Further 

secondary metabolite investigations involving P. 

albocoremium indicated the potential presence of two or 

more taxa. Described here using micromorphology, colony 

characteristics on various media and chemotaxonomic 

profiling are P. albocoremium sensu stricto and two new 

species, P. tulipae and P. carotae, which were resolved 

during a taxonomic survey of P. albocoremium isolates 

contained within the IBT culture collection. Although these 

novel taxa are micromorphologically quite similar, their 

unique secondary metabolite profiles individually 

distinguish them from isolates of P. albocoremium. 

Moreover, these bulb associated species produce the 

following mycotoxins: citrinin, penicillic acid and terrestric 

acid by P. carotae and penitrem A and terrestric acid by P. 

tulipae. 

747 - Cladistic biogeography of Lignincola laevis 

(Halosphaeriales, Ascomycota) 

K.L. Pang 1* , E.B.G. Jones 2 , L.L.P. Vrijmoed 3 & R.Y.C. 

Kong 3 

1 School of Biological Sciences, University of Portsmouth, 

King Henry Building, King Henry I Street, Portsmouth 

PO1 2DY, England, U.K. - 2 BIOTEC, National Centre for 

Genetic Engineering and Biotechnology, 113, Thailand 

Science Park, Paholyothin Road, Klong 1, Klong Luang, 

Pathumthani, 12120, Thailand. - 3 Department of Biology 

and Chemistry, City University of Hong Kong, 83 Tat Chee 

Avenue, Kowloon Tong, Hong Kong, China. - E-mail: kalai.pang@port.ac.uk 

Partial large subunit and the ITS regions of the ribosomal 

DNA of ten Lignincola laevis strains, which were isolated 

from different geographical locations, substrata and habitat, 

were sequenced and subjected to phylogenetic analysis to 

determine their genetic difference. These strains do not 

form distinct geographical groupings. No correlation was 

discovered between strains and habitat as freshwater and 

marine water isolates of L. laevis form a coherent and wellsupported 

group. Strains isolated from mangroves form a 

robust clade while isolates from other substrata constitute 

others. Different genes infer different phylogeny for two L. 

laevis strains. These results suggest that ascospores (asci) 

of L. laevis may be able to disperse over long distances. 

More strains from wider geographical locations, different 

habitats (freshwater, brackish water and marine water) and 

different substrata should be explored to explain the current 

distribution pattern and evolution of aquatic fungi. 

748 - Phylogenetic relationships of Hydnodon Banker 

V. Patiño-Conde 1* , D. González 2 , J. Cifuentes 1 , M. 

Villegas 1 & R. García-Sandoval 1 

1 FCME Herbario, UNAM., PO Box 70-399, Cd. 

Universitaria, CP 04510, DF., Mexico. - 2 Sistemática 

Vegetal, Instituto de Ecología AC, PO Box 63, Xalapa, CP 

91000, Veracruz, Mexico. - E-mail: 

viole_pconde@correo.unam.mx 



The monospecific genus Hydnodon was described by 

Banker (1913), based on Hydnum thelephorum (Léveillé, 

1844) because of the salmon-pinkish spore print and shape 

and ornamentation of the spores. This genus has been 

placed in different families of Aphyllophoralles such as 

Hydnaceae (Banker, 1913; Miller, 1933; Harrison, 1973), 

Thelephoraceae (Donk, 1964), and Hydnodontaceae 

(Jülich, 1981) using different morphological characters. 

Hydnodon has also been related with Scytinopogon because 

the presence of cyanophilic spores in both genera (Maas 

Geesteranus, 1963; Jülich, 1981). This research attempts to 

evaluate the phylogenetic affinities of Hydnodon based on 

recent phylogenetic hypothesis for the 

Homobasidiomycetes (Hibbett, 1997, Hibbett & Thorn, 

2001). DNA sequences of the nuc-SSU and mt-SSU of H. 

thelephorus were added to a data matrix which included 

representatives of each major clade of the 

Homobasidiomycetes. The results corroborated a 

relationship between Hydnodon and Scytinopogon, 

supported by high bootstrap and decay values. This 

hypothesis agree with Maas Geestaranus (1963) and Jülich 

(1981) proposals, and is congruent with the presence of 

inflated and gloeoplerous-like hyphae and a strong 

cyanophilic reaction of the spore ornamentation in both 

genera. Also, the position of Hydnodon and Scytinopogon 

in the general phylogeny of the Homobasidiomycetes 

suggests the presence of an additional clade, however, it is 

necessary to include more evidence to clarify this problem. 

749 - Molecular systematics of Ascochyta spp. infecting 

legumes 

T.L. Peever * , M. Barve & W.J. Kaiser 

Washington State University, Pullman, WA 99164-6430, 

U.S.A. - E-mail: tpeever@wsu.edu 

Host-specific forms of Ascochyta spp. were sampled from 

chickpea (Cicer arietinum), faba bean (Vicia faba), lentil 

(Lens culinaris), pea (Pisum sativum) and hairy vetch 

(Vicia villosa) from various parts of the world and used to 

estimate a phylogeny. Previous results from laboratory 

crosses indicated that the Ascochyta spp. infecting faba 

bean, lentil and chickpea should be considered distinct 

biological species. We tested the hypothesis that these 

biological species were also phylogenetic species and 

extended the phylogeny to include isolates sampled from 

other legumes. We sequenced the nuclear ribosomal 

internal transcribed spacer (ITS) and a glyceraldehyde-3phosphate 

dehydrogenase gene (G3PD) and estimated a 

phylogeny for each dataset. Phylogenies estimated from the 

two genomic regions were highly congruent with G3PD 

revealing significantly more variation. The G3PD 

phylogeny revealed 4 clades, each with high bootstrap 

support. Clade 1 contained isolates of A. rabiei from 

chickpeas, Clade 2 isolates of Phoma medicaginis and 

Mycosphaerella pinodes from pea, Clade 3 isolates of A. 

pisi from pea and A. fabae from faba bean, and Clade 4 

isolates of A. lentis from lentil and Ascochyta sp. from V. 

villosa. These data provide strong support for A. rabiei, A. 

fabae and A. lentis as phylogenetic and biological species. 

The analysis also revealed that A. lentis is most closely 

226 


related the host specific form on V. villosa and that A. 

fabae is most closely related to A. pisi on pea. 

750 - The phylogenetic history of Cyttaria, an obligate 

parasite of Nothofagus 

K.R. Peterson * & D.H. Pfister 

Harvard University, 22 Divinity Ave., Cambridge, MA 

02138, U.S.A. - E-mail: kpeterson@oeb.harvard.edu 

The eleven species of the fungal genus Cyttaria 

(Cyttariales, Ascomycota) are obligately parasitic on 

several species of the Gondwanan angiosperm Nothofagus 

(Nothofagaceae, Hamamelididae). Seven Cyttaria species 

are endemic to southern South America (Chile and 

Argentina), while the other four are endemic to Australasia 

(Australia, Tasmania, and New Zealand). The goals of our 

present investigation are to: 1) identify the closest relatives 

of Cyttaria; 2) infer the phylogenetic relationships between 

Cyttaria species; and 3) infer the phylogenetic relationships 

within Cyttaria species. Thus, we have begun to construct 

molecular sequence data sets from the following markers: 

nuclear ribosomal SSU, LSU, and ITS DNA; mitochondrial 

ribosomal SSU and LSU DNA; and the nuclear protein 

coding gene RPB2. Preliminary results suggest a close 

relationship between Cyttaria and the Helotiales. They also 

suggest two major lineages of Cyttaria: one composed 

entirely of South American species and another composed 

of South American and Australasian species. There also 

seems to be significant intraspecific variation within certain 

of the more widespread species. Ultimately, we will have a 

robust means with which to test the widely accepted 

hypothesis that the species of Cyttaria and Nothofagus 

have cospeciated. We will also be able to examine the 

extent to which vicariance versus dispersal can be invoked 

to explain the modern distributions of Cyttaria species and 

their hosts. 

751 - Diaporthe foeniculacea and its Phomopsis 

anamorph in Portugal 

A.J.L. Phillips 

CREM, Faculdade de Ciências e Tecnologia, Universidade 

Nova de Lisboa, Quinta da Torre, 2829-516 Caparica, 

Portugal. - E-mail: alp@mail.fct.unl.pt 

Niessl described Diaporthe foeniculacea on Foeniculum 

officinale from a collection by Moller at Coimbra, 

Portugal. Wehmeyer considered it to be a species of 

Diaporthopsis and transferred it to Diaporthopsis 

angelicae. He did not mention the anamorph. Saccardo 

described Phoma foeniculina on F. officinale. Camara 

transferred the name to Phomopsis foeniculina based on a 

collection from Sacavém, Portugal. Neither Saccardo nor 

Camara mentioned the teleomorph. In August 2001, a 

fungus with the general characteristics of a Diaporthe sp. 

was found on a plant of F. vulgare on the island of


Madeira. A similar fungus was later found at two separate 

localities near Lisbon. Two morphologically distinct 

Phomopsis species were found on the stems. The 

Diaporthe species were compared with specimens in the 

herbaria of Padova (PAD) and Coimbra (COI). The 

specimen in PAD had unicellular ascospores and appeared 

to be immature. A specimen in COI (285) collected by 

Moller is a typical Diaporthe and was regarded as D. 

foeniculacea. The specimens collected from Madeira and 

near Lisbon correlated morphologically with COI 285 and 

it is recommended that the name Diaporthe foeniculacea 

should be retained. One of the Phomopsis spp. on fennel 

stems correlated with Camaras description of P. 

foeniculina, but it is not the anamorph of D. foeniculacea. 

The teleomorph of P. foeniculina has not yet been found. 

Diaporthe foeniculacea is self-fertile and readily produces 

perithecia in culture. 

752 - Characterization of Pisolithus in Thailand 

C. Phosri 1* , R. Watling 2 , M.P. Martín 3 & A.J.S. Whalley 1 


University, Byrom Street, Liverpool, L3 3AF, U.K. - 

2 Caledonian Mycological Enterprises, Crelah, 26 

Blinkbonny Ave, Edinburgh, EH4 3HU, Scotland, U.K. - 

3 Real Jardin Botnanico de Madrid, CSIC, Plaza de Murillo 

2, 28014, Madrid, Spain. - E-mail: bmscphos@livjm.ac.uk 

A detailed study of Pisolithus, Scleroderma and Astraeus 

from a variety of hosts and geographic areas of Thailand is 

currently being undertaken to assess their potential for 

enhancement of growth in inoculated seedlings of Pinus 

kesiya and Eucalyptus camaldulensis. The results 

demonstrated that basidiospores from most Pisolithus 

isolates are ornamented with well-developed spines. At 

least 2 groups of spore types occur. Most basidiospores 

collected from Eucalyptus are characterized by numerous 

echinulate, blunt spines whilst basidiospores with 

coalesced spines joined at their tip have been found mainly 

in isolates collected from pine forest. This corresponds 

with a recent study of the internal transcribed spacer (ITS) 

DNA sequence. Sequences for 14 Pisolithus isolates 

associated with native Mediterranean hosts, Eucalyptus 

camaldulensis and Pinus kesiya were compared with those 

in GenBank. The phylogenetic analyses grouped Thai 

Pisolithus isolates into 2 distinct groups. Group 1 contained 

Pisolithus isolates collected from Pine forest and Group 2 

contained those collected from Eucalyptus plantation. This 

present study indicates that basidiospore ornamentation, 

together with ITS sequences, can distinguish two 

distinctive Pisolithus groups in Thailand which also exhibit 

clear host association. 

753 - Cercospora species from Venezuela described by 

C. Chupp and A.S. Muller 

N. Pons 1* , U. Braun 2 & P.W. Crous 3 

1 El Milagro, calle F, número 3, Maracay 2101, Venezuela. 

- 2 Martin-Luther-University, Institute of Geobotany and 

Botanical Garden, Neuwerk 21, D-06099, Halle, Germany. 

- 3 University of Stellenbosch, Stellenbosch 7602, South 

Africa. - E-mail: carinipo@cantv.net 

The species Cercospora caracasensis (on Annona 

purpurea, Annonaceae), C. centrosematis (on Centrosema 

virginianum, Leguminosae-Papilionoideae), C. cestri (on 

Cestrum latifolium, Solanaceae), C. chirguensis (on 

Curatella americana, Dilleniaceae), C. conocarpi (on 

Conocarpus erectus, Combretaceae) and C. cyclantherae 

(on Cyclanthera pedata, Cucurbitaceae), described by C. 

Chupp and A.S. Muller, were revised. Based on the 

morphological characteristics of reproductive structures 

exhibited by the type specimens and, on the correlation of 

these criteria with available molecular data, the following 

new species are proposed: Passalora centrosematis, 

Stenella cestri, Pseudocercospora chirguensis and P. 

conocarpi. Cercospora caracasensis is reduced to 

synonymy with Pseudocercospora annonae-squamosae, 

and C. cyclantherae is considered a synonym of C. apii 

s.lat. Descriptions and illustrations of the species treated 

are also provided. 

754 - Morphological characters of Lentinellus cultures 

N.V. Psurtseva 1* & Ph.B. Gannibal 2 

1 Komarov Botanical Institute RAS, Prof. Popov Str., 2, St. 

Petersburg 197376, Russia. - 2 St. Petersburg State 

University, Dept. of Botany, Universitetskaja nab., 7/9, St. 

Petersburg 199034, Russia. - E-mail: 

NadyaPsu@NP1512.spb.edu 

The taxonomy of the genus Lentinellus has recently been 

reevaluated by Dr. R.H. Petersen and his Lab (The 

University of Tennessee, USA) using classical morphology 

of type specimens, mating study and molecular analysis. 

Cultural study of Lentinellus strains received from Dr. 

Petersen's Lab have been undertaken to evaluate the 

diagnostic potential of mycelial characters in the genus. 

Dikaryon cultures from different intersterility groups 

belonging to L. ursinus, L. castoreus, L vulpinus, L. 

montanus, L. flabelliformis and L. micheneri were used. 

The strains were grown on MEA and PDA media. Indexes 

of growth, macro- and micromorphology were described 

using traditional terminology. Mycelial structure was 

studied from scanning electron microscope images. 

Laccase and tyrosinase activities were studied by plate 

diffusion method (Bavendamm reaction) and spot tests 

(substrates: guaiacol, syringaldazine and l-tyrosine). The 

strains' capacity for fructification was evaluated on 

sawdust/bran substrate. It was shown that different species 

were distinguishable by differences in growth indexes, 



texture and color of mycelial mats and in fruitage ability. 

Some of the Lentinellus hyphal structures like clamps, 

gloeocystidiaes, chlamydospores, conidias, hyphal 

swellings, rings and incrustations were species-specific. 

Strains of all studied species could produce laccase, 

whereas tyrosinase was determined only for L. micheneri, 

L. ursinus and L. castoreus. 

755 - Group I intron evolution as mobile and nonmobile 

elements within the lichen-forming fungal 

family Acarosporaceae 

V. Reeb 1* , P. Haugen 2 , D. Bhattacharya 2 & F. Lutzoni 1 

1 Duke University - Biology, Box 90338, Durham NC 

27705, U.S.A. - 2 University of Iowa - Biological Sciences, 

Iowa City IOWA 52242, U.S.A. - E-mail: vreeb@duke.edu 

Autocatalytic group I introns in the nucleus are found only 

in ribosomal DNA genes. These genetic elements are most 

commonly inherited vertically although their widespread 

and often sporadic distribution suggests their ability to 

integrate in novel rDNA sites. Two models exist for group 

I intron spread: 1) intron homing facilitated by an 

endonuclease encoded in the intron, and 2) reversesplicing. 

It seems that many organellar group I introns rely 

primarily on homing for spread, whereas virtually none of 

the rDNA introns contain endonucleases. To date only 

eight endonucleases have been reported in algae, nonlichenized 

ascomycetes, and amoeboflagellates. In this 

paper, we study the evolution of group I introns within the 

lichenized ascomycete family Acarosporaceae. Group I 

introns are well known in the ascomycetes and are 

particularly abundant in the Acarosporaceae (128 in 2.5 Kb 

of rDNA in 35 taxa). The family Acarosporaceae appears 

to be one of the most basal divergences within the 

lichenized ascomycetes, therefore, the study of its introns is 

of particular interest to better understand the evolution of 

lichen introns. We provide evidence for both relatively 

ancient and recently acquired group I intron lineages within 

the Acarosporaceae. Furthermore, we report for the first 

time in lichenized ascomycetes, the presence of a homing 

endonuclease in members of the Acarosporaceae. These 

endonucleases may facilitate the lateral transfer of some 

ascomycete group I introns. 

756 - The rust genus Hemileia - Monograph and 

collection of the characteristics in a DeltaAccess 

database 

A. Ritschel * , R. Berndt & F. Oberwinkler 

Chair of Special Botany/Mycology, University of Tübingen, 

Auf der Morgenstelle 1, D-72076 Tübingen, Germany. - Email: 

anja.ritschel@uni-tuebingen.de 

The genus Hemileia is phytoparasitic mainly on hosts 

belonging to the families Rubiaceae, Apocynaceae and 

Asclepiadaceae. The species are distributed tropically to 

228 


subtropically, probably originally in Africa and Asia. By 

the cultivation of coffee, however, members of the genus 

occur nowadays pantropically. Regarding the life cycle, so 

far only those for uredinia and telia are known. Hitherto, 53 

species of Hemileia have been described. Some of these 

species were described upon the uredinial stage and require 

nomenclatural revision. The type species Hemileia 

vastatrix represents one of the most important pathogens 

world-wide in the cultivation of coffee and thus has great 

economical importance. Although there have been 

numerous studies on the morphology and the parasitism of 

several Hemileia species, no monograph of the genus exists 

up to now. Therefore, an investigation of all Hemileia 

species is carried out by means of light and electron 

microscopy. The work is based on extensive herbarium 

material. The morphological data gathered are entered into 

a database. This database was developed within the 

framework of the GLOPP-project and is designed to 

register rust fungus biodiversity, morphological features 

and to allow the identification of the pathogens. Our final 

aim is to make this database available on the internet. 

757 - Molecular identification of Taphrina species 

parasitic on Prunus 

M.G. Rodrigues * & A. Fonseca 

CREM, SABT, Universidade Nova de Lisboa, Quinta da 

Torre, 2829-516 Caparica, Portugal. - E-mail: 

manuel.jg.rodrigues@clix.pt 

The dimorphic ascomycete genus Taphrina comprises ca. 

100 species recognised by their filamentous parasitic stage 

on several vascular plants. Infection symptoms may 

develop on leaves, fruits, shoots and flowers. Most species 

parasitise a single host and/or cause a specific infection 

symptom. The best known species is Taphrina deformans, 

causal agent of peach leaf curl, a disease with a pronounced 

impact on peach growing areas worldwide. Other 

economically important species of the genus Prunus are 

also potential Taphrina hosts, such as almond, cherry and 

plum trees. The present work focuses on the 

characterisation of Taphrina spp. on Prunus using 

molecular techniques: PCR-fingerprinting (MSP-PCR), 

sequencing of rDNA (ITS and D1/D2 regions) and DNA- 

DNA hybridizations. MSP-PCR proved to be a quick and 

reproducible method for species differentiation, allowing 

the selection of strains for sequencing. The D1/D2 region 

was too conserved to differentiate the Taphrina studied. 

The ITS regions were more useful for this purpose, and 

phylogenetic clustering of Taphrina spp. correlated with 

the host, geographic origin and/or the type of infection 

symptom. Intraspecific variability in the ITS regions was 

evaluated by comparison with the results of MSP-PCR and 

hybridization experiments. The conventional identification 

of the Taphrina species studied was re-evaluated in the 

light of the results of the molecular techniques employed.


758 - Morphotaxonomy and phylogeny of Lentinus and 

Panus from the neotropics 

T. Rolen * , L. Ryvarden & T. Schumacher 

Division of Botany and Plant physiology, P.O. box 1045 

Blindern, 0316 Oslo, Norway. - E-mail: 

tage.rolen@mycoteam.no 

A combined molecular and morpho-taxonomic approach 

was used to delineate taxa and infer main evolutionary 

lineages in members of Lentinus Fr. and Panus Fr. from 

Costa Rica and the neotropics. Previous morpho-taxonomic 

studies have resulted in unresolved delineation of 

(evolutionary) groups and conflicting classifications in this 

diverse and ecologically important group of wood decaying 

fungi of the tropics. ITS nrDNA sequences and the 

combined use of morphology and ITS sequences always 

placed Lentinus and Panus in two distinct clades. Two 

evolutionary lineages of Lentinus, and two strongly 

supported subclades within the Panus clade, were detected. 

The inferred phylogeny suggets that Lentinus is more 

closely related to Polyporus than to Panus. Three species 

assigned to the L. crinitus-complex (L. crinitus, L. bertieri, 

and L. swartzii), e. g. characterized by a continuum of 

morphotypes and not easily assigned to a specific taxon, 

were particularly focused. Sequences from 7 specimens of 

this complex indicated a relatively high level of genetic 

variation, distributed among mostly unresolved groups. In 

some Lentinus and Panus species parts of sequences were 

consistently double-banded, indicating intraindividual ITS 

polymorphism. The ITS phylogeny indicates that lamellate 

hymenophores and similarity in some ultrastructural 

features are not homologous structures in Lentinus and 

Panus. The two divergent groups of taxa justify the 

recognition of two distinct genera, Lentinus and Panus. 

759 - Identification of selected Bipolaris, Curvularia and 

Exserohilum species using cultural features, FOIL and 

Decision Tree(DT) classification tools 

I.H. Rong * , R.P. Rong & O.M. Page 

ARC-PPRI, Private Bag X 134, PRETORIA 0001, South 

Africa. - E-mail: vrehir@plant5.agric.za 

Many closely similar species have been described within 

Bipolaris, Curvularia and Exserohilum. Identification of 

species is further complicated by the variability of some 

morphological features of the group. In other anamorphic 

genera, investigation of cultural and biochemical features 

as taxonomic criteria to supplement morphology, received 

attention for the same reason. The object of this study was 

to investigate the potential of simple measurements on 

solid media as identification tools in some helminthosporic 

fungi. Authentic and representative strains of selected 

Bipolaris, Curvularia and Exserohilum species, including 

the type species of each genus, were obtained form various 

collections. The strains were selected to vary in 

geographical origin and host or substrate or origin. Colony 

diameters of strains were measured on a variety of media 

testing presumptive enzymatic activities, influence of 

inhibitory substrates and utilization of different carbon 

sources. Data accumulated were processed with two datamining 

techniques FOIL and DT. The value of culture 

based tests as diagnostic aid was demonstrated. Using 

FOIL algorithms and 5 of 25 tests, 79% of the strains 

investigated could be placed into the correct species. 

Further refinements to some tests could result in an 

identification tool or the development of selective media 

for some of these species. 

760 - Phylogenetic analysis of cetrarioid lichens with 

globose ascospores 

A. Saag 1* , T. Randlane 1 & A. Thell 2 

1 University of Tartu, Lai 38, 51005 Tartu, Estonia. - 

2 University of Hamburg, Ohnhorsstrasse 18, 22609 

Hamburg, Germany. - E-mail: asaag@ut.ee 

The group of cetrarioid lichens (Parmeliaceae, 

Ascomycota) with globose ascospores includes 37 species 

from eight genera (Ahtiana, Allocetraria, Dactylina, 

Esslingeriana, Nephromopsis, Tuckneraria, 

Tuckermannopsis, Vulpicida). Phylogenetic analysis was 

carried out on two different data sets - morphological (incl. 

anatomical and chemical) and molecular characters (ITS1, 

5.8S and ITS2 rDNA sequences) - to verify the 

correspondence of current taxonomy to the probable 

evolution of the taxa involved. For 17 species both 

morphological and molecular characters were studied while 

14 other species were checked for morphology only. The 

paraphyletic nature of the genus Allocetraria in regard to 

two species of Dactylina is assumed on morphological 

characters. Still, the separation of these genera is strongly 

supported by the analysis of molecular data. The genus 

Tuckermannopsis should be considered paraphyletic in 

regard to Esslingeriana idahoensis and Nephromopsis weii 

according to the analysis based on morphological 

characters. No doubt that Nephromopsis wei does not 

belong to the genus Nephromopsis and a new combination 

for this taxon is needed. The analysis of molecular 

characters reveals the monophyletic origin of all three 

Tuckneraria species included in the analysis. Monophyly 

of the genus Ahtiana is not supported by the analysis of 

morphological data while the analysis based on ITS 

sequences indicates a close relationship between the genera 

Ahtiana and Tuckermannopsis. 

761 - Systematic approach to pyrenomycete taxonomy 

R.A. Shoemaker 

ECORC,Agriculture Canada, Research Br., 930 Carling 

Avenue, Building 49, Ottawa ON K1A 0C6, Canada. - Email: 

shoemakerb@em.agr.ca 



Considerable progress has been made in the elucidation 

and stabilization of the concepts of Families, Orders and 

Classes of Pyenomycetes as reflected in Systema 

Ascomycetum and The Dictionary of the Fungi. From 

Systema Ascomycetum 2001, the Class Dothidiomycetes 

encompasses 7 Orders: Capnodiales (4 Families), 

Dothideales (4), Hysteriales (2), Myriangiales (2), 

Patellariales (1) and Pleosporales (19). These are ultimately 

based on the type species of 32 genera. The Class 

Chaetothyriomycetes has one recognized Order, 

Chaetothyriales, with two families plus 

Rhynchostomataceae without an assigned Order and 

Glyphium not formally assigned to either Family or Order. 

These ultimately are based on 4 species. Forty-three 

Families are provisionally assigned to Dothidiomycetes / 

Chaetothyriomycetes based on 43 generotype species. For 

79 genera, details are provided on the choice of the type 

species and the protologue of the type specimen in the hope 

that future work will focus first on these fundamental 

species rather than those chosen based on easy availability 

from culture collections. 

762 - Basidiome development as a genus level character 

in some species of Dacrymycetales from Mexico 

S. Sierra-Galván 1* , J. Cifuentes 1 & J. Marquéz-Guzmán 2 

1 Herbario, FCME, Fac. de Ciencias UNAM, A.P. 70-399 

Coyoacán C.P. 04510 México, D.F., Mexico. - 2 Desarrollo 

en plantas, Fac. de Ciencias, UNAM, A.P. 70-399 

Coyoacán C.P. 04510 México, D.F., Mexico. - E-mail: 

ssg@hp.fciencias.unam.mx 

The order Dacrymycetales was established by Lindau 

(1897), grouping genera included in the family 

Dacrymycetaceae by Brefeld (1888) such as Calocera, 

Dacrymyces, Dacryomitra and Guepinia. Subsequently, 

other classifications have been proposed, those of Coker 

(1920), Neuhoff (1936), Brasfield (1938), Kobayasi 

(1939), Martin (1952), and more recently, Lowy (1971), 

McNabb in the 60's, Reid (1974) and Oberwinkler (1994). 

One of the commonly debated topic among these works is 

the number of recognized genera, since some authors 

accept 11 genera, whereas others recognize only 2. Jülich 

(1981) established the monotypic family Cerinomycetaceae 

for Cerinomyces. In Mexico this group has been partially 

studied (Lowy, 1968; 1971; 1983; Sierra, 1992; 1993; 

1995; 2000), for this reason the main objectives in the 

undergoing research of the senior author is to monograph 

the order at the national level and to apply new characters 

to its systematic. In order to contribute to clarify genera 

delimitation basidiome development was studied in 9 

species belonging to 5 genera. The methodology proposed 

by Kennedy (1972) was used with modifications in the 

fixation (FAA), staining (PAS, saffranin-fast green, 

toluidin blue, etc.) and infiltrating material (Paraplast). 

230 


763 - Two new phytoparasitic forms of hyphomycete 

genus Mycovellosiella causing foliar spots from India 

R.K. Singh 1 , R.N. Kharwar 1 & S. Narayan 2* 

1 Centre of Advanced Study in Botany, Banaras Hindu 

University, Varanasi-221005, U.P., India. - 2 Dept. of 

Botany, D.D.U., University of Gorakhpur, Gorakhpur- 

273009, India. 

The foliicolous hyphomycete genus Mycovellosiella is now 

well established and defined genus of over 150 species till 

date. Many species of the genus were earlier placed under 

other genera like Cercospora, Cercosporella, 

Cladosporium, Phaeoramularia, Ramularia and Stenella 

etc. Mycovellosiella can be characterised by its superficial 

secondry mycelium forming prostrate and procumbent 

hyphae and forming rope-like structures often ascending 

the leaf hairs, and micronematous to semimacronematous 

conidiophores. Thus fungus produces phragmosporic and 

catenate conidia in simple or branched chains with 

prominent hila. Mycovellosiella produces leaf-spot diseases 

ranging from discolouration to necrosis of the leaf tissues. 

In continuation of the earlier additions from this region, the 

present communication is devoted to the description and 

illustrations of two new species of Mycovellosiella viz., M. 

aegli sp. nov. and M. terminalae sp. nov. occuring on 

leaves of Aegle marmelos (Rutaceae) and Terminalea 

arjuna (Combritaceae) respectively. The concerned 

materials have been deposited in H.C.I.O., New Delhi, 

India. These species have been compared with their allied 

or similar taxa to justify its separate identity. The scrapmounts 

have been prepared in lactofuchsin and glycerine 

separately to observe these structures, under different 

magnifications of compound microscope. 

764 - Fusarium commune, a new species within the 

Gibberella clade identified by morphological and 

molecular phylogenetic data 

K. Skovgaard 1* , K. O'Donnell 2 , H.I. Nirenberg 3 & S. 

Rosendahl 1 

1 Department of Mycology, University of Copenhagen, 

Oester Farimagsgade 2D, 1353 Copenhagen K, Denmark. 

- 2 Microbial Properties Research Unit, National Center for 

Agricultural Utilization Research, U.S. Department of 

Agriculture, Agricultural Research Service, 1815 North 

University Street, Peoria, IL., U.S.A. - 3 Institut für 

Pflanzenvirologie, Mikrobiologie und biologische 

Sicherheit, Biologische Bundesanstalt für Land- und 

Forstwirtschaft, Koenigin-Luise-Strasse 19, 14195 Berlin, 

Germany. - E-mail: kerstins@bot.ku.dk 

Fusarium commune sp. nov. was isolated from soil and 

Pisum sativum in Denmark and several geographically 

widespread locations within the northern hemisphere from 

diverse substrates including white pine, Douglas fir, 

carnation, corn, carrot, barley and soil. Fusarium commune 

is characterised by and distinguished from its putative


sister taxon, the F. oxysporum complex, in having long, 

slender monophialides and polyphialides when cultured in 

complete darkness. Based on the combined DNA sequence 

data from translation elongation factor and the 

mitochondrial small subunit ribosomal DNA, the fifteen 

isolates of F. commune analysed formed a strongly 

supported clade closely related to but independent of the F. 

oxysporum and the Gibberella fujikuroi species complexes. 

765 - Identifying species in the genus Botryosphaeria 

B. Slippers 1* , T.A. Coutinho 1 , B.D. Wingfield 2 , P.W. 

Crous 3 & M.J. Wingfield 1 

1 

Forestry and Agricultural Biotechnology Insititute (FABI), 

Dept of Microbiology and Plant Pathology, University of 

2 

Pretoria, Pretoria, South Africa. - Forestry and 

Agricultural Biotechnology Insititute (FABI), Dept of 

Genetics, University of Pretoria, Pretoria, South Africa. - 

3 

Centraalbureau voor Schimmelcultures (CBS), 

Uppsalalaan 8, 3584 CT Utrecht, The Netherlands. - Email: 

bernard.slippers@fabi.up.ac.za 

Botryosphaeria species are common endophytes and 

opportunistic pathogens of woody hosts, world-wide. 

Approximately 150 Botryosphaeria species have been 

described, but their taxonomy is often confused due to 

limited morphological variation and the wide host range of 

some species. Recent studies have successfully combined 

rDNA sequence and morphological data to define and 

describe species. These characters, however, need to be 

evaluated for their use in defining species boundaries 

between closely related species and species complexes. In 

this study, we combined sequence and PCR-RFLP data 

from the ITS rDNA, β-tubulin and elongation factor-1-α, 

with traditional morphological and ecological criteria, to 

delimit various Botryosphaeria spp. The ITS region was 

sufficiently variable to distinguish all species groups. 

However, some closely related species such as B. ribis and 

B. parva could not be separated. Combined data sets of the 

three sequenced regions, however, clearly separated the 

different species. Morphological characters were found to 

be variable in nature. But under controlled laboratory 

conditions, conidial and cultural morphology could be used 

to recognise most species. Ecological data were also useful 

in defining taxa, as many species are restricted to a 

particular host or environment. The combination of 

morphology, habitat data and DNA sequences produced a 

reliable basis for the characterisation of Botryosphaeria 

spp., both at the phylogenetic and diagnostic levels. 

766 - Phylogenetic position of the Caloplaca aurantia 

group 

U. Søchting 1 & U. Arup 2* 

1 Dept. of Mycology, Botanical Institute, University of 

Copenhagen, O. Farimagsgade 2D, DK-1353 Copenhagen 

K, Denmark. - 2 Institute of Systematic Botany, Lund 

University, Ö. Vallgatan 18, SE-223 61 Lund, Sweden. - Email: 

ulf.arup@sysbot.lu.se 

The Caloplaca aurantia group, consisting of the species C. 

aurantia, C. flavescens and C. thallincola, was formerly 

included in the subgenus Gasparrinia, but is distinguished 

from most other Gasparrinia's by having more or less 

citriform spores. Based on molecular data a phylogenetic 

hypothesis is presented, which places the Caloplaca 

aurantia group apart from most species in Caloplaca 

subgenus Gasparrinia, but close to the Caloplaca velana 

group. 

767 - Characterization of Fusarium proliferatum 

(Matsushima) Nirenberg, the causal agent of bakanae 

disease of rice 

P. Sontirat 1* , P. Aranyanart 2 & S. Hiranpradit 3 

1 

Mycology Group, Plant Pathology and Microbiology 

Division, Department of Agriculture, Paholyothin Road, 

Jatujak, Bangkok 10900, Thailand. - 2 Rice Pathology 

Research Group, Plant Pathology and Microbiology 


Jatujak, Bangkok 10900, Thailand. - 

3 Applied 

Microbiology Group, Plant Pathology and Microbiology 


Jatujak, Bangkok 10900, Thailand. - E-mail: 

wantanee@doa.go.th 

Sixty-four samples of bakanae disease of rice were 

collected from different locations in Thailand during the 

years 1997-2000. Cultural and morphological 

characteristics of 38 isolates were studied and examined 

scrutinizedly using Nelson et al. (1983)'s methods. The 

result revealed that both cultural and morphological 

appearances of all isolates were identical and identified as 

F. proliferatum (Matsushima) Nirenberg, when considering 

the production of polyphialides. Colonies on PDA floccose, 

white when young, and became pinkish orange to reddish 

or bluish purple when old (7-14 days ). Culture were often 

tinged with light blue; reverse pale orange to light or dark 

blue. Some isolates showed blue spots of sclerotia. On 

CLA, microconidia were abundant, formed in chains of 

varying length and in false heads on monophialides and 

polyphialides which often appeared in 'V' shape. They are 

primary single cells, oval to club-shaped with flattened 

base, 6.5 - 11.6 × 2.1 - 3.4 µm. Macroconidia were 

abundant, produced from monophialides on branched 

conidiophores in sporodochia, hyaline, slightly sickleshaped 

to almost straight with basal and ventral surfaces 

parallel. The walls were thin and the basal cells were foot- 



shaped, 3-5 septate, 26.0 - 64.8 × 1.8 - 3.1 µm. 

Chlamydospores absent. Pathogenicity test was conducted 

on rice seeds (RD6 ). It was found that the inoculated rice 

seedlings were elongated in double of the control treatment 

after 6 days of inoculation. 

768 - The nature of the lichen symbiosis: Evidence for 

mutualism 

L.L. St. Clair * & K.B. Knight 

193 MLBM Brigham Young University, Provo, Utah 

84602, U.S.A. - E-mail: larry_stclair@byu.edu 

In 1869 the German botanist Simon Schwendener put forth 

his dual hypothesis proposing for the first time the 

symbiotic nature of lichenized fungi. The structural and 

metabolic complexity of many lichens suggests a longterm, 

synergistic interaction between the symbionts. 

Certainly, the lichenized condition, at least in its most basic 

form, has been in place since the advent of the symbiotic 

partners. Traditionally, lichens have been held up as the 

ultimate example of symbiotic mutualism; however, many 

lichenologists characterize the association as one of 

controlled parasitism or helotism where the mycobiont 

exploits or enslaves the photobiont, extracting organic 

carbon and in some cases organic nitrogen. This 

interpretation has received widespread support; and clearly 

applies to more primitive (poorly organized) lichen species. 

However, more highly evolved species demonstrate a 

complex array of mycobiont adaptations that facilitate and 

accommodate the photobiont and promote the dispersal and 

propagation of the lichen. By definition mutualism requires 

that both symbionts are benefited by the relationship, with 

each partner evolving discernable structural and/or 

metabolic adaptations that enhance the performance of the 

other symbiont. Clearly, many lichen associations meet 

these criteria, and therefore, should be considered 

mutualistic. 

769 - Morphological and ultrastructural characteristics 

of Daldinia species 

M. Stadler 1* , M. Baumgartner 1 , G. Venturella 2 & H. 

Wollweber 3 

1 Bayer AG, Pharma Research, Bayer Pharma Research 

Center, P.O.B. 101709, D-42096 Wuppertal, Germany. - 

2 Universitá di Palermo, Dipartimento di Scienze 

Botaniche, Via Archirafi 38, I-90123 Palermo, Italy. - 

3 Naturwissenschaftlicher Verein Wuppertal, Mykologische 

Sektion, In den Birken 73, D-42113 Wuppertal, Germany. - 

E-mail: marc.stadler@t-online.de 

In our investigation of Daldinia Ces & De Not. 

(Xylariaceae), morphological characters of anamorphs as 

defined in the last world monograph by Ju et al. 

(Mycotaxon 61: 243f., 1997) proved valuable for 

segregation of species. Moreover, SEM was employed to 

232 


study the surface structures and other features of 

ascospores (Mycolog. Progress 1: 31f., 2002). 

Aforementioned characters were employed to validate the 

results obtained by recently evolved PCR methodology and 

chemotaxonomy based on HPLC-UV/visual and HPLC- 

MS metabolite patterns (Mycolog. Res. 105, 1191f., 2001). 

The results of our recent studies of specimens similar to the 

pantropical D. eschscholzii (Ehrenb.: Fr.) Rehm from 

Sicily, the Channel Islands and the Canary Islands suggest 

that the variability within the genus may actually be much 

higher than we had suspected some time ago. The present 

contribution is intended to illustrate characteristic features 

of some well-known and some recently described Daldinia 

spp. Morphological and ultrastructural data also provide 

evidence that a polyphasic taxonomical approach is 

feasible to detect cryptic species and evaluate the 

biological diversity of Daldinia. 

770 - The ITS phylogeny highlights the evolution of 

morphological characters and ecological traits in 

Cordyceps 

Ø. Stensrud 1* , I. Bjorvand Engh 2 , N.L. Hywel-Jones 3 & T. 

Schumacher 1 

1 Div. of Botany and Plant Physiology, Dept. of Biology, 

University of Oslo, P.O.Box 1045, Blindern, 0316 Oslo, 

Norway. - 2 Mycoteam AS, Forskningsvn. 3B, P.O.Box 5, 

Blindern, 0313 Oslo, Norway. - 3 BIOTEC-Mycology, Yothi 

Laboratories, 73/1 Rama VI Road, Rajdhevee, Bangkok 

10400, Thailand. - E-mail: oyvind.stensrud@bio.uio.no 

The phylogenetic relationships of 46 Cordyceps spp. and 

24 anamorphic taxa with known or suspected affinity to 

Cordyceps were inferred from ITS nrDNA sequence data. 

The ITS phylogeny recognized four evolutionary lineages 

and some strongly supported subclades when 

representatives of the four clades were subjected to new 

alignments and analysed separately. Morphological 

characters traditionally used to define subgenera of 

Cordyceps were found to have limited phylogenetic 

information if compared to the ITS phylogenetic tree. The 

ITS tree also questions the present systematics of several 

anamorphic species, e.g. the genera Beauveria and 

Tolypocladium are both polyphyletic. The ITS phylogeny 

confirms the results of some earlier nrDNA studies (SSU, 

LSU) that the plant pathogenic species Claviceps purpurea 

and Epichloë typhina are derived within Cordyceps, 

consequently making the genus Cordyceps paraphyletic. 

We discuss important non-molecular characters, e.g. 

perithecium morphology, host preferences and anamorphic 

affiliation in the light of the inferred ITS phylogeny. We 

conclude that the paraphyletic Cordyceps and its allies 

(anamorphs) deserve a new classification based on a 

combination of molecular and non-molecular evidence.


771 - Are microsporidia really related to fungi ? 

Y. Tanabe 1 , M.M. Watanabe 1 & J. Sugiyama 2* 

1 National Institute for Environmental Studies, Onogawa 

16-2, Tsukuba, Ibaraki 305-8506, Japan. - 2 The University 

of Tokyo, The University Museum; NCIMB Japan, Res. 

Center, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033; Tokai 

Univ. Campus, Orido 3-20-1, Shimizu-shi, Shizuoka, 

Japan. - E-mail: ujsugi@mail.ecc.u-tokyo.ac.jp 

The phylogenetic theory of Microsporidia, which are 

amitochondrial intracellular parasites, has been challenged 

by recent protein sequence-based phylogenies. These 

studies indicate a phylogenetic relationship of 

Microsporidia and Fungi, and in some analyses 

Microsporidia were demonstrated to have evolved from 

within fungal lineages. However, most of these studies did 

not include sequences from basal fungal phyla 

(Chytridiomycota and Zygomycota), and this biased taxon 

sampling could result in an erroneous phylogenetic 

relationship of Microsporidia and Fungi. To further 

investigate the evolutionary origin of Microsporidia, we 

performed molecular phylogenetic analyses with newly 

determined RPB1 and EF-1α sequences from basal fungi. 

Although the phylogenetic position of Microsporidia in the 

EF-1α tree still might be artificially misplaced due to the 

unusually high rate of sequence divergence of the 

microsporidian EF-1α gene, the phylogenies recovered 

based on these two protein sequences do not provide strong 

evidences for a close relationship between Microsporidia 

and Fungi. Moreover, we have identified within EF-1α 

genes a characteristic two amino acid deletion which is 

conserved in all fungal sequences currently available, 

whereas this deletion is absent in microsporidian 

sequences. These results argue against the current view of 

Microsporidia as highly degenerate fungi, suggesting that it 

still remains unresolved whether Microsporidia and Fungi 

are sister taxa. 

772 - The genus Physalacria in Japan 

I. Tanaka 

Lead Discovery Research Labaratories, Sankyo Co., LTD., 

33, Miyukigaoka, Tsukuba-Shi Ibaraki 305-0841, Japan. - 

E-mail: itanak@tsuku.sankyo.co.jp 

The genus Physalacria (Agaricales) forms elliptical to 

subconical and hollow pileus on a central stipe. 

Approximately 30 species have been recognized as 

members of this genus. Berthier (1985) presented a 

worldwide monograph of this genus and three taxa were 

recognized based on their original descriptions from Japan. 

However, their type specimen could not be found 

according to Berthier's monograph. During my 

investigation on minute basidiomycetes in Japan, several 

Physalacria taxa were collected and cultured. The aims of 

the present study are to 1) reexamine the type specimens of 

the 3 Japanese taxa previously described, 2) examine the 

Physalacria flora in Japan and 3) document their cultural 

morphology and mating behavior. In the three taxa, only 

the type specimen of P. orientaliswas preserved in good 

condition and redescribed. The other two taxa were 

considered to be doubtful species. In total, 11 taxa 

including 6, which were not previously described, were 

recognized in Japan. The cultural morphology of 8 taxa out 

of 11 were examined and supported the species 

delimitation in colony appearance, basidiocarp or stroma 

formation, and presence or absence of clamp connections. 

Mating behavior was detected for 5 taxa; two taxa 

exhibited a bifactorial mating system and the other three 

taxa exhibited an amphithallic mating system. 

773 - Pairing test among the strains of Trametes 

versicolor group from different populations from 

Central and South-eastern Europe 

M. Tomsovsky * , L. Homolka & F. Nerud 

Institute of Microbiology, Academy of Science of Science of 

the Czech Republic, Videnska 1083, Prague 4, 142 20, 

Czech Republic. - E-mail: tomsovsk@natur.cuni.cz 

Mating experiments were carried out to investigate genetic 

relationships of different strains of Trametes versicolor 

group. Collections of Trametes versicolor (L.: Fr.) Pilat, T. 

hirsuta (Fr.) Pilat and T. ochracea (Pers.) Gilb. et 

Ryvarden from different localities in the Czech Republic, 

Bulgaria and Montenegro (former Yugoslavia) were 

investigated. Monokaryotic single spore isolates were 

obtained from basidiocarps collected in the field, from 

dikaryotic cultures fruited in the laboratory or by bile-salts 

dedikaryotization. Monokaryons originating from 30 

different dikaryons were mated each other. All pairings 

between different species were indicated by a line of 

demarcation between the paired isolates. Pairing of 

monokaryons belonging to the same species resulted 

mostly in the formation of heterokaryotic mycelium and no 

line of demarcation was observed in any case. No 

intersterility groups were found within the same species. 

Experiments confirmed the validity of contemporary 

species concept of Trametes versicolor group. This work 

has been supported by grant no. 526/02/1216 from the 

Grant Agency of the Czech Republic and by Institutional 

Research Concept no. AV0Z5020903 

774 - Species delimitation in the Russula clelandii 

complex from Australia 

J.E. Tonkin 1* , T. Lebel 2 , T.W. May 2 & C.B. McLean 1 

1 The University of Melbourne, ILFR., Dept Resource 

Management, Burnley Campus, Swan Street, Richmond, 

3121, Victoria, Australia. - 2 Royal Botanic Gardens 

Melbourne, Birdwood Avenue, South Yarra 3141, Victoria, 

Australia. - E-mail: Jenny.Tonkin@rbg.vic.gov.au 



In Australia, the genus Russula (49 listed species) is an 

important widespread mycorrhizal component of forest 

ecosystems. Fruiting bodies are highly coloured and 

conspicuous, and are distinctive components of the 

macrofungal flora that are suitable for ecological 

monitoring. Russula is a taxonomically complex genus and 

contains a number of undescribed taxa. The red-to purplecapped 

species of Russula recorded from southern 

Australia include R. australiensis, Russula clelandii, R. 

lenkunya, R. purpureoflava and R. persanguinea. Various 

Northern Hemisphere names have been used to describe 

members of this group, e.g. R alutacea, R. atropurpurea,R. 

azurea, R.cyanoxantha, R. clusii, R. delica, R. emetica, R. 

expallens, R. fragilis, R. mariae, R. purpurea, R. queletii, 

R. rosacea, R. rubra, and R. sanguinea. Species 

delimitation in this group on the basis of micromorphology 

and macromorphology was investigated using pattern 

analysis (classification and ordination). Nuclear large 

subunit rDNA region was sequenced of representatives of 

each morphological group to determine sequence variation 

in relation to morphological variation. 

775 - Fusarium langsethiae Torp & Nirenberg ined.; a 

new species in section Sporotrichiella 

M. Torp 1* & H.I. Nirenberg 2 

1 National Veterinary Institute, Section of Feed and Food 

Microbiology, Ullevålsveien 68, P.O. Box 8156 Dep, 0033 

Oslo, Norway. - 2 Biologische Bundesanstalt für Land- und 

Forstwirtschaft, Institut für Pflanzenvirologie, 

Mikrobiologie und biologische Sicherheit, Königin-Louise- 

Strasse 19, D-14195 Berlin, Germany. - E-mail: 

mona.torp@vetinst.no 

The new species Fusarium langsethiae Torp & Nirenberg 

ined. has recently been described, illustrated and discussed. 

The species is isolated from kernels of oat, wheat and 

barley in different European countries. Knowledge 

retrieved during surveys on the occurrence of Fusarium 

spp. in grain in Norway, led to the conclusion that the 

regular detection of T-2 toxin and HT-2 toxin in 

Norwegian cereals, especially in oat, is caused mainly by 

this Fusarium species. The combination of observed 

morphological characters and recorded mycotoxin profile 

had not been reported for any other Fusarium species 

before. The fungus has been misidentified mostly as F. 

poae, but it has also been reported as F. sporotrichioides. 

Morphologically it looks similar to F. poae, but it is 

differentiated from F. poae by the powdery appearance on 

PSA and PDA, slower growth, producing less aerial 

mycelium, no odour and the napiform or globose conidia 

are borne on often bent phialides on the agar surface and 

not on the aerial mycelium. No macroconidia are formed. 

Cardinal temperatures for the new species: minimum 7.5 - 

10 °C, optimum 27.5 °C, maximum 35 °C. Based on the 

taxonomic systems of Wollenweber and Reinking (1935), 

Nelson et al. (1983) and Gerlach and Nirenberg (1982) the 

species is placed in the section Sporotrichiella for the time 

being. 

234 


776 - Molecular phylogenetic analyses of the symbionts 

of Costa Rican species of Everniastrum (lichenized 

Ascomycota, Parmeliaceae) 

M.T. Trest * & A. Gargas 

University of Wisconsin - Madison, Department of Botany, 

430 Lincoln Dr, Madison, WI 53706, U.S.A. - E-mail: 

mttrest@students.wisc.edu 

The lichen genus Everniastrum is widespread with over 30 

species inhabiting most tropical and some temperate 

montane regions. Higher species diversity in Andean South 

America and montane Mexico compared to Africa and 

Asia may suggest a New World center of origin and 

relatively recent speciation events. We have collected 

Everniastrum from cloud forest, elfin forest and paramo 

regions of Costa Rica, and are constructing phylogenies 

based on ITS rDNA sequences from both photobionts and 

mycobionts.To investigate hypotheses of codiversification 

of the symbionts we will test for evidence of cocladogenesis. 

Additionally we will assess: 1) whether 

character-based species concepts, including 

chemotaxonomic designations, reflect the evolutionary 

history of the fungal species and 2) whether fungal asexual 

taxa and sexual taxa included in species pairs form 

monophyletic lineages. These preliminary studies will be 

expanded to include multiple-gene analyses of symbionts 

from species of Everniastrum, Cetrariastrum, and 

Concamerella representing their pantropical distribution to 

elucidate phylogeographic relationships of lichens. 

777 - Detection of an anamorph of Tuber dryophilum: 

molecular and immunological evidence 

A. Urban 1* , I. Neuner-Plattner 2 , I. Krisai-Greilhuber 1 & K. 

Haselwandter 2 

1 Department of Mycology and Cryptogamic Botany, 

Institute of Botany, University of Vienna, Rennweg 14, A- 

1030 Wien, Austria. - 2 Department of Microbiology, 

Leopold-Franzens-Universität Innsbruck, Technikerstraße 

25, A-6020 Innsbruck, Austria. - E-mail: 

alexurban@yahoo.com 

We present molecular and immunological evidence that 

Tuber dryophilum, a true truffle belonging to the species 

group of minor white truffles, produces an anamorph of the 

Verticillium type. Nuclear ribosomal sequences were 

amplified from field collected anamorph material with the 

primer pair ITS1F/ITS4. One sequence was nearly identical 

(similarity >99%) to a reference sequence of Tuber 

dryophilum. The identification of this anamorph was 

supported by indirect immunofluorescence using an 

antibody raised in rabbits against Tuber melanosporum 

antigene which has been demonstrated to be valuable for 

the detection of various stages in the life cycle of several 

Tuber species. According to our knowledge, this is the first 

report of an anamorph in the Tuberaceae. The potential 

ecological and biotechnological significance of anamorphs


in the propagation of ectomycorrhizal fungi is not yet fully 

acknowledged. Seedlings inoculated with noble truffle 

species are often infected with less valuable white truffle 

species (e. g. Tuber dryophilum), either in the nursery or in 

the field, sometimes without plausible explanation for the 

way of infection. The existence of an alternative way of 

propagation in the genus Tuber indicates that additional 

precaution against cross contamination in inoculation 

experiments is required. 

778 - Differentiation of Leptographium spp. found in 

Canada 

A. Uzunovic 1* , S. Alamouti 2 , J.-J. Kim 2 & C. Breuil 2 

1 Forintek Canada Corp., 2665 East Mall, Vancouver, B.C., 

Canada. - 2 University of British Columbia, Department of 

Wood Science, Vancouver, Canada. - E-mail: 

adnan@van.forintek.ca 

Leptographium is one of the most commonly found genera 

of fungi causing bluestain in conifers. Some species are 

known to be tree pathogens. Some Leptographium spp are 

linked to specific wood infesting insects, especially bark 

beetles, however their biology, including their potential to 

grow in fresh woody substrates and to cause stain, is not 

fully known. The taxonomy of Leptographium spp is 

complex, and identification to species is often difficult 

when based on the existing taxonomy keys. Molecular 

biology tools are being developed but are not yet available. 

We looked at number of reference Leptographium isolates 

from existing culture collections and we included those 

collected in Canada over the last decade. We investigated 

additional attributes that could assist species delineation. 

This included growth rates on 2% malt extract agar at 

different temperatures and colony morphology at different 

stages of growth. We also compared the ability of some of 

the isolates to colonize freshly cut lodgepole pine phloem 

and to stain the sapwood. The results of these experiments 

and our observations are reported here. 

779 - Lactarius well-represented in West-Africa? Benin 

explored! 

A. Verbeken 1* , A. De Kesel 2 & P. Van Rooij 1 

1 Ghent University, Dpt. Biology, Group Mycology, K.L. 

Ledeganckstraat 35, B-9000 Gent, Belgium. - 2 National 

Botanic Garden of Belgium (BR), Dpt. Thallophyta, 

Domein van Bouchout, B-1860 Meise, Belgium. - E-mail: 

mieke.verbeken@rug.ac.be 

The up to now 80 Lactarius species described from tropical 

Africa are particularly well-represented in the Guineo- 

Congolian rain forests (Congo-Kinshasa) and the 

Zambezian miombo woodlands (Zimbabwe, Zambia, 

Burundi). In these ecosystems the genus Lactarius is one of 

the most important ectomycorrhiza formers. The Sudanian 

woodlands cover large parts of West-Africa and are, 

compared to the Zambezian woodlands, poorly explored. 

They are often dominated by the ectotrophs Isoberlinia and 

Uapaca, while they lack the typical Zambezian trees 

Brachystegia and Julbernardia. Therefore, it seemed logic 

that the species diversity in the Sudanian woodlands would 

be less important than in East-Africa. Some scattered 

records of miombo woodland species in West-Africa 

already suggested an overlap in the Lactarius mycoflora, 

but more studies were needed. Intensive collecting in the 

Isoberlinia and Uapaca savanna woodlands in Benin, 

resulted in at least 24 Lactarius species, including some 

undescribed taxa. In spite of the differences in dominant 

ectotrophs, most Lactarius species from the studied area 

are also known from Zambezian woodlands. This study is 

the first to reveal a marked overlap of the Zambezian and 

Sudanian ectomycorrhizal mycoflora. It also indicates the 

absence of the Sudanian species in the Central African rain 

forests (Guineo-Congolian) and consequently suggests a 

disjunct distribution of the savanna woodland species of 

Lactarius throughout Africa. 

780 - Taxonomy and phylogeny of Septoria 

G.J.M. Verkley, A. Van Iperen & M. Starink-Willemse * 

Centraalbureau voor Schimmelcultures, P.O.Box 85167, 

NL-3508 AD, The Netherlands. 

In the anamorph genus Septoria Sacc. over 2000 names 

have been described. Most taxa cause leaf spot on the host 

plant. Known teleomorphs belong in Mycosphaerella 

Johanson (Dothideales, Ascomycota), a cosmopolitan 

genus with over 20 reported anamorph genera. Crous et al. 

(2001, Mycologia 93: 1081-1101) showed that 

Mycosphaerella is monophyletic, based on ITS rDNA 

sequences. Both strains of S. aceris (teleom. M. latebrosa) 

studied clustered in a clade with Cercospora, separate from 

S. tritici (teleom. M. graminicola), indicating that Septoria 

anamorphs may have evolved in several lineages. We 

further investigated the phylogenetic position of Septoria 

anamorphs using ITS and partial 28S rDNA sequences of a 

larger set of taxa, some with known teleomorphs, isolated 

from a wide array of plant families. An additional eight 

anamorph genera linked to Mycosphaerella were also 

represented in the analyses. The results show that Septoria 

spp. all evolved within Mycosphaerella, but not as a result 

of a single event. The data support a clade comprising S. 

tritici and S. passerini from grasses, close to Ramularia 

spp. Septoria spp. from several deciduous trees represent 

relatively distant lineages. In contrast, many of the Septoria 

spp. from various herbaceous plant families have almost 

identical ITS sequences, and also often very similar 

morphologies in vitro and in planta, indicating that they are 

closely related. They are grouped with Cercospora spp. in 

a large, unresolved cluster. 



781 - Morphological variation of spore ornamentation 

within the genus Gomphus 

M. Villegas * & J. Cifuentes 

Fac. de Ciencias, UNAM, A.P. 70-399 Cuidad 

Universitaria, Del. Coyoacan. C.P. 04510, México, D.F., 

Mexico. - E-mail: mvr@hp.fciencias.unam.mx 

Traditionaly the morphology of the spores has been an 

important source of characters to classify species in 

Gomphus. Neverthless, the understanding of morphological 

variation patterns in spore ornamentation within species is 

misleading. The goal of this research was to evaluate 

ornamentation similarity among differents species 

described as Gomphus and, to identify discrete states to use 

them in systematics. We analyzed the ornamentation in 

sixteen taxa using scanning electron microscopy (SEM). 

Three differents patterns of ornamentation shape and two 

of ornamentation distribution were differentiated. 

Reticulate-lacunose ornamentation was only found in G. 

retisporus, verrucose ornamentation was pretty common 

and it was found in G. clavatus, G. floccosus, and G. 

subclavaeformis; various degrees of verrucae fusion were 

observed. Ornamentation distribution is very characteristic 

in G. grandis and G. retisporus, with a distinct zone 

without ornamentation near hilar appendix. A preliminary 

phylogenetic evaluation, including other available micro 

and macromorphological characters, indicates that spore 

ornamentation provides significant evolutionary 

information within the genus. 

782 - The relevance of anamorphic states for a natural 

classification of Agaricales: results from morphological 

and molecular phylogenetic studies 

G. Walther 1* & M. Weiß 2 

1 University of Jena, Dornburger Str. 159, D-07743 Jena, 

Germany. - 2 University of Tuebingen, Auf der Morgenstelle 

1, D-72076 Tuebingen, Germany. - E-mail: 

Grit.Walther@rz.uni-jena.de 

Single-spore cultures of more than 180 species of 

Agaricales were studied morphologically and by molecular 

phylogenetic methods. The morphological investigations 

revealed anamorphic states in more than 110 species, 

which are often specific at the genus or family level. 

Phylogenetic analysis of the 5' terminal domain of the 

nuclear gene coding for the ribosomal large subunit 

resulted in several well-supported groups that are 

consistent with anamorph morphology. Our results indicate 

that the anamorphic state provides valuable characters for a 

natural classification in the Agaricales. 

236 


783 - Molecular variation among species of Verticillium 

D. Weber 1* , O. Strunnikova 2 & M. Typas 3 

1 Dept. of Botany and Range Science, Brigham Young 

University, Provo, Utah 84602, U.S.A. - 2 Research Institute 

for Agricultural Microbiology, Podbelsky Shoose 3, St. 

Petersburg, Pushkin 8 189620, Russia. - 3 Dept. of Biology, 

University of Athens, Panepistemiopolis, Athens TK 15701, 

Greece. - E-mail: darrell_weber@byu.edu 

The genus Verticillium within fungi imperfecti contains a 

heterogeneous group of asexual species. Many of these 

species are important plant pathogens or entomopathogens. 

The classification is based on morphology of the vegetative 

reproductive organs namely conidiophores, conidia and 

resting structures. Some analyses have been done on 

Verticillium using immunochemical and RFLPs methods. 

We analyzed the variation of the nuclear ribosomal internal 

transcribed spaces (ITS) of the following species: V. alboatrum, 

V. dahliae, V. longisporum, V. lamenicola, V. 

fungicola, V. catenulatum, V. tricopus, V. nigrescens, V. 

chlamydosporium, V. chlamydosporium, V. theobromeae, 

and V. psalliotae. The DNA was duplicated using PCR. 

The DNA sequences were determined using a Perkin Elmer 

ABI PRISM sequencing apparatus. The relationship of the 

different species to each other was determined. A 

concensus tree was determined for the isolates of V. 

dahliae. 

784 - The evolution of cyanobacterial lichens inferred 

by phylogenetic analyses of combined mtSSU rDNA 

and nLSU rDNA sequence data 

E. Wiklund * & M. Wedin 

Dept. of Ecology and Environmental Science, Umeå 


elisabeth.wiklund@eg.umu.se 

Fungi associating with cyanobacteria have been found 

among the oldest verified fungal fossils of the Lower 

Devonian Rhynie chert. The extant cyanobacterial lichen 

associations, particularly of the Lecanorales suborder 

Peltigerineae, are also often claimed to include the most 

primitive extant lichens. It is, however, unclear if the 

association with cyanobacteria has arisen once or several 

times, if the suborder Peltigerineae as currently 

circumscribed is monophyletic, or if it is the sistergroup to 

the rest of the Lecanorales. Here, we will present 

preliminary results from phylogenetic analyses of two 

independent gene loci, including representatives of most 

families in Lecanorales suborder Peltigerineae, and several 

representatives of other Lecanoromycetes. These analyses 

are utilised to test several evolutionary hypotheses 

regarding the evolution and phylogeny of cyanobacterial 

lichen associations.


785 - Phylogeny of the Gymnopus/Marasmiellus group 

based on 28s nrDNA sequence 

A.W. Wilson * & D.E. Desjardin 

Department of Biology, San Francisco State University, 

1600 Holloway Ave. San Francisco, CA 94132, U.S.A. - Email: 

andson@sfsu.edu 

Nuclear ribosomal large subunit sequence was collected for 

89 taxa representing members of the 

Gymnopus/Marasmiellus group, referred to as the 

Lentinula clade in Moncalvo et al. (2002). Tissue was 

extracted from ^70 specimens via PCR and cycle 

sequencing reactions. The remaining sequences were 

obtained from GenBank or the fore-mentioned authors. 

Sequence was aligned and phylogenetic analyses were 

performed using PAUP. Several methods of phylogenetic 

analysis were utilized including weighted parsimony, 

maximum likelihood and neighbor joining. Bootstrap 

values were used to indicate structural soundness of groups 

represented in the tree. The aim of this study is to gather 

molecular evidence that will help define groups within the 

Gymnopus/Marasmiellus cluster that have been 

traditionally recognized using morphological taxonomy. 

786 - Developmental similarities in Echinosporangium 

transversale and Mortierella multidivaricata. Early 

maturity hypothesis 

M. Wrzosek 

Warsaw University, Aleje Ujazdowskie 4, 02-634, Warsaw, 

Poland. - E-mail: mwrzosek@ulmus.bot.uw.edu.pl 

Mortierella multidivaricata and Echinosporangium 

transversale are related genera, and there are classified in 

the family Mortierellaceae, ordo Mortierellales (Voigt, 

Wostemeyer 2001). There are two basic features common 

for both genera and other species of the family 

Mortierellaceae -the presence of anastomoses between 

hyphae and the lack of the columellae. The main 

substantial difference concerns the morphology of the 

sporangia. Echinosporangium has non-typical sporangium 

of sausagous shape with spiny ends while sporangia of 

Mortierella multidivaricata are globose. Phylogenetical 

study (my PhD. thesis, 2000, Voigt and Wostemeyer 2001) 

confirm close affinity of both genera. Echinosporangium is 

located in phylogenetical tree within genus Mortierella. 

Are there any in vivo observations which can help us in 

understanding on one hand the phylogenetic affinity 

resulting from molecular methods and on the other 

morphological dissimilarities between sporangia? Close 

similarities in early ontogenetical development of asexual 

structures in Mortierella multidivaricata and 

Echinosporangium transversale were observed. The 

differences appear at the moment of the spores formation. 

Taking into account these observations and the results of 

molecular studies, we may propose the following 

hypothesis: Small numbers of mutations placed in genes 

for regulation of sporangium development could result in 

apparent morphological changes in asexual structures of 

Echinosporangium. 

787 - Peniophora in Taiwan with simple-septate hyphae 

S.H. Wu 

National Museum of Natural Science, Taichung, Taiwan 

404, Taiwan. - E-mail: shwu@mail.nmns.edu.tw 

In this survey of Taiwanian Peniophora species with 

simple-septate hyphae five species are presented: P. 

bicornis, P. borbonica, P. maliensis, P. reidii, and P. 

taiwanensis. The former four species are newly recorded 

from Taiwan, and the last one is new to science. 

Description and line drawing are given for each species. 

Cultural and cytological studies are also provided for the 

five species. 

788 - Diversity of Sporidesmium and related fungi in 

China 

W.P. Wu * & Y. Zhang 

RD China, Novozymes China, 22 Xinxi Zhong Lu, Shangdi 

Zone, Haidian District, Beijing 100085, China. - E-mail: 

WUWP@novozymes.com 

The Sporidesmium-Endophragmiella complex includes 

dematiaceous hyphomycetes which holoblastically produce 

phragmosporae. This group of fungi is mostly found on 

dead branches or rotting wood. Only one species from 

literatures was known in China. However, this study 

showed that they widely distribute in China and a total of 

147 species in 23 genera have been identified, including 37 

new species and 14 new combinations. All species have 

been fully described and illustrated based on Chinese 

specimens. 

789 - Revision of the genus Septoriella Oudem. 

(Coelomycetes) 

W.P. Wu 1* , B.C. Sutton 2 & Y. Zhang 1 

1 RD China, Novozymes China, 22 Xinxi Zhong Lu, Shangdi 

Zone, Haidian District, Beijing 100085, China. - 2 Apple 

Tree Cottage, Black Heaph, Wenhaston, Near Halesworth, 

IP90 9HD, U.K. - E-mail: WuWP@novozymes.com 

A revision of the coelomycete genus Septoriella Oudem. is 

made and 14 species are accepted. Among them, 5 are new 

species, S. fusiformis sp. nov., S. hendersoniae sp. nov., S. 

intermedia sp. nov., S. latispora sp. nov., and S. petrakii sp. 

nov.; and two are new combinations, S. mastigospora 

(Petr.) comb. nov. (syn. Urohendersoniella mastigospora 



Petr.), and Septoriella panicicola (Petr.) comb. nov. (syn. 

Hendersonia panicicola Petr.). Other accepted species 

include S. canadensis Nag Raj, S. caroliniana Nag Raj, S. 

junci (Desm.) B. Sutton, S. phragmitis Oudem., S. rockiana 

(Petr.) Nag Raj and S. thalassica (Speg.) Nag Raj. All other 

species names described in Septoriella have been excluded 

from the genus and reidentified. This results in 1 new 

combination, Phaeoseptaria philippinensis (Sacc.) comb. 

nov. (S. philippinensis Sacc.). Septoriella atrata Sacc. is 

regarded as a synonym of Septoria heterochora Desm. In 

addition, one new species of the genus Linochorella Syd. & 

P. Syd., L. cylindrospora sp. nov., is also described. 

Related taxonomic problems are also discussed. A key to 

all accepted species in the genera Septoriella and 

Linochorella is provided. 

790 - DNA sequence and IGS RFLP analyses and 

molecular markers for identification of Fusarium poae, 

F. langsethiae, F. sporotrichioides and F. kyushuense 

T. Yli-Mattila 1 , P. Konstantinova 1 , R.L. Mach 2 , I.A. 

Alekhina 3* , S.A. Bulat 3 , C.M. Kullnig-Gradinger 2 , C.P. 

Kubicek 2 & S.S. Klemsdal 4 

1 University of Turku, Lab. of Plant Phys. and Mol. Biol., 

Dept. of Biology, FIN-20014 Turku, Finland. - 2 Technical 

University of Vienna, Inst. for Chem. Engineering, 

Microbial Biochem., Getreidemarkt 9/166, A-1060 Vienna, 

Austria. - 3 Petersburg Nuclear Physics Institute, Lab. of 

Eukaryote Genetics, Dept. of Mol. and Radiation 

Biophysics, Gatchina 188350, Russia. - 4 The Norwegian 

Crop Research Institute, Plant Protection Centre, 

Høgskoleveien 7, N-1432 Ås, Norway. 

Fusarium langsethiae was recently described to 

accommodate 'powdery' isolates of F. poae, which 

morphologically resemble F. poae, but whose metabolite 

profile (including toxins) is similar to that of F. 

sporotrichioides. Sequence analysis of ITS1 and 2 

separated F. langsethiae from F. poae, but was unable to 

separate all F. sporotrichioides strains from F. langsethiae 

strains. Sequence analysis of ß-tubulin distinguished all 

four species but did not resolve them phylogenetically. The 

high intraspecific variability of the IGS sequences were 

found useful to group isolates according to their geographic 

origin. Analysis of all combined sequences reliably 

separated all species studied. Our results are in accordance 

with the results of the UP-PCR hybridization analysis. IGS- 

RFLP analysis also allowed differentiation between the 

closely related species. According to the phylogenetic 

analysis of IGS-RFLP patterns F. langsethiae, F. 

sporotrichioides, F. poae and F. kyushuense strains formed 

four well-supported clades with high bootstrap values. 

Based on the sequence differences in the IGS region, 

species-specific primers were designed for the F. 

langsethiae/F. sporotrichioides group and for F. poae. The 

two phylogenetic subgroups of F. langsethiae, found by 

IGS sequencing analysis, were clearly separated on the 

basis of size differences of the amplification products with 

primers CNL12/PulvIGSr (specific for the F. 

langsethiae/F. sporotrichioides group). 

238 


791 - Molecular, morphological and phylogenetic 

analysis of Fusarium avenaceum / F. arthrosporioides / 

F. tricinctum species complex 

T. Yli-Mattila 1* , S. Paavanen-Huhtala 1 , S.A. Bulat 2 , I.A. 

Alekhina 2 & H.I. Nirenberg 3 

1 University of Turku, Lab. of Plant Phys. and Mol. Biol., 

Dept of Biology, FIN-20014 Turku, Finland. - 2 Petersburg 

Nuclear Physics Institute, Lab of Eukaryote Genetics, 

Dept. of Mol. and Radiation Biophysics, Gatchina 188350, 

Russia. - 3 Inst. fur Mikrobiologie, BBA, Königin-Luise-Str. 

19, D-14195 Berlin, Germany. - E-mail: tymat@utu.fi 

The great variation in the IGS region offers more potential 

to resolve intraspecific phylogenetic relationships, while 

beta-tubulin gives more reliable phylogenetic information 

at species level. The European F. avenaceum strains of 

main group II had identical beta-tubulin sequences with 

one American F. avenaceum strain and four European F. 

arthrosporioides strains, while F. avenaceum strains of 

main group I were closely related to two European F. 

arthrosporioides strains and to one Japanese F. anguoides 

strain. According to the combined beta-tubulin/IGS/ITS 

sequence tree, European F. arthrosporioides strains were 

divided into four groups. F. tricinctum strains formed a 

well-supported cluster, in which two European clusters 

were separated from one African isolate. In the IGS 

sequence tree two European F. acuminatum strains 

together with one American F. acuminatum strain formed a 

cluster, which was separate from another American F. 

acuminatum strain. The F. acuminatum cluster was nested 

within the large F. tricinctum cluster together with one F. 

reticulatum strain in the combined IGS/beta-tubulin tree. 

Several strains may be intermediate between the F. 

avenaceum / F. arthrosporioides / F. anguoides and F. 

tricinctum clusters and represent their own species. These 

results are partially supported by the results of UP-PCR 

hybridization analysis. Thus the molecular results may be 

helpful in future revision in the taxonomy of these species. 

792 - Clarification of the taxonomic relationhips among 

onion neck rot Botrytis spp.: B. allii is an interspecific 

hybrid of B. aclada and B. byssoidea 

D.S. Yohalem * , K. Nielsen & M. Nicolaisen 

Danish Institute of Agricultural Science, Reserach Center 

Flakkebjerg, Slagelse, Denmark. - E-mail: 

david.yohalem@agrsci.dk 

Three species of Botrytis are recognized as associated with 

neck rot of onions. Due to the difficulty of distinguishing 

them by morphological criteria and lack of type material 

associated with two of the names, several synonomies have 

been proposed. Species may be differentiated by conidial 

size and shape, but the character is subtle, variable and 

there is some overlap. Both the smallest spored group and 

the largest spored group have 16 mitotic chromosomes, 

while the intermediate group has 32. Based on significant


differences in Nei's coefficient of genetic differentiation 

derived from universally primed PCR (UP-PCR) 

fingerprints it was possible to recognize distinctions among 

the three nomengroups and B. cinerea and B. squamosa. 

Primers, designed from a sequence characterized UP-PCR 

fragment, were used for direct sequencing of DNA from 

isolates of the 16 chromosome nomengroups. Because of 

apparent ambiguities in the UP-PCR fragment from the 32chromosome 

group, it was cloned and sequenced. 

Clustering show identity with the small-spored B. aclada 

and the large-spored B byssoidea for the two cloned 

molecules from B. allii. Further, the internally transcribed 

spacer rDNA (ITS) amplicons of B. aclada has 2 SphI 

restriction sites; that of B. allii has 1 SphI site. The 

cumulative data suggest that the three groups are 

genetically distinct from each other and that isolates B. 

aclada and B. byssoidea were the ancestors of B. allii. 

793 - Morphological and molecular characterization of 

small-spored Alternaria species 

S.H. Yu * & B.R Kim 

Chungnam National University, College of Agriculture, 

220 Gung-dong, Yuseung-gu, Daejeon 305-764, Korea. - 

E-mail: shunyu@cnu.ac.kr 

The importance and diversity of the genus Alternaria 

highlights the need for accurate identification of species. 

However, many small-spored Alternaria isolates have been 

misidentified due to the use of spore size as the only 

identifying character. In this study eighty one isolates of 

small-spored Alternaria were segregated into 

morphological and pathogenic groups or species as A. 

gaisen, A. mali, A. tenuissima, A. longipes, A. citri, A. 

arborescens, A. infectoria, and A. alternata. Molecular 

characteristics of these isolates were determined using 

universal rice primer (URP)-PCR analysis, sequence 

analyses of nuclear internal transcribed spacer (ITS) and 

mitochondrial small subunit (SSU) ribosomal DNA 

(rDNA). Based on cluster analysis of URP-PCR fragment 

patterns, the eighty one isolates were segregated into 

distinct groups that are morphologically similar but 

identifiable as A. gaisen, A. mali, A. tenuissima, A. 

longipes, A. citri, A. arborescens, A. infectoria, and A. 

alternata. Based on analyses of ITS and mitochondrial 

SSU rDNA sequence data, there was no variability in ITS 

and mt SSU rDNA sequences for species among A. gaisen, 

A. mali, A. tenuissima, A. longipes, A. citri, A. arborescens, 

and A. alternata. However, A. infectoria was differ from 

other species at 21 and 16 nucleotides in ITS and mt SSU 

rDNA, respectively. 

794 - A contribution to the identification of 

Trichoderma species in Iran 

- Zafari 1* , - Ershad 2 , - Zare 2 & - Alizadeh 3 

1 Bu Ali Sina University and Tarbiat Modarres University, 

Bu Ali Sina University Hamadan, Iran. - 2 Plant Pestes and 

Diseases Research Institute, Tehran, Iran. - 3 Tarbiat 

Modarres University, Tehran, Iran. - E-mail: 

zafari_d@yahoo.com 

The aim of the present study is to identify Trichoderma 

species isolated from Iran. Soil samples were collected 

from different parts of Iran focusing on the agricultural 

fields. Trichoderma selective media and malt extract agar 

(MEA) were used to isolate Trichoderma species from the 

soil samples and Petri plates were incubated at 25 °C in the 

dark for the first 24 hours and then 12/12 (dark/ flourescent 

light) with the same temperature. All the cultures were 

purified on 2% water agar by hyphal tip method prior to 

morphological examination. Morphological observations 

were carried out on the cultures grown on 2% MEA at 20 

°C under ambient laboratory conditions. Microscopic 

features of conidiophores and shape and size of conidia 

were studied and recorded 3-5 days after inoculation. Out 

of 350 obtained isolates, using morphological features ten 

species (T. citrinoviride, T. longibrachiatum, T. 

saturnisporum, T. hamatum, T. harzianum, T. inhamatum, 

T. tomentosum, T. virens, T. asperellum, T. koningii) were 

identified. Among the species T. harzianum with 150 and 

T. virens with 57 isolates were the most frequent species. 

795 - An integrated approach to the taxonomy of plantassociated 

Verticillium species 

R. Zare 

Plant Pests & Diseases Research Institute, P.O. Box 1454, 

Tehran 19395, Tehran, Iran. - E-mail: 

simplicillium@yahoo.com 

Molecular approaches were used to re-evaluate the 

morphological criteria used to identify plant-associated 

species of Verticillium Nees. ITS-RFLPs divided the 31 

studied strains of seven Verticillium species, (including the 

type species) into four clusters. Cluster one comprised 

strains of the type species, V. luteo-album (Link: Fries) 

Subramanian, cluster two V. albo-atrum Reinke & 

Berthold, V. dahliae Klebahn, V. nubilum Pethybridge and 

V. tricorpus Isaac; cluster three comprised strains of V. 

theobromae (Turconi) E. Mason & S. Hughes, and cluster 

four comprised strains of V. nigrescens Pethybridge. Betatubulin 

gene RFLPs offered a higher degree of resolution, 

distinguishing all seven species from each other. The 

highest degree of resolution was obtained from 

mitochondrial DNA-RFLPs that divided strains of V. 

theobromae and V. nigrescens into infraspecific groups. 

The beta-tubulin gene digested by Hae III offers a reliable 

way to separate the two economically important and 

controversial species V. albo-atrum and V. dahliae. 



796 - Species of the genus Macentina Vezda 

(Ascomycota, Verrucariaceae) as tertiary relicts of the 

lichen flora Ukraine 

S.D. Zelenko 


2, 01601 Kiev, Ukraine. - E-mail: skondr@botan.kiev.ua 

Natural ranges of the lichens of the genus Macentina are 

concentrated in two distribution centers. The first center is 

located in tropical forests of Africa, where the folicolous 

Macentina species are growing. The second center is 

located in Europe and includes corticolous species. M. 

dictyospora is known from Sweden, Spain, Switzerland, 

the Czech Republic, Slovakia, and Ukraine. M. abscondita 

was registered in Great Britain, Austria, and Ukraine. M. 

stigonemoides is reported from Great Britain, France, 

Spain, Belgium, and Sweden. The natural habitats of 

corticolous species of the genus Macentina are connected 

with suboceanic and mountane regions of Europe. It gives 

evidence that Macentina species prefer humid habitats. In 

Ukraine, species of the genus Macentina are known from 

the Dniper and Podolia heights. These geomorphological 

structures have never been covered by glaciers during the 

Quarternary, and thus acted as refugia for species of forest 

vegetation, and the Tertiary flora as well. European species 

of the genus Macentina are corticolous lichens, they 

probably were widespread in broad-leaved forests during 

the Neogene. In our opinion, species of the genus 

Macentina are Tertiary relicts of the lichen flora of 

Ukraine. This conclusion is based on the analysis of their 

geographic distribution, ecology of Macentina species, and 

data of geological and palaeobotanical studies. 

797 - Gibberella konza, a new species from prairie 

grasses 

K.A. Zeller 1 , B.A. Summerell 2* , J.F. Leslie 1 & S. Bullock 2 

1 Kansas State University, Manhattan, Kansas, U.S.A. - 

2 Royal Botanic Gardens, Sydney, Australia. 

The Gibberella fujikuroi species complex (Fusarium 

section Liseola and allied taxa) is composed of an 

increasingly large number of morphological, biological, 

and phylogenetic species. Most of the known species in 

this group have been isolated from agricultural ecosystems, 

or have been described from only a small number of 

isolates. We have been sampling Fusarium communities 

from native prairie grasses in Kansas and have recovered a 

large number of isolates that superficially resemble F. 

anthophilum. We have used a combination of 

morphological, biological, and molecular characters to 

describe a new biological species, Gibberella konza 

(Gibberella fujikuroi mating population I [MP-I]), from 

native prairie grasses in Kansas. Although female fertility 

for field isolates of this species appears to be low, G. konza 

is heterothallic, and we have generated reliably female 

fertile mating population tester strains for this biological 

240 


species. The F. konzum anamorph is differentiated from F. 

anthophilum and from other Fusarium species in section 

Liseola by mating compatibility, morphology, AFLP 

fingerprint profile, and the DNA sequence of the ß-tubulin 

gene. 

798 - Floristic composition of rust fungi in Tibetian 

Everest Himalaya 

J.Y. Zhuang 

Institute of Microbiology, Academia Sinica, P.O. Box 2714, 

Beijing 100080, China. - E-mail: zhuangjy@sun.im.ac.cn 

The rust flora of Tibetan Everest Himalaya mainly consists 

of temperate genera and species. Among the 22 known 

genera (excluding form genera), Puccinia, Uromyces and 

Phragmidium are cosmopolitan; Chrysomyxa, 

Coleosporium, Cronartium, Gymnosporangium, 

Hyalopsora, Melampsora, Melampsoridium, 

Pucciniastrum, Thekopsora and Uredinopsis are mainly 

north temperate; Blastospora, Miyagia and Pucciniostele 

are typically eastern Asian; Endophyllum, Phakopsora, 

Physopella, Pileolaria, Ravenelia and Trochodium are 

tropical. Of the 155 known species, 20 (sharing 13% of the 

total) are endemic in Himalaya. Apart from cosmopolitan 

species (12%) and species restricted to China (3%) and 

Himalaya, the predominant floral elements are eastern 

Asian (27%), northern temperate (20%) and Old World 

temperate (8%). Nine species (6%) are disjunctive between 

eastern Asia and North America. No typical Central Asian 

species is found. The arid and frigid plateau steppe and 

desert in northern Tibet impede intermigration of rusts, 

resulting in the separation of the rust flora from that of 

Central Asia. Only about 10% of the total known species 

are tropical in affinities with the majority of these (8%) 

identifiable with species of general distribution in Indo- 

Malaysia. The lower percentage of tropical species 

suggests that the connexion between the rust flora and that 

of the tropics seems weak.

IMC7 Main Congress Theme III: PATHOGENS AND NUISANCES, FOOD AND MEDICINE Posters 

799 - Monitoring of patulin production of Penicillium 

expansum using Fungi static 2-Deoxy D-Glucose 

N.F. Abo-Dahab 

Botany & Microbiology Department, Faculty of Science, 

Al-Azhar University, Assuit, 71524, Egypt. - E-mail: 

abo_dahab@hotmail.com 

Three isolates of Penicillium expansum which known as 

patulin producer were treated with three concentrations of 

2-deoxy-D-Glucose. The 2-deoxy-D-Glucose which used 

as a post harvest disease control of P. expansum. The High 

Performance Liquid Chromatography (HPLC) data 

revealed that the 2-Deoxy-D-Glucose reduced patulin 

production on the pure culture but in the main time 

enhanced the other metabolites production like citrinin and 

ochratoxin A. 

800 - Bacteriological and mycological study of external 

otitis in patients referred to ENT cilinic in sari in 1999 

M. Ahanjan & T. Shokohi * 

Microbiology Dept., Medical school khazar boulved sari 

Iran, Iran. 

We decided to study the etiological agents of external otitis 

in summer in 78 sampling were collected from the patients 

who were diagnosed clinically as external otitis. The 

samples was performed by two sterile swabs wetted in 

sterile saline and transferred to laboratory as soon as 

possible to culture in media. The sample was inoculated on 

blood agar and Macconky agar incobated for 24 hours and 

was followed by the biochmical tests to identify the species 

of bacteria. Meanwhile the same sample was inoculated on 

saborauds dextrose agar incubated at for 48 hours to 

identify fungi. A wet mount with 10% KOH for 

morphological observation. In this study out of 101 (59 

females and 42 males) cases suspected to clinical external 

otitis 39 (38.6%) and 23 (22.8%) cases were of bacterial 

and fungal respectively and 16 (15.2%) cases were of both. 

x test showed significant relationship between the 

occurrence of otomycosis and gender of the patients 

(p


803 - Black spores Aspergilli in Turkish raisin 

T. Askun 1* & R. Eltem 2 

1 Balikesir University, Faculty of Science and 

LiteratureBiyoloji Böl., Balikesir, 10100, Turkey. - 2 Ege 

University, Faculty of Bioengineering, Department of 

Bioengineering, Ýzmir, 35100, Turkey. - E-mail: 

taskun@balikesir.edu.tr 

In this study, black spores Aspergillus strains isolated from 

fresh grapes and raisins, yielded in 1999, in 20 different 

vineyards which contain 55 different stations in Izmir and 

Manisa province, were given in their detailed descriptions. 

These are follows: A. aculeatus, A. awamori, A. 

carbonarius, A. ficuum, A. foetidus, A. foetidus var. acidus, 

A. foetidus var. pallidus, A. heteromorphus, A. japonicus, 

A. niger, A. olivaceo-fuscus, A. phoenicis, A. pulverulentus 

and A. tubingensis. 

804 - Computer tomograph for the living trees internal 

inspection 

V. Bahyl 1 , M. Pavlik 2* , T. Phan Van 1 & J. Vyboh 1 

1 Dept.of physics and applied machanics, Technical 

university, Masarykova 24, Zvolen 960 53, Slovakia. - 

2 Dept.of forest protection and game management, 

Technical university, Masarykova 24. Zvolen 960 53, 

Slovakia. - E-mail: mpavlik@vsld.tuzvo.sk 

There was been realised the computer tomograph (CT) 

construction at the Technical Univerity in Zvolen which 

allows to inspect the internal structure of trees. This 

construction allows inspect the trees up to 18 cm in 

diameter but the tomograph going to the 70 cm in diameter 

is under construction. The CT is working in the fan beam 

geometry and the radioisotope source of radiation is used. 

The parallel beam geometry is also possible under the 

special conditions. The detector system is based on the 

CdZnTe semiconductor detector arrays. The picture of 

internal structure can be obtained up to five minutes. 

According to the state authorities approval the CT is not 

ecologically dangerous. The CT is portable (meanwhile 

120 kg of weight) and it is working under 24 volts of DC. 

The reconstruction algorithms are of our own realisation 

and the communication software is very user friendly. With 

our CT we can inspect the internal structures of the living 

trees and we are able to detect the internal cracks, red 

decay, rot, root of knots and so on. The future of our 

construction we see in the possibilities to detect the healthy 

conditions of the internal structures of the tree far before 

their consequences are visible in outside. Next it is possible 

to inspect the wood quality before cutting and to be able to 

take special care of the high quality wood material. 

242 


805 - A chemical comparison of two types of antifungal 

defence: reaction zones and branch protective zones in 

Eucalyptus globulus 

K.M. Barry 1* , N.W. Davies 2 & C.L. Mohammed 1 

1 School of Agricultural Science, University of Tasmania, 

GPO Box 252-12 Hobart Tasmania 7001, Australia. - 

2 Central Science Laboratory, University of Tasmania, 

GPO Box 252-74 Hobart Tasmania 7001, Australia. - Email: 

Karen.Barry@ffp.csiro.au 

While reaction zones are actively formed in response to 

fungal infection of sapwood, protective zones form 

passively during the process of branch death. In Eucalyptus 

globulus, the protective zone appears to form across the 

whole branch and is rarely surpassed by invading decay 

fungi after branches are pruned. When living branches are 

pruned before the protective zone forms, invading decay 

often spreads into the sapwood, where it is constricted by a 

reaction zone. Samples of branch wood, protective zones, 

stem sapwood near the branch and reaction zones were 

prepared from 19 living or dead branches (from 5 different 

trees) and extracted with 70% acetone. Total phenol levels 

of reactions zones were almost double that of the protective 

zones, but both were significantly increased from the stem 

sapwood and branch sapwood. Phenolic compounds were 

identified by liquid chromatography - mass spectrometry 

and included hydrolyzable tannins (pedunculagin, 

casuarinin, methyl-di-galloylglucose, tri- and tetragalloylglucose), 

epicatechin-3-O- gallate and a flavonone 

glycoside. This poster will present the trends in semiquantitative 

levels of these compounds between the four 

tissues. 

806 - Acacia mangium heartrot: preliminary studies of a 

poor defence 

K.M. Barry 1* , N.W. Davies 2 , R.S.B. Irianto 3 & C.L. 

Mohammed 1 

1 School of Agriculture, University of Tasmania, GPO Box 

252-12 Hobart Tasmania 7001, Australia. - 2 Central 

Science Laboratory, University of Tasmania, GPO Box 

252-74 Hobart Tasmania 7001, Australia. - 3 Forest and 

Nature Conservation Research and Development Centre, Jl 

Gunung Batu No. 5 Bogor, Indonesia. 

Acacia mangium is the major plantation tree species in 

Indonesia. Recent surveys in Indonesia (Irianto et al., 

unpublished) have shown that heartrot is of high incidence 

in areas of Sumatra and Java. Heartrot has been previously 

reported in other SE Asian countries as a major problem 

for solid-wood production. Previous studies of wounded A. 

mangium branches indicated that defence responses were 

poorly developed - for example no tyloses were formed. To 

investigate this we have studied both the anatomy and 

phenol chemistry of various A. mangium tissues. We have 

studied samples of heartwood (decayed, discoloured and 

sound) and sapwood from naturally-infected harvest-age A.


mangium from two sites in Sumatra. Total phenol levels 

were highest in inner heartwood and discoloured wood. 

Analysis by liquid chromatography - mass spectrometry 

revealed that condensed tannins and a range of flavonoids 

(e.g. tetrahydroxyflavonone) were mainly responsible for 

this increase. Studies of the antifungal nature of these 

extracts in bioassays will be presented as well as 

anatomical investigations. 

807 - Study of of onychomycosis in Tehran 

Sh. Bassiri * & A.A. Khaksar 

Pasteur Institute of Iran, Tehran Pasteur Ave. Kargar St. 

No.69 (13164), Iran. - E-mail: 

pooyan20002000@yahoo.com 

Fungal infection of the nail, also known as onychomycosis, 

is a worldwide problem. It is estimated that onychomycosis 

constitutes 15-22% of all nail disorders. It is also known 

that 2-5% of the adult population in Europe have 

onychomycosis. This problem is quite common in the 

dermatology clinic of the countries in the Midle East, India 

and the Far East. The infection rates and types of fungi 

involved in onychomycosis vary with conditions such as 

age, sex, occupation, hygiene, foot wear and several 

enviromental and climatic factors. This study reports the 

causative agents of onychomycosis in the presented 

patients in the section of Medical Mycology, Pasteur 

Institute of Iran. The 1985 patients were examined for 

onychomycosis. Diagnosis was confirmed by 

demonstration of fungi in direct (KOH 20%) and cultured 

examination. Onychomycosis was proven in 601 patients. 

350 patients was femailes (59%) and ranging in age from 3 

mounths to 84 years old. In the present study, 

onychomycosis was observed to be more common in 

females than in male. In this study Dermatophytes was 

51%, Candida 46% and other molds was 3%. 

808 - Effects of selected Penicillium mycotoxins in food 

waste on pig lymphocytes 

A. Bernhoft 1* , M. Keblys 1 , A. Flåøyen 1 & H.J.S. Larsen 2 

1 National Veterinary Institute, P.O. Box 8156 Dep., N- 

0033 Oslo, Norway. - 2 Norwegian School of Veterinary 

Science, P.O. Box 8146 Dep., N-0033 Oslo, Norway. - Email: 

aksel.bernhoft@vetinst.no 

The six Penicillium mycotoxins citrinin, cyclopiazonic 

acid, ochratoxin A, patulin, penicillic acid, and 

roquefortine C may be present in mouldy food and are 

known or suspected to be immunomodulators. The aims of 

our study were to determine the effect on lymphocyte 

proliferation of individual toxins and their combinations. 

We used an in vitro method that expresses the direct 

mycotoxin effect on T-lymphocytes and implies a 

simplification of the immunotoxic effect elicited from in 

vivo exposure. Blood samples were collected from 

clinically healthy pigs of 3-4 months age. The mitogen 

concanavallin A and pure mycotoxins were added to 

isolated lymphocytes and incubated, before further 

incubation with tritiated thymidine. Cell proliferation was 

measured as radioactivity ingested by the cells. Ochratoxin 

A and patulin were the most potent inhibitors of 

lymphocyte proliferation. Based on molar concentrations, 

these toxins were 15, 30, 40 and 65 times more potent 

inhibitors than penicillic acid, citrinin, cyclopiazonic acid, 

and roquefortine C, respectively. Examination of the 

cellular response of toxin combinations revealed that 

ochratoxin A and citrinin elicited synergistic effect. Other 

pair combinations showed additive, independent or 

antagonistic effects. As occurring mycotoxins may elicited 

very different kinds of interactive effects, the sum effect of 

mouldy food may be difficult to predict. 

809 - Occurrence of fine root pathogens in native scots 

pine forests of North Scotland 

W.J.A. Bodles 1* , S. Woodward 1 & C. Leifert 2 

1 University of Aberdeen, Department of Agriculture & 


AB24 5UA, Scotland, U.K. - 2 University of Newcastle, 

Tesco Centre for Organic Agriculture, King George VI 

Building, Newcastle, England, U.K. - E-mail: 

w.bodles@abdn.ac.uk 

Soil samples were removed from around Scots pine trees in 

five semi-natural and plantation forests in the north of 

Scotland. Pathogens were isolated by apple baiting from 

the soil samples and identified to the genus level using 

microscopy. Genera identified included species of Pythium, 

Phytophthora, Fusarium (Nectria) and Verticillium. 

Further characterization of isolates using ITS PCR and 

sequencing identified several potential pathogens, 

including Pythium ultimum var. ultimum, Pythium 

undulatum, Phytophthora cinnamomi var. cinnamomi, 

Nectria gliocladiodes, Nectria galligena and Verticillium 

rexianum. Different pathogen species were not present in 

all forest areas sampled. In tests of virulence on Scots pine 

seedlings, isolates had varying effects on root system 

development in both in vitro and potted plants. The 

apparently widespread occurrence of fine root pathogens in 

native Scots pine forest areas suggests that these organisms 

may have a role in fine root turnover dynamics in forest 

ecosystems and could impact on overall tree health and 

regeneration potential in different microclimates. 

810 - Acute toxicity of two toxigenic strains of fungi 

isolated from poultry feed on locally produced saudi 

arabian fertilized eggs and chicks 

F. Bokhari 

King Abdu Aziz University, P. O. Box 12161, Jeddah 

21473, Saudi Arabia. - E-mail: fmbokh@kaau.edu.sa 



Acute toxicity assays were performed for two toxigenic 

strains of fungi, Aspergillus parasiticus and Penicillium 

viridicatum isolated from poultry feed (sorghum, millet) on 

fertilized eggs and chicks. Two to three day old chicken 

embryos were inoculated with either standard toxin or 

spore suspensions of the toxic fungi. The most frequent 

malformations were observed after administrating the spore 

suspension. These malformations were characterized by 

vascular fragility, hemorrhage, and defects of the limbs. 

Results demonstrate the possibility of toxigenic fungi 

growth inside the egg, and consequent production of 

mycotoxins at incubation temperature. Heavy fungal 

growth took place in the air sac area, spreading to the shell 

membrane, and, to a lesser extent, the surrounding 

albumen. In CHEST assays, four-week-old chicks were 

given feed contaminated with A. parasiticus, or water 

containing a fungus filtrate. Chicks receiving a 1:1 ratio of 

contaminated to normal feed had the highest mortality rate 

(100%). Postmortem examination showed that all 

individuals who died suffered hemorrhages of the liver and 

liver enlargement. 

811 - The distribution of trichothecenes in shell and 

kernel of oats 

N.B. Bremnes * , P.-E. Clasen & A. Bernhoft 

National Veterinary Institute, P.O. Box 8156 Dep., N-0033 

Oslo, Norway. - E-mail: nanna.bremnes@vetinst.no 

Twenty-nine samples of oats were collected from a mill in 

Moss, Norway (Regal Mølle) from the harvest of 2000. 

This mill produces mainly products for the food industry. 

All the samples were divided into shells and kernels. The 

grinded samples (25 g) were extracted with acetonitrile / 

water (84:16, v/v). After filtration, the extracts were 

cleaned on a Mycosep #225 column (Romer Labs), 

evaporated and derivatized with pentafluoropropionic acid 

(PFPA) before they were analysed on GC-MS. In the 

shells, HT-2 toxin was found in 90% and T-2 toxin in 66% 

of the samples. The median concentrations of HT-2 and T- 

2 toxin were 181 and 56 µg/kg, respectively. The highest 

concentrations of HT-2 and T-2 toxin were 711 and 196 

µg/kg, respectively. Deoxynivalenol (DON) was detected 

in a few samples, with the highest level 220 µg/kg. In the 

kernel samples, no trichothecenes were detected (detection 

levels for all trichothecenes in the trial was 20 µg/kg). The 

results indicate that the main amounts of trichothecenes are 

connected to the shell part of the oat. As oat shell is of 

minimal use, the problem with mycotoxin contamination in 

oats seems to be a smaller problem than first assumed. The 

investigation will be continued. 

244 


812 - Exploitation of the mycoparasitic fungus 

Trichoderma harzianum against plant pathogens 

* 

J. Brozová , J. Hýsek & M. Vach 

Research Institute of Crop Production, Drnovská 507, 

Praque 6, 161 06, Czech Republic. - E-mail: 

brozova@vurv.cz 

Trichoderma harzianum is a worldwide soilborne fungus 

belonging between anamorphic fungi. It is a facultative 

parasite of a wide spectrum of fungi, but it is able to live as 

saprophyte too. The manifestation of its antagonism 

display as support competition, parasitism and antibiosis. 

Biofungicide Supresivit conteining konidia of T. harzianum 

was registrated in the Czech republic in 1994. It is licenced 

for protection of ornamental plants, ornamental and forest 

trees and peas. It could be applicated on seeds before 

sawing, on roots of sedlings or as watering or spraying. 

Commercial prepared mixture the biopreparation and 

granulated mineral fertilizer presents the new way of its 

introduction to plant enviroment. This method of 

application had the positive effect on 5% rising of yields of 

significant field crops (winter wheat, spring barley, winter, 

maize, and potatoes). The qualitative parametres of crop 

yield was slightly better (gluten conteins of winter wheat, 

oiliness of winter, starchiness of potatoes) or coincident 

(oily acid conteins of winter cole-seed, protein conteins of 

spring barley, gluten index of winter wheat) in comparison 

with yields on check parcels. The important result is, that 

occurrence of mycotoxines is decreased. The Supresivit 

suppress grey mould too. The regulal spraying of 

strawberries increased weight of healthy fruits and 

decreased the attack of grey mould even by strong infection 

press in 2001. 

813 - Impact of essential oils on growth and ochratoxin 

production by Aspergillus ochraceus and Penicillium 

verrucosum on a wheat-based substrate 

V. Cairns * & N. Magan 

Applied Mycology Group, Biotechnology Centre, Cranfield 

University, Silsoe, Bedford MK45 4DT, U.K. - E-mail: 

v.cairns.s00@cranfield.ac.uk 

The effect of water activity (a w; 

0.901-0.999) on growth of 

the ochratoxigenic species Aspergillus ochraceus and 

Penicillium verrucosum (3 strains) was evaluated on a 

wheat-based substrate. Water relation profiles were similar 

for all strains and species. Optimum growth was observed 

at sub-optimal a w levels (0.95-0.98). An in-vitro initial 

screen of a range of 24 essential oils on inhibition of spore 

germination and mycelial growth was made for all strains 

at 25 °C and 0.995 a w. 

Of the essential oils screened, bay 

leaf (west Indian), cinnamon leaf, clove bud, clove leaf 

(cleaned), lemongrass and thyme were found to be the most 

effective. A detailed screen of the most effective essential 

oils was made using a 2% wheat agar modified with 

glycerol to a range of water activities (0.901-0.999).


Essential oils were incorporated at different concentrations 

(0; 50; 100; 250; 500; 1000 ppm). Three isolates of each 

species were inoculated and colony growth was 

periodically measured for 21 days at two temperatures. At 

sub-optimal concentrations, some essential oils were found 

to stimulate growth. Ochratoxin production was modified 

by the different treatments. Acknowledgement: This work 

is part of the EU project 'Prevention of ochratoxin A in 

cereals' (FAIR QLK1-CT-1999-00433). 

814 - Degradation of lignocellulose by acellulolytic 

strains (c-) of Pleurotus ostreatus 

H. Camaño Heredia * , H. Leal Lara & R. Ramirez Carrillo 

Department of Food Science and Biotechnology, Faculty of 

Chemistry, National University of Mexico (UNAM), 04510 

Ciudad Universitaria, Col Copilco, Mexico D.F., Mexico. 

The main obstacle for bioutilization of lingocellulose byproducts 

is lignin but It can be degraded by white-rot fungi 

like the edible mushroom Pleurotus ostreatus. This fungus 

is cultivated on agricultural and forestry wastes but wild 

strains produce a simultaneous degradation of 

polysaccharides and lignin. Acellulolytic strains (C-) of 

Pleurotus ostreatus were bred from acellulolytic mutants to 

promote a selective degradation of lignin. Commercial and 

acellulolytic Pleurotus sp. strains were cultivated on a 

commercial substrate prepared with fermented straw. 

Changes in substrate composition were followed during the 

whole production cycle. Lignin, glucan, xylan and ash 

content were determined at spawning (t=0), at the end of 

the incubation period (t=1) and after the first flush of 

mushrooms (t=2) Ash content in substrate increased with 

all strains when substrate was fully invaded and after the 

first crop of mushrooms indicating substrate degradation. 

Highly significant differences in lignin, glucan and xylan 

degradation were observed. Acellulolytic strains (C-) 

produced a larger degradation than commercial strains. 

However, acellulolytic strain 71x512(C-) showed identical 

lignin and xylan degradation as commercial strain K8501 

and its glucan degradation was also identical to that of 

another commercial strain, K1508. Remarkably, it was 

observed with all strains that production of mushrooms 

increases as xylan degradation decreases. 

815 - Virulence of soil borne pathogenic fungi isolated 

from forest areas and nurseries in north of Scotland on 

scots pine and common alder 

D. Chavarriaga * & S. Woodward 

University of Abedeen, Department of Agriculture & 


AB24 5UA, Scotland, U.K. - E-mail: 

d.chavarriaga@abdn.ac.uk 

Soil borne fungi isolated from forest areas and nurseries in 

North east of Scotland using baiting techniques, were 

identified using classical taxonomy and molecular methods 

(PCR amplification of ribosomal ITS regions; restriction 

digestion; sequencing of PCR products) as Fusarium 

lateritium, Fusarium tricinctum, Phytophthora cinnamomi, 

Pythium ultimum var. ultimum and Rhizoctonia binucleate 

(Ceratobasidium sp.). Virulence was tested in vitro on 

young seedlings of Pinus sylvestris and Alnus glutinosa, 

and Koch's postulates fulfilled through reisolation of the 

pathogens and confirmation of fungal penetration into host 

tissues. Root growth was measured using the Winrhizo 

program, and dry weights recorded. Symptoms on aerial 

parts were assessed 8, 12 and 48 days after inoculation 

using a categorical scale from 0 (healthy) to 5 (damage > 

76%). Fusarium spp. caused significantly different (P< 

0.01) symptom intensity on both host plants. However, no 

significant difference in root growth was found between 

treatments and control (P < 0.05). Highest mean root 

length, numbers of root tips and forks, volume and surface 

area on Alnus glutinosa occurred in plants inoculated with 

P. cinnamomi and Pythium ultimum. 

816 - Antifungal activity of natural antimicrobial 

compounds incorporated in to cyclodextrin for slow 

release in food packaging systems 

D. Dorph-Petersen, K.I. Suhr & P.V. Nielsen * 

Technical University of Denmark, Sølvtofts plads,DK-2800 

Lyngby, Denmark. - E-mail: pvn@Biocentrum.dtu.dk 

The antifungal effect of spices has been known for 

centuries. Examples are the use of hobs in beer making and 

the use of myrrh and thyme in mummification. Recently 

attention on using essential oils from spices as 

antimicrobial agent in food has grown. The effects of 

essential oils are strongly depended on application method, 

storage conditions, source and age of the oils and of course 

on the microorganisms likely to colonise the food product. 

In this research project we are investigating if antimicrobial 

compounds from spices and herbs can be used in food 

packaging systems either as a surface coat or integrated 

into a film. These compounds are often too volatile and 

insufficiently heat stable to be used directly. Incorporation 

of these active compounds into cyclodextrin (CD) can 

radically improve the heat stability of these compounds and 

to some degree control their release into the packaging 

atmosphere. We will here report our findings from studies 

of antifungal effect of selected active agent from essential 

oils incorporated into cyclodextrin, as compared to the 

effect of the corresponding pure essential oils. The active 

agents that were used were allyl-isothiocyanate (AITC) 

from mustard oil, citral from lemongrass oil and thymol 

from thyme oil. The moulds, Penicillium commune, P. 

roqueforti and P. nalgiovense, all associated with cheese, 

were used as test organisms. 



817 - Dimmeric naphtoquinone metabolite aurofusarin 

- new mycotoxin produced by Fusarium graminearum 

J. Dvorska 

Sumy State Agrarian University, Kirova Str 160/5 Fl 91, 

Sumy 40021, Ukraine. - E-mail: ppdvorska@rs.net.ua 

Fusarium species of fungi produce a range of mycotoxins 

(trichothecene, zeralenone and fumonisins). There is a 

group of pigments which are produced by Fusarium 

species and they have been ignored until recently. A 

comprehensive work conducted by Kotyk (1999) with such 

a pigment, aurofusarin, clearly has shown that this 

compound could be included into the list of important 

Fusarium mycotoxins. A series of our experiments on 

effect of aurofusarin on quails showed that there was no 

toxicity signs when the compound was fed at a level of 

26.4 mg/kg. Egg production was decreased, but body 

weight egg weight and yolk/albumin proportions did not 

change. There were negative changes in immune system of 

quails (titre antibody to Newcastle disease significantly 

decreased and the spleen was reduced in size). Antioxidant 

system of the adult quails was compromised as a result of 

aurofusarin consumption. Histochemical studies revealed 

degenerative changes in kidney (degeneration and 

resorption of the glomeruli). The ovary was brown in 

colour with a decreased weight and number of follicles, 

and liver was enlarged with haemorrhages. The egg yolk 

colour was changed from yellow-orange to browngreenish. 

Aurofusarin decreased DHA proportion in egg 

yolk lipids and increased linoleic acid concentration. 

Concentrations of vitamin E, carotenoids and vitamin A in 

the egg yolk also decreased as a result of aurofusarin 

supplementation. Fertility and hatchability of aurofusarin 

enriched eggs was decreased. 

818 - Traumatic oil glands induced by pruning in the 

wound-associated phloem of Eucalyptus globulus: 

chemistry and histology 

A. Eyles 1* , N.W. Davies 2 & C. Mohammed 3 


Forestry., GPO Box 252-12 Hobart, Tasmania, 7001, 

Australia. - 2 Central Science Laboratory, University of 




Alieta.Eyles@ffp.csiro.au 

The natural occurrence of oil glands in various organs such 

as bark and leaves is well established as a characteristic of 

Myrtaceae, but this is the first reported case of traumatic oil 

glands induced in response to wounding. The new phloem 

enveloping the wound, which was formed in the two years 

following pruning in 5-year-old Eucalyptus globulus, was 

morphologically distinct from healthy stem phloem. 

Histological examinations revealed this wound-associated 

phloem to be largely composed of secretory cavities similar 

246 


in appearance to oil glands. Subsequent analysis of the 

wound-associated phloem extracts by GC-MS confirmed 

the presence of volatile terpenes and phenols. The total oil 

content determined for wound-associated phloem extracts 

was significantly higher (>50 times) than for healthy stem 

phloem extracts. The identities of twenty-six components 

of the oil extracts and their approximate relative 

abundances are presented. Implications of the role of 

terpenes as an inducible secondary metabolite in tree 

wound responses are discussed. 

819 - Host response to natural infection by a stem 

canker (Cytonaema sp.) in the aerial bark of Eucalyptus 

globulus: a preliminary report 

A. Eyles 1* , N.W. Davies 2 , Z.Q. Yuan 1 & C. Mohammed 3 


Forestry., GPO Box 252-12 Hobart, Tasmania, 7001, 

Australia. - 2 Central Science Laboratory, University of 




Alieta.Eyles@ffp.csiro.au 

The chemical and anatomical host responses to natural 

infection by a Cytonaema sp. in the aerial bark of 3-yearold 

E. globulus plantation trees were examined for the first 

time. The lesion margin (LM) of the canker-infected bark 

was characterized by the formation of a layer of dark 

extractives visible to the naked eye. Chemical analysis of 

the LM by GC-MS, HPLC-UV and HPLC-MS using 

negative ion electrospray ionisation indicated the presence 

of a range of compounds including hydrolyzable tannins, 

proanthocyanidins, flavonone glycoside, formlyated 

phloroglucinol compounds and volatile terpenes. These 

compounds were either undetectable in healthy tissue or 

else present at significantly lower concentrations than in 

the LM. The LM of the canker-infected bark was 

morphologically distinct from healthy phloem, its 

characteristics varying depending on severity of canker 

infection. In superficial infections (in which only the 

phloem was affected), three types of LMs were observed i). 

a continuous ligno-suberized boundary zone of multiple 

layers positioned between polyphenolic parenchyma cells 

ii). a boundary zone comprising of only polyphenolic 

parenchyma cells derived from the de-differentiation of 

pre-existing parenchyma cells iii). a combination of i and 

ii. In cases of severe canker infections in which the 

vascular cambium had been killed, the new phloem formed 

subsequently contained traumatic oil glands in addition to 

the responses observed for superficial canker infections. 

820 - Breeding bread wheat with multiple resistance 

and high yield for Ethiopia 

H. Fehrmann 1* , S. Assefa 2 & A. Badebo 2 

1 

Institute für Pflanzenpathologie und Pflanzenschutz, 

University of Göttingen, Germany. - 

2 

Ethiopian


Agricultural Research Organisation, Addis Abeba, 

Ethiopia. 

Yield of bread wheat (Triticum aestivum) in Ethiopia is 

poor. Five leaf diseases of fungal origin are most harmful: 

stripe rust (Puccinia striiformis), stem rust (P. graminis), 

leaf rust (P. dispersa), Septoria tritici leaf blotch, and tan 

spot (Pyrenophora tritici-repentis). 'Bread for the World' 

subsidized a project for the development of new genotypes 

with better resistance, higher yield and regional adaptation. 

Aegilops tauschii, a natural diploid progenitor of hexaploid 

bread wheat, is employed as alien source for new 

resistances against the different diseases. A. tauschii 

derived resistances are exploited by bridge crosses with 

tetraploid T. durum, embryo rescue culture and several 

bread wheat crosses. - For multiple resistance, a single 

plant selection until F7 is indispensable. Further 

development is based on a shuttle between greenhouse 

work (resistance selection and further crossing) and field 

tests. Top lines are multiple resistant, and yield is increased 

by 20 to 30% (average from 5-6 replicated experiment 

station trials). Regional differences in yield are striking. 

Two new varieties were already released. 

821 - Saxicolous lichens and rock-inhabiting fungi on 

different medieval monuments in Aosta Valley (Italy) 

S. Florio, E. Savino * , M. Brusoni, I. Carulli, A. Vaccari & 

M. Valcuvia 

Dep. Ecologia Territorio & Ambienti Terrestri, Via S. 

Epifanio, 14 - 27100 Pavia, Italy. - E-mail: 

esavino@et.unipv.it 

Microorganisms colonize all types of cultural artefacts 

causing biodeterioration, but little is known about the 

species involved. Among these, it seems nowadays that 

dematiaceous meristematic fungi are more widespread than 

previously thought. Saxicolous lichens and rock-inhabiting 

fungi on three different Italian medieval monuments have 

been investigated. The collecting sites are located in Aosta 

Valley (Western Alps), an area with a somewhat 

continental climate, at an altitude of about 1000m above 

sea-level. The monuments were built in XI-XIV century, 

with local different stones belonging to magmatic and 

metamorfic rocks, weathered and deformed to different 

extent. The stones, with the organisms to sample, were 

observed directly. Specific cultural medium for 

meristematic fungi was used. Statistical analysis has been 

performed. Six different lichens were identified, two of 

them rare for this mountain area. All of them grow on 

subneutral or acid substrata, in sunny, rather xerophytic, 

sites with weak eutrophication. A total of 10 micromycetes 

taxa was identified; besides white yeasts, mycelium with 

clamps, dematiaceous fungi with meristematic growth and 

mycelia sterilia were present. Eleven different types of 

stone were recognized. This preliminary data would 

suggest that lithic substrate is not selective for fungal 

communities living on the rocks. At the moment an 

association between lichen and fungal population does not 

seem to be significant. 

822 - Does the plant defensin-like peptide SPI1 from the 

gymnosperm Norway spruce protect seeds and 

seedlings against pathogenic fungi? 

C.G. Fossdal * & N.E. Nagy 

Norwegian Forest Research Institute, Høgskoleveien 12, 

N-1432 Ås, Norway. - E-mail: carl.fossdal@skogforsk.no 

Plant defensins are thought to protect germinating seeds, 

seedlings and plants against pathogenic fungi. However, 

the detection, developmental accumulation and cellular 

location of the corresponding protein in a gymnosperm 

have not been reported before. On this background and 

based on the predicted SPI1 amino acid sequence, 

antibodies were raised to detect and localize the putative 

plant defensin protein in the conifer Norway spruce [Picea 

abies (L.) Karst.]. The purpose was to detect and localize 

the 5kD SPI1 protein in seeds and during development, and 

to study the local and systemic effect on its accumulation 

after infection with three different pathogens, in young 

Norway spruce seedlings. The pathogenic fungi used for 

the infection studies were Heterobasidion parviporum, 

Ceratocystis polonica and the pathogenic oomycete 

Pythium dimorphum. The accumulation of the SPI1 protein 

during root development and after infection is described 

using immunoblotting. We also report that the putative 

plant defensin SPI1 is detected by antibodies on Pythium 

dimorphum hyphae invading the root cortex, suggesting a 

specific interaction between the SPI1 protein and hyphae. 

823 - Morphological and pathological variations of the 

Indonesian Cochliobolus heterostrophus 

A. Gafur 1* , S. Mujim 1 , T.N. Aeny 1 , B. Tjahjono 2 & A. 

Suwanto 3 

1 Department of Plant Protection, University of Lampung, 

Jalan S. Brojonegoror 1, Bandar Lampung 35145, 

Indonesia. - 2 Department of Plant Pests and Diseases, 

Bogor Agricultural University, Jalan Raya Pajajaran, 

Bogor 16144, Indonesia. - 3 Department of Biology, Bogor 

Agricultural University, Jalan Raya Pajajaran, Bogor 

16144, Indonesia. - E-mail: gafur@uwalumni.com 

Morphological variations of Cochliobolus heterostrophus 

(Drechsler) Drechsler collected from different places of 

Indonesia including corn areas in Bali, Java, Kalimantan, 

Sulawesi, and Sumatera were investigated microscopically. 

This mainly included observation on colony colour, 

conidial size and septa, and ascal size although variations 

in hyphae and conidiophores were also examined. All of 

the fungal isolates were grown in Petri dishes containing 

complete medium. Pathogenicity tests of the isolates were 

subsequently established. The experiments that involved 

different corn varieties, both local and introduced, were 

conducted by exposing all corn varieties to each of the 

different isolates of C. heterostrophus. Disease symptoms 

were recorded on a daily basis. Data collected from the 

present study indicated that morphological variations, 



including number of septa and size of conidia, among 

Indonesian isolates of C. heterostrophus existed, an 

indication that Indonesian C. heterostrophus might have 

originated from different ancestors. In contrast, 

pathogenicity of each isolate measured by disease 

incidence on several corn varieties was not significantly 

different. Disease severity of the corn varieties inoculated 

with each isolate was also relatively the same. The results 

imply that the corn varieties tested possess potential risks 

of southern corn leaf blight outbreaks. 

824 - Diagnosis of cryptic microbes based on ITS2 

rRNA secondary structure information 

A. Gargas 1* & P.T. DePriest 2 

1 Dept. of Botany, University of Wisconsin, Madison, 132 

Birge Hall, 430 Lincoln Dr, Madison, WI 53706, U.S.A. - 

2 Dept. of Botany, National Museum of Natural History, 

Smithsonian Institution, Washington, DC 20560, U.S.A. - 

E-mail: agargas@facstaff.wisc.edu 

Conventional techniques of isolating and identifying fungi 

or other cryptic microbes are time-consuming and 

expensive, limiting their utility for rapid and widespread 

surveys. We invented a novel method for detection and 

identification of microbes including those never before 

isolated or identified. This approach relies on analyses of 

the internal transcribed spacer 2 (ITS2) rDNA sequences - 

highly variable regions flanked by the 5.8S and 28S 

ribosomal subunits within nuclear DNA. After determining 

secondary structures of ITS2 rRNAs we identify nucleotide 

motifs between 20 and 50 nt in length within each primary 

sequence. These signature sequences, readily defined and 

present in high copy number, allow taxonomic diagnoses of 

known and unknown microbes from various substrates 

including agricultural, medical and ecological samples. 

Such sequences are readily adapted for use with probing 

techniques including PCR, microarray assays or other 

molecular detection methods. 

825 - First report of Coniothyrium stem canker on E. 

camaldulensis in Ethiopia 

A. Gezahgne * , J. Roux & M.J. Wingfield 

Department of Plant Pathology and Microbiology, Tree 

Pathology Co-operative Programme (TPCP), Forestry and 

Agricultural Biotechnology Institute (FABI), University of 

Pretoria, Pretoria 0002, South Africa. - E-mail: 

alemu@fabi.up.ac.za 

During a survey of Eucalyptus diseases in Ethiopia, a 

serious stem canker disease was discovered on E. 

camaldulensis trees at several localities in the South and 

South Western parts of the country. The disease is 

characterised by the presence of discrete necrotic lesions, 

stem cankers, cracking of the stems, production of kino 

pockets in the wood, as well as malformation of stems. 

248 


These symptoms are similar to those caused by 

Coniothyrium zuluense in South Africa. The aim of this 

study was to positively identify the causal agent of the 

disease in Ethiopia. This was achieved by sequencing the 

ITS region of the rRNA operon for a representative set of 

isolates. Sequences for the Ethiopian isolates were 

compared with those from authenticated isolates collected 

in South Africa, Thailand and Mexico. Based on these data, 

the Ethiopian isolates were shown to group more closely 

with those from South Africa, than with those from other 

areas. This study represents the first report of C. zuluense 

and its associated disease in Ethiopia. Currently the disease 

is causing considerable losses in yield and quality of timber 

and it also impacts negatively on the lives of subsistence 

farmers. We are currently studying additional isolates of 

the fungus and utilising sequences of different genes to 

study the phylogenetic relationships between C. zuluense, 

from a wide range of countries. 

826 - Enzyme mediated infection of host Pinus short 

lateral roots by Tricholoma matsutake 

W. Gill 1 , K. Suzuki 2 , M. Brown 3 & C. Mohammed 4* 

1 Tasmanian Institute of Agricultural Research, 13 St Johns 

Ave, New Town, Tasmania 7008, Australia. - 2 Laboratory 

of Forest Botany, Graduate School of Agricultural and Life 

Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, 

Tokyo 113-8657, Japan. - 3 Huon Valley Mushrooms, Main 

Road, Glen Huon, Tasmania 7109, Australia. - 4 School of 

Agricultural Science, University of Tasmania, GPO Box 

252-54, Tasmania 7001, Australia. - E-mail: 

caro.mohammed@utas.edu.au 

Tricholoma matsutake (Ito et Imai) Sing. (Matsutake) is an 

edible mycorrhizal basidiomycete highly prized in Japan 

for its culinary and medicinal properties and historical 

social significance. However, the natural supply of 

Matsutake in Japan is in decline and domestic demand 

exceeds supply. Consequently, the development of a viable 

commercial artificial cultivation system for this mushroom 

is of prime concern. In both natural conditions and artificial 

culture, Matsutake forms morphologically atypical 

ectomycorrhizas on host Pinus short lateral roots. 

Furthermore, aspects of the mushroom's nutrition are 

unusual and it is this atypical interaction which must be 

understood in order to devise an artificial cultivation 

system. Here, we present evidence of host infection and the 

parallel ability of T. matsutake to form a range of plant cell 

wall degrading hydrolytic enzymes in artificial culture. The 

hydrolytic enzymes tested were detected over a range of 

activities but the majority peaked after 2 or 3 weeks of 

incubation. Conversely, xylosidase and glucosidase 

enzymes were detected in comparatively high activities 

which did not peak but rather increased in activity over the 

duration of the assay. The significance of the hydrolytic 

enzymes produced in terms of Matsutake ectomycorrhizal 

establishment, development and observed morphology is 

discussed.


827 - Molecular probe development for tree root 

associated Rhizoctonia sp. and Suillus bovinus and 

detection by Southern dot blot and liquid hybridization 

H. Grönberg 1* , L. Paulin 2 & R. Sen 1 

1 Department of Biosciences, University of Helsinki, 

Viikinkaari 9, P.O.POX 56, Helsinki- 00014, Finland. - 

2 Institute of Biotechnology, University of Helsinki, 

Viikinkaari 9, P.O.POX 56, Helsinki- 00014, Finland. - Email: 

henrietta.gronberg@helsinki.fi 

Specific rDNA ITS (internal transcribed spacer) targeted 

probes for detection of different conifer root associated 

Rhizoctonia strains and Suillus bovinus were developed by 

comparing two different Southern (dot blot and liquid) 

hybridization methods. Probes (124-151bp) specific to six 

different Rhizoctonia strains (uninucleate strain 263, 

binucleate strains 268, AV-2, 251, 266, and 269 and 

multinucleate R. solani) and one specific to Suillus bovinus 

and a positive control probe were developed using pure 

culture DNA. Shorter (20-25 bp) oligonucleotide probes 

specific to strains 263, 268 and Suillus bovinus were 

developed in order to obtain more specificity and for used 

in liquid hybridizations. Total fungal DNA was used as a 

target with longer DIG (digoxigenin) labeled probes, but in 

biotin labeled oligonucleotide probes the target DNA was 

PCR amplified ITS (ITS1, 5.8S, ITS2). The optimal 

hybridization temperatures obtained from dot blots also 

gave the strongest signals in liquid hybridization protocol, 

although low temperature washing (30-35 °C) ocassionally 

affected to the specificity. We found the liquid 

hybridization protocol more useful when probing unknown 

target DNA, as it resolves targets using combined probe 

hybridization and ITS length polymorphism by resolving 

fragments differing by 5 bp in a sequencing gel (ALF 

express). 

828 - Ethnomycological knowledge and ecology of 

Phlebopus sudanicus in Burkina Faso 

K.M.L Guissou * , Ph. Sankara & S. Guinko 

Université de Ouagadougou, UFR/SVT, Département de 

Biologie et Physiologie Végétales, 03 BP 7021 

Ouagadougou 03, Burkina Faso. - E-mail: 

laure_guissou@univ-ouaga.bf 

The use and the ecology of Phlebopus cf sudanicus is 

examined in this study undertaken in the Reserve of 

Hippopotamus pound of Bala (Burkina Faso) and its 

vicinities. Fixed questions regarding ethnomycological 

knowledge were submitted to 180 native informants from 6 

villages. Collecting field works toward the forest were 

organised to recognise the mean habitats of this fungus. It 

appears that a total of 98% of the informants recognise this 

species as being edible. Two kinds of habitats of Phlebopus 

sudanicus have been found: under Piliostigma thonningii 

(Caesalpiniaceae) and Mitragina inermis (Rubiaceae) 

during the fieldwork. 

829 - Genetic diversity of fungi associated with seeds of 

tropical forest trees (Podocarpus falcatus and Prunus 

africana) 

A. Gure * , K. Wahlström & J. Stenlid 

Department of Forest Mycology & Pathology, Swedish 

University of Agricultural Sciences, Box 7026, 750 07 Ulls 

vag 26A, Uppsala, Sweden. - E-mail: 

Abdella.Gure@mykopat.slu.se 

The association and interactions between plants and fungi 

have long been observed and studied. However, the 

knowledge about the diversity of the fungal flora and the 

nature of their interactions with their hosts has been very 

scarce in the area of tropical forest tree seeds. In this study, 

matured fruits of Podocarpus falcatus and Prunus africana 

were collected directly from the crowns and from the 

ground just under the mother trees in March 2000 and May 

- June 2001. Seed collection was carried out at different 

forest sites in Ethiopia. Fungi were isolated from fresh 

fruits, and from seeds with and without seed coat. In all 

cases, surface sterilised and unsterilised seeds were plated 

on standard growth media. All in all 150 fungal isolates 

belonging to 25 genera have been identified by 

morphological and molecular methods (internal transcribed 

spacer- ITS sequence analysis) at least to the genus level. 

Most of the isolates were ascomycetes and a few were 

basidiomycetes. Some of these fungi are putative 

seedborne pathogens that will be studied further. 

830 - Biocontrol of the Canada thistle (Cirsium arvense) 

with fungal pathogens 

S. Guske 1 , B. Schulz 2* & C. Boyle 2 

1 Biologische Bundesanstalt, Messeweg, D-38104 

Braunschweig, Germany. - 2 Institut für Mikrobiologie, 

Technische Universität Braunschweig, Spielmannstr. 7, D- 

38106 Braunschweig, Germany. - E-mail: b.schulz@tubs.de 

In order to develop a mycoherbicide for biocontrol of 

Cirsium arvense (L.) Scop. (Canada thistle), fungal 

pathogens were tested, both singly and in combination, in 

semi-field pot experiments in the course of two vegetation 

periods. Isolates were selected from fungi previously 

isolated both as pathogens and endophytes from C. arvense 

and subsequently tested for their virulence. The fungi 

inoculated singly were Phoma destructiva, Phoma 

hedericola, Phoma nebulosa, M. sterila isolate 1.4a-4 and 

the rust, Puccinia punctiformis. Disease severity was 

evaluated according to disease symptoms (degree of 

chloroses, necroses, macerations) and various parameters 

of growth and development, i.e. length and death rates of 

main and secondary shoots, inflorescences, seed production 

and germination, wet and dry weights of shoots and roots 

after harvest. With the exception of Puccinia punctiformis 

(local infections) all of the isolates applied singly, 

negatively influenced all measured parameters, those of 



wet and dry weights of roots and shoots as well as the 

number of inflorescences, significantly. A combined 

inoculum of Puccinia punctiformis and P. hedericola was 

less effective than P. hedericola inoculated singly. The 

most effective inoculum was an inoculum mixture of the 

four perthotrophs. Further experiments should concentrate 

on formulation of the pathogens and application in field 

experiments. 

831 - Real-time PCR procedure to reliably detect and 

quantitate the pathogen Heterobasidion parviporum in 

infected spruce tissue culture clones 

A. Hietala, M. Eikenes, H. Kvaalen, H. Solheim & C.G. 

Fossdal * 

Norwegian Forest Research Institute, Høgskoleveien 12 N- 

1432 Ås, Norway. - E-mail: carl.fossdal@skogforsk.no 

One of our main interests is to learn about the molecular 

basis of host defense responses, using the coniferous host 

Norway spruce infected with the pathogen Heterobasidion 

parviporum as the experimental system. This 

basidiomycete and the closely related pathogen H. 

annosum are the major root rot causing pathogens in 

conifers. To screen host material for differential resistance 

towards H. parviporum, it is a necessity to quantify the 

fungal colonization of the host tissues. Therefore, we 

aimed to develop and compare the sensitivity of a real-time 

PCR to an ergosterol based method for determining the rate 

of colonization. We developed a quantitative multiplex 

real-time PCR procedure that reliably detecting down to 

1pg H. parviporum DNA and 1ng host DNA. There was a 

very high correlation between the fungal-biomass/totalbiomass 

and fungal-DNA/total-DNA rankings obtained 

with ergosterol and real-time PCR, strengthening the 

credibility of both methods. The results indicate that this 

real-time procedure can be a useful method to screen 

different spruce material for their relative resistance to the 

pathogen H. parviporum. 

832 - A TaqMan TM real-time PCR assay for detection 

and quantification of T-2/HT-2 toxin producing 

Fusarium species in cereal matrices 

A. Holst-Jensen * , A. Løvseth, A.K. Knutsen, M. Torp & 

K.G. Berdal 

National Veterinary Institute, Section of Feed and Food 

Microbiology, Ullevålsveien 68, P.O.Box 8156 Dep., 0033 

Oslo, Norway. - E-mail: arne.holst-jensen@vetinst.no 

Fusarium sporotrichiodes and Fusarium langsethiae Torp 

& Nirenberg ined. are known producers of T-2/HT-2 toxin. 

However, these taxa may easily be confused with other 

taxa of Fusarium, e.g. F. poae, that very rarely have been 

reported to produce the toxin. The toxin is one of the most 

important mycotoxins in cereal products in the Norhern 

part of Europe, and it is therefore considered to be 

250 


important to detect and quantify the presence of producers 

of the toxin. Based on partial translation elongation factor 

1á gene sequences and phylogenetic analyses, we have 

identified diagnostic sequence motifs and successively 

developed a quantitative real-time PCR assay for detection 

of these two taxa in cereal matrices, using TaqMan TM 

chemistry. The assay and its performance will be 

presented. 

833 - A multi-target environmental approach to control 

growth and mycotoxin production by Fusarium species 

using essential oils and antioxidants 

R. Hope * & N. Magan 


University, Silsoe, Bedford MK45 4DT, U.K. 

This study investigated how Fusarium culmorum and F. 

graminearum isolates from the UK are affected by aw, 

temperature, and fungicidal agents. The combined effects 

of these different variables were also investigated. In 

addition to how the morphology and ecology of the species 

is affected by the environmental conditions, the effect on 

mycotoxin production (in particular deoxynivalenol) was 

also determined. Essential oils and antioxidants were 

screened for antifungal properties in vitro using clearing 

zone assays and temporal growth studies. The five most 

effective agents identified by the in vitro screen were tested 

further in situ on irradiated wheat grain under different 

environmental conditions and a w levels. Growth rates of F. 

culmorum and F. graminearum were significantly effected 

by temperature,a w and antifungal agent. Efficacy of 

essential oils and antioxidants agents at inhibiting fungal 

growth was dose and type dependent. At least 500 ppm was 

required for significant inhibition of growth. Cinnamon and 

clove essential oils were the most effective growth 

inhibitors of the five agents across all the conditions. Of 

antioxidants, PP and BHA were most effective. The effect 

on DON and Nivalenol mycotoxins was variable with 

effects better at 500 ppm of essential oils than 100 

regradless of aw level. The potential for using these 

alternative food grade preservatives in stored cereals will 

be discussed. 

834 - A study of the trypsin genes in Metarhizium 

G. Hu * & R.J. St. Leger 

University of Maryland, Rm 4112 Plant Sciences Building, 

college park, MD 20742, U.S.A. - E-mail: 

gh70@umail.umd.edu 

Metarhizium anisopliae var. anisopliae produces 3 trypsins 

among a variety of other cuticle degrading enzymes. As 

well as animals, the trypsin family of Metarhizium has 

homologs in streptomycetes and four related pathogenic 

pyrenomycetous ascomycetes. The limited distribution of 

trypsins in a small group of related fungi is consistent with


their being derived from an animal or streptomycete source 

via horizontal gene transfer. To investigate trypsin 

distribution, fragments from genomic DNA of 43 

representative fungal species were amplified using 

degenerate primers. The fragments contain a highly 

conserved and diagnostic area for trypsins that includes the 

active site His and Ser residues. Trypsin-like genes were 

absent from most ascomycetes and had a very patchy 

distribution in zygomycete and basidiomycete fungi. The 

results were analyzed using sequence composition, 

maximum parsimony, maximum likelihood and distance 

methods. They show that there are several lineages of 

trypsins in fungi and that multiple gain and loss events 

happened during the evolution of this gene family. 

835 - Detect and diagnosis Monosporascus 

cannonballus, the causal agent of muskmelon root 

rot/vein decline, by PCR method 

K.S. Huang 1* , P.-H. Wang 2 & Y.S. Lin 1 

1 Department of Plant Pathology, National Chung Hsing 

University, 250, Kuo-Kuang Rd., Taichung, Taiwan. - 

2 Department of Microbiology, Soochow University, 70, 

Lin-Hsi Rd., Taipei, Taiwan. - E-mail: 

pathogen@ms17.hinet.net 

In Taiwan, root rot and vine decline became a growthlimited 

factor of muskmelon and watermelon since 1994. 

The causal agent was Monosporascus cannonballus, a 

soilborne pathogen. This species is notoriously difficult to 

identify, it lacks a conidial stage, perithecia form only after 

3 to 4 weeks in culture, and the ascospores do not 

germinate under standard laboratory conditions. The 

isolates often fail to produce perithecia in culture, making 

their identification impossible. We detected the pathogen 

using a PCR-based method, which developed by Lovic et 

al. A 430-bp DNA product was amplified from M. 

cannonballus isolates of Taiwan, and the product was 

obtained from symptomatic muskmelon and other 

cucurbitaceous plants collected from fields. No PCR 

product was obtained from the plants without symptoms. 

The fungus could be detected during the growing seasons. 

The method is being used to detect and monitor the 

pathogen on different cucurbitaceous plants and varieties 

for the selection of the disease resistance/tolerance 

rootstock. 

836 - The influence of the biopreparation Supresivit 

(Trichoderma harzianum) in the mixture with inorganic 

fertilizers on cereals 

J. Hýsek * , J. Brozová & M. Vach 

Research Institute of Crop Production, Drnovská 507, 

Praque 6, 161 06, Czech Republic. - E-mail: 

hysek@vurv.cz 

The influence of the treatment with the biopreparation 

Supresivit (Trichoderma harzianum) in the mixture with 

mineral fertilizers (NPK, LAV - ammonium nitrate with 

limestone ) was studied. The optimal dose of the 

biopreparate was 0.5 g per 1 kg of the fertilizer. The 

infestation of leaf and ear pathogen of winter wheat 

(septorioses and fusarioses) and on spring barley 

(rhynchosporioses and helminthosporioses) decreased. The 

yield was also increased (approximately about 5-8%). It 

was due to the decrease of leaf and ear spot fungal 

diseases. 

837 - Mycotoxines of the genus Fusarium on spring 

barley and the protection against them 

J. Hýsek 1* , J. Brozová 1 , M. Vánová 2 , J. Hajslová 3 & S. 

Sykorova 1 

1 RICP, Drnovská 507, Praque 6, 161 06, Czech Republic. - 

2 Agricultural Research Institute, Ltd., Havlíckova 2787, 

Kromeríz, Czech Republic. - 3 Institue of Chemical 

Technology, Technická 3, Praque 6, 166 28, Czech 

Republic. - E-mail: hysek@vurv.cz 

Spring barley (varieties: Akcent, Amulet, Kompakt, 

Nordus, Olbram and Tolar) was cultivated after pre-crops: 

sugar-beet, cereals and Maize. We infected artificially 

named varieties with the isolate of Fusarium culmorum 

which produced deoxynivalenol and other trichothecene 

mycotoxines. After artificial infection we studied the 

spectrum of the genus Fusarium. Introduced Fusarium 

culmorum was isolated only in little amount. The genus of 

Fusarium tricinctum prevailed, less occurrence had the 

genus of Fusarium poae. Some strains of Fusarium 

culmorum differed in the production of mycotoxines in the 

mycelium and in young plants (age 5 years). Maximal 

yields were evaluated after pre-crop of sugar beet (6.2 t/ha) 

the second one was the maize (5.25 t/ha) and the last there 

were cereals (4.39 t/ha). The treatment with the fungicide 

Folicur BT increased the yield about 25.7%, other one with 

Charisma increased the yield about 19.8 (application in the 

phase of flowering). The lowest occurrence of 

contaminated grain after pre-crop of sugar beet (20.5%), 

the highest one was after pre-crop maize (34%). 

838 - Diversity of Fusarium spp. isolates from the wheat 

and rice growing areas of rice-wheat cropping system of 

Punjab, Pakistan: Isolation, pathogenicity and RAPDs 

analysis 

S. Iram 1* , J. Cullum 1 & I. Ahmad 2 

1 University of Kaiserslautern, Kaiserslautern, Germany. - 

2 National Agriculture Research Center, Islamabad, 

Pakistan. - E-mail: iramshazia@hotmail.com 

Surveys were conducted for the assessment of leaf and root 

diseases of wheat and rice crops in main rice-wheat 

cropping areas, namely Gujranwala, Sheikhupura, Sialkot 



and Narowal districts of Punjab. Assessment of prevalence 

and disease intensity of leaf and root diseases was done. 

Leaf and root fungi were isolated and identified. From leaf 

and root samples Fusarium spp. were isolated. 

Pathogenicity experiments were conducted on the two 

commercial varieties of rice and wheat. Fusarium spp. 

were pathogenic on wheat and rice varieties. Spore 

suspension method was used for the evaluation of virulent 

isolates of Fusarium spp. After 30 days the diseased roots 

were platted and fungi were re-isolated and compared with 

mother culture for the confirmation of Koch Postulates. 

After pathogenicity the Random Amplified Polymorphic 

DNA (RAPD) technique was used for identification and 

differentiation of Fusarium isolates. For plant pathogenic 

fungi, RAPD analysis has been used to analyze genetic 

variations or distinguish races in Fusarium isolates. 100 

strains were studied using a RAPD analysis with different 

decamer primers. All the primers produced polymorphic 

amplification patterns. The results were analyzed by Phylip 

programme. From this survey it was concluded that 

Fusarium was more prevalent fungi. 

839 - Protection of Lepidium sativum against 

phytopathogenic fungi Pythium ultimum by inoculation 

of nonpathogenic Fusarium isolates 

H. Ishimoto 1* , Y. Fukushi 2 & S. Tahara 2 

1 INRA, 17 rue Sully, 21065 Dijon Cedex, France. - 

2 Division of Applied Bioscience, Graduate School of 

Agriculture, Hokkaido University, kita-9 Nishi-9,Kita-ku, 

060-8589 Sapporo, Japan. - E-mail: 

ishimotoh2001@yahoo.com 

Two nonpathogenic Fusarium strains, Ls-F-in-4-1 and Rs- 

F-in-11, induced the resistance in L. sativum against 

pathogenic fungi, Pythium ultimum. These Fusarium 

strains caused the increase of the content of antifungal 

compounds and its precursor, isothiocyanate and 

glucosinolate, in roots of the host plants. The resistance of 

L. sativum against P. ultimum may be due to the increase of 

isothiocyanate content induced by the inoculation of 

Fusarium fungi. The increase of the content of 

isothiocyanate occurred regardless of the abilities of 

inoculated fungal strains whether they could hydrolyze 

glucosinolate to isothiocyanate or not, so these changes 

seemed to be the defense responses of host plant. Ls-F-in- 

4-1, a Fusarium isolate which showed myrosinase activity, 

inhibited the growth of P. ultimum on agar medium 

containing glucosinolate. This isolate would hydrolyze 

glucosinolate to generate isothiocyanate in tissue or 

rhizosphere of host plant and inhibit the growth of 

pathogenic fungi consequentially. This isolate may have 

another protection mechanism in rhizosphere of L. sativum, 

in addition to the ability to cause the increase of antifungal 

compounds in the roots of host plant. 

252 


840 - Opportunistic mold development in controlled 

environment agriculture 

O.A. Jejelowo 1 , D.J. Barta 2 & K.P. Anthony 1* 

1 Texas Southern University, 3100 Cleburne Avenue, 

Houston, Texas, U.S.A. - 2 NASA, Johnson Space Center, 

2101 NASA Road One, Mail Code EC3, Houston, Texas, 

U.S.A. - E-mail: kanthony@mail.arc.nasa.gov 

Hydroponically grown crop plants are being considered by 

NASA for air revitalization and food production in space. 

Hydroponic cultures can be infected by opportunistic 

molds, which proliferate due to the humid environment and 

warm temperature inside the growth chambers. This paper 

reports mold development in different hydroponic cultures. 

Fungal isolates include Pythium aphanidermatum, Pythium 

dissotocum, Alternaria species, Cladosporium herbarium, 

Penicillium species, Aspergillus species, Rhizoctonia 

species and Fusarium species. The results are discussed in 

terms of impact of pathogenic and opportunictic molds on 

plant growth. 

841 - Inhibition of sexual mating of Ustilago scitaminea 

by bacterial extracts 

Z.D. Jiang * , L. Ji, J.L. Xu & L.H. Zhang 

Institute of Molecular Agrobiology, 1 Research Link, 

National University of Singapore, Singapore 117604, 

Singapore. - E-mail: zide@ima.org.sg 

Ustilago scitaminea is an important pathogenic fungus of 

sugarcane which causes culmicolous smut disease and 

results in severe losses every year. Pathogenic infection of 

the fungus is caused by dikaryotic mycelium which is 

produced after mating of non-pathogenic, yeast-like 

sporidia that derived from germination of teliospore on the 

germinating nodes of sugarcane. Sexual mating is a short 

period that separates the yeast-like sporidia and the 

dikaryotic mycelium growth phases and is a potential target 

for developing new approaches for disease control. To 

explore the possibility, we isolated teliospores from the 

infected sugarcane and confirmed that the sporidia of U. 

scitaminae can be classified into two mating types. We 

have screened more than 1400 bacterial isolates for matinginhibition 

phenotype. Among them 16 were found to 

secrete substances which inhibit mating or hyphae growth. 

Preliminary taxonomic analysis suggests that they belong 

to 10 genera, including Bacillus sp., Chryseobacterium sp. 

and Filibacter sp. The inhibitors from the three species 

have been extracted and showed to be heat-resistant and are 

of low molecular weight, suggesting the non-enzymatic 

nature of the inhibitors. Conventional and electron 

microscopy indicate that the inhibitors have no effect on 

morphology of both mating types of U. scitaminae 

sporidia, except on hyphae development. Studies are in 

progress to understand the chemical nature of these 

inhibitors and their molecular targets.


842 - Gliotoxin production by Aspergillus fumigatus 

E. Jonsson 1* , A. Stepinska 1 , V. Wenehed 2 , P. Haggblom 1 & 

A. Forsby 2 

1 Dept of Animal Feed, National Veterinary Institute, 751 

89 Uppsala, Sweden. - 2 Dept of Neurochemistry and 

Neurotoxicology, Stockholm University, 106 91 Stockholm, 

Sweden. - E-mail: Eva.Jonsson@sva.se 

It is well known that various moulds can grow on animal 

feed and produce a range of toxic metabolites 

(mycotoxins). To study production of mycotoxins, 

Aspergillus fumigatus, that is one of the most common 

fungi in animal feed, was chosen. A. fumigatus is known to 

produce, among other toxins, gliotoxin, a highly toxic 

mycotoxin. The purpose of the study is to develop a 

screening in vitro method to identify toxicological 

components in feed, which may help to minimise 

application of animal tests also for other purposes. A. 

fumigatus CCUG 17460 were grown on Czapek-Dox Broth 

with 30% glucose (37 °C; 10 samples). The Czapek-Dox 

Broth was extracted and purified by a solid-phase 

purification method. The extracts were run at HPLC, and 

gliotoxin was detected in 9/10. The maximum 

concentration of gliotoxin was produced after 87 hours of 

inoculation. The purified extracts were applied to the 

human neuroblastoma SH-SY5 cells, and the general 

cytotoxicity was determined. The extracts showed to be 

cytotoxic to the SH-SY5 cells. This study proves that it is 

possible to produce gliotoxin in vitroby means of 

inoculating Czapek-Dox Broth with A. fumigatus. The 

effects of the in vitro produced gliotoxin agree with the 

effects of pure gliotoxin. 

843 - Expressed sequences in the basidiomycetaus tree 

pathogen Heterobasidion annosum during early 

infection of Scots pine 

M. Karlsson * , Å. Olson & J. Stenlid 

Forest Mycology & Pathology, Box 7026, SE-750 07, 

Uppsala, Sweden. - E-mail: 

Magnus.Karlsson@mykopat.slu.se 

The basidiomycete fungus Heterobasidion annosum is the 

causal agent of annosum root rot and is economically the 

most important disease of coniferous forests in northern 

temperate regions. H. annosum is a wood-decaying 

pathogen which can utilize a variety of carbon sources. The 

purpose of this research project is to investigate the 

molecular mechanisms behind the pathogenicity of H. 

annosum and to identify key factors, which enables the 

fungus to infect and cause disease. A cDNA library was 

constructed from mycelia that had been challenged with 

pine seedling roots. Fragment lengths are spanning from 

300 bp to 3000 bp with an average of 850 bp. Individual 

cDNA fragments were sequenced once from the 5' end to 

generate expressed sequence tags (ESTs) which were used 

for similarity searches against already available sequence 

data. In this way we can get an idea about the putuative 

function of the corresponding genes. So far 1150 fragments 

have been sequenced, corresponding to 337 different 

contigs from which 228 have significant similarity to other 

genes. The largest fuctional groups are those involved in 

information pathways (including protein synthesis) and 

basic metabolism but genes involved in electron transport 

and transmembrane transport are also common. In order to 

pinpoint the genes that are important for the pathogenicity 

of the fungus the cDNA fragments are screened for 

differential expression during different stages of the 

infection process. 

844 - Fungal infection for the recipients of living donor 

liver transplantation 

N. Kawagishi * , N. Ohkohchi & S. Satomi 

Division of Advanced Surgical Science and Technology, 

Tohoku University, 1-1 Seiryoumachi Aobaku Sendai 980- 

8574, Japan. - E-mail: kawan@mail.cc.tohoku.ac.jp 

(Introduction) Fungal infection is potentially fatal 

complication in patients with liver transplantation. The aim 

of this study was to review the single-center experience 

with fungal infections in living donor liver transplantation 

and to report its management for the early diagnosis and 

treatment. (Methods) From July 1991 to December 2001, 

52 cases of LDLTx were performed in our institution. 

Immunosuppression consisted of tacrolimus or 

cyclosporine, corticosteroids and in some cases 

azathioprine or mycophenolate mofetil. The diagnosis of 

fungal infection was based on clinical findings, culture and 

serological examinations such as β-D glucan, Candida Ag, 

Aspergillus Ag and Cryptococcus Ag. For the prophylaxis 

of the fungal infection, administration of fluconazole and 

gargle by amphotericin B were performed before LDLTx. 

Active fungal infection was treated by fluconazole, 

amphotericin B and flucytosine. (Results) Mycoses were 

cultured for 41 cases (78.8%) and 23 cases(44.2%) were 

detected before LDLTx. C. albicans was accounted for 

more than 60% of the recipients and the greatest number of 

fungal isolates were obtained from pharynx and stool. 

There was no recipient who died directly because of 

invasive fungal infection. Detection of β-D glucan was 

effective for the prevention from severe invasive fungal 

infection. (Conclusion) For the management of fungal 

infection in LDLTx, it is important to notify the role of 

prophylaxis, empiric and pre-emptive therapy. 

845 - Phyllachora epidemic on common ragweed 

(Ambrosia artemisiifolia): a unique natural control 

phenomenon in Hungary in 1999 

L. Kiss 1* , L. Vajna 1 , Gy. Bohar 2 , D. Magyar 1 , U. Paksiri 3 & 

S. Takamatsu 4 

1 Plant Protection Institute, Hungarian Academy of 

Sciences, H-1525 Budapest, P.O. Box 102., Hungary. - 

2 Bioved Bt., 2310 Szigetszentmiklos, Ady Endre u. 10., 



Hungary. - 3 Faculty of Agriculture, Chiang Mai University, 

Chiang Mai 50200, Thailand. - 4 Faculty of Bioresources, 

Mie University, 1515 Kamihama, Tsu 514-8507, Japan. - 

E-mail: LKISS@NKI.HU 

Since the early 1990s, common ragweed (Ambrosia 

artemisiifolia) has become the most widespread and most 

important allergenic weed in Hungary. Our annual surveys 

of fungal diseases of ragweed revealed that this plant, 

introduced to Europe from North America, is one of the 

healthiest weeds in the Carpathian basin. Only nine species 

of fungal pathogens were found on ragweed in Hungary 

that caused only minor infections in the field between 

1995-1999. However, in summer 1999, a serious epidemic 

developed on ragweed caused by Phyllachora ambrosiae. 

This holobiotrophic pathogen has not been reported from 

Europe prior to 1999 [1]. The identification of the pathogen 

was based on both morphological and molecular data. 

From mid-September, all plants examined in all regions of 

the country were infected and exhibited dead leaves and 

inflorescences. Thus, P. ambrosiae reduced the fitness of 

ragweed and also the period of production of the allergenic 

pollen in Hungary in 1999. According to the data of the 

pollen monitoring service in Budapest, much less ragweed 

pollen occurred in the air in September and October 1999 

than in the previous ten years. This natural epidemic 

clearly demonstrated the capacity of a pathogenic fungus to 

reduce the harmful effects of a noxious weed. Its main 

lesson is that biological control using fungal pathogens 

could be a possibility to suppress ragweed populations in 

Hungary. [1] Vajna, L., Bohar, Gy. Kiss, L.: Plant Dis 84, 

489 (2000). 

846 - AFLP of cheese contaminating Penicillium strain 

C.F. Kure 

Matforsk, Osloveien 1, 1430 Ås, Norway. - E-mail: 

cathrine.finne.kure@matforsk.no 

Amplified fragment length polymorphism (AFLP) analysis 

was performed on isolates of Penicillium commune and P. 

palitans originating from cheese and indoor environment in 

four cheese factories. The AFLP method was found to be a 

useful tool for identification of P. commune and P. palitans 

on, as well as below, species level. Specific P. commune 

and P. palitans strains were found in the same factories 

over a period of more than a year and showed that the 

cheese factories have contaminating strains that are well 

established. Several of cheese contaminating Penicillium 

strains could be related to air in the wrapping room, which 

must be considered to be a critical point for contamination 

of cheese. 

254 


847 - The telomeric repeat sequence of Aspergillus 

oryzae consists of unique dodeca-nucleotides 

K. Kusumoto * , S. Suzuki & Y. Kashiwagi 

National Food Research Institute, 2-1-12 Kannnondai, 

Tsukuba, Ibaraki 305-8642, Japan. - E-mail: 

kusumoto@nfri.affrc.go.jp 

The telomeres of the chromosomes of Aspergillus oryzae 

NFRI1599 were cloned and identified. They were sensitive 

to BAL31 exonuclease digestion, compared with the 

chromosome internal control, 18S rDNA, and thus were 

proved to be located at the most terminal region of the 

chromosomes. The telomeric repeat sequence of A. oryzae 

consisted of unique dodeca-nucleotides, 

TTAGGGTCAACA. The former six nucleotides TTAGGG 

in the repeat sequence was conserved as telomeric repeat 

sequence in several filamentous fungi, including 

Aspergillus nidulans (Bhattacharyya and Blackburn 1997). 

The remaining sequence TCAACA in the repeat is not 

reported as telomeric sequence in any other organisms. 

Therefore the newly identified A. oryzae telomere is unique 

in this organism. Length of the telomeric repeat region was 

114-136 bp. Considering that the length of the telomere 

was similar to that of A. nidulans, the rigid regulation of 

the telomere length might be conserved among Aspergillus 

species. 

848 - Assessment of air-borne fungi present in a 

biomedical plastics factory 

C.P. Larralde-Corona * , M.R. Santiago-Mena, C. Jacques- 

Hernandez, D. Resendez-Perez & H.A. Barrera-Saldaña 

Centro de Biotecnologia Genomica - IPN, Apartado Postal 

152, Reynosa 88710, Tamaulipas, Mexico. - E-mail: 

patyla@mail.cbg.ipn.mx 

Fungi can be found in high concentrations in the bioaerosol 

of northeast Mexico, where we have a subtropical climate 

and agriculture is the main activity. Because our city is in 

the border with the United States, there are hundreds 

factories. One of them, dedicated to injecting biomedical 

application plastics, had a fungal contamination problem. 

We were asked to evaluate the issue, and accordingly the 

production zone and warehouse were carefully sampled. 

Media used were: Potato-Dextrose Agar, 8-Vegetables 

Agar and Lennox Broth supplemented with agar. By the 

pattern of distribution of genera, we found that the entrance 

of fungi was the ventilation system, and that this income of 

bioaerosol was accummulating on horizontal high zones 

and then being slowly distributed, specially over the 

packing area. We found 20 genera of fungi, being the most 

important in terms of frequency: Cladosporium (3 species), 

Aspergillus niger, Penicillium (four species), Rhizopus and 

Alternaria (2 species). A close examination of the cleaning 

policies and ventilation system of the factory showed that, 

althought being efficient for the environment where they 

were designed, they were not the best options for a region


like ours, because its high average annual temperature (25 

centigrades) and relative humidity (50% or more). Among 

suggestions were a much more frequent filter change, 

sealing of all porous surfaces and a tight control of relative 

humidity inside the production zone. 

849 - A new Phytophthora infestans molecular marker 

O.I. Lavrova * & Y.T. Dyakov 

Moscow Lomonosov State University, 119899, Vorob'evy 

gory, MSU, Biology Faculty, Mycology and Algology 

Department, Russia. - E-mail: ilinir@mail.ru 

Short interspersed elements (SINEs) or short retroposons 

are 80-400 bp repetitive DNA sequences that proliferate in 

eukaryotic genomes via transcription followed by reverse 

trancription. Recently SINEs have been used as molecular 

markers for intraspecies differentiation. Most SINEs have 

an internal promoter for RNA polymerase III composed of 

two boxes (A and B) spaced by 30-35 bp similar to tRNA 

genes. To detect and clone novel tRNA-derived SINEs in 

Phytophthora infestans genome we have used PCRamplified 

total genomic DNA as a template and primers 

specific to boxes A and B for PCR amplification. This 

reaction, designated A-B PCR, amplifies the region 

between boxes. The resulting DNA fragments have been 

cloned and sequenced. The sequences obtained of 8 

fragments (45-51 bp) are similar but not identical 

suggesting that they may have been amplified from 

individual copies of a single SINE famile. All 8 fragments 

have 24 bp conservative sequence. On the basis of this 

sequence we constructed one primer to use it as a marker 

for P. infestans intraspecies differentiation. This marker is 

more reliable than RAPD because it is specific sequence 

and it gives the same results in the several experiment 

repeatings. Acknowledgements: This work was supported 

by the grant from the International Science and Technology 

Center No. 1640. 

850 - Plant and microorganisms interactions mediated 

by Pythium oligandrum 

G. Le Floch 1* , P. Rey 1 , N. Benhamou 3 , M.I. Salerno 2 & Y. 

Tirilly 1 

1 Laboratoire de Microbiologie et Sécurité Alimentaire, 

ESMISAB technopôle Brest Iroise 29 280 Plouzane, 

France. - 2 Laboratorio de Proteccion Forestral, CISAUA- 

Facultad de Ciencias Agrarias y Forestales UNLP 60 y 

119 CC31 (1900) La Plata, Argentina. - 3 Département 

Recherche en Sciences de la Vie et de la Santé, Pav. Ch.E. 

Marchand, Université Laval Sainte-Foy Québec GIK7P4, 

Canada. - E-mail: gaetan.lefloch@univ-brest.fr 

Over the past few years, experiments performed by our 

group have demonstrated that the role played by antagonist 

fungus, Pythium oligandrum, on plant development and 

protection involves a tripartite interaction between the 

biocontrol microorganism, the pathogen agent and the 

plant. Such a complex process includes indirect effects 

through the control of pathogens in the rhizosphere and/or 

direct effects mediated by plant-induced resistance. 

Interaction of P. oligandrum with soil-borne pathogens to 

control their development in the rhizosphere and even in 

planta consists in a series of events involving mainly 

mycoparasitism and/or antibiosis. However, antagonism is 

a multifaceted process dependent on the involved target 

host; for example, by forming a thicken wall barrier 

Rhizoctonia solani and Phytophthora cinnamomi limit P. 

oligandrum antagonistic activity. We also showed 

hyperparasitism on various sclerotia. Plant-induced 

resistance mediated by P. oligandrum is a key event in this 

kind of fungus-plant interaction. Evidence of plant 

sensitisation by P. oligandrum to respond more rapidly and 

efficiently to pathogen attacks was also provided. It can 

protect plants from B. cinerea, Fusarium oxysporum f.sp. 

radicis lycopersici, Phytophthora parasitica, P. ultimum 

attacks by either inducing local protection or triggering 

systemic resistance. 

851 - Selection of baiting materials for Phytophthora 

palmivora (Butler) Butler., the causal agent of coconut 

nut falling disease 

S. Likhitekaraj * & W. Ouvanich 

Plant Pathology and Microbiology Division, Department of 

Agriculture, Chatuchuk, Bangkok 10900, Thailand. 

Coconut nut falling disease is caused by Phytophthora 

palmivora (Butler) Butler. A serious outbreak of this 

disease was found on 'Yellow Dwarf' cultivar of coconut 

grown at Surat Thani Rubber Research Center. Since P. 

palmivora is a soil-borne fungus, to isolate the fungus 

needs to apply baiting techniques to trap the fungus from 

the soil. Leaves of nine crop plant species, namely 

pararubber, cacao, citrus, durian, coffee. Black pepper, oil 

palm, and coconut cvs. 'yellow Dwarf' and F1 Hybrid 

Sawee I. The leaves were immersed in the zoospore 

suspension for 1, 3, 6 and 24 hours. It was found the leaves 

of all tested plant species excluding pararubber could trap 

up to 95% of the fungal spores when immersed in the 

zoospore suspension for 24 hr. An experiment was 

conducted to study on the optimum duration of fungal 

existiance in the soil to be best baited by using cacao 

leaves. Among 1, 15, 30, 45, and 60 days of the durations, 

it was found that 90% of the fungus could be trapped from 

the soil at 60-days duration. Suitable methods in using 

leaves of nine plant species to bait P. palmivora from the 

soil was also performed. The three methods include placing 

the leaves on the soil surface, placing the leaves one inch 

below the soil surface, and immersing the leaves in the 

infected soil suspension. The results revealed that 

immersing the leaves in the infected soil suspension was 

best in baiting the fungus from the soil. 



852 - Inhibition of microconidia of F. oxysporum var. 

vasinfectum with antibiotic substances from Bacillus 

subtilis strains 

R.N. Mannanov * & R.K. Sattarova 

Department of Phytopathology and Plant Physiology, 

Tashkent State Agrarian University, Tashkent 700140, 

Uzbekistan. - E-mail: abdukahar.kadyrov@syngenta.com 

Previously obtained results from comparative 

physiological, biochemical and antimicrobal investigation 

of endophyte and soil borne cultures of Bacillus subtilis N 

and 23 had allowed to suggest that both strains produce the 

same antibiotic substances active against cotton 

phytopathogens. To additionally prove the hypothesis, we 

carried out phase-contrast microscopy of phytopathogens' 

cells in the inhibition zone. The use of phase-contrast 

microscopy had shown inhibitory effect of antibiotic 

substances from 23 and N strains of B. subtilis on growth 

of microconidia which was different depending on the 

diffusion of antibiotic substances from the 0.5 cm well on 

agar in the centre with antibiotic filtrates from cultural 

liquids of B. subtilis strains towards edges of Petri dishes. 

Basing on the analysis of comparative influence of filtrates 

of 23 and N strains in the following dilutions: 1:1, 1:2, 1:4, 

1:9 and 1:99, we had systemized investigated microconidia 

of F. vasifectum by the character of inhibition to four 

conventional groups. Obtained results allowed to determine 

the unit of biological activity of filtrates of 23 and N strains 

which was equal to 85 mg of extracted antibiotic 

substances, whereas 1 ml of filtrates of each strain 

contained 11.8 units of antibiotic activity. Further 

experiments will be directed on studies of mechanisms of 

biological action of antibiotics produced by 23 and N 

strains on cells of different groups of fungal and bacterial 

microorganisms. 

853 - Bee pollen as a substrate for fungal development 

and mycotoxin production 

R. Mateo 1* , G. González 2 , J.M. Sáez 2 , A. Medina 2 , A. 

Llorens 2 & M. Jiménez 2 

1 

Unidad de Analisis de Sanidad Animal (Conselleria de 

Agricultura, Pesca y Alimentación), Av. Manuel Soto 18, 

2 

E-46024, Valencia, Spain. - Departamento de 

Microbiologia y Ecología (Univ. de Valencia, Dr. Moliner 

50, E-46100, Burjasot, Valencia, Spain. - E-mail: 

rufino.mateo@uv.es 

Pollen is a basic food for bee larvae development due to its 

high content of proteins, which contain all the amino acids. 

Pollen also contains minerals, vitamins, enzymes, growth 

regulators, fatty and organic acids and flavonoids. It is 

considered by the FAO a source of essential nutrients for 

the daily intake. Pollen consumption has increased in the 

last years as a diet complement in cases of fatigue, 

undernourishment and in vegetarian diets. Bees gather 

pollen in flowers, mix it with honey and nectar to make 

256 


pellets and carry it to the beehive to serve as a food for 

larvae. Beekeepers catch pollen in traps put at the hive 

entry. Pollen remains in traps for some time, then it is taken 

and carried to stores where it is cleaned, fumigated, stored 

and marketed. During this stage pollen can be 

contaminated by several fungal species, among them those 

that are mycotoxin producers. We have evaluated for the 

first time the capacity of bee pollen to serve as a substrate 

for the development of Fusarium and Aspergillus species 

that are potential producers of zearalenone, 

deoxynivalenol, fumonisins and aflatoxins. The levels of 

the mycotoxins produced in pollen have been determined 

and a comparative study between pollen and other 

especially susceptible substrates, like cereal grains has 

been made. The results obtained point out that the toxin 

levels found in pollen are significantly higher than those 

produced by the same isolates in cereal grains under the 

same incubation conditions. 

854 - Analysis of gene expression in targeted infection 

structures of obligate parasites, Blumeria graminis f. sp. 

hordai by intracellular RT-PCR 

Y. Matsuda * , T. Nonomura & H. Toyoda 

Laboratory of Plant Pathology and Biotechnology, Faculty 

of Agriculture, Kinki University, 3327-204 Nakamachi 

Nara 631-8505, Japan. - E-mail: 

ymatsuda@nara.kindai.ac.jp 

To detect specific mRNAs in postinfectional differentiation 

of obligate fungal parasites Blumeria graminis f. sp. 

hordai, we applied an intracellular RT-PCR to conidia 

inoculated onto barley coleoptile epidermis. The RT-PCR 

primers for target genes were constructed using a EST 

library of powdery-mildewed Italian ryeglass leaves. In the 

present study, the chitin synthase gene (chs1) was used as a 

target gene expressed specifically during the differentiation 

of fungal infection structures. The primers were 

constructed on the base of nucleotide sequences between 

translation and 3'-untranslation regions. The first strand 

cDNA was conducted in the fungal structures (conidia, 

appressoria and secondary hyphae) by microinjecting the 

reaction mixture (primers, reverse transcriptase, rTaq 

polymerase and dNTPs) and subsequently amplified by 

PCR using the primers amplifying the internal region of the 

chs1. The nucleotide sequence of the amplified DNA was 

determined and confirmed to be consistent with the chs1 

gene of B. graminis f. sp. hordai. Thus, the present work 

provides a new molecular tool for analyzing gene 

expression during the differentiation process of infection 

structures of obligate fungal pathogens, especially 

emphasizing that cell-specific gene expression in 

individual infection structures of the pathogen can be 

monitored with respect to host and parasite interactions.


855 - Evaluation of the occurrence of toxigenic fungi in 

barley and malt in Spain 

A. Medina 1 , M. Jiménez 1 , J.M. Sáez 1 , M.J. Hinojo 1 & R. 

Mateo 2* 

1 

Departamento de Microbiología y Ecología (Univ. de 

Valencia), Dr. Moliner 50 E-46100, Burjasot, Valencia, 

2 

Spain. - Unidad de Análisis de Sanidad Animal 

(Conselleria de Agricultura, Pesca y Alimentación), Av. 

Manuel Soto 18, E-46024, Valencia, Spain. - E-mail: 

rufino.mateo@uv.es 

The presence of some fungal species in malt produced from 

contaminated barley may pose health risks due to the 

production of mycotoxins. The predominant mycobiota can 

vary due to several factors, including barley growing 

location or climatic conditions. Heavy fungal 

contamination can affect the malting process, the malt 

quality and final beer attributes, but the presence of some 

fungal species requires especial attention due to the 

unacceptable risk of mycotoxin contamination in barley 

and malt. In Spain there is little information about the 

occurrence of mycotoxin-producing species in barley and 

malt destined to breweries. The potential mycotoxinproducing 

mycobiota occurring in barley and malt taken in 

Spanish breweries and stores is studied in this work. The 

nature and extent of fungal contamination were performed 

using direct plating techniques with different culture 

media. After the incubation period, fungal colonies were 

examined, identified and transferred to appropriate culture 

media for mycotoxin production. The results show that 

both barley and malt were contaminated at different levels 

with Penicillium, Aspergillus and Fusarium. Other fungi 

such as Alternaria and Cladosporium were found but their 

frequencies were significantly lower than those reported in 

other countries. Various fungal species involved in the 

production of ochratoxins, deoxynivalenol and aflatoxins 

were found whereas species implicated in production of 

other mycotoxins showed low occurrence level. 

856 - Fungi of indoor environs of occupational 

importance in India: Are they significant in causing 

allergy 

J.K. Misra 

Mycological Research Unit, Sri J.N.P.G. College, Botany 

Department, Sri J. N. P. G. College, Lucknow-226001, 

India. - E-mail: jitrachravi@sify.com 

Occupational environs such as cobbler's shops, ginneries, 

flour and sawmills were surveyed for their fungi suspended 

in the air. Forty fungal forms were recovered from cobbler 

shops. The flora of the cobbler shops was dominated by 

Aspergilli. Some fungal forms tested for their allergenic 

behaviour showed positive reactions among the patients in 

varying degrees. From the ginneries only 17 fungi were 

isolated but their load in the environment was very high. 

Almost 50% of the workers of the ginneries interviewed 

had allergenic and respiratory disorders. But no testing 

could be done for the fungi from the ginneries. Forty-one 

fungi were isolated from flourmills while fifty from 

sawmills. No significant variation was noted between the 

indoor and outdoor fungal flora of flour and sawmills. 

Spores of various fungi were also observed through the 

samplers in varying frequencies. In these environs too 

Aspergilli dominated the flora. Thus, at all the places 

surveyed, species of the genus Aspergillus were the 

dominant ones. Aspergillus flavus, however, was greater in 

occurrence in flourmills than saw mills. Distribution and 

dominance of fungal forms were correlated with ecological 

factors and the nature of occupation. Most of the fungal 

forms were found to have allergenic potentialites. 

857 - Fungi, paper materials and preservation 

conditions 

A. Montemartini 1* & V. Salvo 2 

1 DIP.TE.RIS. sect. Botanical, C.so Dogali 1M -cod 16136, 

Italy. - 2 DIP.TE.RIS. University of Genoa, C.so Europa 26 

- cod 16132, Italy. - E-mail: amontem@unige.it 

In several libraries and museums of Genoa, in which the 

thermohygrometric conditions were not optimum to the 

preservation, were found on books and maps mouldy spots. 

The determinations pointed out some species widespread in 

Genoa and moulds correlated to the micro-environmental 

conditions of the surrounding. In fact on eleven species, 

that were analysed in the library inside the Botanical 

Garden, ten were soil fungi whereas smaller amount was 

found in a book-warehouse located in a built-up area. The 

fungal species could be introduced by frequenters of near 

gardens and this species could grow with suitable 

conditions. Therefore this suggest limit the entry of people 

or at least to avoid they to introduce spores. In addition 

some books and maps showed rusty-red spots of foxing: a 

typical paper infection. Twenty paper samples of different 

centuries were examined to devise a no destructive method 

for taking samples from spots of museum's maps. The 

stains were examined by fluorescence and was checked 

that colonies no develop from areas without fluorescence 

therefore samples were taken using swabs from spots 

fluorescent. Moreover was proved that on the spots not 

clearly visible by naked eyes defined 'discoloring', but 

fluorescent, develop fungi if placed in humidity chamber. 

We should recommend the early identification of 

'discoloring' by fluorescence in order to arrange suitable 

thermohygrometric conditions for preventing development 

of infection. 



858 - Specific toxigenic, biological and molecular traits 

of a Gibberella moniliformis (Fusarium verticillioides) 

population from banana 

A. Moretti 1* , G. Mulè 1 , A. Susca 1 , M.T. González-Jaén 2 & 

A. Logrieco 1 

1 Institute of Science of Food Production (CNR), Viale 

Einaudi, 51, 70125, Bari, Italy. - 2 Faculdad de Biologia. 

Departamento de Genética. UCM, Avenida Complutense, 

s/n, Madrid, Spain. - E-mail: moretti@area.ba.cnr.it 

Gibberella fujikuroi includes at least 8 mating populations 

(MP), corresponding to biological species. Each MP 

generally possess a specific toxicological profile and a 

preferential host. Fusarium verticillioides and F. 

thapsinum, anamorphs of MPs A (G. moniliformis) and F 

(G. thapsina), shared the identical morphological traits, but 

they have a different preferential host (maize and sorghum) 

and only members of MP A can produce fumonisins. 

Strains morphologically identified as F. verticillioides were 

isolated from banana fruits, analysed for fumonisin 

production, none being a producer, and crossed by tester 

strains of MPs A and F. Although they produced fertile 

perithecia of MP A, time for obtaining perithecia and size 

of them differed significantly from usual fertile crosses 

among strains from maize. Pathogenicity tests with both 

banana and maize strains were also performed on banana 

fruits showing higher ability of strains isolated from 

banana to cause infection. Finally, banana and maize 

strains were compared by using AFLP and sequence of 

IGS, ITSs and calmodulin regions, allowing to split in two 

different clusters. Finally, strains of MP A from banana 

showed peculiar traits in toxin production, pathogenicity 

and molecular profiles. This could reflect important 

differences in ecology and natural history and triggers 

further investigations on the mechanisms of toxin 

production and pathogenicity in the same MP. Support of 

EU Fifth Framework (Detox fungi QLK1-CT-1999- 

001380). 

859 - Gnomonia fragariae - a causal agent of strawberry 

root and crown rot in Latvia 

I. Morocko 1* , J. Fatehi 2 & B. Gerhardson 2 

1 Pure Horticultural Research Station, Abavas 2, Pure, LV- 

3124, Latvia. - 2 Plant Pathology&Biocontrol Unit, SLU, 

P.O. Box 7035, S-750 07, Uppsala, Sweden. - E-mail: 

pures_dis@tukums.parks.lv 

In a survey on possible causes for an observed ongoing 

decreased productivity of strawberry plantations in Latvia 

about 200 severely stunted and wilted strawberry plants 

from 16 plantations were assessed for fungal root rot 

pathogens. An often encountered, and also the most clearly 

pathogenic fungus isolated was identified as Gnomonia 

fragariae. Agar plating on PDA and PCA were used for 

isolation and identification, based on morphological 

characters. Isolates of this pathogen have so far been 

258 


obtained from five fields in three locations. It was isolated 

from roots and crowns of stunted plants, showing wilt, 

reddish color in the older leaves and crown necrosis. 

Pathogenicity was confirmed using a detached leaf assay 

where wounded and unwounded leaves of the strawberry 

cultivar 'Honeoye' were inoculated by agar plugs with 

fungal colonies. Clear necrotic lesions were observed 

around agar plugs 2-3 days after inoculation on wounded 

leaves, and after 3-4 days on unwounded leaves. After few 

weeks the fungus also formed perithecia with long necks 

on infected leaves. The fungus was re-isolated from surface 

sterilized infected tissues. Based on the results obtained we 

conclude that Gnomonia fragariae is an important 

pathogen in strawberry root rot complex in Latvia and this 

respect also is of great economic importance. 

860 - Population dynamics and mycotoxin production of 

filamentous fungi in winter wheat grown in different 

tillage systems in eastern Germany 

M. Mueller 

Center for Agricultural Landscape and Land Use Research 

(ZALF), Institute of Primary Production and Microbial 

Ecology, Eberswalder Str.84, D-15374 Müncheberg, 

Germany. - E-mail: mmueller@zalf.de 

The effect of different wheat cultivation systems on 

mycoflora in wheat plants during the vegetation period 

from April to the harvest in August was investigated in 

long time field trials. Systems with silage maize or potatoes 

as preciding crops and conservation soil tillage with 

cultivator or conventional soil tillage with plough were 

compared concerning the population dynamics of 

filamentous fungi and the mycotoxin accumulation in 

plants or in harvested kernels. Special attention was given 

to the widely distributed genera Fusarium and Alternaria. 

Many species of them are plant-pathogens and able to 

synthesize toxic metabolites: trichothecenes and 

zearalenone by Fusarium species and alternariols and 

tenuazonic acid by Alternaria spp. Alternaria toxins were 

determined by reversed phase HPLC with fluorescence or 

DAD detection. The Fusarium toxins were analysed by 

means of ELISA test kits. The four different cultivation 

variants showed similar quantitative fungal colonization 

with an increase of the total fungal population during 

vegetation period. At each sampling date, the plants grown 

in conservation soil tillage with maize as preceding crop 

had the greater population of the genera Fusarium and 

Alternaria. In some cases Fusarium toxins were already 

analysed in wheat ears six weeks before the harvest 

independent of the tillage system. Alternaria toxins were 

only rarely detected in all variants. The influence of abiotic 

driving factors (temperature, rain fall) is discussed.


861 - Monitoring of compost-derived volatiles in the 

surrounding of composting facilities 

T. Müller, G. Fischer * , R. Thißen, S. Braun & W. Dott 

Institute for Hygiene and Environmental Medicine, 

University Hospital RWTH, Pauwelsstr. 30, D-52074 

Aachen, Germany. - E-mail: Guido.Fischer@post.rwthaachen.de 

Composting plants are known to emit malodorous 

compounds due to microbial activity. The objective of this 

study was to evaluate the emission of (M)VOC and their 

occurrence in the surrounding of composting plants. Data 

on the amount of windblown losses of malodorous 

compounds should be obtained. The spectrum of (M)VOC 

was compared with those of fungal cultures on compost as 

substrate in laboratory experiments. Samples were analysed 

by thermal desorption and GC/MS. Sampling locations 

were set up in downwind direction in distances of 50 m up 

to 800 m from the composting plants, and within these. 

Three compost plants varying in the annual turnover and 

type of process engineering were investigated. Laboratory 

experiments were carried out to characterise the speciesspecific 

production of (M)VOC by microfungi. (M)VOC 

were detected in distances up to 800 m, but the amount did 

not seem to be correlated with the type of process 

engineering, but rather with the activity within the 

facilities. The occurrence of (M)VOC coincided with the 

sensory perception of typical compost-odour on the 

sampling locations and increased numbers of microfungi in 

the air. Emission of (M)VOCs during composting cannot 

be avoided even if more sophisticated techniques of 

process engineering are used. It turned out that detection of 

certain VOC may be used as indicator for the occurrence of 

plant-related emissions of bioaerosols including microfungi 

in the surrounding of composting plants. 

862 - Fungal diseases of cultivated mushrooms in 

Armenia 

S.G. Nanagulyan 1* & A.G. Yesayan 2 

1 Yerevan State University, Department of Botany, 

A.Manoogyan str.1, Yerevan, 375025, Armenia. - 2 Yerevan 

State University, Department of Ecology and Nature 

protection, A.Manoogyan str.1, Yerevan, 375025, Armenia. 

- E-mail: snanagulyan@ysu.am 

Nowadays only two species - Agaricus bisporus and 

Pleurotus ostreatus - are cultivated in special cultivated 

areas in Armenia. In this report we are providing 

summarized information about cultivated areas of Armenia, 

data about inspection of some cultivated areas, and 

information concerning detection and occurrence of 

mushroom diseases in these areas. We have collected all 

possible data from various governmental and private 

organizations about mushrooms cultivated areas in 

different regions of Armenia. We have interviewed the 

local population concerning approaches to the cultivation, 

for example the carpophores of Agaricus bisporus were 

infected mostly by Wet Bubble Disease (Mycogone 

perniciosa), which has been recorded in very severe form. 

The percentage of infected areas was very high, 

approximately 80-90%. The preliminary examination has 

shown that now the small scale mushrooms' farms are 

attacked by several species of moulds and bacteria: 

Trichoderma, Penicillium, Aspergillus; Papulospora 

byssina; Chaetomium olivaceum; agaricoid fungi from 

genera Coprinus; Pseudomonas tolaasii, Dactylium 

dendroides. The results of investigation show that these 

diseases are observed often in the farms with insufficient 

ventilation, increased air and compost humidity, 

infringement of compost preparation technology, overuse 

of nitrogen fertilizers, wrong pasterization. New 

approaches to the investigation and further theoretical 

study in this field are required. 

863 - Anti-fungal activity of natural preservatives from 

plant extracts depends on the application method 

P.V. Nielsen * & K.I. Suhr 

BioCentrum-DTU, Building 221, DTU, DK-2800 Lyngby, 

Denmark. - E-mail: pvn@biocentrum.dtu.dk 

Antimicrobials from species and herbs are interesting 

alternatives to traditional preservatives, as they may act 

through the gas phase and have complementary 

antimicrobial spectra. Traditionally antimicrobial 

compounds are either added to the product or applied on 

the surface. For foods like bread and cheese addition to the 

packaging gas may be more effective. Ten plant essential 

oils were investigated by addition either 1) directly to a rye 

bread media or 2) to the surface or 3) as volatiles to the 

packaging atmosphere. Oils were from: bay, cinnamon 

leaf, clove, lemongrass, mustard, orange, sage, thyme and 

rosemary (two formulations). Test organisms were the 

common bread spoilage fungi Penicillium roqueforti, P. 

corylophilum, Erotium repens, Aspergillus flavus and 

Endomyces fibuliger. Smaller volatile compounds, such as 

allyl isothiocyanate, limonene, (and citral), were most 

efficient when applied through the gas-phase, whereas 

larger phenolic compounds, such as thymol and eugenol, 

worked better in direct contact, e.g.100 times more allyl 

isothiocyanate had to be added to the media to get the same 

effect as observed through gas phase addition. The optimal 

mixture of essential oils on the surface varied by the total 

amount added. A combination of cinnamon bark and lemon 

grass gave the best results. These results showed that 

application method is crucial and that, for active packaging 

applications, small highly volatile compounds are most 

interesting. 



864 - Simultaneous detection of GFP- and GUS-marked 

fungi of different formae speciales of Fusarium 

oxysporum on plant roots 

T. Nonomura * , Y. Matsuda & H. Toyoda 


of Agriculture, Kinki University, 3327-204 Nakamachi 


nonomura@nara.kindai.ac.jp 

In our attempt to visualize infection behavior of the fungal 

wilt pathogens inoculated onto plant roots, the fungi were 

genetically marked with two reporter genes. F. oxysporum 

f. sp. lycopersici (FOL) and F. o. f. sp. melonis (FOM) 

were transformed with the green fluorescence protein gene 

(GFP) and the β-glucuronidase gene (GUS), respectively. 

In the present study, we focused mainly on the attachment 

and subsequent hyphal elongation by microconidia 

inoculated onto roots of tomato and melon seedling. In 

addition, we attempted to directly distinguish different 

formae speciales of F. oxysporum onto the same plant roots 

by expression of different marker genes. Microconidia of 

GFP-marked FOL (KFOL-001) and GUS-marked FOM 

(KFOM-002) were inoculated onto roots of cotyledonal 

seedlings, and inoculated roots were first observed under a 

fluorescence microscope to detect KFOL-001 and then 

stained with X-gluc (substrate for GUS assay) to detect 

KFOM-002 under a light microscope. Consequently, both 

transformed pathogens could be clearly distinguished at the 

same site of inoculation. These results suggest that dual 

transformation of F. oxysporum is useful for analyzing 

behavior of nonpathogenic F. oxysporum challengeinoculated 

with pathogenic F. oxysporum. 

865 - Biscogniauxia and Daldinia; latent pathogens of 

deciduous trees 

L.K. Nugent * , G.P. Sharples & A.J.S. Whalley 

School of Biomoleculr Sciences, Liverpool John Moores 

University, Byrom Street, Liverpool L3 3AF, U.K. - E-mail: 

beslnuge@livjm.ac.uk 

Biscogniauxia Kuntze and Daldinia Ces. and De Not. are 

two wood inhabiting xylariaceous genera. Biscogniauxia 

species are frequently linked with canker diseases in 

stressed hosts e.g. B. mediterranea causes coal canker in 

Quercus suber (cork oak) and B. nummularia canker in 

Fagus (beech) while D. concentrica (Bolt. ex. Fr.) causes 

calico wood in Fraxinus (ash). Studies on ascospore 

germination and development of the anamorphs in culture 

in response to host extracts is presented. Biscogniauxia 

nummularia and Daldinia concentrica. have been isolated 

from their respective host leaves and branches and there are 

frequency of isolation maybe linked to ascospore 

production. The presence of Daldinia in leaves and in 

wood has been investigated microscopically, chemically 

and by molecular techniques, in addition to traditional 

isolation techniques following surface sterilisation. The 

260 


presence of latent pathogens and their relationship to stress 

of the host is presented. Experimentation on conditions 

leading to the latent invasion and subsequent development 

of teliomorphs are being undertaken in both field and 

laboratory. 

866 - Establishment of the first Karnal Bunt testing 

laboratory in South Africa 

O.M. O'Brien, I.H. Rong * & E.J. van der Linde 

ARC-Plant Protection Research Institute, Private Bag 

X134, Pretoria 0001, South Africa. 

The fungal wheat disease Tilletia indica, commonly known 

as Karnal Bunt (KB), was detected in a limited area of one 

of South Africa's wheat producing regions during 1999. 

Aimed at managing T. indica, a national survey to 

determine the occurrence of the disease was initiated by the 

Directorate Plant, Health and Quality, National Department 

of Agriculture. To this end, the Mycology Unit, ARC-Plant 

Protection Research Institute, Pretoria was tasked with 

setting up a laboratory for the analyses of seed and grain 

samples. Due consideration was given to the geographical 

distance of the laboratory from the main wheat producing 

areas of the country. The KB protocol, as recommended by 

the USDA/APHIS, was followed with some adaptations. 

Analyses were conducted for two consecutive years, 

providing valuable experience in managing a quarantine 

analytical facility of this nature. Protocols and procedures 

representing different phases of the process were devised 

for each workstation. These phases included: reception and 

registering of samples, sub-sampling, washing and sieving, 

centrifuging and preparing of microscope slides, detection 

of T. indica, data processing and reporting, and waste 

management. Laboratory procedures, problems 

encountered and the development of novel techniques, as 

well as the management and maintenance of the quarantine 

facility, are discussed. 

867 - Aflatoxins in the weaning food of Kenyan children 

S.A. Okoth 

Botany Department, University of Nairobi, P. O. Box 

30197, Nairobi, Kenya. - E-mail: dorisokoth@yahoo.com 

Cereal grains run a high risk of mycotoxin contamination 

yet they form the basis of gruels used in weaning children 

in Kenya. To these grains (maize, sorghum, millet) 

supplements such as cassava, groundnuts, beans, and fish 

are added and ground together, depending on the means 

and education of the parents. Among the mycotoxins, 

aflatoxins have been implicated in human diseases, 

including kwashiorkor. Sampling for aflatoxin 

contamination was done in Kisumu District, Kenya, an area 

with high relative humidity and temperatures, high 

incidence of kwashiorkor and the highest prevalence of 

absolute poverty, 63%, in the country. A total of 180


samples of the weaning flour were collected from Winam 

Division of Kisumu District, Kenya. These samples were 

analysed for aflatoxins in order to find out whether there 

exists a relationship between the high incidence of 

kwashiorkor and aflatoxin contamination. The samples 

were analysed by thin layer chromatography. Fifty two 

samples (29%) were positive for aflatoxins (concentration 

range 2-82 µg/kg), some exceeding the advisory limit. This 

indicates that aflatoxin is likely to present a major health 

hazard to weaned children in this part of Kenya. 

868 - Prevention of ochratoxin A in cereals - an EU 

project within Quality of Life and Management of 

Living Resources 

M. Olsen 1* , N. Jonsson 2 , N. Magan 3 , J. Banks 4 , C. Fanelli 5 , 

A. Rizzo 6 , A. Haikara 7 , A. Dobson 8 , J.C. Frisvad 9 , S. 

Holmes 10 , E. Pajunen 11 , S.-J. Persson 12 & A. Jonsson 13 

1 

National Food Administration, PO Box 622, SE-751 26 

Uppsala, Sweden. - 2 Swedish Institute of Agricultural and 

Environmental Engineering, P.O. Box 7033, SE-750 07, 

Sweden. - 3 Cranfield Biotechnology Centre, Cranfield 

University, Barton Road, Silsoe, Bedfordshire MK45 4DT, 

U.K. - 4 Central Science Laboratory, Sand Hutton, York 

YO41 1LZ, U.K. - 5 Laboratorio di Micologia, Università 

"La Sapienza", Largo Cristina di Svezia 24, I-00165 Roma, 

Italy. - 6 National Veterinary and Food Res. Inst., P.O. Box 

45, FIN-00581, Helsinki, Finland. - 7 VTT Biotechnology, 

P.O. Box 1500, FIN-02044 Espoo, Finland. - 

8 

Microbiology Department, University College Cork, 

National University of Ireland, Cork, Ireland. - 

9 

BioCentrum-DTU, Technical University of Denmark, 

Building 221, Søltofts Plads, DK-2800 Kgs. Lyngby, 

Denmark. - 10 ADGEN Ltd, Nellies Gate, Auchincruive, Ayr 

KA6 5HW, U.K. - 

11 Oy Panimolaboratorio- 

Bryggerilaboratorium AB, P.O. Box 16, FIN-02150 Espoo, 

Finland. - 12 Akron Maskiner, SE-531 04 Järpås, Sweden. - 

13 Svenska Lantmännen, Östra hamnen, SE-531 87 

Lidköping, Sweden. - E-mail: mool@slv.se 

Ochratoxin A (OTA) is a mycotoxin of considerable 

concern for human health and is classified as a possible 

human carcinogen. Cereals normally account for 50-80% 

of average consumer intake of OTA. Consequently, 

prevention of OTA formation by specific moulds in cereals 

would have a significant impact on levels of human 

exposure. The aim of this multidisciplinary project is to 

identify the key elements in an effective HACCP 

programme for OTA in cereals, and provide tools for 

preventative and control procedures. The project includes 

the whole food chain from field to the final processed 

product. The project objectives are divided into 4 different 

tasks, all important steps in a HACCP managing 

programme for ochratoxin A in cereals. Task 1: 

Identification of the critical control points (CCP). The 

ochratoxin producing species are being identified and 

characterised in order to identify the sources of 

contamination. Task 2: Establishment of critical limits for 

the CCP's. New knowledge concerning the microbial 

ecology of the ochratoxin producing species is provided 

and critical limits are being specified. Task 3: Developing 

rapid monitoring methods. Rapid detection methods for 

ochratoxin A and for the producing fungi are being 

developed. Task 4: Establish corrective actions. Reduction 

during cereal processing (milling etc) and during malting 

and brewing, is being investigated. 

869 - Biological efficacy of Trichoderma harzianum 

against wilt disease of sugarcane 

W. Ouvanich 

Plant Pathology and Microbiology Division, Department of 

Agriculture, Paholyothin Road, Jatujak, Bangkok 10900, 

Thailand. - E-mail: wantanee@doa.go.th 

Biocontrol efficacy of Trichoderma harzianum isolate Th1 

and Th2, against sugarcane wilt disease caused by 

Fusarium subglutinans and Cephalosporium sacchari were 

determined. Result from pot experiment show that both 

isolates of T. harzianum were able to significantly reduced 

wilt pathogen population in soil and wilt infection 

percentage of sugarcane roots. Their effect on wilt disease 

has been further studied in field experiment. Each isolate of 

T. harzianum was grown on sorghum seeds and then mixed 

the infested seeds with rice polish and manure. The mixture 

of each isolate of T. harzianum was applied to wilt 

pathogens infested field soils. Population of wilt pathogens 

in soil, percentage of infected root and yield of sugarcane 

were determined in all treatments. Population of wilt 

pathogens in infested field soil were significantly reduced 

by both T. harzianum Th1 (2.1 × 103 cfu/g soil ) and T. 

harzianum Th2 (2.8 × 103 cfu/g soil), as compare with 

control (5.5 × 103 cfu/g soil). Percentage of wilt infected 

root only was reduced by T. harzianum Th2 (20.4%). 

Significant difference in percentage of wilt infected root 

was not observed between T. harzianum Th1 (25.3%) and 

control (26.3%) treatments. Yield of sugarcane in T. 

harzianum Th2 treatment was also higher than in T. 

harzianum Th1 and control treatments. T. harzianum 

isolate Th2 has shown to be the effective biocontrol agent 

of wilt disease of sugarcane. 

870 - Natural flavonols fungitoxic to formae speciales of 

Fusarium oxysporum pathogenic to some ornamentals 

C. Pasini * , P. Curir, G. Brofiga & F. D'Aquila 

Istituto Sperimentale per la Floricoltura, Corso Inglesi, 

508 - 18038 Sanremo (Im), Italy. - E-mail: 

difesa@istflori.it 

Three constitutive flavonols from as many carnation 

(Dianthus caryophyllus) cultivars, resistant to Fusarium 

oxysporum f. sp. dianthi pathotype 2, have been assayed, 

alone or in combination, against the six following formae 

speciales of F. oxysporum affecting ornamental plants: f. 

sp.dianthi pathotypes 2 and 8, f. sp. asparagi, f. sp. 

ranunculi, f. sp.cyclaminis, f. sp. lilii. The fungitoxic effect 

was tested both as mycelial growth and as spore 

germination inhibition. The results show that some 



constitutive flavonols of carnation are fungitoxic towards 

the tested fungal formae speciales. Besides other fungitoxic 

mechanisms, such as the mitochondrial oxidative 

phosphorylation inhibition, flavonols may be antifungal 

due to their ability to associate as polydentate ligands with 

pathogen proteins: only a low solubility ensures a strong 

association via phenolic groups, and the low soluble 

flavonol aglycones could be therefore more effective, as 

complexing agents, than the respective soluble glycosides. 

In effect, among the flavonol aglycones, the 4'methoxylated 

kaempferide was particularly effective, while 

the glycosylated forms were less active than the 

corresponding aglycones. The inhibitory activity of the 

assayed flavonols was different depending on the fungal 

forma specialis, and a common physiochemical basis to 

explain the effect of the tested compounds was not found. 

871 - Broad spectrum herbal antifungal active against 

onychomycosis 

M. Patra 1* , S.K. Shahi 1 , K.G. Singh 2 & A. Dikshit 1 

1 

Biological Product Lab, Botany Department, University of 

Allahabad, Kuchery PO, PB No. 2026, Allahabad-211002, 

India. - 

2 

MLN Medical College, Department of 

Dermatology, Allahabad, India. - E-mail: 

patraindia@rediffmail.com 

During antifungal evaluation of some essential oils, 

Eugenia caryophyllata Thunb. was found to be most 

effective antifungal. The minimum inhibitory 

concentrations were found 0.1 µl ml -1 against E. floccosum, 

T. mentagrophytes, T. rubrum and 0.4, 0.5 µl ml -1 against 

Candida albicans, Scytalidium dimidiatum respectively. 

The oil inhibited potency against heavy doses of inoculum 

at 1.0 µl ml -1 concentrations. The oil also have broad 

fungitoxic spectrum as it also killed some other fungi at the 

range of 0.1-1.0 µl ml -1 concentrations. Our tests proved 

that the oil did not cause any adverse effect on human nail 

as well as on skin upto 5 µl ml -1 concentration. Further, we 

have formulated the oil in the form of antifungal lotion and 

ointment (2 µl ml -1 ) and subjected to topical testing on 

patients attending Out Patient Department (OPD) of 

M.L.N. Medical College, Allahabad. 10 patients were 

selected on the basis of KOH positive results and 

diagnosed as fungal nail infection. After 4th week of 

treatment, 60% patients were mycological cure (KOH 

negative). At the end of 10th week, all patients cured 

mycologically. No KOH negative cases of relapse were 

observed when patients were re-examined after two months 

following the end of 10th weeks. Thus, the oil in the form 

of lotion can be exploited as commercially after 

undergoing successful multicenter clinical trials, which is 

in progress. 

262 


872 - Microwave irradiation: a tool to improve the 

staining with fluorochromes of Aspergillus fumigatus 

conidia 

V. Prigione * & V. Filipello Marchisio 

Dipartimento di Biologia Vegetale Università di Torino, 

viale Mattioli 25 - 10125 Torino, Italy. - E-mail: 

valeria.prigione@unito.it 

Fungal conidia are ubiquitous in the air, where they can be 

the major pollutant and sources of infections, allergic 

reactions and toxicoses. Microscopy is the only currently 

available method for evaluating the total load of fungus 

aerosol. This kind of analysis could be made easier by the 

use of fluorochromes but, unfortunately, propagules of 

most fungi are not stainable or only a small percentage of 

the total number fluoresce, owing to their resistant wall. In 

this study we tested the use of chemical and physical 

treatments, respectively oxidation by the use of sodium 

hypochlorite and microwave irradiation, to improve the 

staining of A. fumigatus conidia with six different 

fluorochromes. We calculated the percentage of stained 

conidia respect to the total ones, comparing the percentages 

obtained after each treatment with those of the treatment 

free samples (control samples). In the control samples 

percentage of stained conidia was always lower than 44%; 

only Auramine O gave higher percentage (99%). The 

treatment with sodium hypochlorite gave high percentages 

of stained conidia (up to 99% with DAPI) but more than 

30% of them were lost during the following two 

centrifugations required to remove the reagent. On the 

contrary microwave irradiation always gave percentages 

higher than 98% and did not require further treatment. In 

addition fluorescence intensity appeared enhanced, making 

easier the observations by epifluorescence microscopy. 

873 - Flow cytometry, a novel approach to quantitative 

assessment of airborne fungal propagules 

V. Prigione 1* , G. Lingua 2 & V. Filipello Marchisio 1 

1 Dipartimento di Biologia Vegetale - Università degli Studi 

di Torino, viale Mattioli 25 - 10125 Torino, Italy. - 

2 Dipartimento di Scienze e Tecnologie Avanzate Università 

del Piemonte Orientale, corso Borsalino 54 - 15100 

Alessandria, Italy. - E-mail: valeria.prigione@unito.it 

Current limitations in the methodology for enumeration of 

airborne fungal propagules compromise the precision and 

accuracy of bioaerosol exposure assessment. Flow 

cytometry (FCM) is a tool with great potential for use in 

environmental microbiology because of the quantity and 

quality of data it provides in a timely fashion. There have 

been no reports of the application of FCM to the study of 

airborne mycoflora. In this study, FCM was used to count 

fungal propagules in laboratory suspensions with 

concentrations ranging from 10 3 to 10 6 conidia/ml and in 

field samples collected by an impinger device. To verify 

the accuracy and the precision of this technique, fungal


propagules counts made by flow cytometry were compared 

with counts by direct observation using epifluorescence 

microscopy. Flow cytometric counts of laboratory 

suspensions were performed by the use of forward angle 

light scattering, a parameter related to particle size. Field 

samples were stained with propidium iodide after 

microwave irradiation, to discriminate the biological 

particles, while forward angle light scattering was used for 

identifying and counting fungal propagules population. A 

close agreement was found between FCM and 

epifluorescence microscopy counts. 

874 - A 6 methylsalicylic acid synthase (6MSAS) 

homologous gene isolated Byssochlamys nivea is 

expressed during patulin production 

O. Puel 1* , A. Foures 1 , R. Benaraba 1 , M. Delaforge 2 & P. 

Galtier 1 

1 I.N.R.A. Laboratoire de Pharmacologie et Toxicologie, 

180 Chemin de Tournefeuille BP3 31931 Toulouse cedex 9, 

France. - 2 CEA Saclay DSV/DRM/ SPI, Bat136, F91191 

Gif sur Yvette Cedex, France. - E-mail: 

opuel@toulouse.inra.fr 

The Byssochlamys nivea NRRL 2615 strain can produce 

mycophenolic acid and patulin. We used degenerate PCR 

primers matching a ketosynthase nucleotide motif for a 

RACE-PCR strategy and isolated a polyketide synthase 

gene from B. nivea. The deduced amino acid sequence 

(1778 residues) displays 74% identity with Penicillium 

patulum 6-methylsalicylic acid synthase (6MSAS). Two 

6MSAS homologous fragments located respectively on the 

5 and 3 extremities were isolated from the mycophenolic 

acid producer P. brevicompactum genome. After 

translation, these fragments display 87 and 93% identity 

with P. patulum 6MSAS. B. nivea and P. brevicompactum 

cultures were monitored for PKS transcription kinetics by 

RT-PCR. The B. nivea messenger is expressed throughout 

the first 10 days of culture with a maximum observed level 

between day 2 and 5. Using the HPLC/DAD and LC/MS, 

the patulin precursor 6- methylsalicylic acid (day1-5), 

patulin and mycophenolic acid (2-10) were detected in B. 

nivea cultures. On the other hand, the mycophenolic acid 

was detected in P. brevicompactum cultures, but not 

patulin and 6-methylsalicylic acid. The P. brevicompactum 

messenger was not expressed during culture. The B. nivea 

amino acid sequence does not contain any methyl 

transferase site required in the 5-methylorsellinic acid 

synthesis (first precursor mycophenolic acid). The results 

strongly support the identification of a new 6MSAS 

involved in patulin and not in acid mycophenolic acid 

synthesis in B. nivea. 

875 - Comparison of genotypes and pathobiological 

phenotypes of environmental and commensal isolates of 

Candida albicans 

M.G. Quick 1 , J. Xu 2 , T.G. Mitchell 2 & D.E. Padgett 1* 

1 

Biology Dept., Univ. of NC at Wilmington, 601 S. College 

Rd. Wilmington, NC 28403, U.S.A. - 

2 

Dept. of 

Microbiology and Immunology, Duke Univ., Medical 

Center, Durham, NC 27710, U.S.A. - E-mail: 

Padgett@uncwil.edu 

The patterns of genetic variation, production of 

phospholipase, and growth rate were analyzed among 129 

isolates of the human pathogenic yeast, Candida albicans. 

Two populations of C. albicans were studied: 71 strains 

were collected from aquatic sources and 58 control strains 

were isolated from oral samples of healthy human 

volunteers. The phenotypes and genotypes were compared 

to determine if environmental isolates represent a greater 

risk to human health than commensal isolates. Genetic 

analysis, which was performed by PCR fingerprinting, 

revealed these two populations were genetically similar and 

agree with previous studies based on codominant genetic 

markers that the population structure of C. albicans is 

predominately clonal. There was no difference in the 

percent of phospholipase positive cultures between the two 

populations. However, among the isolates that were 

positive for enzyme production, the commensal isolates 

secreted significantly more phospholipase than the 

environmental isolates. Growth rate studies revealed that 

the environmental isolates replicated at approximately the 

same rate, as did commensal isolates. The DNA genotypes 

and the phospholipase results also were similar for the 

environmental and commensal isolates. 

876 - Ultrastructure of pycnidial cells and 

conidiogenesis of Septoria hyperici in dead tissues of 

host plant 

E. Rakhimova 

Institute of Botany & Phytointroduction, 36 D Timiryazev 

Str., Almaty, Kazakhstan. - E-mail: envirc@nursat.kz 

The techniques of light and electron microscopy were used 

to elucidate the details of pycnidial cells and conidium 

ontogeny in Septoria hyperici Desm., causal agent of 

foliage disease (leaf blotch) of common St. John's wort 

(Hypericum perforatum L.). The pycnidia were present in 

the intercellular spaces of host leaf tissues. The wall of 

subspherical pycnidium, visible under the light microscope, 

was composed of 1-3 layers of hyphae with normal 

structure. The hyphal cell protoplast was highly vacuolated, 

with well defined nucleus, numerous mitochondria, 

ribosomes, short cisternae of ER, and concentric bodies. 

Pycnidial wall septa had typical ascomycetous structure. 

The conidiophores with electron-dense cytosol, numerous 

mitochondria and large nucleus were found in inner layer 

of pycnidial wall. Conidiophores were less vacuolated, than 



pycnidial cells. In the apical region of conidiophore 

numerous vesicles were found. Ontogeny of conidia was 

holoblastic. Contents of the conidiophore and conidium 

were continuous during migration of full complement of 

organelles into conidial cell. Then a delimiting septum 

formed. Conidia were liberated schizolytically. Fusiform, 

septate conidia with numerous mitochondria, ribosomes 

and large lipid droplets were observed in the mucilagefilled 

central cavity of pycnidium. The conidium organelles 

were difficult to distinguish. Conidial wall consisted of two 

layers. The outer layer was moderately electron-dense wirg 

fibrillar surface. 

877 - Breeding of Pleurotus and Lentinula hybrids by 

pairings of neohaplonts for commercial cultivation 

R. Ramirez Carrillo * & H. Leal Lara 

Department of Food Science and Biotechnology, 04510 

Ciudad Universitaria, Col. Copilco, Mexico D.F., Mexico. 

- E-mail: rebecarc@servidor.unam.mx 

Cultivation of Lentinula edodes is slow and rather 

inefficient and aggressive strains, able to grow on a wide 

range of substrates, like hybrids of Lentinula with 

Pleurotus is a promising option. Interspecies hybrids 

cannot be obtained by conventional breeding, they have 

been produced only by protoplasts fusion, but such hybrids 

are sterile and highly unstable. Dedikaryotization yields the 

2 monokaryotic components of a strain and it has been 

used for improving strains of the same species. Therefore, 

7 strains of Lentinula and 3 of Pleurotus were 

dedikaryotized, Lentinula neohaplonts were classified in 4 

incompatibility groups and Pleurotus neohaplonts were 

also classified in 4 groups. Surprisingly, matings of 

neohaplonts of Lentinula and Pleurotus yielded 73 hybrids 

and only 5 negative pairings. From 19 hybrids cultivated 

on fermented straw, 2 produced fruit bodies with Lentinula 

morphology in the first flushes while 15 hybrids yielded 

Pleurotus morphology with BE around 160%. Parental 

strains of Lentinula and Pleurotus yielded fruit bodies with 

the typical morphology for each species; BE values were 

around 148 to 182% for Pleurotus and 24 to 261% for 

Lentinula. In the meiotic progeny from 3 hybrids, 4 mating 

types were present in 1:1:1:1 ratio (p 95%), confirming 

fertility of the hybrids and inheritance of tetrapolar 

heterothallic compatibility. Separation of these organisms 

into 2 different strains has to be reconsidered in view of 

these results. 

878 - Influence of environmental factors and fungicides 

on growth and deoxynivalenol production by Fusarium 

graminearum 

M.L Ramirez 1* , S.N. Chulze 2 & N. Magan 1 

1 Applied Mycology Group, Cranfield Biotechnology 

Centre, Cranfield University, Silsoe, Bedford MK45 4DT, 

U.K. - 2 Dpto. Microbiologia e Inmunologia, Universidad 

Nacional de Rio Cuarto, Ruta 36 Km 601, Rio Cuarto, 

264 


Cordoba., Argentina. - E-mail: 

m.l.ramirez@cranfield.ac.uk 

The impact of five fungicides (prochloraz, propioconazole, 

epoxiconazole, tebuconazole and azoxystrobyn, 0.5-50 

ppm) on growth of F. graminearum was evaluated in 

relation to water activity (aW, 0.99, 0.97, 0.95) and 

temperature (15 and 25 °C) on wheat-based media (in 

vitro). All fungicides reduce growth rates when compared 

to the control, and this reduction increased as the fungicide 

concentration increased. In general, none of the isolates 

was able to grow in the presence of any fungicide at 

concentrations higher than 15 ppm, regardless of the 

a W/temperature regime. The same fungicides were used in a 

second study on wheat grain (in situ), in order to evaluate 

the effect of two concentrations (0.5 and 5 ppm), three a W 

levels (0.995, 0.99 and 0.97) and two temperatures (15 and 

25 °C) and their interaction on growth rate and 

deoxynivalenol (DON) production by F. graminearum. All 

fungicides showed inhibition of growth at both 

concentrations in most conditions. The fungicides tested 

were less effective on grain in controlling growth than in in 

vitro studies. All fungicides showed DON stimulation or 

reduction in at least one of the conditions assayed. Our 

results show that stimulation or reduction in DON 

production in the presence of fungicides is influenced by 

complex interactions between aW, temperature, fungicide 

concentration and time of incubation in both strains 

studied. Such information is critical for effective fungicide 

control of Fusarium head blight of wheat. 

879 - Relationship between osmotic and matric 

potential, growth and endogenous accumulation of 

polyols and sugars in colonies of Fusarium 

graminearum 

M.L Ramirez 1* , S.N. Chulze 2 & N. Magan 1 

1 Applied Mycology Group, Cranfield Biotechnology 

Centre, Cranfield University, Silsoe, Bedford MK45 4DT, 

U.K. - 2 Dpto. Microbiologia e Inmunologia, Universidad 

Nacional de Rio Cuarto, Ruta 36 Km 601, Rio Cuarto, 

Cordoba., Argentina. - E-mail: 

m.l.ramirez@cranfield.ac.uk 

The effect of ionic (NaCl) and non-ionic osmotic potential 

(glycerol) and matric potential (PEG 8000) in the range - 

0.7 to -11.2 MPa, on growth of F. graminearum at 15 and 

25 °C on 2% wheat flour agar has been studied. The effect 

of stresses on endogenous accumulation of low and high wt 

polyols (glycerol, erythritol, arabitol and mannitol) and 

sugars (glucose and trehalose) in whole colonies of this 

fungus were also quantified. At 25 °C and high water 

potential growth rates on matric-modified media were 

higher than on osmotic-modified media. However, in the 

same range of water potentials on osmotically modified 

media, F. graminearum grew faster on NaCl than on 

glycerol-amended media. In general, the concentration of 

total polyols in whole colonies of F. graminearum 

increased as osmotic and matric potential was reduced to - 

8.4 MPa. Also marked differences in the ratios of low and 

high mol wt polyols in mycelial colonies were found. 

There was an increase in the glycerol content with a


progressive decrease in mannitol, arabitol and erythritol in 

colonies as osmotic potential decreased to -8.4 MPa on 

glycerol and NaCl amended media. However, on matrically 

modified media there was an increase in arabitol with a 

progressive decrease of glycerol and erythritol in colonies 

as the matric potential was decreased. There were also 

marked changes in the content of trehalose and glucose in 

colonies of F. graminearum in relation to osmotic and 

matric potential modification. 

880 - Inhibition of biosinthesis of different mycotoxins 

by using antioxidants alone or in association with food 

preservatives 

A. Ricelli 1 , P. Trionfetti-Nisini 2 , A.A. Fabbri 2 , M. Jestoi 3 , 

A. Rizzo 3 , L. Corazza 4 & C. Fanelli 2* 

1 ISPA, CNR, Viale Einaudi 51, 70125 BARI, Italy. - 

2 Università Roma "La Sapienza", Largo Cristina di Svezia 

24, 00165 Roma, Italy. - 3 Nat. Veterinary Food Research 

Institute, Hameentie 57, 00231 Helsinki, Finland. - 4 Ist. 

Patologia Vegetale, MIPAF, Roma, Italy. - E-mail: 

corrado.fanelli@uniroma1.it 

The efficacy of different antioxidant compounds both 

synthetic and of natural origin, alone or in combination, has 

been tested in order to verify their action both on the 

growth and on toxins production in different 

mycotoxigenic fungi. In particular we have tested some of 

the toxigenic fungi that often contaminate food and feed: 

Aspergillus ochraceus and Penicillium verrucosum 

(ochratoxin A producers), Fusarium graminearum 

(Zearalenon producer), F. moniliforme and F. proliferatum 

(fumonisins producers). The experiments have been 

performed on wheat and maize seeds moistened up to a 

water activity of 0.85 and 0.95 and incubated at 28 °C (A. 

ochraceus and P. verrucosum) or at 25 °C (Fusaria strains) 

for 30 days after the inoculum. The tested compounds were 

BHA, BHT 0.02% w/v, K sorbate 0.2% w/v, Na propionate 

0.2% w/v, resveratrol 10-4 and 10-3 M, ellagic acid 10-4 

and 10-3 M, Lentinula edodes 4% w/v cultural filtrates 

(LCF). Concerning both the ochratoxin A and fumonisin 

producing fungi, the best results, both on the growth and on 

toxin production (more than 80% inhibition) have been 

observed with BHA and resveratrol on maize seeds and 

with K sorbate or resveratrol on wheat seeds. In some cases 

it has been detected a very high inhibition of ochratoxin A 

production on wheat seeds also by using LCF Other 

experiments are in progress in order to obtain a long-lasting 

inhibiting effect. This work has been sponsored by grants 

PL-QLRT-1999-433 and 966 of EC. 

881 - Effect of fungicides commonly used for 

Podosphaera leucotricha control on the powdery mildew 

hyperparasite, Ampelomyces quisqualis 

J.C. Russell 1 , X. Xu 2 & P. Jeffries 1* 

1 Horticulture Research International, East Malling, Kent, 

ME19 6BJ, U.K. - 2 University of Kent at Canterbury, 

Canterbury, Kent, CT2 7NJ, U.K. 

Apple powdery mildew, caused by Podosphaera 

leucotricha, can significantly reduce crop yield. Severity 

may increase with successive seasons, resulting in stunting 

and distortion of leaves and general decline in tree health. 

Current strategy for apple powdery mildew control relies 

mainly on fungicide application at intervals of 

approximately ten days throughout the growing season. A 

biological control agent, using the hyperparasite 

Ampelomyces quisqualis, is one possible way to reduce this 

intensive chemical input. For it to be incorporated into the 

current management practice the hyperparasite must 

tolerate most commonly used fungicides. Eight isolates of 

A. quisqualis, representing each of the eight ITS groups 

(based on analysis by SSCP), and the commercial 

formulation AQ10, were tested for their tolerance to 

Systhane, Dorado, Nimrod, Topas, and Sulphur. The 

effects of fungicide on spore germination and colony 

growth were investigated. Dorado had no significant effect 

on the rate of germination of isolate DSM 2223, but 

otherwise all fungicides had a suppressive effect on spore 

germination of the isolates examined. This suppression 

varied between 15.28% for Systhane on isolate CBS 

130.79 and 89.63% for Topas on isolate 265. Fungicide 

application to established colonies seemed to have little 

initial effect, but by 21 days post application hyphal growth 

had been arrested. Sulphur and Systhane had the least and 

the greatest effect on hyphal growth rate respectively. 

882 - The main wheat leaf spot pathogens in the Czech 

Republic in the last years 

J. Sarova 

Research Institute of Crop Production, Drnovska 507, 

Prague 6 - Ruzyne, 161 06, Czech Republic. - E-mail: 

sarova@vurv.cz 

Leaf spot diseases of wheat are caused by different 

pathogenic fungi. This study was focused on relatively new 

wheat leaf spot diseases in the Czech Republic especially 

on tan spot caused by Pyrenophora tritici-repentis (anam. 

Drechslera tritici-repentis). The occurrence of wheat leaf 

spot diseases was monitored in the Czech Republic in 2000 

and 2001. The samples were collected from different 

localities of the Czech Republic. Fungal pathogens were 

determined with microscopic methods. Frequency of the 

pathogens varied in both years because the occurrence of 

fungal leaf spot pathogens depends on climatic factors 

especially on the temperature, the humidity and the 

precipitation. The most prevalent fungal leaf spot 



pathogens of wheat were Pyrenophora tritici-repentis and 

Phaeosphaeria nodorum (anam. Stagonospora nodorum) 

in 2000. Mycosphaerella graminicola (anam. Septoria 

tritici) was the most common fungus isolated from the 

wheat leaf spot samples collected in 2001. Didymella 

exitialis (anam. Ascochyta sp.) was one of the most 

important pathogens in 2001. Probably it has not been 

described in the Czech Republic yet. This study was 

supported by the Grant Agency of the Czech Republic 

(project No. 521/00/0226). 

883 - The grass endophyte gene for the first step in 

ergot alkaloid biosynthesis 

C.L. Schardl 1* , C. Machado 1 , J. Wang 1 & D.G. 

Panaccione 2 

1 University of Kentucky, Dept. Plant Pathology, Lexington, 

KY 40546, U.S.A. - 2 West Virginia University, Div. Plant & 

Soil Sci., Morgantown, WV 26506, U.S.A. - E-mail: 

schardl@uky.edu 

The endophytic Neotyphodium spp. grow in aerial tissues 

of grass hosts where they cause no symptoms and exhibit 

no signs; yet, often they help protect against insects, 

nematodes and vertebrate herbivores, increase drought 

tolerance, and improve plant growth and nutrient 

acquisition. Several endophyte alkaloids are variously 

active against insects and/or vertebrates. Among these are 

ergot alkaloids, including lysergic acid and its peptine 

derivative, ergovaline. Ergovaline is closely related to 

ergotamine, which is produced by the infamous ergot 

fungus (Claviceps purpurea), and is a potent mycotoxin 

(but a useful pharmaceutical at low doses). Ergovaline is 

thought to be responsible for toxicoses and reproductive 

problems in livestock that ingest certain endophyteinfected 

fescues and ryegrasses. The dmaW gene, encoding 

the probable first step in ergot alkaloid biosynthesis 

(dimethylallyltryptophan synthase), was cloned from 

Claviceps fusiformis and C. purpurea, then from the tall 

fescue endophyte, N. coenophialum, and a Neotyphodium 

sp. (Lp1) from perennial ryegrass. There were two dmaW 

genes in N. coenophialum, but only one in Lp1. Markerexchange 

mutagenesis of dmaW in Lp1 eliminated 

production of ergovaline as well as simpler ergot alkaloids, 

confirming the role of the dmaW gene. Mutagenesis of 

toxin-production genes in this manner presents 

opportunities to develop forage grass cultivars with 

protective endophytes that lack anti-livestock activities. 

884 - A polyphasic study of Fusarium species in the 

section Sporotrichiella - different approaches for 

distinguishing closely related fungal species 

H. Schmidt 1* , A. Holst-Jensen 2 , S.S. Klemsdal 3 , C.M. 

Kullnig-Gradinger 4 , C.P. Kubicek 4 , R.L. Mach 4 , L. 

Niessen 1 , H.I. Nirenberg 5 , U. Thrane 6 , M. Torp 3 , T. Yli- 

Mattila 7 & R.F. Vogel 1 

266 


1 TU München, Lehrstuhl für Technische Mikrobiologie, 

Weihenstephaner Steig 16, 85350 Freising, Germany. - 

2 National Veterinary Institute, Section of Food and Feed 

Microbiology, P. O. Box 8156 Dep, 0033 Oslo, Norway. - 

3 The Norwegian Crop Research Institute, Plant Protection 

Centre, Høgskoleveien 7, N1432 Ås, Norway. - 4 Institute 

for Chemical Enginering, Microbial Biochemistry and 

Gene Technology Group, Technical University of Vienna, 

Getreidemarkt 9/166, A1060 Vienna, Austria. - 5 Institute of 

Blant Virology, Microbiology, and Biological Safety, 


Forestry, Königin-Kuise-Str. 19, 1195 Berlin, Germany. - 

6 Technical University of Denmark, Department of 

Biotechnology, Bldg 221, DK-2800 Lyngby, Denmark. - 

7 Laboratory of Plant Physiology and Molecular Biology, 

Department of Biology, University of Turku, FIN-20014 

Turku, Finland. - E-mail: Holger.schmidt@bl.tum.de 

An integrated systematic study was carried out to clarify 

the taxonomic position and relationship of Fusarium 

langsethiae Torp and Nirenberg, ined. within the Fusarium 

section Sporotrichiella. Different isolates of this species 

were compared with strains of the closely related species F. 

poae and F. sporotrichioides using a composite data set. 

This set consisted of DNA sequences derived from the 

intergenic spacer region (IGS), the internal transcribed 

spacer region (ITS) and partial sequences of the b-tubulin 

and the elongation factor-1 alpha (EF-1a) gene, AFLP 

fingerprints, chromatographic data on secondary 

metabolites and morphology. From these combined data a 

consensus matrix was calculated. This was used as the 

basis for the construction of an UPGMA dendrogram and a 

multidimensional scaling which both revealed a clear 

separation of these three taxa. The discriminatory power of 

the different approaches varied significantly. The 

correlation of the clustering obtained by analysis of the 

combined data and that one obtained by an individual 

experiment was worst with ITS sequences, whereas AFLP 

appeared to reflect the composite data matrix best. 

885 - Development of SCAR-PCR primers for the 

detection of ochratoxinogenic Aspergilli 

H. Schmidt * , L. Niessen & R.F. Vogel 

TU München, Lehrstuhl für Technische Mikrobiologie, 

Weihenstephaner Steig 16, 85350 Freising, Germany. - Email: 

Holger.schmidt@bl.tum.de 

The production of ochratoxin A (OTA) is described for 

Penicillium verrucosum, P. nordicum and a wide variety of 

Aspergillus species belonging to different sections. Besides 

the taxonomic inhomogenity of OTA producing fungi the 

ability to synthesize OTA is highly variable even within a 

species. AFLP (amplified fragment length polymorphism) 

typing revealed, that no clear correlation of the ability of a 

strain to produce the toxin and the taxonomic position can 

be found. As a consequence it was decided to develop 

group specific PCR primers for potential OTA producing 

Aspergillus species. The strains were screened for 

polymorphic DNA markers using AFLP. Bands that were 

considerd to be characteristic for a specific group were cut 

out from silverstaind polyacrylamide gels. The DNA was


eluted from the gel, reamplified and the fragments were 

sequenced. Based on these results SCAR-PCR primers 

(sequence characterized amplified region) were 

constructed, the PCRs were optimized and tested against 

DNA of different fungal strains. The results show that this 

approach is valuable for the rapid early detection of 

potential ochratoxinogenic fungi in food. 

886 - Identification of Malassezia species in patients 

with pityriasis versicolor submitted to the Razi Hospital 

in Tehran 

M. Shams 1* , M. Rasaee 2 , M. Moosavi 1 & M. Razzaghi 

abyaneh 3 

1 Dept.of Mycology, Faculty of Medical Sciences,Tarbiat 

Modares University, P.O.Box:14115-111, Tehran, Iran. - 

2 Dept.of Biotechnology, Tarbiat Modares University, 

P.O.Box:14115-111, Tehran, Iran. - 3 Dept.of Myclogy, 

Pasteur Institute of Iran, P.O.Box:13164, Tehran, Iran. - 

E-mail: shamsm@modares.ac.ir 

Lipophilic yeasts of the genus Malassezia (Pityrosporum) 

belong to the normal flora of human skin and many warmblooded 

animals. These fungi can produce a diverse range 

of diseases that most frequent of them is pityriasis 

versicolor, a chronic recurrent skin disease occurring 

primarily in tropical regions. The genus Malassezia has 

been recently enlarged to include seven distinct species. 

This survey was undertaken to present a practical approach 

for differantiation of all Malassezia yeasts isolated from 

clinical materials of patients with pityriasis versicolor for 

the first time in Iran. The presence of disease was 

confirmed on the basis of the observation of budding yeast 

cells and short curved hyphae in skin specimens by direct 

microscopy. Malassezia yeasts were isolated after culturing 

the samples on modified Dixon (mDixon) agar. A 

combination of different characteristics includes yeast cell 

morphology, ability to growth on sabouraud dextrose agar, 

catalase test and ability to utilize individual Tweens 

(20,40,60,80) were used for identification of species. In 

general, 138 patients with pityriasis versicolor includes 

52.2% male and 47.8% female were identified. Direct 

microscopy and culture results were positive in 94.4% and 

63% of patients, respectively. Totally, 91 isolates of 

Malassezia belonging to four different species, M. globosa 

(66 isolates), M. furfur (18 isolates), M. obtusa (5 isolates) 

and M. sympodialis (2 isolates) were identified. 

887 - Allergic fungal sinusitis (AFS) among the 

candidates of functional endoscopy sinus surgery 

(FESS) 

T. Shokohi 1* & M. Maddani 2 

1 Department of Medical Mycology and Parasitology 

Mazandaran University of Medical Sciences, po box 483, 

Iran. - 2 Ear, Nose,and Throat, Mazandaran University of 

medical Sciences, po box 483, Iran. - E-mail: 

taherehshokohi@yahoo.com 

This study was designed to determine the rate of AFS 

among the patients with chronic sinusitis who failed to 

respond to repeated courses of antibiotic and were 

candidated for FESS in Sari. Allergic mucin and Sinus 

lavage were collected during FESS from 100 Patients 

meeting diagnostic criteria for AFS and were submitted for 

mycology and pathology investigations. The specimens 

were centrifuged and the sediment was mounted in KOH 

and Gram stained for direct examination. Multiple 

fragments of mucosa were removed at surgery and stained 

with H&E and PAS for the pathology evaluation. In this 

study we report 9 proven cases of AFS (with demonstration 

of fungal hyphae by direct exam and culture) and 8 

suspected cases (confirmed by direct exam or culture). The 

patients' age ranged 12 to 62 years with a mean age of 24.5 

years with female predominance. All of the patients were 

immunocompetent. 47% of the patients had a history of 

atopy. Histopathologically, hyphae were not seen. The 

genera of the fungus identifiable in all but in one patient it 

was not identifiable. 53% of isolated fungus are of hyaline 

hyphomycete particularly Aspergillus and Penicillium and 

47% of them are members of dematiaceous family 

particularly Cladosporium and Nigrospora. In eleven out 

of the seventeen, fungal hyphae were noted and in all of the 

seventeen, there were positive fungal culture. During 6 

months follow up; we identified recurrence in 2 patients 

clinically without any microscopy and culture evidence. 

888 - Identification of virulence variety in ascochyta 

blight fungus [Ascochyta rabiei (Pass) Lab.] using 

differentials lines 

F. Shokoohifar 1 , A. Bagheri 2 & M.F. Rastegar 2* 

1 Research Center For Plant Science, 2 Sardadvar av.; 

Bahar St.; Zip Code:91747484815; Mashhad., Iran. - 

2 Ferdowsi University of Mashhad, College of Agriculture, 

P.O.Box;91775-1163; Mashhad., Iran. 

The study was conducted to distinguish the pathogenecity 

variety of the fungus population in the country. To achieve 

this goal, 26 isolates were choosing as candidates for 16 

provinces that were the most important chickpea 

production areas. Pathogenecity test was achieved using of 

16 differential lines in the greenhouse condition. It was 

used current methods for classification of isolates as well 

as a new method on the base of analysis of variance. The 

later method was better than other methods in classification 

of variety because of statistical base. In this method 

differential lines have been grouped in six levels of 

resistance and isolates have been grouped in six 

pathotypes. Also their distribution pattern has been 

identified in Iran. Results of this method are more useful 

for resistance selecting in national mass. As whole, the 

study revealed that there were too vigorous pathogenic 

group in west and north west of the country, that none of 

tested lines didn't show any resistance an this study. It 

seems that it is better to design breeding projects for 

screening of resistance line regarding to identify pathotype 

groups in each region. 



889 - Performance of Pinus roxburghii inoculated with 

pure culture of four indigenous ectomycorrhizal fungi 

G. Shresta 

Royal Nepal Academy of Science & Technology (RONAST, 

P.O.Box. No. 3323, Kathmandu, Nepal. - E-mail: 

geetashrestha1@hotmail.com 

Many ectomycorrhizal fungi were isolated from the 

sporocarps of different places such as (Dadheldhura, 

Makwanpur and Palpa district etc.) of pine forest of 

Nepal.Out of the isolated ectomycorrhizal fungi, four 

different ectomycorrhizal fungi namely Astraeus 

hygrometricus, Boletus sp., Lycoperdon pyriforme and 

Pisoolithus arrijus. These four fungi were subjected to pure 

culture synthesis. These fungi were selected from the pot 

culture experiment. Then these four indigenous pure 

ectomycorrhizal fungi were inoculated in nursery Pinus 

roxburghii for their performance at two nursery sites, 

Godavari forest nursery and Chalnakhel forest nursery 

sites. Their performance were observed after six, nine and 

twelve months of seedlings. Inoculated seedlings attained 

better shoot height and had better root length. more lateral 

roots, more mycorrhizal roots, more weight of shoot and 

more weight of root per plant. The inoculated transplants 

had higher survival rate in the field as compared to those 

which were not inoculated. 

890 - Effect of immunization against candidiasis after 

ultrasonic treatment of fungal keratoconjunctivitis 

D.S. Silin 

Odessa State Agrarian University, Department of animal 

pathology, of.11, 65, Kanatnaya str., Odessa, 65012, 

Ukraine. - E-mail: silin12@hotmail.com 

Ocular candidiasis was diagnosed in six 12-weeks old 

Persian kittens after four-weeks unsuccessful treatment of 

chronic keratoconjunctivitis by Sofradex. Bilateral corneal 

erosions and conjunctival infiltrations were detected. 

Examination of corneal scrapings revealed oval, yeast-like 

cells attributable to Candida albicans. Examination of 

conjunctival fluid samples by ELISA revealed decreasing 

of Immunoglobulins A and G quantity with absence of 

specific IgA and IgG to Candida whole-cell antigens. 

Ultrasound treatment was applied on right eyes of all cats 

for determination of antifungal activity and 

immunostimulation of ultrasound in course of 10 

applications daily by 10 minutes with 0.2 W/cm2 dosage. 

Almost full healing was occurred in all, even in 

contralateral eyes after treatment. Candida cells were 

absent, specific IgA to Candida whole-cell antigens were 

detected in high titers. Probably, ultrasound destroyed 

Candida cells and delivered antigens through epithelium to 

immune representative cells followed by local humoral 

immune response formation. Local immunity was 

stimulated by ultrasound as well. This immune response 

was not limited in exposed eyes, but was expanded to 

268 


contralateral eyes due to specific immune cells exchange. 

Thus, ultrasound can be useful for some transmucous 

vaccine delivery systems and for specific treatment without 

infectious agents determination, particularly, in 

parasiteocenotic cases. 

891 - Role of salicylic acid in systemic resistance 

induced by Pseudomonas spp. against Macrophomina 

phaseolina in chickpea 

T. Singh * & A. Dilip 

Banaras Hindu University, Department of Botany, B.H.U., 

Varanasi, India. - E-mail: singhtanu71@rediffmail.com 

Pseudomonas fluorescens isolate 4-92, applied to roots, 

induced systemic resistance against charcoal rot disease in 

chickpea roots caused by Macrophomina phaseolina 

(Tassi) Goid. Salicylic acid (SA) from bacterial culture on 

plant tissues was quantified by high performance liquid 

chromatography. P. fluorescens produced SA in king's B 

broth and also induced chickpea roots to accumulate 

endogenous SA one day after bacterial inoculation. Using a 

split root system, more SA accumulated in roots treated 

with bacteria than in distant roots on the opposite side of 

the root system in the first two-three days, but this 

difference disappeared after 5-6 days. SA levels were 

significantly higher in plants treated with bacteria 

compared to the split control, from one to six days after 

bacterization. SA did not inhibit sclerotial growth of M. 

phaseolina at 100-200 µg ml -1 , invitro, but higher levels 

inhibited sclerotial growth. Exogenously applied Sa failed 

to induce local or systemic resistance against a challenge 

infection by the pathogen in planta. The results of this 

study show that exogenous applied SA does not induce 

systemic resistance against charcoal rot disease in chickpea 

root caused by M. phaseolina, but endogenous SA 

accumulation in chickpea roots may be involved in induced 

systemic resistance. 

892 - Investigations of Phytophthora infestans oospores 

in Moscow region 

A.N. Smirnov 1* , S.A. Kuznetsov 1 & S.N. Elansky 2 

1 Moscow Agricultural Academy, Lystvennichnaya al. 2, 

Moscow 127550, Russia. - 2 Moscow State University, 

Vorobjevi gory Moscow 119899, Russia. - E-mail: 

nik.smi@g23.relcom.ru 

Our purpose was to investigate naturally formed oospores 

in Moscow region. On the presence of oospores, 1099 

blighted potato leaves, 123 tomato leaves, and 188 tomato 

fruits were checked. The frequency of oospores in potato 

leaves was 5.6%. In tomato fruits and leaves the oospores 

were found more often. Their frequencies were 22.8 and 

42.0% respectively. This can make the problems for tomato 

cropping in Moscow region. The oospores had a quite 

different morphology. They varied in size (their diameter


was from 18 µ to 48 µ), thickness of the wall, as well as in 

the presence of antheridium and the form of oogonium. 

According to these features, in 1999 and 2000 45 

morphological types were established among detected 

oospores. They corresponded to different stages of oospore 

maturation. The oospores can be formed as a result of 

cross-fertilization between mycelia of A1 and A2 mating 

types (hybrid oospores), as well as of self-fertilization, and 

of parthenogenesis (non-hybrid oospores - from one 

mycelium). Studying the distribution of mating types and 

morphology of oospores it was possible to assess their 

origin. It was found that in 1999 and 2000 non-hybrid 

oospores occurred much more often than hybrid oospores. 

The estimated frequency of hybrid oospores was about 

15%. The hybrid and non-hybrid oospores may have 

different role in the infection cycle of P. infestans. 

893 - The needle blight fungus Delphinella abietis 

attacks Abies species in western Norway 

H. Solheim 

Norwegian forest research institute, Høgskoleveien 12, 

1432 Ås, Norway. - E-mail: halvor.solheim@skogforsk.no 

During the last decade the needle blight fungus Delphinella 

abietis has been attacking various Abies species in western 

Norway. A clonal trial with subalpine fir (Abies 

lasiocarpa) was seriously attacked in 2000. It was 

significant differences between clones in the attack. Some 

clones were heavily attacked while others were free for 

symptoms. In a greenery trial of Nobel fir (Abies procera), 

most branches had good quality until 1994 when 31% of 

the branches were discarded after attack by D. abietis. 

Since then a high level of losses has been noted and 87% of 

the branches were discarded in 2000. It seems that frost did 

not initiate or maintain the disease in the stand, but 

generally low temperatures may have been important. 

894 - Toxicological characterisation of animal feed 

using in vitro methods 

A. Solyakov 1* , E. Jonsson 1 , V. Wenehed 2 , P. Haggblom 1 & 

A. Forsby 2 

1 Dept of Animal Feed, National Veterinary Institute (SVA), 

751 89 Uppsala, Sweden. - 2 Dept of Neurochemistry and 

Neurotoxicology, Stockholm University, 106 91 Stockholm, 

Sweden. - E-mail: Alexey.Solyakov@sva.se 

Mould growth on animal feed is a worldwide problem that 

can reduce the quality of the feed. It is known that moulds 

can produce mycotoxins, which can be mutagenic, 

carcinogenic but also neurotoxic. Poisoning by mycotoxins 

can cause suffering for animals, massive economical losses 

and a possible transfer of toxins to humans via the 

foodstuffs. Therefore there is a need for identification of 

feed of bad quality that may possess hazards to animals 

and/or humans. Aspergillus fumigatus, which is known to 

produce, among other toxins, gliotoxin, was chosen for 

development of a screening method for various toxic 

substances in animal feed. Three matrixes - maize, 

commercial animal feed and wheat - were adjusted to water 

content 12, 21 and 32%, and inoculated with A. fumigatus 

for 3 days, 2 and 4 weeks at 37 °C. The samples were 

extracted and purified by solid-phase technique. HPLC and 

LC-MS was used to detect gliotoxin as a marker of toxic 

substances produced by A. fumigatus. No gliotoxin was 

detected in the samples. The purified extracts were also 

applied to the human neuroblastoma SH-SY5Y cells, and 

the general cytotoxicity was determined. The extracts 

showed to be cytotoxic to the SH-SY5Y cells. The results 

indicate that the screening method based on the SH-SY5Y 

cells can be used for screening of toxic metabolites of A. 

fumigatus. Further research is in progress to investigate if 

the method can be applicable for other toxic substances. 

895 - A nucleic acid probe detects different species of 

Pestalotiopsis spp. isolated from five fruit crops 

K. Sridhara Gupta 1* , R.D. Rawal 2 & A. Vani 2 

1 Department of Plant Pathology, University of Agricultural 

Sciences, GKVK, Bangalore - 560 065, India. - 2 Division of 

Plant Pathology, Hessaraghatta Lake Post, Hessarghatta, 

Bangalore - 560 089, India. - E-mail: 

sridharagupta@hotmail.com 

Fruit crops play a major role in the economy of the 

country. Pestalotiopsis species causes leaf spot, leaf blight 

and fruit rot diseases in many fruit crops. The reports in the 

literature say that the Pestalotiopsis species are named after 

the host it is isolated from. To overcome this discrepancy, 

in the present studies, the pathogen was isolated from the 

leaves of mango grapes, guava, sapota and macadamia, 

from the berries and canes of grapes and also from fruits of 

guava. The pathogen was cultured on Potato Dextrose Agar 

medium. Total Nucleic Acid of the fungus was isolated 

from each isolate. A 5.3 kb EcoRI fragment generated from 

genomic DNA of the mango isolate was labeled with DIG 

and was used as a probe. Genomic DNA of the fungus 

isolated from mango, sapota and grape leaf bound to the 

probe strongly while that from grape berries, macadamia 

and guava bound to the probe only at low stringency. The 

isolate from grape cane did not bind to two other pathogens 

commonly found on fruit trees namely Greenari uvicola 

and Colletotrichum capsici. Morphological differences in 

the spores and cross-pathogenicity of these isolated were 

also found. 

896 - Enhancement of seedling vigour and biological 

control of Alternaria leaf spot of chili (Capsicum annum 

L.) using Trichoderma harzianum 

A. Srivastava * & J. Mishra 

PG Department of Botany, SMM Town PG College, Ballia, 

277 001, U.P., India. - E-mail: aloksrivastva@hotmail.com 



Isolates of Trichoderma harzianum enhanced seedling 

vigour in Chili (Capsicum annum) germinated in potting 

soil. When chili seeds were sown in the potting soil 

inoculated with Trichoderma harzianum 25-92 or 29-92 the 

germination rate increased by 15 and 20%, respectively, 

compared with water treatment control. After 5 days of 

growth, the average plant height increased by 25 and 39%, 

respectively; the average fresh weight increased by 33 and 

36%, respectively. The level of nitrogenase, peroxidase, 

glucanases, chitnases and average phenolic content in the 

roots was evaluated as key indicators of vigour. 1.5 to 3 

fold increase in nitrogenase, peroxidase and chitinase was 

recorded, whereas, the phenolic contents increased by 18 

and 17%, respectively. There was no any significant 

increase in glucanses in roots. Total chlorophyll in first, 

second and third leave was increase by 35-56%. The plants 

were exposed to Alternaria solani by spraying the conidial 

suspension on leaves of 3-week-old plants previously 

assesed for enhanced vigour. No disease was recorded on 

the plants indicated that pretreatment of chili seedlings at 

nursery level with Trichoderma isolates induced resistance 

in the plants. 

897 - Use of a microcosm system for biological 

screening against Botrytis cinerea on Pinus sylvestris 

seedlings 

E. Stenström * , K. Capieau & J. Stenlid 

Department of Forest Mycology and Pathology, Swedish 

University of Agricultural Sciences, P.O. Box 7026, 

Sweden. 

Botrytis cinerea, the causal agent of grey mould, is the 

most common and serious pathogen in all Swedish forest 

nursery production. Most nurseries make use of fungicidal 

spraying programs to prevent grey mould infections. 

Biocontrol has so far not been tested in forest nurseries 

even if biological control agents (BCA) are used 

internationally in agriculture. We set up and standardised 

an inoculation method in a microcosm system, which 

would allow us to screen fungal and bacterial isolates 

against B. cinerea. In this microcosm system small P. 

sylvestris seedlings are heat stressed before inoculated with 

a spore solution containing pathogen spores, BCA or a 

combination of those. Results are scored ten days after 

inoculation by examining the seedlings for the presence of 

B. cinerea's conidia. To test the reliability of this 

microcosm system method, the performance of three 

commercial biocontrol products, in suppressing B. cinerea 

on pine seedlings was tested. Those results were then 

compared with two other testing systems, laboratory 

experiments and field trials using the same commercial 

products. In all three testing systems we found that the 

commercial products Binab TF.WP TM and GlioMix TM 

performed better than Mycostop TM and we concluded that 

the similarity of the results between the different testing 

systems was a strong indication for the usefulness of the 

microcosm system in screening isolates against B. cinerea. 

270 


898 - Effect of Lactobacillus rhamnosus and a fermented 

milk on the growth of Aspergillus and Penicillium 

species 

J. Stiles 1* , V. Carter 2 & L.B. Bullerman 3 

1 University of Nebraska, 319 FIC, Lincoln 68583-0919, 

U.S.A. - 2 University of Nebraska, 318 FIC, Lincoln 68583- 

0919, U.S.A. - 3 University of Nebraska, 349 FIC, Lincoln 

68583-0919, U.S.A. - E-mail: jstiles2@unl.edu 

This study was done to investigate the effect of 

Lactobacillus rhamnosus VT1 and skim milk fermented by 

this strain on the growth of Aspergillus niger and 

Penicillium commune. Initially, live bacterial cells (0.1% 

v/v, 1% v/v, 10% v/v), as well as skim milk fermented by 

L. rhamnosus VT1 (0.1% v/v, 1% v/v, 10% v/v), were 

added to modified MRS broth, which was simultaneously 

inoculated (1% v/v) with Aspergillus niger or Penicillium 

commune. The cultures were incubated at 30 °C for 10 

days. The mycelial weight and the pH of the broth were 

determined on days 0, 2, 4, 6, 8, and 10. All concentrations 

of Lactobacillus rhamnosus VT1 as well as the fermented 

skim milk were found to significantly reduce the growth 

rate of both Aspergillus niger and Penicillium commune in 

modified MRS broth. However, the L. rhamnosus VT1 

treatment was slightly more effective than the fermented 

milk treatment at controlling mycelial growth of both P. 

commune and A. niger. Lactobacillus rhamnosus VT1 at 

1% caused a 50% reduction in mycelial weight of P. 

commune and a 36% reduction in mycelial weight of A. 

niger at 10 days of incubation, while the fermented milk 

caused a 27% reduction in mycelial weight of P. commune 

and a 34% reduction in mycelial weight of A. niger for the 

same time period. These results suggest that L. rhamnosus 

VT1 has potential as a biological control agent of molds in 

the food industry. 

899 - Research and education in specialty gourmet and 

medicinal fungi in Tasmania, Australia 

K. Stott 1 , W. Gill 2 & C. Mohammed 3* 

1 Specialty Gourmet and Medicinal Fungi Research, 

Tasmanian Institute of Agricultural Research, 13 St. Johns 

Avenue, New Town, Tasmania, 7008, Australia. - 2 Huon 

Valley Mushrooms, Main Road, Glen Huon, Tasmania, 

7109, Australia. - 3 School of Agricultural Science, 

University of Tasmania, GPO Box 252-54, Hobart, 

Tasmania, 7001, Australia. - E-mail: 

caro.mohammed@utas.edu.au 

The research group at the Tasmanian Institute of 

Agricultural Research is the main research and euducation 

provider for the specialty mushroom industry in Australia. 

Current research projects investigate the edible fungi: 

Grifola frondosa (Maitake), Morchella species (Morel) and 

Tricholoma matsutake (Matsutake). The edible fungus 

Morchella has been collected in Tasmania, in situations 

ranging from forest through to garden soil and bark chip


mulch. Twenty six cultures were obtained and, on the basis 

of fruitbody morphological characters, placed into one of 

the three following species: M. elata, M. deliciosa, M. 

esculenta var. crassipes/angusticeps. Maitake has been 

grown, using the Japanese bag method, to assess the 

response of this fungus to eucalypt substrate ameliorated 

with different percentages of rice bran, maize meal and 

wheat bran. Findings indicate that only relatively small 

quantities of additive will be required for successful 

cultivation. The development of an artificial cultivation 

system for Matsutake (Tricholoma matsutake) is 

commercially desirable and necessary to ensure the 

survival of this highly prized mushroom. Interaction 

studies between the fungus and host Pinus lateral roots are 

being undertaken to elucidate the infection process and to 

understand the exact nature of this atypical symbiosis. In 

collaboration with the University of Tasmania, the 

researchers are offering on-line and residential courses on 

specialty gourmet and medicinal fungi. 

900 - The inmpact of Phytophthora cinnamomi in 

National Parks in New South Wales, Australia 

B.A. Summerell 1* , K.D. McDougall 2 & J.L. Walsh 1 

1 Royal Botanic Gardens, Sydney, Australia. - 2 National 

Parks and Wildlife Service, Queenbeyan, ACT, Australia. 

Although Phytophthora cinnamomi is regarded as having a 

significant impact on native vegetation in many parts of 

southern Australia, the pathogen has been considered 

benign and possibly endemic in New South Wales. The 

evidence for the different behaviour in NSW has included 

that 1) P. cinnamomi is extremely widespread and easy to 

detect in soils, 2) generally susceptible genera such as 

Banksia are unaffected when it is present (suggesting a 

long host/pathogen interaction), 3) multiple plant deaths 

associated with P. cinnamomi are extremely rare and 4) P. 

cinnamomi has been recovered in remote areas (suggesting 

that it is endemic). Recent surveys of National Parks in 

eastern NSW have found that although P. cinnamomi is 

widespread it cannot be detected in some areas despite 

extensive soil sampling. Although most Banksia spp. seem 

to be relatively resistant to symptoms of infection, other 

taxa (especially some Xanthorrhoea spp.) are very 

susceptible. The loss of Xanthorrhoea cover may adversely 

affect threatened animals such as the Smoky Mouse and 

Southern Brown Bandicoot, which use the plants for cover 

and nesting. Glasshouse susceptibility trials have shown 

that a number of rare taxa are very susceptible to infection 

and molecular analysis has shown that while some 

variation occurs in populations this variation is limited. 

Phytophthora cinnamomi may be widespread in NSW, 

however it is doubtful that it is endemic and is certainly not 

always benign. 

901 - The survey of spring barley and rye 

contamination by Fusarium mycotoxins (artificial and 

natural infection) 

S. Sykorova * , L. Papouskova, V. Sip, J. Chrpova & J. 

Hysek 

Research Institute of Crop Production, Drnovska 507,161 

06 Prague 6, Czech Republic. - E-mail: sykorova@vurv.cz 

Majority of wheat and barley varieties are susceptible to 

Fusarium spp. infection, and there is no host specificity in 

the species F. graminearum and F. culmorum. Using 

quantitative ELISA method the DON content was 

determined in the samples of spring barley artificially 

infected by F. culmorum. 12 varieties were tested in 2000 

and 19 in 2001. The other 32 spring barley and 15 rye 

samples were earned from 32 districts of the Czech 

Republic. The DON content in artificially infected barley 

in 2000 varied from 5,1 to 19,1 ppm (average 9,7 ppm); in 

2001 from 1,3 to 8,2 ppm. In 2000 the correlation between 

DON content and TGW-values was close (r = -0,89). The 

comparison of DON content and TGW values in 2000 and 

2001 was performed. DON content in 2001 was lower than 

in 2000 and TGW- values were in 2001 higher than in 

2000. Monitoring of DON content and mycological control 

in barley samples confirmed DON appearance in all 

samples (0,03 to 3,77; average 0,4 ppm). The limit value 

was exceeded in the one case. F. graminearum was in 2001 

prevailing toxinogenic species. Its occurrence is consistent 

with higher levels of DON. It confirmed that mycological 

control of all studied samples is need. There is also evident 

that toxinogenic species F. culmorum was replaced in 2001 

by F. graminearum which was strong producer of DON 

too. The DON content found in tested rye samples was 

very low ( 0,02 to 0,33 ppm) and therefore the mycological 

control was not done. 

902 - Pyrenopeziza betulicola and Marssonina betulae 

leaf spot fungi in birch: Monitoring the disease 

development 

L. Syrjälä * & M. Poteri 

Finnish Forest Research Institute, Juntintie 40, 77600 

Suonenjoki, Finland. - E-mail: leena.syrjala@metla.fi 

In late summer and autumn birch (Betula sp.) leaves are 

full of spots caused by fungi. Often these fungi are also a 

reason for premature yellowing and defoliation, which may 

reduce growth and frost hardening of trees. In Finland 

common necrotic leaf spots causing species are 

Pyrenopeziza betulicola and Marssonina betulae. Though 

these fungi have been characterized and found to be 

common, there are very few studies of their infection 

biology or disease development in trees. We have studied 

leaf spot fungi - Betula pendula - interaction. Because 

global climatic change is assumed to increase atmospheric 

ozone and CO 2 concentration in the future, we have also 

followed the effect of these gases on the development of 



leaf spot disease. For monitoring the disease development 

we have used both image analyses technique and visual 

observation. According to the results the ability of P. 

betulicola strains to infect birch leaves as also the 

resistance of birch clones to the disease varied. There were 

also differences between strain-clone -combinations in the 

intensity and rate of premature yellowing and falling of 

leaves. The ability of M. betulae to cause leaf spots was 

affected by leaf age. In the older leaves necrotic spots 

developed faster and the diseased leaf area was also larger. 

Under ozone and CO2 fumigation treatments the gases 

alone didn't explain the disease development, but the 

disease was dependent also on birch clone, age of leaves 

and performance year of the experiments. 

903 - Genetic diversity in Ampelomyces hyperparasites 

inferred from SSCP analysis 

O. Szentivanyi 1 , J.C. Russell 2 , L. Kiss 1* , J.P. Clapp 2 & P. 

Jeffries 2 

1 Plant Protection Institute, Hungarian Academy of 

Sciences, H-1525 Budapest, P.O. Box 102., Hungary. - 

2 Department of Biosciences, University of Kent, 

Canterbury, Kent, CT2 7NJ, U.K. - E-mail: 

LKISS@NKI.HU 

SSCP analysis (Single Strand Conformation 

Polymorphism) allows a rapid and sensitive detection of 

sequence differences between nucleic acid samples. The 

method relies on the different migration rates of single 

stranded nucleic acid fragments. It is known that even a 

single point mutation can cause a change in the secondary 

structure and thus a changed running behaviour of DNA or 

RNA samples in the gel. SSCP can thus be used to rapidly 

screen a range of individuals for intraspecific genomic 

variation. This method has not yet been applied widely in 

plant pathology, but we have recently used it to assess 

variation in a world-wide collection of isolates of the 

mycoparasitic genus Ampelomyces. These fungi are 

common intracellular hyperparasites of powdery mildews. 

To study their genetic diversity, 29 isolates were studied by 

SSCP analysis of the rDNA ITS region. Based on SSCP 

profiles, the isolates were included in eight different 

groups. These results largely supported earlier data [1, 2], 

but revealed additional genetic differences, too. 

Interestingly, a new group of isolates were delineated that 

consisted of all Ampelomyces isolates obtained from apple 

powdery mildew in Hungary. 1. Kiss, L.: Mycol Res 101, 

1073 (1997). 2. Kiss, L., Nakasone, K.K.: Curr Genet 33, 

362 (1998). 

904 - Mixed infection of Trichosporon cutaneum and 

Candida parapsilosis in a white piedra case from Qatar 

S.J. Taj-Aldeen 1* & H.I. Al-Ansari 2 

1 Hamad Medical Corporation, Dept. Laboratory Medicine 

& Pathology, P.O. Box 3050, Doha, Qatar. - 2 Hamad 

Medical Corporation, Dept. of Dermatology & Venerology, 

272 


Rumala P.O. Box 3050, Doha, Qatar. - E-mail: 

saadtaj@hotmail.com 

White piedra is a rare fungal infection of the hair shaft 

characterized by small, firm, irregular white-brown 

nodules. The infection is due to Trichosporon cutaneum (= 

T. beigelii). The disease occurs in tropical and subtropical 

areas and reported in Saudi Arabia and Kuwait. Until now, 

where a 28 year old female patient acquired the infection, it 

was not reported in Qatar. In this case the scalp was the 

only site affected but in a very extensive manner. The hair 

had yeast odour and appeared beaded with nodules. Under 

the microscope, the nodules were light-brown made of a 

compact mass of arthrospores arranged perpendicularly 

around the hair shaft and varying in size, measuring up to 

1.5 mm. Sabouraud`s culture developed rapidly T. 

cutaneum accompanied by Candida parapsilosis along the 

infected hair shaft at room temperature and 37 °C. Colonies 

of T. cutaneum were wrinkled, tan and cycloheximide 

negative. Assimilation profile was consistent with the 

organism identification. Microscopic examination showed 

hyphae which fragmented into rectangular arthrospores and 

budding cells. The patient treated by daily application of 

Econazol (shampoo & cream) followed by Ketoconazole 

shampoo, completely cured (clinically & mycologically) 

after two months. White piedra infection in this patient is 

caused by mixed infection with C. parapsilosis. It is not 

clear if the disease was due to the synergistic action 

between T. cutaneum and C. parapsilosis. 

905 - Allergic Aspergillus flavus rhinosinusitis: A case 

report from Qatar 

S.J. Taj-Aldeen 1* , A.A. Hilal 2 & A. Chong-Lopez 1 

1 

Hamad Medical Corporation, Dept. Lab. Med. & Path., 

2 

PO box 3050, Doha, Qatar. - Hamad Medical 

Corporation, ENT Section, Rumala, PO box 3050, Doha, 

Qatar. - E-mail: Saadtaj@hotmail.com 

Fungal involvement of the rhinosinusitis is classified into 

four major forms: mycetoma, invasive, allergic and 

fulminant form. It can become life-threatening if not 

diagnosed and treated properly. The preliminary diagnosis 

is usually by nasal endoscopy and computed tomography 

(CT) imaging but tissue biopsy and culture is of vital 

importance in confirming the disease and in planning 

treatment. We present a case of allergic fungal 

rhinosinusitis caused by Aspergillus flavus. Clinical 

manifestation of the disease was the presence of an 

extensive left nasal polyp. Allergic workup revealed 

systemic eosinophilia (11.7%), High serum IgE level (1201 

IU ml -1 ) and positive skin test for Aspergillus. CT scan 

showed a total opacification and expansion of the left nasal 

cavity and sinuses with secondary inflammatory reaction 

on the right side. There was no bony erosion beyond the 

sinus walls. The patient has been operated by endoscopic 

approach (polypectomy and ethmoidectomy) where an 

abundant amount of allergic fungal mucin and dark crusts 

were found filling the sinuses. Fungal hyphae and spores 

were evident by direct lactophenol mount and by 

histopathological sections of the removed polypi. Culture 

of the debris resulted in growth of Aspergillus flavus. The


findings were highly suggestive of allergic fungal 

rhinosinusitis. The patient received a full course of 

systemic steroids and had no recurrence after 9 months 

follow up. 

906 - Aflatoxin B1 in Rice bran: Sources of 

contamination 

V. Theraisnathan * & I. Kalyanasundaram 

Centre for Advanced Studies in Botany, University of 

Madras, Chennai-25, 10/4, L & T Colony, Phase-II, 

Ramakrishnapuram, Virugambakam, Chennai-600 092, 

India. - E-mail: wincente@yahoo.com 

A wide survey on the prevalence of Aflatoxin B1 (AFB1) 

in rice bran was conducted in South India. Sixty two per 

cent of the samples contained AFB1 and the levels far 

exceeded the permissible limit of 50 ppb, with 60% of the 

positive samples having 50-500 ppb and 30% up to 2000 

ppb. The mycoflora of bran included over 20 osmophilic 

species, of which Aspergillus flavus Link showed the 

highest frequency (75%) as well as abundance (37.6%). 

The high frequency and high levels of AFB1 showed no 

correlation with moisture content of the samples, climatic 

variations, or with the raw or parboiled status of the paddy, 

but with the maintenance of sanitation in the rice mills, 

suspected to be a major source of contamination of bran 

with aflatoxigenic fungi/aflatoxin. This was done by 

subjecting specific lots of rough rice (paddy) to parboiling 

in the mill followed by drying through different 

procedures; milling of the raw and parboiled paddy 

samples separated in commercial mills and in the 

laboratory, and comparing the mycoflora with reference to 

A. flavus - and AFB1. The mill environment did act as a 

source of contamination of bran, but only to enhance the 

toxin which had already formed earlier in the paddy. It was 

thus a source of secondary contamination, and its actual 

contribution to the prevalence of AFB1 in rice bran should 

be determined only by making a survey of the prevalence 

of AFB1 in paddy in the region and comparing it with that 

of bran. 

907 - Aflatoxin in conidia of airborne fungi and its 

relevance for occupational hygiene 

R. Thißen, G. Fischer * , T. Müller, S. Braun & W. Dott 

Institute of Hygiene and Environmental Medicine, 

University Hospital RWTH, Pauwelsstr. 30, D-52074 

Aachen, Germany. - E-mail: Guido.Fischer@post.rwthaachen.de 

At work places in the waste-handling industry mycotoxinproducing 

fungi can be found in high numbers in air. In the 

present study, the amount of mycotoxins associated with 

airborne conidia was investigated. Species from the A. 

flavus group known to produce aflatoxins were tested as a 

model system. The production of mycotoxins was 

investigated for fresh isolates from a composting plant. 

Conidial suspensions of known aflatoxin producers (A. 

flavus, A. parasiticus) were investigated qualitatively and 

quantitatively for their aflatoxin content. Analysis was 

done using both HPLC-FLD and ELISA. Extracts of 

additional species (A. niger, A. versicolor, A. fumigatus, E. 

nidulans) and mycotoxin standards were tested for cross 

reactivities. The amount of aflatoxin correlated with the 

number of conidia extracted. A. parasiticus conidia 

contained significantly more aflatoxins (amounts of pg per 

10,000 conidia) than those of A. flavus. A. versicolor 

showed a slight unspecific reactivity that may be due to 

sterigmatocystin, a precursor in the aflatoxin biosynthesis. 

The results show that aflatoxin can be detected in relatively 

small amounts of conidia. As species of the A. flavus gr. 

occur in numbers of >1000 cfu per cbm air in composting 

plants, they may be of toxicological relevance. Further 

investigations are necessary to evaluate the incidence of 

toxinogenic strains in situ. 

908 - Attraction of phorid and sciarid pest flies during 

cultivation of Agaricus bisporus 

L. Tibbles 1* , L. Boddy 1 , M. Jervis 1 & D. Chandler 2 

1 Cardiff University, PO Box 915, Cardiff, CF10 3TL, U.K. 

- 2 Horticulture Research International, Wellesbourne, 

Warwick, CV35 9EF, U.K. - E-mail: Tibbles@cf.ac.uk 

The cultivation of Agaricus bisporus is a worldwide, multimillion 

pound industry. It is susceptible to many pests and 

pathogens, the dipterans Lycoriella castanescens (sciarid) 

and Megaselia halterata (phorid) are most serious causing 

losses of 7% of annual revenue in the UK. Pest control is 

largely by chemicals and there is a need for alternative 

modes of pest control. Determining the source of attraction 

for these pest flies could provide a method for preventing 

their infestation of mushroom houses. A static air 

olfactometer was developed to quantify the behavioural 

response of female sciarid and phorid flies to mushroom 

cultivation substrates: pasteurised compost, 4 d spawned 

compost, fully spawned compost and button mushrooms. 

Phorid response varied depending on substrate, with 

greatest attraction to fully spawned compost, those that best 

support development of this obligate mycelial feeder. 

Sciarids were attracted to all substrates. Phorid oviposition 

increased with mycelial growth through the substrate, 

whilst sciarids exhibited high oviposition on all substrates. 

Emergence of both species was substrate dependent. 

Sciarid emergence was highest from pasteurised compost, 

decreasing with mycelial growth through the substrate 

indicating that mycelial growth is deterimental to 

development. Greatest emergence of phorids was from the 

4 day spawned compost. Results are consistent with the use 

of volatiles in detection of oviposition sites for both 

species. 



909 - Ochratoxin producing Aspergillus species in 

organic raisins and sultanas 

M. Torp 1* , P. Kruse 1 , P.-E. Clasen 1 & F.J. Cabañes 2 

1 National Veterinary Institute, Department of Food and 

Feed hygiene, Ullevålsvn. 68, P.O.Box 8156 Dep., N-0033 

Oslo, Norway. - 2 Universitas Autònoma de Barcelona, 

Departament de Sanitat i d'Anatomia Animals, Grup de 

Micologia, Bellaterra E-08193, Spain. - E-mail: 

mona.torp@vetinst.no 

On behalf of the Norwegian National Food Control 

Authority, a study on potential mycotoxin producing fungi 

in organically grown raisins and sultanas was conducted. 

The 61 tested samples were bought in stores in Oslo during 

spring and summer 2001. The dried fruits were cut in 

halves and plated out on Dichloran Glycerol agar and 

incubated at 25 °C for seven days. Potential mycotoxin 

producers were isolated from all the 15 samples of 

sultanas, while 44 samples (95%) of raisins were affected. 

Fungi belonging to the Aspergillus niger aggregate were 

the most dominant, infesting all samples of sultanas and 

95% of the raisins, while Aspergillus carbonarius was 

isolated from 93% of the sultanas and 2% of the raisins. 

The high recovery of these fungi in raisins and sultanas is 

of concern, since they have been reported as producers of 

the highly toxic Ochratoxin A (OTA). In this study, 18 

strains were analysed by HPLC for OTA production in 

pure culture on Yeast Extract Sucrose agar. Nine out of 12 

tested strains of A. carbonarius and four out of six tested 

strains of the A. niger aggregate produced OTA. The ITS- 

5.8S rDNA RFLP patterns of the OTA-producing isolates 

in the A. niger aggregate were also determined. All four 

isolates were classified as type N [Accensi et al. 1999], 

corresponding to the RFLP group 'A. niger'. The results 

indicate that OTA analyses of organically grown raisins 

and sultanas would be of interest to the consumers. 

910 - A monitoring system for green fluorescence 

protein gene-transformed Fusarium oxysporum in 

melon seedlings 

H. Toyoda * , T. Nonomura & Y. Matsuda 


of Agriculture, Kinki University, 3327-204 Nakamachi, 


toyoda@nara.kindai.ac.jp 

Using melon seedlings at the cotyledon stage and 

genetically marked fungi, a system for monitoring 

pathogenic and nonpathogenic Fusarium oxysporum was 

devised in the present study. Protoplasts were prepared 

from three formae speciales (melonis, radicis-lycopersici 

and fragariae) of F. oxysporum and transformed with a 

synthetic gene for green fluorescence protein. 

Transformants were primarily isolated in the presence of 

hygromycin B and then screened by the emission of bright 

green fluorescence. Roots of melon seedlings were 

274 


inoculated with fluorescing microconidia of these fungi, 

and fungal infection behavior was traced. Using 

fluorescence microscopy, we directly observed not only the 

fungus at the root surface, but also the mycelia elongating 

in the trachea of roots. Both pathogenic and nonpathogenic 

fungi germinated and hyphae elongated superficially on the 

surface of root. Only pathogenic fungi caused root necrosis 

at the inoculation site. Hyphae grew within the stem to 

induce constriction or cracking of lower hypocotyls, then 

causing wilting of the seedlings. Infection behavior of 

genetically marked pathogenic and nonpathogenic F. 

oxysporum could be successfully monitored after 

inoculation of cotyledons of seedlings. 

911 - Biological control of bluestain in logs using an 

albino bluestain fungus 

A. Uzunovic * & A. Byrne 

Forintek Canada Corp., 2365 East Mall, Vancouver, B.C., 

Canada. - E-mail: adnan@van.forintek.ca 

Bluestain causes significant economic losses to the 

Canadian forest industry. We examined the feasibility of 

using an albino bluestain strain (Cartapip TM , now renamed 

Sylvanex TM ) of Ophiostoma pilferum to control bluestain in 

lodgepole pine logs. Cartapip and four challenge bluestain 

fungi were sprayed alone, and in sequence, on two types of 

wounds artificially produced on pine billets. When 

inoculated alone, Cartapip colonized fresh lodgepole pine 

and did not cause stain. The challenge fungi alone caused 

significant stain. However, when the challenge fungi were 

applied 2 or 10 days after Cartapip, the stain was negligible 

in most cases and Cartapip outcompeted the fungi. Field 

work using commercial size logs was done in 2000 and 

2001 in Alberta, Canada. Cartapip suspension was sprayed 

on logs and the bluestain development quantified and 

compared with logs sprayed only with water. Statistical 

analysis of the data after 12-13 weeks of storage showed 

that Cartapip significantly reduced the amount of bluestain. 

After 6 weeks of summer storage in 2001, Cartapip-treated 

logs remained almost spotless compared to the 

considerable stain found on non-treated logs. After 13 

weeks of storage there was some stain development in 

Cartapip-treated logs but the amount was significantly less 

than in non-treated logs. The product, and the concept of 

using albino isolates to control stain, therefore have 

potential for industrial use. 

912 - Expression of fruit body color in Pleurotus spp. 

hybrids derived from pairings of compatible 

neohaplonts 

G. Valencia del Toro 1* & H. Leal Lara 2 

1 Department of Chemistry, UPIBI, IPN; Research Division, 

FESI, UNAM;, ENEP Iztacala, Av de los Barriois S/N, 

54090 Los Reyes Iztacala, Tlalnepantla, Estado de México, 

Mexico. - 2 Department of Food Science and Biotechnology, 

Faculty of Chemistry, National University of Mexico


(UNAM), Cd. Universitaria, 04510 México D.F., Mexico. - 

E-mail: tavaltor@servidor.unam.mx 

The monocaryotic components (neohaplonts) of 10 

Pleurotus spp. strains (ECS127, ECS187, IB67, 1E202, 

IE200, IE201, INI8, POROS, RP y P15) producing fruit 

bodies of contrasting colors were obtained by chemical 

dedicaryotization. By pairing these neohaplonts, 5 

intersterile groups were recognized. Four strains belong to 

the first group, IB67 (gray), IE200 (white), PORO (pink) y 

RP (pink) and their neohaplonts showed compatibility 

factors AmBm, and AnBn, respectively. In the second 

group, a white (IE201) and a pink (IE202) strain were 

found; their nehohaplonts could have either completely 

different compatibility factors AoBo and ApBp or the 

complementing factors of the first group, AmBn and 

AnBm. In the third group, 2 gray strains, INI8 and 

ECS127, were found with compatibility factors AqBq and 

ArBr, while a yellow strain, ECS187, was found in the 

fourth group with compatibility factors AsBs and AtBt. 

Finally, a gray strain, P15, was found in the fifth group 

with compatibility factors AuBu and AvBv. Thirteen 

hybrid dikaryons were obtained by cross mating 

neohaplonts from groups 1 and 2 and they were grown on a 

commercial Pleurotus sp. substrate (feremented straw). 

Gray fruit bodies were obtained from 11 hybrids and, 

unexpectedly, yellow fruit bodies were produced by two 

hybrids resulting from matings of neohaplonts from a white 

strain (IE201) and a pink one (IE202). Regulation of gene 

expression or extrachromosomal factors to explain such 

results is discussed. 

913 - New plant pathogenic fungus from the sterile 

white basidiomycete complex 

O. Vinnere * , J. Fatehi & B. Gerhardson 

Plant Pathology and Biocontrol Unit, SLU, P.O. Box 7035, 

S-75007, Uppsala, Sweden. - E-mail: 

olga.vinnere@vpat.slu.se 

An isolate of Sterile White Basidiomycete (SWB) was 

obtained from the visually healthy roots of buffalo grass 

(Pennisetum landistinum), growing in a natural bush land 

in Perth, Western Australia. The greenhouse pathogenicity 

tests revealed that the fungus is able to infect 12 different 

plant species. This isolate was compared with a strain of 

SWB, obtained from the American Type Culture 

Collection (ATCC 28344), originated from Florida, United 

States, which has been reported as an aggressive pathogen 

on 16 genera of agricultural crops. Mycelial features, 

growth rate, as well as pathogenicity pattern and inoculum 

potential of these two pathogens were distinctly different. 

In addition, sequence analysis of the Internal Transcribed 

Spacer (ITS) regions, 5.8S and the first 800 bp of the 25S 

ribosomal RNA genes strongly supported that the 

Australian fungus was clearly different from the American 

isolate. The sequence data confirmed the previous report by 

another research group that the American SWB strain 

belonged to Marasmius graminearum. Further studies are 

carried out in order to resolve the taxonomic position of the 

Australian SWB. 

914 - The biodiversity of microfungi from selected 

organically produced fruit and vegetable commodities 

from Norway 

N.W. Waipara 1* & M. Torp 2 

1 HortResearch, Canterbury Research Centre, P.O.Box 51, 

Lincoln, New Zealand. - 2 National Veterinary Institute, Box 

8156 Dep., 0033 Oslo, Norway. - E-mail: 

nwaipara@hortresearch.co.nz 

The mycofloras of four organically produced fruit and 

vegetable commodities; dried fruit, strawberry, corn and 

carrot, for sale in Norway were examined. The species of 

fungi obtained from each food commodity were mostly 

saprophytic. Xerophilic fungi such as Eurotium spp., 

Aspergillus spp., and Wallemia sebi were isolated in many 

dried fruit samples along with fungi capable of food 

spoilage such as, Rhizopus spp. and Zygosaccharomyces 

rouxii. Mycotoxin- producing Aspergillus flavus was also 

isolated, and aflatoxins were also detected in low levels in 

these samples. The fruit spoilage pathogen, Botrytis 

cinerea, was frequently isolated from all strawberry 

samples along with two other strawberry pathogens Idriella 

lunata and Colletotrichum gloeosporioides, which were 

less frequently isolated. Most fungi obtained from corncob 

kernels were aerial contamination species such as 

Cladosporium and Epicoccum, Zygomycetes and yeasts. 

The fungi obtained from carrot samples were common 

saprophytic species frequently reported from soil and roots, 

although carrot spoilage pathogens such as Thielaviopsis 

basicola and Rhizoctonia carotae were also isolated in low 

numbers. 

915 - Physiological regulation of biomass and 

polysaccharides production by medicinal mushrooms 

S.P. Wasser 1 , V.I. Elisashvili 2* , M. Stajic 1 & M. Didukh 1 

1 Institute of Evolution, University of Haifa, Haifa 31905, 

Israel. - 2 Institute of Biochemistry and Biotechnology, 

Tbilisi 380059, Georgia. - E-mail: 

velisashvili@hotmail.com 

Agaricus nevoi, Inonotus levis, Phellinus igniarius, Ph. 

robustus, Pleurotus ostreatus, and Tremella mesenterica 

biomass and polysaccharide production in dependence on 

the carbon and nitrogen sources have been investigated. All 

strains grew well in the presence of glucose, maltose or 

mannitol in the medium yielding in shake flasks 

experiments up to 13-18.9 g l -1 of crude biomasspolysaccharide 

ethanol precipitate. Among carbon sources 

studied, glucose appeared to be the best for the 

polysaccharide production by I. levis, Ph. igniarius, Ph. 

robustus, P. ostreatus, whereas T. mesenterica produced 

the highest level of polysaccharide during growth in the 

presence of sucrose and mannitol. It has been shown that 

the yield of polysaccharides correlated with carbon source 

concentration in the medium. Among nitrogen sources 

studied, organic compounds and ammonium sulfate 



stimulated fungi growth and polysaccharides production. 

The best polysaccharide producers, I. levis and T. 

mesenterica, were cultivated in BioFlo Fermentor 2000 

(New Brunswick, USA). In this case yield of crude proteinpolysaccharide 

products reached 23 and 27 g l -1 , 

respectively. Data on the dynamic of fungi growth in 

fermentor indicate that in the culture of T. mesenterica 

biosynthesis of polysaccharide parallels to fungus biomass 

growth, while in the case of I. levis polysaccharide 

synthesis mainly occurs in the later logarithmic phase of 

growth. 

916 - Mechanism of wilting in oak mortality in Japan 

caused by Raffaelea quericivori 

T. Yamada 1* , M. Yamato 2 , D. Sakaue 1 & K. Suzuki 2 

1 Experimental Station at Tanashi, The University Forests, 

The University of Tokyo, Midori-cho 1-1-8, Nishitokyo, 

Tokyo 188-0002, Japan. - 2 Graduate School of Agricultural 

and Life Sciences, The University of Tokyo, Yayoi 1-1-1, 

Bunkyo-ku, Tokyo 113-8657, Japan. - E-mail: 

yamari@uf.a.u-tokyo.ac.jp 

Mass mortality of oaks (mainly Quercus serrata and Q. 

crispula) has been appeared along the Japan Sea since 

1980's. Blockage of xylem sap ascent induced by infection 

of a fungus Raffaelea quericivori, which is vectored by 

ambrosia beetle Platypus quercivorus, is considered to 

have close relationship with this mortality of oaks. Lower 

stems of Q. serrata and Q. crispula seedlings were 

inoculated with R. quercivori. Dye treatment revealed that 

blockage of sap-flow occurred only around the inoculation 

points. Symptoms during wilting after the R. quercivori 

inoculation and those after cutting of lower stem were 

similar. Further, supplying water into stem lengths cut from 

upper half of the seedlings just after the appearance of 

wilting also delayed the further development of wilting in 

both of them. Water conductivity in the stem was measured 

with a high pressure flowmeter (HPFM) after the fungus 

inoculation onto Q. crispula seedlings, which survived 

after the inoculation. A parameter of chlorophyll 

fluorescence, Fv/Fm, decreased slightly in several 

seedlings. Conductivity was not changed in the upper part 

of the seedlings, though conductivity aroud the inoculation 

points was conspicuously reduced in fungus-inoculated 

seedlings, especially ones with lowered Fv/Fm. Our results 

suggested that upper part of the seedlings was not directly 

affected by the fungal inoculation, and that wilt was 

induced by the blockage of sap ascent around the 

inoculation point. 

917 - Penicillium fungi from Picea glehnii seeds protect 

the seedlings from damping-off 

K. Yamaji * , Y. Fukushi, Y. Hashidoko & S. Tahara 

Department of Applied Bioscience, Faculty of Agriculture, 

University of Hokkaido, Kita-ku, Sapporo, 060-8589, 

Japan. - E-mail: Keiko.Yamaji@joensuu.fi 

276 


We have postulated that germinating seeds and seedlings of 

Picea glehnii might be protected from pathogens by seedepiphytic 

fungi that produce antimicrobial compounds. We 

screened seed-epiphytic microorganisms that produced 

antibiotics against pathogenic Pythium vexans by the agaron-paper 

method. Of the 149 isolates, 13 fungi showed 

high antifungal activities and they were identified as 

Penicillium fungi. Three isolates (P. cyaneum, P. 

damascenum, and P. implicatum) showed high antifungal 

activity and they produced patulin, citrinin, palitantin and 

frequentin, respectively. In the co-inoculation test of three 

Penicillium species and P. vexans to P. glehnii seedlings, 

the number of surviving seedlings which were inoculated 

with P. vexans 5 days after pre-inoculation with P. 

damascenum increased compared with those inoculated 

with P. vexans alone. Dense mycelial growth of P. 

damascenum was observed microscopically around the 

roots of the inoculated seedlings. Furthermore, the 

antifungal and phytotoxic compound citrinin produced by 

P. damascenum was detected as a component released 

around the roots of the normally growing fungusinoculated 

seedlings. We proposed two possible 

mechanisms for the protection of P. glehnii seedlings by P. 

damascenum from P. vexans; first, chemical protection by 

an antibiotic agent, citrinin produced by P. damascenum, 

and second, the occupation of the space around the P. 

glehnii roots by the P. damascenum mycelia. 

918 - Growth inhibition for wood decaying fungi by 

lichen mycobionts 

Y. Yamamoto 1* , M. Komine 1 , K. Hara 1 , S. Doi 2 & K. 

Takahashi 3 

1 

Akita Prefectural University, 241-7, Kaidobata-nishi, 

2 

Shimoshinjo-nakano, Akita, Akita, Japan. - Akita 

Prefectural University, 11-1, Kaieizaka, Noshiro, Akita, 

Japan. - 3 Meiji College of Pharmacy, 2-522-1, Noshio, 

Kiyose, Tokyo, Japan. - E-mail: yyamamoto@akitapu.ac.jp 

About 400 strains of lichen-derived cultures induced 

natural thalli and spores are maintained in our laboratory 

and we previously indicated that they showed several 

biological activities. In natural conditions, lichens may 

oppose mushroom on the bark. Screening of growth 

inhibition of two wood decaying fungi, Trametes 

versicolor (L. ex Fr.) Pilat and Fomitopsis palustris (Berk. 

et Curt.) Gilbn & Ryv. in 46 strains of lichen mycobionts 

cultured. Two cell-aggregates of each mycobiont strain 

were placed oppositely on agar-plates of malt-yeast extract 

medium, glucose peptone medium and potato dextrose 

medium in a Petri dish and pre-incubated at 20C in the 

dark. After a month, one agar block of malt-yeast extract 

medium on which a wood decaying fungus grew was 

inoculated in the center of the agar-plate on which its 

mycobiont grew and maintained at 20C in the dark. After a 

week, inhibitory effect on growth of a fungus by lichen 

mycobionts was judged from the extent of its inhibition 

circle. Three mycobionts of Acarospora fuscata, Arthonia 

cinnabarina and Ramalina exilis inhibited the growth of 

both fungi. The growth of T. versicolor was inhibited by 

mycobionts of Stereocaulon sorediiferum and Dibaeis


absoluta, on the other hand that of F. palustris was done by 

those of Cladia aggregata, Haematomma puniceum and 

Xanthoria elegans. 

919 - Wood stain and causal fungi on Canadian 

hardwood species 

D.Q. Yang * , M.-C. Bisson, M. Gignac & T. Lihra 

Forintek Canada Corp., 319 rue Franquet, Sainte-Foy, 

Quebec, G1P 4R4, Canada. - E-mail: dianqing.yang@qc.forintek.ca 

In most countries, trees are harvested into logs and stored 

in a log yard for a certain time before sawing into lumber. 

Logs left lying on the ground during warm weather are 

vulnerable to attack by sapstaining fungi. Wood sapstain is 

caused by several groups of fungi that penetrate deeply into 

wood with dark pigmented hyphae. Hardwood species are 

used to a greater extent in value-added wood products, 

reducing sapstain in these species has a significant 

economical impact. This study investigated the speed of 

sapstain development in Canadian hardwood species and 

the major fungal species involved in this discoloration. 

Trees of sugar maple and yellow birch were felled in June 

2001. Logs were transported to and stacked in a sawmill 

within two weeks after felling. The evaluation was 

conducted every 2 weeks. Logs were sampled into discs, 

stain development on these discs was examined, and 

isolation and identification of staining fungi were followed 

up. The results showed that stain was detected on logs 3 

weeks after harvesting and logs were stained less than 5% 

of wood in 5 weeks. After 13-week storage, most sapwood 

of logs was stained. Yellow birch was more susceptible to 

fungal stain than sugar maple, and log ends were more 

stained than the internal sections. The frequently isolated 

sapstaining fungi were Ophiostoma piceae, O. piliferum, 

Leptographium sp., Aureobasidium pullulans, Alternaria 

alternata, and Cladosporium cladosporioides. 

920 - Production, formulation and application of a 

bioprotectant for wood protection 

D.Q. Yang * , M. Gignac & M.-C. Bisson 

Forintek Canada Corp., 319 rue Franquet, Sainte-Foy, 

Quebec, G1P 4R4, Canada. - E-mail: dianqing.yang@qc.forintek.ca 

Moulds, stain and decay cause serious problems on wood 

utilization. Though a growing volume of wood is kilndried, 

the market for green exports remains significant, and 

environment-friendly wood protection is required to 

replace traditional chemicals. As a solution to this problem, 

Forintek Canada Corp. developed a bioprotectant for 

protecting logs and green lumber from moulds, stain and 

decay, which was granted a US patent. The method relies 

on an albino fungus, Gliocladium roseum. One objective of 

the project was to assess the technical feasibility of large- 

scale production and utilization of this bioprotectant. 

Fermentation in 150L fermentors was achieved within 

three days. The spores and mycelia of the fermented fungus 

were formulated into a powder form. A shelf-life study 

indicated that this bioprotectant did not lose any of its 

vigour after one-year storage at -20 °C. Large-scale 

application and efficacy tests were carried out on 2400 

boards in a sawmill; the lumber tested was a mix of 2"x 3" 

x 8' black spruce and balsam fir boards. The treatment was 

applied in two different manners: a) with a spray system 

normally used for chemical application, and b) by dipping 

in a tank. After three months storage at the mill, 100 

percent of the treated board were found to be acceptable as 

compared to only 36 percent for untreated controls. 

921 - Medicinal mushrooms as inhibitors of 

angiogenesis 

M. Yassin 1* , J. Mahajna 2 & S.P. Wasser 1 

1 Institute of Evolution University of Haifa, Mount Carmel, 

Haifa 31905, Israel. - 2 Galilee Institute for Applied 

Research, P.O. Box 415, Nazareth 16103, Israel. - E-mail: 

majedyassin22@hotmail.com 

Medicinal mushrooms have been an important source of 

therapeutic substances for the treatment of various human 

illnesses including cancer. In Japan, few polysaccharide 

antitumor agents have been developed such as lentinan, 

krestin, and schizophyllan. Angiogenesis is an essential 

component of the body's physiology and contributes to the 

pathogenesis of a variety of diseases such as psoriasis, 

rheumatoid arthritis, and cancer. Our objective is to select a 

medicinal mushroom extracts with anti-angiogenic and 

antitumor activities, monitoring ability of selected 

mushroom extract to interfere with the proliferation of 

endothelial cells and other tumor cell lines. Our interest is 

on selecting mushroom extracts that promote apoptosis of 

the appropriate cell lines. We developed rapid, accurate and 

reliable assays that allowed us to detect different events of 

apoptosis. More than thirty species of edible and medicinal 

higher basidiomycetes were evaluated. Screening data of 

selected medicinal mushroom extracts will be presented. 

Selected mushroom extracts or fractions that show 

promising activity in the screening assays will be further 

evaluated in animal models. 

922 - Identification of viruses from diseased edible 

mushrooms, Pleurotus ostreatus, Pleurotus eryngii and 

Flammulina velutipes 

H.J. Yu, J.S. Lee, Y.C. Kwan, N.J. Lee & H.S. Lee * 

Division of Life Science, college of Natural science, and 

Genetic Engineering Institute, Gyeongsang Natl. Uni. 660- 

701 Chinju, Korea. - E-mail: hslee@nongae.gsnu.ac.kr 

Natural viral epidemic was observed in the commercial 

farms of edible mushroom, Pleurotus ostreatus, Pleurotus 



eryngii, Flammulina velutipes in Korea. The disease 

always accompanied the presence of isometric virus 

particles. Using purification procedures involving Tris- 

EDTA buffer extraction, PEG-NaCl precipitation, 

differential centrifugation, and equilibrium centrifugation 

in CsCl gradient (1.585 g/cm3), we have obtained three 

isometric viral particles, one of 27 and two of 34 nm in 

diameter, with different densities from P. ostreatus and P. 

eryngii. The 27 nm particle, denser than 34 nm particle, 

encapsidated ssRNAs as its genome. There were two kinds 

of lighter particles with the same size of 34 nm which 

encapsidated dsRNA. Coomassie Brilliant Blue stained 

polyacrylamide gel (12%) electrophoresis showed different 

polypeptides of coat proteins of Mr 29, 71, and 62 kD in 

viral coat proteins, respectively. These results 

demonstrated that there are at least 3 different kinds of 

isometrc viruses whose genomes were ssRNA or dsRNA in 

P. ostreatus and P. eryngii. The Flammulina velutipes 

harbored different spherical ssRNA viruses. In an attempt 

to prove that the viral complex is the causative agent for 

the disease symptoms we cured the viruses from diseased 

spawn. The virus-cured spown formed normal mushroom 

and recovered from the epidemic. Therefore, the 

phenonena of mushroom malformation and reduced harvest 

in the mushroom farms could be of viral nature. 

923 - Comparison of the aflR gene sequences of strains 

in Aspergillus section Flavi 

G.-F. Yuan * , C.-Z. Lee & G.-Y. Liou 

Food Industry Research and Development Institute, P. O. 

Box 246, Hsinchu, Taiwan, R.O.C., Taiwan. - E-mail: 

gfy@firdi.org.tw 

Aflatoxins are polyketide-derived secondary metabolites 

produced by Aspergillus parasiticus, A. flavus and A. 

nomius. Human health and economic benefit are most 

affected by the toxic effects of aflatoxins. The aflR gene is 

a regulatory gene of aflatoxin biosynthesis and encodes a 

protein containing a zinc cluster DNA binding motif. 

Although A. oryzae and A. sojae, used in fermented food 

and ingredient manufacture, never have a record of 

producing aflatoxin, they have shown the presence of aflR 

gene. In this study, 34 strains belong to the Aspergillus 

section Flavi were examined and the aflR gene of 23 

strains of these strains were successfully amplified and 

sequenced. No PCR products of aflR were found in five 

strains of A. sojae and six strains of A. oryzae, these results 

suggest that the aflR gene might not exist or significantly 

different in some strains of A. sojae and A. oryzae. The 

sequenced aflR genes of the 23 strains have 96% similarity, 

especially the zinc finger DNA-binding domain are highly 

conserved. The aflR gene of A. sojae has conspicuous 

character, an extra CTCATG fragment inserted and a C to 

T transition change cause earlier termination of the 

encoded AFLR protein. The differences between A. 

parasiticus/A. sojae and A. flavus/A. oryzae were also 

found in some bases of aflR gene. Although the aflR genes 

have no obvious difference between A. flavus and A. 

oryzae, some differences may exist in aflatoxin producing 

and no producing strains. 

278 


924 - Sixty-one alleles of the ALS7 open reading frame 

in sixty-six Candida albicans strains: ALS7 is a 

hypermutable contingency locus 

N. Zhang 1 , A.L. Harrex 2 , B. Holland 3 , R.D. Cannon 2 & J. 

Schmid 1* 

1 IMBS,Massey University, Palmerston North, New 

Zealand. - 2 Dept. of Oral Sciences and Orthodontics, 

University of Otago, Dunedin, New Zealand. - 3 IFS, 

Massey University, Palmerston North, New Zealand. - Email: 

J.Schmid@massey.ac.nz 

The Candida albicans ALS (agglutinin-like sequences) 

family of genes encodes agglutinin-like proteins believed 

to play a role in adherence of the yeast to endothelial and 

epithelial cells and is discussed as a virulence factor of this 

important fungal pathogen of humans. Using 66 C. 

albicans isolates, representing the major phylogenetic 

lineages in a worldwide collection of 266 infection-causing 

isolates, we discovered >= sixty-one different alleles of the 

ALS7 open reading frame. Differences between alleles were 

largely caused by polymorphisms in two nonadjacent 

repeats, the so-called tandem repeat domain (21 different 

types occurred) and the so-called VASES domain (21 

different types). C. albicans is diploid, and combinations of 

ALS7 alleles generated >=50 different genotypes. All 

alleles tested were expressed in human patients; parts of 

some open reading frames were transcribed in both 

directions. Isolates representing a more pathogenic generalpurpose 

genotype cluster (Schmid et al., 1999, 

Microbiology 145: 2405-2414) had a higher number of 

tandem repeats than other strains; of the 21 types of 

VASES region found, two were largely exclusive to these 

strains, the remaining almost exclusive to non-cluster 

strains. Our results suggest that ALS7 is a hypermutable 

contingency locus, important for the success of C. albicans 

as an opportunistic pathogen of humans.

IMC7 Main Congress Theme IV: POPULATION DYNAMICS AND ECOLOGY Posters 

925 - Arid element in the lichen flora of Kugitangtau 

(Turkmenistan) 

Z.H. Abdurahimova 

National Institute of Desert, flora and fauna Minister of 

Nature Protection of Turkmenistan, Bitarap Turkmenistan 

Avenue 15, Ashgabat 744000, Turkmenistan. - E-mail: 

zar_zuw.ru@rambler.ru 

Kugitangtau is the South-West ridge of the Pamir and Alai 

mountains and is a constituent part of the Central Asian 

mountain province. In the flora compositions of lichens 

have been revieled 240 species, 5 varieties and 12 forms. 

The analysis of Kugitangtau lichen flora according to 

geographical element have been done and 9 elements have 

been singled out: arid (68 species - 28,3% of total species, 

multiregional (36-15%), eurogolarktical (30-12,5% boreal 

(18-7,5%), hypoarkto-montane (12-5,2%), arkto-highland 

(11-4,5%), mediteranean (5-4,5%), montane (12-5%) and 

non-moral. Among them the species of arid element prevail 

which refer to 21 genera of 10 families. They include 45 

species epilities, 19-epigeides, 8-epiphytes and 2 migrating, 

but some of them (6 species) can be observed both on the 

soil and rocks. Turklmenistan is situated in the arid zone, 

therefore it is natural that the highest percent of 

Kugitangtau lichen flora includes the arid species. The area 

spectrum of the arid element accounts for 18 names. This 

element is mainly represented by xeromeridional, desert 

and steppe species.The bulk of the arid element is formed 

by species with central asian (12), european and asian (9), 

asian(8), xero-merideonal-golarktical (7) and golarktical 

(6) type of area. 

926 - Population genetics of Marasmius oreades in a 

Norwegian sand dune ecosystem analyzed by DNA 

amplification fingerprinting 

E. Abesha, G. Caetano-Anollés & K. Høiland * 

Department of Biology, P.O. Box 1066 Blindern, N-0316 

Oslo, Norway. - E-mail: klaus.hoiland@bio.uio.no 

The population genetics and spatial structure of Marasmius 

oreades was studied at the molecular level. Basidiocarps 

were collected from fairy rings from the sand dunes on 

Peninsula Lista in SW Norway. Samples were collected 

after mapping of fairy rings. DNA amplification 

fingerprinting (DAF) with arbitrary oligonucleotide 

primers was used to study genetic relationships between 

the basidiocarp samples. The fungal populations contained 

a high number of genotypes and each fairy ring generally 

represented a separate genet. Both cluster and phylogenetic 

analyses of the DAF products established relationships 

between fairy rings and showed that genetically similar 

basidiocarps were found close to each other. Overall results 

showed, however, only a weak correspondence between 

genotype and spatial distribution and no correspondence 

between genotype and composition of the surrounding 

vegetation. 

927 - Impact of monsooning of coffee on fungal 

community structure, enzyme profiles and toxins 

R. Ahmad 1* , N. Magan 2 & V. Sanchis 3 

1 Department of Applied Botany, Mangalore University, 

Mangalagangotri 574199, India. - 2 Applied Mycology 

Group, Biotechnology Centre, Cranfield University, Silsoe, 

Bedford MK45 4DT, U.K. - 3 Food Technology Department, 

Lleida University, 25198 Lleida, Spain. - E-mail: 

rasheeds_2K@yahoo.co.uk 

Monsooning of cofee beans is a semi-solid state 

fermentation process occurring over a 8-10 week period 

under high humidity conditions of South-west peninsular 

India. The moisture content of beans undergoing 

monsooning increases by up to 20%, consequently 

significant variation in the microbial populations were 

observed between the non-monsooned and monsooned 

beans. Total populations of bacteria and fungi varied 

significantly in these two treatments in both cultivar 

Arabica and Robusta. Dominant species included 

Aspergillus niger and A. ochraceus. Hydrolytic enzyme 

assays showed that there were significant differences 

between coffee types indicative of physiological activity of 

different fungal communities. The level of ochratoxin in 

naturally and controlled monsooneing was less than 5 ppb. 

The implications of these results for developing controlled 

monsooning systems are discussed. 

928 - Promoted growth of cabbage seedlings inoculated 

with arbuscular mycorrhizal fungus 

M. Akiyama 1* , T. Yoshino 1 , T. Maeda 2 , H. Kakuta 2 & K. 

Oosawa 1 

1 

Department of Horticultural Science, Graduate School of 

Agriculture, Hokkaido University, Nishi-9, Kita-9, Kita-ku, 

2 

Sapporo, 060-8589, Japan. - Plant Ecochemicals 

Research Center, 3-1-1, Megumino-kita, Eniwa City, 061- 

1371, Japan. - E-mail: acky@res.agr.hokudai.ac.jp 

Arbuscular mycorrhizal (AM) colonization promotes the 

growth of host plants. Non-host plants (Brassicaceae, 

Chenopodiales and Polygonaceae etc.) are not infected and 

do not build symbiosis with AM fungus. However, the 

growth of cabbage seedlings (Brassica oleracea L. var. 

capitata L., cv. Earlyball) was significantly promoted by 

the inoculation of AM fungus (Gigaspora margarita), 

though no fungal infection of the root was recognized. The 

factor of the phenomenon was investigated in terms of the 

experiment process, the phosphorus condition of the soil 

and the species of AM fungus. The growth of seedlings 

was the same to control (no addition of inoculum) in spite 

of addition of autoclaved inoculum to rhizosphere. This 

observation shows that the extender included in inoculum 

did not play a role of source of the nutrients. By addition of 

P fertilizer to soil, the growth of seedlings at high-P-soil 

was more vigorous than at low-P-soil and growth 

promotion by inoculation of AM fungus was additively 



maintained. All the three AM fungus species (Gigaspora 

margarita, Glomus fasciculatum and Glomus mosseae) 

used in experiment promoted the growth of cabbage 

seedlings significantly. However, the level of the 

promotion varied among the three species. These results 

suggest that the existence of AM fungus in the rhizosphere 

improves the soil condition for plant growth, and it 

provides a new approach for the plant-AM fungus 

symbiotic system. 

929 - Interactive effects of nutrient applications and 

arbuscular mychorrhizal colonization on little bluestem 

grass (Schizachyrium scoparium) 

H. Antonsen 1* , R.C. Anderson 2 , S.A. Juliano 2 & S.S. 

Dhillion 1 

1 Department of Biology and Nature Conservation, 

Agricultural University of Norway, Postbox 5014, N-1432 

Aas, Norway. - 2 Department of Biological Sciences, Illinois 

State University, Campus box 4120, Normal, Illinois 

61790-4120, U.S.A. - E-mail: hilde.antonsen@ibn.nlh.no 

A manipulative field study was conducted in 1985 to 

evaluate the impacts of supplemental nutrients (N, P, K and 

Ca+Mg) on little bluestem grass (Schizachyrium 

scoparium) with a high (20.2 ±1.6%) or low (2.6 ±0.7%) 

level of colonization by arbuscular mycorrhizal fungi 

(AMF). Low-colonized plants were grown in autoclaved 

soil and outplanted into fumigated soil in the field, while 

high-colonized plants were grown in unsterilized soil and 

outplanted into unfumigated field soil. Originally, data 

were analyzed by multiple univariate analyses of variance. 

Previous conclusions drawn from these analyses were that 

bases (Ca+Mg) were the limiting nutrients in this system. 

A reanalysis of the data was performed by means of 

multivariate statistics. New interpretations suggest that P 

was the limiting nutrient in this system, since application of 

P increased growth of S. scoparium when AMF 

colonization was low. However, no effect of P application 

on growth was found for plants with high levels of 

colonization. This interaction is explained by the cost of 

having a fungal partner. Thus, in this sand prairie system 

AMF restricts growth of S. scoparium. In addition, 

application of bases enhanced AMF colonization under 

natural conditions. 

930 - Phytopathological characteristics and 

environmental impact on the different fungi of the 

Gaeumannomyces/Phialophora (G/P) complex 

C. Augustin * , K. Ulrich & A. Werner 

ZALF, Dep. of Land Use Systems and Landscape Ecology, 

Eberswalder Str. 84, D-15374 Muencheberg, Germany. - 

E-mail: caugustin@zalf.de 

The G/P complex contains fungi that cause take-all 

diseases on cereals as well as apathogenic fungi. To study 

280 


the interactions of different groups, about 2000 isolates 

from numerous geographic locations in Germany were 

differentiated into species and varieties as well as on the 

intravarietal level using RAPDs. Based on this strain 

collection, the phytopathological behaviour of 

representative isolates was evaluated on several cereal 

varieties. Beside differences between species and varieties 

we found a high variation in aggressiveness, especially to 

wheat, among isolates of var. tritici. Whereas one RAPD 

group of the var. tritici consisted exclusively of high 

pathogenic isolates, the other one contained pathogenic, 

apathogenic and even growth promoting isolates. In order 

to predict the potential risk of take-all it is necessary to 

analyse both the composition of the G/P fungi population 

and the impact of different environmental factors on the 

abundance of respective fungal groups. In our study we 

monitored spatial occurrence and distribution of species, 

varieties and subgroups of the G/P-complex as a function 

of soil variables, weather conditions and crop rotation over 

4 years. Data show that the various fungi analysed 

displayed clearly different claims concerning 

environmental factors. Actual weather and several soil 

parameters were determined as the most important 

discriminating influences. Results may provide a useful 

basis for detailed improvement of prognostic models. 

931 - The direct effect of nitrogen as a mechanism for 

change in ectomycorrhizal fungal communities 

P.G. Avis, D.J. McLaughlin * , I. Charvat & P. Reich 

University of Minnesota, Plant Biological Sciences 

Graduate Program, 220 Biosciences Center, 1445 Gortner 

Avenue, St. Paul MN 55108, U.S.A. - E-mail: 

davem@tc.umn.edu 

The mechanisms causing shifts in ectomycorrhizal fungal 

(EMF) communities exposed to increased nitrogen (N) by 

atmospheric deposition or fertilization are not understood. 

The effects of N increase could be direct, or could be due 

to ancillary effects such as altered pH or a change in 

limiting nutrients. This study examines the direct effects of 

increased N by examining the response of EMF 

communities to a 16-year fertilization experiment in an oak 

savanna. The experiment consists of two levels of nitrogen 

fertilization (5 and 17 g N m -2 yr -1 ) and unfertilized plots. 

Each fertilization treatment also receives equal background 

levels of P, K, Ca, Mg, and S added to offset ancillary 

effects of N. Fertilization has increased soil N, held pH 

constant and not caused limitation by other nutrients. 

Meanwhile, fertilization 1) decreased the abundance of 

aboveground EMF sporocarps for all species except 

Russula amoenolens (which increased 7 fold); 2) held 

belowground EMF richness constant as measured by 

morphotype and PCR-RFLP methods; but 3) altered the 

dominant fungi in the EMF communities: Unfertilized 

EMF communities are dominated by species with extensive 

external hyphae, notably Cortinarius species, while heavily 

fertilized communities are dominated by those that lack 

external hyphae, primarily R. amoenolens. These data show 

that the direct effect of N addition may be the driving 

mechanism behind the dramatic shifts in EMF communities 

in ecosystems that experience N deposition.


932 - Ligninolytic enzymes and biodegradation during 

the interactions of white-rot fungi and soil 

microorganisms 

P. Baldrian 

Institute of Microbiology AS CR, Videnska 1083, CZ- 

14220, Prague 4, Czech Republic. - E-mail: 

baldrian@biomed.cas.cz 

White-rot fungi are able to degrade lignin and related 

compounds. Under natural conditions, the process occurs in 

the presence of other microorganisms. Introduction of 

microorganisms to liquid cultures of Trametes versicolor 

led to the increase of activity of the ligninolytic enzyme 

laccase. High increase was achieved with soil fungi 

Trichoderma harzianum (2810% of control), Penicillium 

rugulosum (1940%), Fusarium reticulatum (1690%), 

Cephalosporium sphaerospermum (840%), and Humicola 

grisea (480%). The increase was lower after addition of 

bacteria - Escherichia coli (310%) and Bacillus subtilis 

(190%), or the yeast Endomyces magnusii (220%). After 

one-week cultivation with Trichoderma harzianum, the 

mycelium of T. versicolor was killed, which was 

accompanied by the decrease of Mn-peroxidase activity. 

Increase of laccase activity is a common response - it was 

found also in the white-rot fungi Abortiporus biennis, 

Coriolopsis occidentalis, Pleurotus ostreatus, Pycnoporus 

cinnabarinus and Trametes hirsuta. It might be involved in 

active defence, since some products of laccase exhibit 

antimicrobial activities. Increase of laccase activity 

correlated with the increase of decolorization of the 

synthetic dye Remazol brilliant blue R. It seems, that 

interspecific interactions can affect the biodegradative 

activity of white-rot fungi in situ. This work was supported 

by the Grant Agency of the Czech Academy of Sciences 

(B5020202). 

933 - Population structure of Ceratocystis fimbriata 

from Congo, Colombia and Uruguay, determined using 

microsatellite markers 

I. Barnes 1* , J. Roux 1 , B.D. Wingfield 2 & M.J. Wingfield 1 

1 University of Pretoria, Forestry and Agricultural 

Biotechnology Institute, Department of Microbiology and 

Plant Pathology, 74 Lunnon Road, FABI, University of 

Pretoria, Pretoria, 0002, South Africa. - 2 University of 

Pretoria, FABI, Department of Genetics, 74 Lunnon Road, 

FABI, University of Pretoria, Pretoria, 0002, South Africa. 

- E-mail: irene.barnes@fabi.up.ac.za 

Ceratocystis fimbriata is a haploid ascomycete that causes 

serious diseases on a wide range of plants, world-wide. 

Very little is known regarding the population biology or 

origin of this important pathogen. The aim of this study 

was to use 11 polymorphic PCR-based microsatellite 

markers previously designed for C. fimbriata, to examine 

the population structure and genetic diversity for different 

populations of C. fimbriata. Populations consisting of 32 

isolates from each of Congo, Colombia and 22 from 

Uruguay were studied. High genetic diversities for all 

populations were observed (H = 0.48-0.31; G = 24-48%). 

Colombia was the most diverse population consisting of 

82% of the total, and 53% of the unique, alleles. Genetic 

differentiation between populations was great (GST= 0.39) 

and minimal gene flow was observed (Nm = 0.77). I A, 

PTLPT, and linkage disequilibrium tests to determine 

mode of reproduction showed little evidence for 

recombination within populations. C. fimbriata appears to 

reproduce primarily without outcrossing. UPGMA 

dendrograms showed that the Colombian population was 

more distantly related to the Congo and Uruguay 

populations than they were to each other. Some of the 

groups in the Colombian and Uruguay populations were 

genetically similar to isolates from the Congo. Results of 

this study show that African isolates of C. fimbriata 

originated in Latin America. Moreover, evidence indicates 

that Latin America is also a likely area of origin for C. 

fimbriata. 

934 - Studies in the life cycle of Puccinia glechomatis 

J.B. Boellmann & M.S. Scholler * 

Purdue University, Department of Botany & Plant 

Pathology, Arthur & Kriebel Herbaria, Lilly Hall, West 

Lafayette, IN 47907, U.S.A. - E-mail: scholler@purdue.edu 

Puccinia glechomatis DC. is a microcyclic rust fungus on 

Glechoma spp. (Lamiaceae) forming only telia and basidia 

(spore states III, IV). The species is a native of Eurasia but 

naturalized in North America where it is spreading since 

the early 1990s. We studied the life cycle of the species by 

propagating the fungus on G. hederacea (ground-ivy) in 

the greenhouse, by various inoculation and germination 

techniques and by using the light microscope. Nuclear 

conditions were documented by epifluorescence 

microscopy and DAPI as DNA specific fluorochrome. The 

complete life cycle requires c. 20 d. After karyogamy in 

teliospores both cells germinate with a phragmobasidium. 

In basidia meiosis produces four nuclei, which move in the 

developing basidiospores. Further mitosis in the 

basidiospore may result in up to four nuclei. All but one 

nucleus degenerates during germination of the 

basidiospore. The germ tube penetrates the epidermis on 

the upper leaf surface and develops a rich haploid 

mycelium especially around stomata. Fusion of haploid 

hyphae occurs could not be documented. Our studies 

indicate that the fungus is homothallic. 

935 - Preliminary study of NE Portugal's macrofungal 

communities 

S.M. Branco * & A.P. Rodrigues 

Parque Natural de Montesinho/Instituto da Conservação 

da Natureza, Apartado 90, 5301 Bragança, Portugal. - Email: 

sarabranco@yahoo.com 



In Portugal, as in many other countries, there are few 

studies concerning macrofungi and there is much to be 

known. The NE Portugal is no exception, and habitats like 

oak forests and serpentine soil sites have not yet been 

explored. The present study takes place in two different 

habitats, a Quercus pyrenaica stand and a Quercus 

rotundifolia in serpentine soil stand. The objective is to 

characterize the macrofungal communities present in both 

habitats and relate them with soil and vegetation variables. 

We expect to find different mycota and that the influence 

of environmental factors is different in the two habitats. 

936 - Interspecific interactions between saprotrophic 

basidiomycetes: Effect on gene expression and enzyme 

activity 

A.W. Branney * , L. Boddy & H. Rogers 

Cardiff University, School of Biosciences, Cardiff 

University, PO Box 915, Cardiff, CF11 3TL, U.K. - E-mail: 

abranney@hotmail.com 

Saprotrophic basidiomycetes rarely form monospecific 

populations in the soil or organic substrata that they 

inhabit, therefore interspecific mycelial interactions 

continually occur. Gross outcome can either be deadlock 

(where neither species gains headway) or replacement 

(where one species wrests territory from the other). 

Irrespective of the outcome, dramatic morphological 

changes may occur in both mycelia. Morphological 

changes are correlated with differences in physiology and 

enzyme production, and may occur distant from the site of 

interaction. Currently, there is little or no information on 

spatial and temporal changes in gene expression and in 

enzyme production in mycelia in soil, either when growing 

alone or during interactions. In interactions between 

Hypholoma fasciculare and Phlebia radiata, we test the 

hypothesis that: (1) there is spatial and temporal variation 

even in mycelia growing alone, related to morphological 

differentiation and presence of substrates; (2) massive 

changes occur in the vicinity of interactions; (3) changes 

also occur elsewhere in support of the interaction front. 

937 - Genetic sieves in sexual reproduction: sexual 

spores of Aspergillus nidulans have lower mutation load 

than asexual spores 

J. Bruggeman * , P.J. Wijngaarden, A.J.M. Debets & R.F. 

Hoekstra 

Genetics Department, Wageningen University, 

Arboretumlaan 4, 6703 BD Wageningen, The Netherlands. 

- E-mail: judith.bruggeman@genetics.dpw.wau.nl 

Whatever the evolutionary forces are for the widespread 

occurrence of sexual reproduction, Mendelian transmission 

is interwoven with it. This system ensures maximal 

uncertainty for the transmission of genetic information, 

thereby preventing parasitic genes or sequences to gain a 

282 


transmission advantage. A disadvantage of this system is 

that both low quality and high quality alleles have equal 

chances of being transmitted to the progeny. Clearly, 

evolution should favour processes within an individual that 

alleviate this disadvantage either by selection on gamete 

producing cells, on gametes or on support-relying zygotes. 

We introduce the term 'genetic sieves' for these processes. 

The haploid homothallic ascomycete fungus Aspergillus 

nidulans is excellently suited to study genetic sieves due to 

important properties, which we will explain on our poster. 

We predict that 'genetic sieves' in the sexual reproduction 

cycle prevent or slow down the accumulation of deleterious 

mutations. We tested this in a mutation accumulation 

experiment for both sexually selfing and asexually 

reproducing lines. In this poster we present the result of 

this mutation accumulation experiment that support our 

prediction. From the fitness data we calculated mutation 

rate and mean fitness effect of a mutation using the 

Bateman-Mukai equations. This gives us insight in the 

operation of this sieve. 

938 - Armillaria species and the legacy of forest 

disturbance in Ozark Highlands landscapes of the 

central U.S.A. 

J.N. Bruhn 1* , R.P. Guyette 2 , J.D. Mihail 1 & J.M. Kabrick 3 

1 Dept. of Plant Microbiology & Pathology, University of 

Missouri, 108 Waters Hall, Columbia, Missouri, 65211, 

U.S.A. - 2 School of Natural Resources, University of 

Missouri, 203 ABNR, Columbia, Missouri, 65211, U.S.A. - 

3 Missouri Department of Conservation, 1110 South 

College Avenue, Columbia, Missouri, 65201, U.S.A. - Email: 

bruhnj@missouri.edu 

Relationships between disturbance regimes and diverse life 

forms are being studied to understand forest landscapes and 

their responses to management. Disturbance regimes were 

characterized by abrupt ring-width reductions in Pinus 

echinata and fire frequency. Disturbance indices are 

correlated with topographic roughness and the abundance 

of certain bird, reptile, soft mast fruit, Armillaria, and tree 

species. In turn, Armillaria spp. contribute to forest 

decline, functioning both as long-lived disturbance factors 

and as catalysts for further disturbance, in a feedback 

manner. Three Armillaria spp. occur regionally with 

different distributions. Nearly ubiquitous, A. mellea often 

occurs alone on thermally exposed aspects; A. tabescens 

and A. gallica occur typically on ridges vs. protected 

aspects in more rugged landscapes, respectively. 

Behaviorally, A. gallica fruits infrequently, produces most 

rhizomorphs, and causes least disease. Most root disease is 

caused by A. mellea, which fruits prolifically but produces 

fewer rhizomorphs. Less abundant, A. tabescens produces 

the most limited rhizomorph systems, fruits well, and 

causes some disease. Armillaria spp. take advantage of 

successive stress events to gradually invade infected root 

systems. Tree mortality, mostly Quercus coccinea, Q. 

velutina, and Cornus florida, is often associated with root 

crown invasion by A. mellea or A. tabescens. Greater 

mortality on exposed aspects is associated with the 

exclusive presence of A. mellea.


939 - Mycology within the Georgia Coastal Ecosystems 

Long-Term Ecological Research Project (GCE/LTER), 

subproject Decomposer Consortia Experiments (DCE) 

A. Buchan 1 , J.I.L. Moreta 1 , S.Y. Newell 2* & M.A. Moran 1 

1 Department of Marine Sciences, University of Georgia, 

Athens, GA 30602-3636, U.S.A. - 2 Marine Institute, 

University of Georgia, Sapelo Island, GA 31327-0032, 

U.S.A. - E-mail: newell@uga.edu 

The GCE/LTER focuses on potential effects of alteration 

of the quantity and quality of freshwater flow from the 

Altamaha River into Georgia (USA) saltmarshes. 

Ascomycetes are major secondary producers within 

Georgia saltmarshes (about 550 g m -2 y -1 ). One goal of the 

DCE subproject is seasonal measurement of ascomycetous 

and prokaryotic biodiversity in/on the decaying shoots of 

smooth cordgrass (Spartina alterniflora), using both direct 

microscopy (ascospores) and DNA technology (base 

sequences in rDNA/ITS, using ascomycete-specific PCR). 

The DCE subproject also includes examination of genes for 

lignocellulose-degrading enzymes of prokaryotes and 

ascomycetes - the ascomycetous enzyme targeted is 

laccase. Selected findings follow. 1) We found no rDNA of 

major, cryptically occurring ascomycetes - the same three 

species that are the principal ascomatal producers were also 

the major rDNA sources (Phaeosphaeria spartinicola, 

Mycosphaerella sp.2, P. halima). 2) The species with the 

highest rates of ascospore expulsion (P. spartinicola) may 

not be the species with the highest biomass yield - 

Mycosphaerella sp.2 exhibited the greatest rDNA-detection 

peaks (T-RFLP). 3) All major and minor ascomycete 

species examined had one or more laccase genes (15 

distinct types total), and laccase genes of the three 

predominant ascomycetes (+ two unidentified laccase 

genes) were found within naturally decaying leaf blades, 

with the greatest % of clones belonging to laccases of P. 

halima. 

940 - The diversity of mycophagous diptera and their 

macrofungi hosts 

B.A. Bunyard 1* & B.A. Foote 2 

1 Ursuline College, Dauby Science Center, Pepper Pike, 

Ohio 44124, U.S.A. - 2 Kent State University, Department of 

Biological Sciences, Kent, Ohio, U.S.A. - E-mail: 

bbunyard@ursuline.edu 

Despite their ubiquity in nature, few studies have been 

conducted worldwide to determine the ecological 

importance of mycetophagous diptera (fungi-feeding flies). 

For this study 134 species from 30 families of 

Basidiomycetous fungi and 19 species from 11 families of 

Ascomycetous fungi were collected from different sites in 

northeastern Ohio. Different sites were selected to obtain a 

diversity of mushroom substrate, as well as biotic and 

abiotic conditions, and consisted of mature forest, mixed 

mesophytic forest, urban forest, and urban residential. 

Adult flies were reared from 87 different fungal 

collections. Families of Diptera that seem to include 

mycophagous species are: Drosophilidae, Chloropidae, 

Phoridae, Mycetophilidae, Sciaridae, Tipulidae, 

Cecidomyiidae, and Platypezidae. Several other dipteran 

families are probably scavengers (Anthomyiidae, 

Sarcophagidae), occurring only infrequently in decaying 

mushrooms. Many mycetophagous flies are poorly known; 

several have larval stages that remain completely 

undocumented. While some fungal species seem to host a 

single fly species per mushroom, most do not. How 

mycophagous dipteran species can avoid competition is 

uncertain. Unless factors are in operation to prevent it 

(niche partitioning, predation, parasitism), mycophagous 

diptera may pose a challenge to the competitive exclusion 

principle. Preliminary evidence suggests parasitism by 

species of parasitic wasps and predation by ants and beetles 

may play a role. 

941 - r DNA ITS-sequence analysis of ericoid 

endophytes from Australia and the USA 

S.C. Burns 1 , C.B. McLean 1* , M.C. Starrett 2 & A.C. 

Lawrie 3 

1 University of Melbourne, Institute of Land and Food 

Resources, School of Resource Management, Burnley 

College 500 Yarra Boulevard, Ricmond, Victoria, 3121, 

Australia. - 2 University of Vermont, College of Horticulture 

and Life Sciences University of Vermont, Burlington, 

Vermont, U.S.A. - 3 RMIT University, Department of 

Biotechnology & Environmental Biology, GPO Box 71, 

Bundoora, Vic, 3083, Australia. - E-mail: 

cmclean@unimelb.edu.au 

One hundred and seventeen slow-growing, sterile root 

endophytes were isolated from Leucopogon parviflorus 

Lind. (Epacridaceae) collected at two coastal sites in 

southeastern Australia in Autumn, 2001. Twelve isolates 

were then selected for the sequencing study along with four 

root endophytes isolated from Pieris floribunda (Ericaceae) 

collected in the southeast of the United States. All sixteen 

sequences along with sequences from the Northern 

Hemisphere mycorrhizal fungi Hymenoscyphus ericae and 

Oidiodendron maius and the closest GenBank matches 

were aligned using programs made available by the 

Australian National Genomic Service. Most Australian 

sequences from this study clustered with unnamed, sterile, 

isolates from previous studies of the Epacridaceae, not 

related to H. ericae. Three of the Australian sequences 

matched closely (96%) with O. maius and conidia 

produced confirmed them as Oidiodendron spp. Two of the 

fungi isolated from P. floribunda were 99% similar to H. 

ericae sequences in the GenBank the other two fungi 

clustered with Australian dark sterile isolates. 



942 - Effect of arbuscular mycorrhiza (AM) on 

seedlings of six endemic Mimosa species from semiarid 

south-central Mexico 

S.L. Camargo-Ricalde 1* , S.S. Dhillion 1 , C. Jiménez- 

González 2 & R. González-Gómez 2 

1 Agricultural University of Norway (NLH), Dept. Biology 

and Nature Conservation (IBN), P.O. Box 5014, N-1432, 

Ås, Norway. - 2 Universidad Autónoma Metropolitana- 

Iztapalapa (UAMI), Dept. Biología, Apdo. POstal 55-535, 

09340, D. F., Mexico. - E-mail: sara.camargo@ibn.nlh.no 

We report on the effects of arbuscular mycorrhiza (AM) on 

the growth of seedlings of six endemic Mimosa species: M. 

adenantheroides, M. calcicola, M. lacerata, M. luisana, M. 

polyantha and M. texana var. filipes. Seeds were collected 

directly from plants. In the greenhouse, germination was 

carried out in pots containing the soil of the localities 

where the species occur. Ten seedlings per treatment per 

species were examined. Treatments were applied once per 

week: control, benomyl (B), phosphorus (P) and B+P. 

Shoot length was measured twice per week. Plants were 

harvested after seven months. Both root and shoot length 

were measured, dry biomass quantified and shoot 

phosphorus content; and AM root colonization percentage. 

Seedlings treated with B had a significant lower growth 

and less biomass than the other seedlings. Phosphorus 

content was higher in the seedlings treated with P and P+B. 

The highest AM colonization percentage was registered on 

the seedlings treated with P and in the control. We found a 

higher number of nodules in the seedlings treated with P. 

Based on other parallel studies, after germination, which 

takes place during the wet season, a fast growth favored by 

AM symbiosis, may increase the survival probability of 

seedlings in a harsh environment such as semiarid 

ecosystem. 

943 - Molecular ecology of root endophytes in an alpine 

Bistorta vivipara - Kobresia myosuroides tundra plant 

community 

T.A. Carlsen * , K. Høiland & T. Schumacher 

Div. of Botany and Plant Physiology, Dept. of Biology, 

University of Oslo, P.O. Box 1045 Blindern, 0316 Oslo, 

Norway. - E-mail: torac@bio.uio.no 

A total of 210 endophyte axenic cultures from root tips of 

22 plant species, including the ectomycorrhizal herb 

species Bistorta vivipara and Kobresia myosuroides were 

grouped according to culture morphology and HinfI and 

HaeIII restricted ITS-RFLP groups. nrDNA ITS sequences 

(98) from representative isolates of the 23 ITS-RFLP 

groups were obtained. More than 90 % of the sequences 

had affinities to the Helotiales (Ascomycota) and were 

subjected to phylogenetic analyses in PAUP* with 47 

sequences from GenBank. Altogether 39.5 % of the total 

sample of root endophytes were referable to 

Leptodontidium orchidicola. Phialophora finlandia was 

284 


recorded in four herbaceous plant species, i.e. Bartsia 

alpina, Carex atrata, K. myosuroides and B. vivipara, and 

most often in association with the ectomycorrhizal root tips 

of B. vivipara and K. myosuroides. Axenic cultures of 

Phialocephala fortinii were obtained from ectomycorrhizal 

root tips of B. vivipara and K. myosuroides only. The L. 

orchidicola ITS genotypes (31) were unevenly distributed 

in four plots investigated. Hypotheses for host specificity 

for fungal endophytic strains and possibilities of a common 

fungal network are proposed. 

944 - Effect of binucleate and multi-nucleate 

Rhizoctonia spp. on seed germination and growth of 

Anoectochilus formosanus Hayata 

D.C.N. Chang * , L.C. Chou & G.C. Lee 

National Taiwan University(NTU), Dept. of Horticulture, 

Taiwan. - E-mail: hyha@ccms.ntu.edu.tw 

Anoectochilus formosanus Hayata is a medical and 

ornamental orchid in Taiwan. It was considered as a 'King 

medicine'. Asymbiotic germination of A. formosanus in 

Hyponex # 3 agar medium (H3A) was 65%. While 

symbiotic germinations inoculated with binucleate 

Rhizoctonia sp. (R02) or multi-nucleate Rhizoctonia sp. 

(R04) in oat meal agar (OMA) medium were 42% and 31% 

respectively. But the growth of symbiotic seedlings were 

faster than those in asymbiotic medium (H3A). Both 

binucleate (R02) and multi-nucleate (R04 and R05 ) 

Rhizoctonia spp. could enhance the growth of A. 

formosanus plantlets in vivo. The binucleate Rhizoctonia 

sp. R02 could be Rhizoctonia callae. By hyphal 

anastomosis tests, the results showed that multi-nucleate 

Rhizoctonia spp. (R04, R05, R06, R07, R09) belong to 

AG-6 group. From RAPD data, using UPGMA clustering, 

and light microscopy observation showed that that R03 and 

R08 were Fusarium spp., and the other multi-nucleate 

Rhizoctonia spp. were Rhizoctonia solani. It was suggested 

that the inoculation of binucleate or multi-nucleate 

Rhizoctonia spp. for A. formosanus plantlets in vivo could 

promote their growth in practical use. 

945 - Ecology of micro-fungi in mangrove sediments of 

the Ganges river estuary in India 

S. Chaudhuri * & A. Sengupta 

Department of Plant Pathology,BCKV(Agriculture 

University), Kalyani-741235, West Bengal, India. - E-mail: 

mycolab@yahoo.com 

Tidally inundated riverine sediments in the tropics at 

outfall are inhabited by mangroves, the climax formation of 

hydro-halophytes. Population dynamics and ecology of 

micro-fungi in surface sediments of four floristically 

different, nutrient limited mangrove succession stages at 

the Ganges river estuary in India are reported. Population 

density and species diversity of micro-fungi were inversely


related to salinity and inundation levels and positively with 

species diversity and nitrogen content of the sediments. 

None of the 46 species of micro-fungi isolated was 

halophile, only adaptive halotolerance was observed in 

most of them. While population of most halotolerant 

species declined with increasing salinity, the same for some 

remained constant and for a few increased. Although ED50 

salinity concentration for in vitro growth retardation of the 

isolates varied between 8.2 - 14.5 dSm-1, they differed in 

growth dynamics from their mesic counterparts between 

3.0-5.5 dSm-1, suggesting a lower salinity tolerance limit 

for physiological function than for their ecosystem 

presence. Comparison of the population fraction of 

cellulose decomposer and insoluble phosphate solubilizer 

fungi, their diversity and distribution, in vitro 

decomposition-solubilization ability and limits of salinity 

tolerance with that of sediment bacteria indicated that the 

fungi were no less significant as a population group of 

carbon and phosphorus cycling microorganisms in the 

extreme saline environment. 

946 - Comparison of the post-fire dynamics of the 

ectomycorrhizal community in two Quercus ilex stands 

in northern Spain 

V. Clavería * , A.M. de Miguel & M. de Román 

Dep. Botany. University of Navarra, C/Irunlarrea s/n 

31080 Pamplona, Spain. - E-mail: 

vclaver@alumni.unav.es 

A comparative study of the post-fire recolonization of 

ectomycorrhizae in a evergreen oak stand (Quercus ilex L. 

subs. ballota (Desf.) Samp.) has been carried out in two 

Quercus ilex stands located in Nazar and San Cristóbal 

(Navarra, Spain) respectively. The research in our group 

has been focused during the last three years on the 

description and identification of ectomycorrhizae in burnt 

Quercus ilex stands, and in adjacent areas which were not 

affected by fire and act as control plots for the study. In 

1993 a stand in Nazar burnt, but it was not until 1998 that 

the study started. On the contrary, the study in San 

Cristóbal started immediately after the stand had burnt (in 

the year 2000), thus enabling us to analyse the mycorrhizal 

communities in the first stages after the fire. Therefore we 

have been able to compare the regeneration in both stands 

and the species composition five years after the fire and 

immediately after the fire. The sampling of 

ectomycorrhizae has been done in Nazar seasonally for 

three years while in San Cristóbal the monitoring of the 

mycorrhization has been done according to the same 

methodology during one and a half years so far. The 

description of several of the most important morphotypes 

of ectomycorrhizae found in both stands are provided, 

including photographs of different macroscopic and 

microscopic features of the mycorrhiza. 

947 - Influence of mycorrhizal inoculum on clonal 

propagation in Epacris impressa Labill. 

M. Conomikes 1 , C.B. McLean 1* , M.C. Starrett 2 & A.C. 

Lawrie 3 

1 University of Melbourne, Institute of Land and Food 



Australia. - 2 University of Vermont, College of Horticulture 

and Life Sciences University of Vermont, Burlington, 

Vermont, U.S.A. - 3 RMIT University, Department of 




Members of the Epacridaceae are traditionally difficult to 

propagate and are in decline in parts of Australia. Attack by 

soil-borne fungi has led to some species of Epacridaceae in 

Western Australia being listed as endangered. Seed is 

nearly impossible to germinate and cuttings often have a 

strike rate as low as 10%. Previous studies have shown that 

introduction of soil collected from beneath adult plants 

showed improved health and survival of cuttings of several 

epacrid species. In this study cuttings were grown in either 

potting mix alone or potting mix containing either 

mycorrhizal inoculum or soil from beneath adult plants 

collected in the wild. Plants were grown under glasshouse 

conditions for 20 weeks and monitored for health and 

survival before harvesting. Strike rate and mycorrhizal 

status was then determined. Statistical analysis of results 

showed no significant difference between treatments and 

no mycorrhizas present in the roots of any cutting in any of 

the treatments. 

948 - The development and use of co-dominant SNPs 

and SSRs in the study of gene flow in Scottish late 

blight populations 

D.E.L. Cooke * , A.K. Lees, S. Hussain, L. Sullivan, N.A. 

Williams & J.M. Duncan 

Host Parasite Co-evolution, Scottish Crop Research 

Institute, Invergowrie, Dundee, DD2 5DA, U.K. - E-mail: 

dcooke@scri.sari.ac.uk 

Potato blight is the most serious diseases of the potato 

industry worldwide. Infecting both the foliage and tubers, it 

reduces yield and quality of ware and seed potato crops. A 

survey of Scottish potato blight (Phytophthora infestans) 

populations from 1995-1997 yielded 500 isolates from over 

80 disease outbreaks. The isolates were characterised by 

mating type, fungicide sensitivity and AFLP fingerprinting. 

These data were examined alongside cropping details to 

determine the population structure in the context of 

existing disease management. Both mating types were 

found and considerable AFLP diversity observed. Three 

sub-populations were identified with higher ratio of the A2 

mating type within one group. Evidence of occasional 

sexual recombination, and thus gene flow was observed. 



Since the production of long-lived sexual oospores has an 

epidemiological and evolutionary significance that impacts 

disease management, it is important that we understand P. 

infestans population biology. At SCRI we have developed 

a range of co-dominant SSR and SNP markers for this 

purpose. They are being used to examine the fine-scale 

diversity of Scottish populations, form the basis of a 

'molecular toolkit' for examining populations worldwide 

and have utility in tracking isolates in epidemiological 

studies. 

949 - ITS-sequence study of root endophytes isolated 

from Alpine Epacridaceae 

P.W. Davies * , S.C. Burns & C.B. McLean 

University of Melbourne, Institute of Land and Food 



Australia. - E-mail: cmclean@unimelb.edu.au 

Three hundred and forty sterile fungi were isolated from 

the roots of five species of Epacridaceae (Epacris 

paludosa, E. microphylla, E. petrophila, Leucopogon 

montanus and Richea continentis) collected in Jan 2002 

from Mt Kosciuszko National Park, NSW, Australia (- 

36°27', 148°15', Elevation 2228 m). Fifty isolates were 

selected for sequencing studies and each was grown in 

liquid culture, harvested and DNA extracted. The universal 

primers ITS1 and 4 were used to amplify the ITS 1, 5.8s 

and ITS2 regions of the rDNA. Sequences were edited and 

used to search the database for similar matches. All fifty 

sequences plus the closest GenBank matches were aligned 

using programs made available on the Australian National 

Genomic Information Service and the resulting alignment 

analysed using PAUP. The phylogenetic tree created was 

similar to those produced in previous studies of epacrid 

endophytes isolated from plants growing at sea level. None 

of the fifty fungi isolated had similar ITS sequences to the 

known ericoid endophyte Oidiodendron maius. 

950 - Life forms of Rhizoplaca baranowii (Poelt) 

Golubkova 

E.A. Davydov 

Altai State University, Lenina Str. 61, Barnaul, 656099, 

Russia. - E-mail: davydov@bio.asu.ru 

The Project presented aims in comparing the 2 

morphotypes or life forms of the lichen R. baranowii. 

Studies are based on published data as well as herbarium 

materials and field observations in Altai Mts (Russia, 

Mongolia). R. baranowii is endemic to Altai Mts and is 

known from the few localities in high mountain steppe 

habitats. All specimens can be divided into 2 groups based 

on their morphology 'subplacoid' (P) and 'spheroid' (S). 

Most specimens in majority of sites belong to the 'S', and 

this is the morph that falls into original description of the 

286 


species by Poelt. 'P' is characterized by the unattached 

plates up to 12 mm in diam with down-turned margins. The 

main peculiarity of 'P' is its dorsiventral structure. Further 

with time the open part of the black hypothallus is 

becoming smaller and finally disappearing. Transitional 

forms between 'P' and 'S' are found, but both forms can't be 

treated as subsequent stages in the life span as they are 

reproducing by fragmentation independently giving rise to 

the same form respectively. Reproduction by apothecia is 

known only for 'S'. The series of transitional forms between 

'P' and 'S' illustrates in our opinion the evolutionary process 

of evolving of spheroid thallus from unattached plates. 

Ecologically both morphotypes discussed are the life forms 

induced by the differing environmental conditions. 

Population analysis and comparison between the habitats of 

'S' and 'P' proves that sheep grazing largely induces the 

latter form. 

951 - Morpho-productive and genetic diversity of a 

collection of Pleurotus eryngii isolates 

T. de Gioia 1* , D. Sisto 1 , G. Figliuolo 2 , G.L. Rana 2 & R. 

Marino 2 

1 Dipartimento di Biologia e Patologia Vegetale, Università 

degli Studi di Bari, via G. Amendola 165/A, 70126 - Bari, 

Italy. - 2 Dipartimento di Biologia, Difesa e Biotecnologia 

Agro-Forestale, Università degli Studi della Basilicata, 

C.da Macchia Romana, 85100 - Potenza, Italy. - E-mail: 

teresa.degioia@agr.uniba.it 

The evaluation of phenotypic and genetic variation of P. 

eryngii var. eryngii, P. eryngii var. ferulae and P. eryngii 

var. nebrodensis germplasm is a prerequisite to correctly 

distinguish groups for traits of interest and to identify 

strains with high yield potential. In this study, a population 

of 142 strains, belonging to the above varieties and having 

different Italian geographic origins, was studied for 

quantitative, qualitative and molecular traits. An 

experimental design with three blocks and three replicates 

for each strain within block, was used to estimate trait 

parameters. High significant differences were observed 

among strains for number and weight of sporocarps, while 

no significant differentiation was observed among 

geographic origins and taxonomic groups. By using 

qualitative traits, a degree of differentiation was only 

observed with the Sardinian and nebrodensis strains 

respectively. Some non-commercial strains performed 

better than the commercial ones in terms of yield. On 

average, yield per strain (basidiocarp weight) was more 

correlated with number of fruit bodies than with their size. 

The most stable traits across replicates were basidiocarp 

'number' and 'weight' per strain. It follows that such traits 

can be used for an efficient genetic selection of the best 

yielding strains. Both minisatellites and RAPD markers 

performed very well in identifying each strain and in 

discriminating the different taxonomic groups.


952 - Ecological specificity of Laboulbenia 

(Laboulbeniales, Ascomycetes) through transmission 

experiments and artificial infections 

A. De Kesel 

National Botanic Garden of Belgium, Domein van 

Bouchout, B-1860 Meise, Belgium. - E-mail: 

dekesel@br.fgov.be 

Host specificity of Laboulbenia slackensis, and congeneric 

taxa, was studied on living hosts and under controlled 

conditions. From abiotic factors, soil composition and 

relative humidity have the strongest impact on the fungus 

population growth. The best growth conditions are close to 

natural conditions. Transmission of L. slackensis is affected 

by copulation, host population density and spore longevity. 

Direct transmission is promoted by host copulation and 

increasing host population density. It surpasses indirect 

infection as spores are short-lived and soilborne infections 

rare. Both transmission types enhance isolation and 

promote specialization as they reinforce intraspecific and 

obstruct interspecific transmission. Successful artificial 

infections with L. slackensis, L. flagellata, L. giardii, L. 

vulgaris and L. pedicellata were obtained on new carabid 

hosts, providing that optimal growth conditions are met. 

Hosts from non-carabid families could not be infected, 

suggesting that physiological barriers are present, but 

active on higher taxonomic levels. The specialization of 

Laboulbenia species, with a narrow natural carabid host 

range, is explained by (1) a physiological dependence and 

obligate ectoparasitism to Carabidae, (2) a certain degree of 

isolation due to the lack of interspecific transmission (time 

and space) between potentially suitable carabid hosts and 

3) the adaptation of the ectoparasite to a specific 

environment which is rigorously chosen by the host. 

953 - Fungal biodiversity in truffle beds in Navarra 

(Spain): Ectomycorrhizae and hypogeous fungi 

A.M. de Miguel * , M. de Román, M.L. Etayo & V. Clavería 


31080 Pamplona, Spain. - E-mail: amiguel@unav.es 

Since 1993 we have been studying the ectomycorrhizae of 

truffle beds in order to know more about the evolution of 

mycorrhizal colonization in plantlets artificially inoculated 

with Tuber melanosporum. The richness of the 

ectomycorrhizal community increased considerably during 

the first sampling years, and we realised soon how 

important the process of natural colonization was. Apart 

from monitoring the permanence of the inoculated 

mycorrhizae of Tuber melanosporum in field conditions, 

we have recorded other Tuber mycorrhizae, as well as 

mycorrhizal morphotypes belonging to the genera Genea, 

Scleroderma, Hebeloma, Pisolithus and many Tomentellalike 

morphotypes. Although the identity of most of the 

mycorrhizal morphotypes is still unknown, the detailed 

morphological and anatomical descriptions carried out will 

probably enable us to identify the fungus forming the 

mycorrhiza in future studies. On the other hand, we have 

also studied the sporocarps, paying special attention to 

hypogeous fungi and gasteromycetes due to their special 

importance as ectomycorrhizal species. The 

ectomycorrhizal morphotypes characterized and the 

sporocarps collected in truffle beds will be presented, 

together with some comments about the possible link 

between them. 

954 - Above- and below-ground views of tomentelloid 

fungi in a Quercus ilex stand in northern spain 

M. de Román * , A.M. de Miguel & V. Clavería 


31080 Pamplona, Spain. - E-mail: mderoman@unav.es 

Since autumn 1998 we have carried out seasonal samplings 

of ectomycorrhizae in a Quercus ilex stand located in 

Navarra (N Spain). The ectomycorrhizal morphotypes 

found have been described and identified when possible, 

yielding a total of 43 morphotypes. After three years of 

study, one of the most outstanding facts is the great amount 

of Tomentella-like ectomycorrhizae found, which account 

for ca. 30% of the total mycorrhized root tips analysed. 

Apparently, the situation changes considerably when we 

consider the above-ground community. Few studies have 

paid attention to resupinate fungi when collecting 

ectomycorrhizal carpophores, thus inducing a mismatch 

between the species composition above- and below-ground. 

Nevertheless, a closer look at the ectomycorrhizal 

carpophores, considering especially the resupinate 

basidiomata, shows there is not such a big mismatch in our 

study site. Tomentelloid fungi seem to be well-adapted to 

survive in dry environments, such as the Mediterranean 

region in which our study site is located, thus occurring 

abundantly both as ectomycorrhizae and as carpophores. A 

preliminary catalogue is presented, including the list of 

tomentelloid fungi found both in the above- and belowground 

communities as well as descriptions and pictures of 

the ectomycorrhizal morphotypes and the basidiomata. 

955 - Biosorption of copper by Auricularia polytricha 

F. Di Mario * , E. Galli & P. Rapanà 

IBAF-CNR, Via Salaria Km.29,300 00016 Monterotondo 

Scalo (RM), Italy. - E-mail: francesca.dimario@mlib.cnr.it 

The biosorption of copper ions has been studied using 

lyophilised mycelium ofAuricularia polytricha (SMR 0055 

of International Bank of Edible Saprophytic Mushrooms of 

National Council of Research) in a batch system. This 

isolate was chosen for its ability of producing large 

amounts of biomass able to adsorbe metal ions. 

Experiments were performed in a pH range 2 - 6 at 

temperature within 15 - 35 °C, using different amounts of 

mycelium (100, 200, 300, 400 mg) in the presence of 1, 5, 



10, 30, 60 ppm of copper. Biosorption equilibrium was 

established in 5 minutes at pH 3 and 25 °C. The 

biosorption is pH-dependent but indipendent from 

temperature. The results comfirm the previous ascertained 

ability of A. polytricha to chelate heavy metals. The 

optimal condition for copper biosorption were determined 

at pH 3, when 100 mg of biomass were incubated with 30 

ppm of copper. Fungal biomass was also immobilised on a 

PVA matrix for further applications on heavy metal 

polluted streams. 

956 - The co-introduction of Australian 

ectomycorrhizal fungi with eucalypt plantations in the 

Mediterranean. Potential environmental risks 

J. Diez * , G. Moreno & J.L. Manjon 

Dpto. de Biología Vegetal, Universidad de Alcalá, E-28871 

Alcalá de Henares, Spain. - E-mail: jesus.diez@uah.es 

A growing evidence supports the recent dispersion of 

Australian ectomycorrhizal fungi with plantations of 

eucalypts and Australasian acacias worldwide. The effects 

of such introduction on the natural ecosystems have been 

poorly studied. In the Iberian Peninsula, such cointroduction 

seems to facilitate the adaptation of eucalypts 

to the soils of plantation sites. Consequently, the 

replacement of eucalypts for native sclerophyllous species 

is difficult and causes high costs in areas where eucalypts 

were extensively planted, such as the Natural Park of 

Monfragüe in Cáceres (Spain). In eucalypt plantations 

close to Monfragüe, we have found fruit-bodies of 

ectomycorrhizal fungi specific to Australasian trees, such 

as Hydnangium carneum, Hymenogaster albus, 

Hysterangium inflatum, Setchelliogaster rheophyllus and 

eucalypt-specific strains of the Pisolithus species complex. 

Other exotic ectomycorrhizal fungi, unable to fruit outside 

its natural geographic range, can have gone undetected 

belowground. Whether these exotic fungi extend beyong 

the eucalypt plantations will depend on their compatibility 

with native hosts and their ability to compete with native 

mycosymbionts; i.e. of the Australian species Laccaria 

fraterna (=L. lateritia), which was found in Mediterranean 

shrublands under ectomycorrhizal Cistus species. Further 

studies will be necessary, if we are to predict potential risks 

involved with the large-scale use of eucalypts in the 

Mediterranean. 

957 - Development of extraradical mycelia of 

ectomycorrhizal fungi and interaction with 

saprotrophic and root pathogenic fungi in soil 

microcosms 

D.P. Donnelly 1* , J.R. Leake 2 & L. Boddy 1 

1 Cardiff School of Biosciences, Cardiff University, PO Box 

915, Park Place, Cardiff, CF10 3TL, U.K. - 2 Department of 

Animal and Plant Sciences, Alfred Denny Building, 

University of Sheffield, Sheffield, S10 2TN, U.K. - E-mail: 

donnellyd@cardiff.ac.uk 

288 


Our recent studies of mycelial systems in soil microcosms 

using combined image analysis, radioisotope tracers and 

digital autoradiography, have provided new insights into 

their structure and function. Nutrient foraging and transport 

by mycorrhizal mycelia, together with the patterns of 

carbon allocation from plants to mycorrhizal mycelia have 

been visualised and quantified using 33P and 14C tracers. 

Fungal morphology (mycelial fanning and cord formation) 

and area cover of ectomycorrhizal mycelial systems of 

Paxillus involutus and Suillus bovinus varied between 

isolates and species over 27d. Encounter with pine litter 

patches significantly increased mycelial cover to 230% of 

controls. Extraradical mycelial morphology also influenced 

ingress by saprotrophic fungi. Dense mycelial systems of 

P. involutus resisted invasive cords of Phanerochaete 

velutina, but led to allocation of host-derived carbon away 

from the interaction zone. In contrast, the more aggregated 

mycelia of Suillus bovinus had less mycelial density, 

vigour and allocation of host derived carbon to the entire 

extraradical mycelium when over grown by P. velutina 

mycelia, compared to controls. We are currently 

investigating the mechanisms by which extraradical 

mycelia may also provide protection from migratory and 

non-migratory root pathogenic fungi. We hypothesise that 

differences in extraradical mycelial morphology and 

function will determine degree of protection of host roots 

from pathogen infections. 

958 - Amplified Fragment Length Microsatellites 

(AFLM): A simple method to develop microsatellite 

markers in fastidious microorganisms applied to AM 

fungi 

G.W. Douhan * & D.M. Rizzo 

University of California, Davis, Department of Plant 

Pathology, One Shields Avenue, Davis, CA 95616, U.S.A. - 

E-mail: gwdouhan@ucdavis.edu 

Microsatellites are abundant, highly mutable arrays of 

simple DNA sequence repeats which are powerful 

molecular markers to study population and evolutionary 

biology. Developing microsatellite markers for organisms 

that cannot, or are not, easily manipulated under laboratory 

conditions can be difficult because sequence information is 

needed. To overcome this problem in the arbuscular 

mycorrhizal (AM) fungi Glomus etunicatum and 

Gigaspora gigantea, global amplification of the genomes 

of each species were performed using linker-adaptor-PCR 

from single spores. Amplified fragments were enriched for 

microsatellite motifs using biotinylated oligonucleotides 

and recovered by magnetic streptavidin beads. The 

recovered fragments were reamplified, run on denaturing 

polyacrylamide gels, and sixteen selected bands were 

excised, cloned, and sequenced. Seven microsatellite 

motifs were detected from six clones (efficiency rate of 

43.8%). Primers were designed for all putative 

microsatellite loci and most were successfully amplified 

from three single spore preparations and from pools of 5, 

10, and 20 spores after global amplification. We term this 

technique amplified fragment length micosatellites and 

propose that they can be useful markers for fastidious 

microorganisms, such as Glomalean fungi. However, the


technique can also be used to isolate microsatellite loci in 

any organism. 

959 - Componets and techniques of spore sampling 

from determining presence to molecular biology 

M.G. Eversmeyer 1* , C.L. Kramer 2 & R.L. Bowden 1 

1 USDA-ARS, Throckmorton Hall, Manhattan, Kansas 

66506, U.S.A. - 2 Division of Biology, Ackert Hall, 

Manhattan, Kansas 66506, U.S.A. - E-mail: 

epidem@ksu.edu 

Spore samplers, from a simple petri dish or glass slide to 

complex volumetric samplers, have been used to determine 

the presence, concentration, and identification of airborne 

fungal spores since the beginning of mycological studies. 

The simplest of spore traps that collect spores by gravity 

deposition, have been used primarily for identification of 

spores present in the air. Volumetric samplers provide the 

added advantage of measuring the concentrations of fungal 

spores under existing wind conditions within a given air 

mass. Flow meters that provide variable adjustments in the 

rate of air intake of the sampler may be used to obtain isokinetic 

volumetric samplers. Small personal samplers have 

been developed for use by individuals with specific 

allergies to obtain levels of allergens that individuals may 

encounter on a daily basis. Techniques for identification of 

fungal spores collected by volumetric samplers using 

molecular markers are being developed. This presentation 

identifies various types of samplers that have been used in 

studies of identification, production, release, local dispersal 

and long distance transport, survival, and deposition of 

mycological material important in disease epidemics. 

Photos or sketches of many of the samplers used in 

research in our laboratory are shown including photos of 

the initial samplers used in private aircraft or on board 

ships. Major advantages and disadvantages of each type 

sampler are discussed. 

960 - Monitoring a non-pathogenic strain of Fusarium 

oxysporum on tomato plant using Green Fluorescence 

Protein gene marker 

J. Fatehi * , A. Lindberg & B. Gerhardson 

Plant Pathology & Biocontrol Unit, SLU, P.O. Box 7035, 

Uppsala 750 07, Sweden. - E-mail: 

jamshid.fatehi@vpat.slu.se 

Non-pathogenic strains of Fusarium are known organisms 

to effectively reduce the incidence of wilt diseases caused 

by the pathogenic isolates of this genus. Competition for 

nutrition in soil and rhizosphere, induced systemic 

resistance and competition for root colonization have been 

suggested as modes of action involved in such antagonistic 

effects. Here a non-pathogenic strain of Fusarium 

oxysporum, previously demonstrated to be an effective 

inhibitor of the tomato wilt pathogen caused by Fusarium 

oxysporum f.sp. lycopersici, was genetically transformed 

with both hygromycin B resistance and a gene for green 

fluorescence protein (GFP). Stable transformants with 

bright green fluorescence and resistant to hygromycin B 

were obtained. Several transformants with similar growth 

rate and colony morphology to the wild-type isolate were 

further used for the inoculation of tomato seedlings, in 

order to study their behavior on the host and the pattern of 

colonization of the roots. The growth of fungal mycelia on 

the root surface, penetration into the root cortex and 

colonization of this tissue were observed using a 

fluorescence microscopy. Re-isolation of the transfromants 

from surface sterilized root and stem tissues of inoculated 

tomato plants, 4 weeks after inoculation, indicated that the 

fungus was able to extensively colonize its host tissues. 

961 - Dynamics of microbial communities associated 

with submerged Typha angustifolia litter: effects of 

litter manipulations on microbial decay processes 

A.J. Foley * , M.D. Collins, R.K. Neely & K.A. Kuehn 

Department of Biology, Eastern Michigan University, 316 

Mark Jefferson, Ypsilanti, Michigan 48197, U.S.A. - Email: 

afoley@online.emich.edu 

In wetlands, most of the plant biomass enters the detrital 

pool to be decomposed by microorganisms. Prior studies of 

wetland plant decay have often used litter that has been 

prematurely harvested and/or manipulated (e.g, oven-dried) 

prior to submergence. We examined the effects of litter 

manipulations, such as oven drying and litter age 

(senescent vs. standing-dead), on microbial assemblages 

associated with decaying litter of Typha angustifolia. Plant 

litter of T. angustifolia was harvested after senescence and 

after a period of standing-dead decay. Litter of both age 

groups were air-dried and oven-dried (80 °C), placed into 

litter bags (1 mm mesh), and submerged in the wetland. 

Litter bags were retrieved periodically and analyzed for 

mass loss, microbial biomass, fungal production rates, and 

rates of microbial respiration. Several microbial parameters 

differed markedly between the litter treatments. Fungal 

biomass (ergosterol) associated with oven-dried litter was 

62% lower than air-dried litter during the first 114 days of 

decay (i.e., 83 ±53 and 220 ±43 µg ergosterol g -1 AFDM, 

respectively). Rates of fungal production and microbial 

respiration exhibited similar trends. Initial rates of 

microbial respiration showed a 5-fold difference between 

oven-dried and air-dried treatments (ca. 10 vs. 70 µg CO2- 

C g -1 AFDM h -1 , respectively). These results point to 

contrasting patterns in microbial dynamics during emergent 

macrophyte decay as a result of litter manipulations. 



962 - Community structure and diversity of below 

ground ectomycorrhiza fungi in Danish beech (Fagus 

sylvatica) forest with different cropping history 

K. Føns * & R. Kjøller 

Department of Mycology, Botanical Institute, University of 

Copenhagen, Øster Farimagsgade 2D, Dk-1353 

Copenhagen K, Denmark. - E-mail: kirstenf@bot.ku.dk 

The aim of this work is to compare the below ground 

community of ectomycorrhiza fungi in Danish beech forest 

sites with different cropping history. Six locations in Grib 

Skov, Northern Zealand, Denmark were studied. A 

monoculture of 100 years old beech trees are growing on 

all six locations, and the cropping histories of these 

locations are well described since early 1800s. At this time 

200 years old naturally regenerated beech were growing in 

the locations. This tells us that beech have been continuous 

at least for 400 years, and probably for many more 

hundreds years at the locations. At three of the locations 

the continuity of beech was broken from about 1830 to 

1900 with Norway Spruce (Picea abies). All the six 

locations were replanted with beech again in 1900. The 

different cropping histories give us a unique chance to 

study, whether broken continuity after 100 years can have 

an impact on the ectomycorrhizal community. Previously, 

the under story herbal communities at these same locations 

have been shown to differ between the continous and noncontinous 

plots (Flemming Rune, Danish Forest and 

Landscape Research Institute, unpublished data) In 2001, at 

late spring and early autumn soil cores were taken in the 

six plots. From the samples ectomycorrhiza fungi on the 

roots are sorted by a combination of morhotyping and ITS- 

RFLP and identified by sequencing. The first results on the 

ectomycorrhizal community structure at these sites will be 

presented in this poster. 

963 - Seasonal distribution of higher filamentous fungi 

in a subtropical estuary 

M. Fraser 1 , S. Schatz 2 , H. Laubach 3 , A. Rogerson 1 & C. 

Vogel 1* 

1 

Nova Southeastern University, College of Oceanography, 

8000 North Ocean Drive, Dania FL 33004, U.S.A. - 2 Nova 

Southeastern University, Colleges of Optometry and 

Oceanography, Davie FL 33314, U.S.A. - 

3 Nova 

Southeastern University, College of Medical Sciences, 

Davie Fl 33314, U.S.A. - E-mail: vogelc@nova.edu 

Little is known of the distribution of higher marine fungi 

along tropical and subtropical estuarine salinity gradients 

and how species composition may change seasonally. To 

examine these trends, five stations were set up along the 

New River Estuary in south Florida, USA. Salinity, 

dissolved O2 and pH were measured bimonthly. Substrates 

for collection were submerged panels of white oak 

(Quercus alba) and Douglas fir (Pseudotsugu menziessi). 

One pair per station was collected every three months, 

290 


incubated and observed for fungi. Twelve species were 

identified (4 Ascomycetes, 8 Deuteromycetes). The 

Ascomycetes Halosphaeria quadricornuta and Verruculina 

enalia were the dominant species by frequency of 

occurrence. Some species showed a physiological 

preference for higher salinity waters. The terrestrial species 

Alternaria sp., Aspergillus sp. and Penicillium sp. were 

isolated from the low salinity station and Trichocladium 

achrasporum was isolated only from the high salinity 

location. Despite no apparent patterns of seasonal 

distribution, successional patterns were discernible. 

Maximum species diversity occurred during the first three 

months which is consistent with previous findings. 

Compared to temperate studies species diversity was low 

suggesting interference behavior. Many species observed at 

low frequencies are cosmopolitan or temperate. Our 

observations indicate that salinity and temperature directly 

influence species composition and abundance in 

subtropical estuaries. 

964 - The impact of global warming on the mycorrhizal 

fungal community of a Canadian High Arctic site 

K.E. Fujimura * & K.N. Egger 

University of Northern British Columbia, 3333 University 

Way, Prince George, BC V2N 4Z9, Canada. - E-mail: 

fujimurk@unbc.ca 

The impact of global climate change is hypothesized to be 

more prevalent in arctic than temperate ecosystems. 

Experiments have shown that increase of temperature by 

the use of open-top chambers (OTCs) has impacted plant 

growth and phenology. However, data on the impact of 

global warming on the mycorrhizal fungal community are 

scant. The objective of this study is to examine the impact 

of increased air and soil temperature on the mycorrhizal 

fungal community. Three sites, each containing three plots, 

were chosen at Alexandra Fiord, Nunavut Canada. Air and 

soil temperatures were increased using 1-m diameter 

OTCs, which have been on the sites for at least five years. 

Plots within 2 m of the OTCs were used as ambient 

controls. Dominant plant species Cassiope tetragona, 

Dryas integrifolia, Salix arctica, and Saxifraga 

oppositifolia were harvested from warmed and control 

plots. Root tips were randomly selected for DNA extraction 

and morphological analysis. Mycorrhizal fungal 

communities were assessed with terminal restriction 

fragment length polymorphism (T-RFLP) analysis, a 

technique that has previously been applied primarily to 

bacterial community analysis. Representative fungal 

isolates were sequenced to determine phylogenetic 

affiliations. Comparisons between warmed and ambient 

plots will be presented.


965 - Mycorrhiza of blueberry and control of 

Verticillium wilt on strawberry: Two examples of 

rhizosphere microbial steady states in agricultural land 

use 

J. Golldack 1* , P. Schubert 2 , H. Schwärzel 2 , P. Lentzsch 1 & 

B. Münzenberger 1 

1 

Centre for Agricultural Landscape and Land Use 

Research, Eberswalder Str. 84, D-15374 Müncheberg, 

Germany. - 

2 

Landesamt für Verbraucherschutz und 

Landwirtschaft, Eberswalder Str. 84, D-15374 

Müncheberg, Germany. - E-mail: jgolldack@zalf.de 

The roots of blueberry are mycorrhizal, but on sandy 

agriculturally used soils, the mycorrhizal symbiosis has to 

be stabilized by a specific management of the mulch layer 

which provides organic C- and N-compounds. In long-term 

plot studies, a high sensitivity of this symbiosis depending 

on the mulch layer, particularly tree species of wood chips 

and time of application was found. Thus, the microbial 

population structure of the rhizosphere of variety 'Reka' 

varied considerably. In one plant rhizosphere, the microbial 

community was similar to the rhizosphere community of a 

natural peaty soil which was correlated with the highest 

fruit mass. This leads to the assumption that the 

mycorrhizal symbiosis of highbush blueberry depends on 

an appropriate fungus-bacteria-rhizosphere community 

which can be influenced by the management of the mulch 

layer. Strategies to achieve an appropriate microbial steady 

state can be used to control the Verticillium wilt on 

strawberry. The effect of different Verticillium strains and 

antagonistic bacteria is going to be analysed under 

controlled climatic conditions and in field trials. On 

different agriculturally used sites, Verticillium strains will 

be isolated to select the regionally best adapted 

antagonistic bacteria to control the Verticillium wilt. 

966 - Influences of T4-lysozyme producing potato plants 

on the endophytic fungi of the roots studied by a 

classical and a molecular approach 

M. Götz 1* , J. Lottmann 2 , G. Berg 2 , H.I. Nirenberg 3 & K. 

Smalla 1 

1 Federal Biological Research Centre for Agriculture and 


Biosafety, Messeweg 11/12, 38104 Braunschweig, 

Germany. - 2 University of Rostock, Gertrudenstraße 11a, 

18051 Rostock, Germany. - 3 Federal Biological Research 

Centre for Agriculture and Forestry, Institute for Plant 

Virology, Microbiology and Biosafety, Königin-Luise- 

Straße 19, 14195 Berlin, Germany. - E-mail: 

m.goetz@bba.de 

T4-lysozyme expressed in potato plants seems to be a 

promising strategy to enhance the plants' resistance against 

phytopathogenic bacteria like Erwinia carotovora. 

However, lysozyme has unspecific effects on other bacteria 

and on fungi. The monitoring of bacterial and fungal 

rhizosphere communities of transgenic and non-transgenic 

plants with a cultivation-independent molecular 

fingerprinting technique (PCR-DGGE) revealed in no cases 

a plant line dependent influence. The lysozyme is secreted 

into the apoplastic space where its concentration should be 

higher than in the rhizosphere. Therefore, the influence on 

endophytes which live inside of the apoplast could be 

stronger than that on the microorganisms in the 

rhizosphere. Endophytic fungi were isolated from roots of 

the parental and a transgenic line after surface sterilisation. 

The isolates were cultivated, identified and characterised 

by molecular methods. Here significant differences 

between the parental and the transgenic line could be 

found: the roots of the parental line were colonised with 

Verticillium dahliae around three times higher than those 

of the transgenic line. Additionally the surface sterilised 

roots were examined by PCR-DGGE. Fingerprints of the 

endophytic fungi could be shown for the first time. To 

associate the bands to fungal isolates clones were generated 

and sequenced. The results of the two methods of 

investigation are compared and discussed. 

967 - Substrate selectivity of corticioid fungi from 

Nothofagus forests in Patagonia (Argentina) 

A.G. Greslebin 


Patagónico, CC 14, 9200, Esquel, Chubut, Argentina. - Email: 

alina@ciefap.cyt.edu.ar 

Corticioid fungi (Aphyllophorales, Basidiomycetes) grow 

on almost every type of woody substrate, but species 

usually show preferences on certain features of the 

substrate such as the decay stage and/or the part of the tree 

involved. These preferences determine the spatial and 

temporal arrangement of these organisms along the 

degradation process. During an intensive floristic survey of 

Corticiaceae growing on 3 Nothofagus spp. in the 

Patagonian Andes forests, the habitat preferences and the 

degree of selectivity of 42 species in relation to three 

substrate features (decay stage, part of the tree and 

diameter of the detritus) were evaluated. Decay stage was 

classified into six categories according to hardness and 

other features. Part of the tree was classified as bark, 

sapwood, hardwood or the combination of bark/sapwood or 

sapwood/hardwood. Diameter was classified as 0-5, 6-10, 

11-20, 21-30 or >30 cm (from small branches to big 

trunks). The differential distribution (not aleatory) of the 

species according to the characteristics of the substrate was 

statistically tested (X 2 ). 95% of the species showed a 

differential distribution in relation to the decay stage, 64% 

in relation to the part of the tree and 54% in relation to the 

diameter. In order to evaluate the degree of selectivity, the 

range of categories where a species was recorded for each 

variable was analyzed. Species with a range ≤ 3 were 

considered highly selective for that variable. 



968 - Adaptive spatiotemporal distribution of soil 

microfungi in 'Evolution Canyon' II, Lower Nahal 

Keziv, Western Upper Galilee, Israel 

I. Grishkan * , E. Nevo, S.P. Wasser & A. Beharav 

Institute of Evolution, University of Haifa, Mount Carmel, 

Haifa 31905, Israel. - E-mail: 

grishkan@research.haifa.ac.il 

We describe and interpret spatiotemporal micromycete 

community structure and adaptive complexes to xeric and 

mesic contrasting microclimates in the soils of 'Evolution 

Canyon' II, western Upper Galilee, Israel. A total of 192 

species belonging to Zygomycota (9 species), Ascomycota 

(13 species), and mitosporic fungi (170 species) were 

isolated. The fungal communities on the xeric south-facing 

slope (SFS) demonstrated significantly greater diversity 

(species richness, Shannon index, and evenness) than on 

the mesic north-facing slope (NFS). Seasonally, winter 

slope communities were less heterogeneous. Forest 

localities on the NFS and in all seasons and the shady 

locality on the SFS in the winter were overwhelmingly 

dominated by mesophilic Penicillium species. The sunny 

locality on the SFS was characterized by a dominance of 

melanin-containing fungi that was most pronounced in the 

summer and by high occurrence and abundance of 

thermotolerant Aspergillus and Fusarium species. 

Ascomycetes and zygomycetes were the minor components 

in all local mycobiota studied; sexual ascomycetes, being 

stress-selected fungi, were more abundant than 10 fold in 

the soil of the SFS, with the peak of abundance in the 

sunny summer community. The results demonstrated a 

microscale adaptive spatiotemporal inter- and intraslope 

divergence in soil mycobiota structure. Microclimatic 

natural selection appears to be the major factor affecting 

soil fungi diversity patterns and spatiotemporal dynamics. 

969 - Structural diversity of fungal populations in soil 

under the influence of different farming management 

systems 

A. Hagn * , K. Pritsch & M. Schloter 

GSF-Research Centre for Environment and Health, Insitute 

of Soil Ecology, Ingolstädter Landstraße 1, 85758 

Munich/Neuherberg, Germany. - E-mail: 

alexandra.hagn@gsf.de 

Although it is known that fungi play a vital role for 

ecosystems, only few studies were published so far 

showing fungal diversity and dynamics in soil. Mainly in 

agricultural ecosystems fungi are essential for the soil 

quality, as many species are able to degrade complex 

organic compounds, supply plants with nutrients and serve 

as biocontrol organisms against plant pathogens. However 

some strains can also act as plant pathogens. Therefore we 

investigated the influence of precision farming (a new 

agricultural technique, focusing on the heterogeneity of a 

field site and adapting the amount of fertilizer given to the 

292 


yield expected in a particular plot). For comparison 

samples were also taken from plots, where fertilizers were 

applied using conventional techniques. To access the 

different fungal populations culture-dependent and cultureindependent 

methods were used. Fungal cultures were 

isolated on three different media and examined by means 

of microscopy and a genetic fingerprinting method 

(interLINE, PAGE). Additionally DNA was directly 

extracted from the soil samples and clone libraries of the 

18S rDNA were established. Yet the data of the culturedependent 

and the culture-independent methos show a 

seasonal influence as well as an influence of the high- and 

low-yield areas. Besides also a direct influence of the 

agricultural management systems on the structure of the 

fungal populations was detected. 

970 - Microbial companions of an introduced tree: 

needle endophytes of Siberian larch (Larix sibirica) in 

Finland 

J. Hantula 1* , M. Kauhanen 1 , E.J. Vainio 1 & P. Niemelä 2 

1 Finnish Forest Research Institute, PO Box 18, 01301 

Vantaa, Finland. - 2 University of Joensuu, Faculty of 

Forestry, PO Box 111, 80101 Joensuu, Finland. - E-mail: 

jarkko.hantula@metla.fi 

Endophytes are organisms inhabiting plant tissues without 

causing symptoms. The distinction between pathogens and 

endophytes is not clear as some endophytes are pathogenic 

for other plants. Some pathogens also show periods of 

latency. In single hosts pathogenicity or endophytic 

behavior may be determined by only one gene. Plants are 

frequently introduced to novel geographic areas in the 

modern world. It is not known how commonly endophytes 

are co-introduced with their plant hosts. This information 

should, however, be available as nonindigeneous plants 

may transport fungal endophytes pathogenic to native 

plants. We analysed needle endophytes of Siberian larch in 

its natural distribution area in Russia and in old or young 

stands in Finland, where the tree does not occur naturally. 

880 isolates were collected, and identified to species level 

based on colony morphology and DNA-analyses. The cointroduction 

frequency of endophytes turned out to be high; 

only one common (in Russia) endophyte was missing from 

one Finnish stand. In contrast, several rather common 

endophytes observed in Finland lacked from the natural 

stand in Russia. We also analysed genetic variation in one 

endophyte species. 

971 - Interspecific fungal interactions and recognition 

in decayed wood 

J. Heilmann-Clausen 1* & L. Boddy 2 

1 Danish Forest and Landscape Research Institute, 

Hørsholm Kongevej 11, 2970 Hørsholm, Denmark. - 

2 Cardiff University, School of Biosciences, Museum 

Avenue, PO Box 915, Cardiff CF10 3TL, Wales, U.K. - Email: 

jhc@kvl.dk


We compared interspecific interactions in fresh beech 

wood and beech wood partly decayed by the primary decay 

agents, Eutypa spinosa, Fomes fomentarius, Stereum 

hirsutum and Trametes versicolor. The fungi used in the 

experiments included five secondary decay agents, 

common on beech wood in advanced decay, as well as the 

aforementioned primary decayers. The experiments were 

carried out using sterilised, recolonised wood and lasted for 

seven months. Interactions involving secondary decayers 

were affected by wood type, with outcomes changing from 

complete replacement to deadlock in at least three species 

combinations. Most of the secondary decayers were able to 

replace some of the primary decayers. However, several of 

the secondary decayers were unable to colonise sterilised 

wood decayed by Trametes versicolor or Stereum 

hirstutum. This lead to a new series of experiments in 

which the effects of exudates from fresh and partly decayed 

wood were investigated with respect to hyphal extension 

rates in a larger range of secondary decay fungi. Sterilised 

wood pieces were placed on 0,5% malt agar, opposite to 

small plugs containing the test fungi, which showed very 

variable growth responses to the various wood types. Most 

species increased hyphal extension rates as response to 

wood decayed by Eutypa spinosa and Fomes fomentarius, 

while wood decayed by Stereum hirsutum resulted in 

reduced or completely lacking growth in a majority of 

species. 

972 - Physiological characteristics of symbiotic fungi 

associated with the seed germination of Gastrodia elata 

I.P. Hong 1* , S.X. Guo 2 , T.S. Lee 3 & M.W. Lee 4 

1 Division of Seri-Culture and Api-culture, NIAST, Suwon, 

441-707, Korea. - 2 Institute of Medicinal Plant, Chinese 

Academy of Medical Sciences, P. R., China. - 3 Department 

of Biology, Incheon University, 177, Dohwa-dong, Nam- 

Ku, 402-749, Korea. - 4 Department of Biology, Dongguk 

University, 26, Pil-Dong 3ga, Chunggu, Seoul, 100-715, 

Korea. 

This study was carried out to investigate the possibility for 

seeds germination of Gastrodia elata using symbiotic 

fungi. Since seeds of G. elata are very small and lack an 

endosperm and other nutrients, their germination is 

difficult without requirement for external nutrients. Out of 

twenty six isolates collected from protocorms of G. elata 

and roots of native orchids inhabited in wild, two strains 

(H-2 and H-21) were observed to stimulate the seed 

germination of G. elata. The seed germination of G. elata 

was excellent on oak tree leaves medium. The optimal 

conditions for mycelial growth of symbiotic fungi were 25 

°C and pH 6.0, respectively. The mycelial growth of H-2 

strain was excellent on YMA medium, while h-21 was 

poor on PDA medium. In case of carbon sources, the 

mycelial growth of H-2 and H-21 was good on media 

supplemented with glucose and dextrin, respectively. 

Calcium nitrate was good for mycelial growth of H-2 strain 

as a nitrogen sources, whereas urea was effective to H-21 

strain. 

973 - Classification of wood decay fungi isolated from 

durable wood species, ekki 

S. Horisawa 1* , R. Ito 2 , Y. Sakuma 3 & S. Doi 1 

1 Institution of Wood Technology, Akita Prefectural 

University, 11-1 Kaieizaka, Noshiro, Akita 016-0876, 

Japan. - 2 Akita Wood Technology Transfer Foundation, 11- 

1 Kaieizaka, Noshiro, Akita 016-0876, Japan. - 3 Biological 

Resources Division, Japan International Research Center 

for Agricultural Sciences (JIRCAS), 1-1 Ohwashi, Tsukuba, 

Ibaraki 305-0074, Japan. - E-mail: horisawa@iwt.akitapu.ac.jp 

Ekki (Azobe, Bongossi; Lophira alata Banks), a native 

African wood species, is known as a large durable species 

and has been utilized without preservative treatment for 

outdoor architectures such as bridges in EU. In Japan, 

however, several bridges of ekki wood are decayed in a 

short time after construction. In this study, as the first step 

of the elucidation of decay in ekki wood, basidiomycetes 

isolated from damaged members of ekki bridges were 

classified by PCR-RFLP. Decayed wood pieces and fruit 

bodies from ekki bridges were incubated on potatodextrose-agar 

plates containing antibiotics. Twenty-six 

isolates of basidiomycetes were obtained from different 

areas in Japan. One of the isolates was identified 

morphologically as Perenniporia tephropora (Mont.) Ryv. 

(Loweporus tephroporus). Genomic DNAs were extracted 

from isolates using the benzyl chloride method. The whole 

inter specific transcribed (ITS) regions (ITS1, ITS2, and 

5.8S) were amplified by PCR. The PCR products were 

digested with 4 restriction enzymes to obtain RFLPs. 

Restricted fragments were separated by electrophoresis in 

8% of polyacrylamide gel. Comparing fragment patterns, 

14 isolates were identified as P. tephropora. To obtain 

further molecular information on the decay basidiomycetes, 

we undertake to determine sequences of 18S rDNAs. 

974 - Aspergillus section Flavi in the United States: 

characterization of soil populations from agricultural 

fields 

B.W. Horn 

National Peanut Research Laboratory, United States 

Department of Agriculture, Agricultural Research Service, 

P.O. Box 509, Dawson, Georgia 31742-0509, U.S.A. 

Species belonging to Aspergillus section Flavi often 

produce aflatoxins and cyclopiazonic acid, mycotoxins that 

contaminate preharvest peanuts, corn and cottonseed. Soil 

populations of A. flavus, A. parasiticus, A. nomius, A. 

tamarii and A. caelatus were examined over a large 

geographic area within the United States and in greater 

detail within a single field. A transect was established from 

New Mexico to Virginia, and species from section Flavi 

showed distinct patterns in their geographic distribution. A. 

flavus (large sclerotial morphotype or L strain) was the 

most widely distributed species, but A. flavus var. 



parvisclerotigenus (small sclerotial morphotype or S strain) 

was primarily confined to eastern Texas and Louisiana 

where cotton is intensively grown. A. parasiticus was most 

common along the southern Atlantic coast, particularly in 

areas where peanuts are cultivated. A. tamarii and A. 

caelatus occurred at low incidences along most of the 

transect, and A. nomius was detected only in the 

Mississippi River Delta region (Louisiana and Mississippi). 

Individual vegetative compatibility groups of A. flavus and 

A. parasiticus were widely distributed along the transect. In 

a single peanut field from southwestern Georgia included 

in the transect, soil populations of A. flavus and A. 

parasiticus were genetically diverse, as indicated by the 

large number of vegetative compatibility groups. 

975 - Isolated Pinus contorta seedlings in coastal dunes 

are supported primarily by fungi with high host 

specificity whose spores are carried in deer feces 

T.R. Horton * , S. Ashkannejhad & C. Hazard 

SUNY-ESF, Syracuse, NY 13210, U.S.A. - E-mail: 

trhorton@esf.edu 

Pinus contorta seedlings are establishing in sand dunes on 

the coast of Oregon, U.S.A. in a primary succession 

pattern. The seedlings establish at least 100 m away from 

the continuous forest and thus establish without the benefit 

of an existing ectomycorrhizal network. Over 120 species 

of fungi in 37 genera have been recorded fruiting along the 

edges of the continuous forests and spores from these fungi 

may disperse to the isolated seedlings via wind, water or 

mammals. We identified EM fungi associated with the 

roots of the isolated seedlings and found that they are 

colonized almost exclusively by `suilloid' fungi: Suillus 

brevipes, S. umbonatus, S. tomentosus, Rhizopogon 

fuscorubens, R. occidentalis, and R. vulgaris. Deer 

commonly traverse the dune system from the surrounding 

forest past the isolated seedling areas and their fecal pellets 

contain large amounts of spores. Spore slurries made by 

bringing up individual fecal piles with variable pellet 

numbers in 100 ml of water and were found to contain 

between 1 x 10 6 and 1 x 10 8 spores/ml. In a bioassay, 

seedlings inoculated with 20 ml of slurries made from fresh 

fecal piles became colonized by the same fungi observed 

on the field-collected seedlings. These results suggest that 

1) deer can vector suilloid spores to areas where 

mycorrhizal networks are non-existent, 2) suilloid spores 

may lie dormant in soils until pine seedlings germinate, and 

3) in contrast to other early succession work, pine seedlings 

establish and are supported by fungi with high host 

specificity. 

294 


976 - Effect of root endophytes on the germination of 

seed of the Australian terrestrial orchid Caladenia 

formosa G. W. Carr 

T.T. Huynh 1 , C.B. McLean 1* & A.C. Lawrie 2 

1 

University of Melbourne, Institute of Land and Food 



Australia. - 

2 RMIT University, Department of 




This study investigated two seed germination methods to 

determine which would maximise the production of plants 

of the endangered Australian orchid Caladenia formosa for 

use in re-establishment in situ. Root endophytes were 

isolated from adult plants in the wild at various stages in 

the orchid life cycle (budding, leafing, flowering, capsule 

production and senescence). Seed was germinated on 

minimal (Oatmeal Agar) and complex (PA5 containing 

coconut water) media with and without root endophytes 

under axenic conditions. Germination and subsequent 

growth was recorded at monthly intervals for a period of 

twelve months. Mycorrhizal status of the seedlings was 

determined by SEM. Seed grown on minimal media 

inoculated with mycorrhizal fungi from the leafing, 

budding and flowering stages gave fastest (within one 

month) and maximum (>50%) germination rates. Seed 

grown on the complex media did not germinate in the first 

month. Symbiotic seedlings grown on the minimal medium 

were defasked and survived to produce tubers under 

glasshouse conditions. 

977 - Effects of in vivo sub-culturing on changes in 

morphology and virulence of the entomogenous fungus 

Metarhizium anisopliae 

L. Ibrahim 1* , T.M. Butt 2 & P. Jenkinson 1 

1 Harper Adams University College, Newport, Shropshire, 

U.K. - 2 University of Wales Swansea, Singleton Park, 

Swansea, U.K. - E-mail: librahim@harper-adams.ac.uk 

One of the major drawbacks with fungal biocontrol agents 

is that they can lose virulence if maintained on artificial 

culture media. This attenuation of virulence has been 

reported by several workers and is a major concern in the 

development of insect-pathogenic fungi for pest control. 

Understanding the factors responsible for this phenomenon 

will contribute important information for development of 

media to stabilise or increase virulence. The effects on 

pathogenicity of Metarhizium anisopliae (Metsch.) Sor. 

isolate V208 following one passage through the 

heterologous insect-host Myzus persicae and 11 in vitro 

successive subcultures were investigated. Attenuation of 

virulence as measured by LT 50 (median lethal time) was 

manifested in rapid decline with recovery and slight 

decline after recovery. Similarly, sporulation of attenuated


subcultures was poor with some subcultures ultimately 

becoming sterile. Significant decline in germination was 

observed in subculture 3 with slow recovery by subculture 

11. Most studies to date report a gradual or rapid decline in 

virulence with recovery only being achieved by passaging 

through an insect. Here recovery was shown in the absence 

of passaging. Bioassays alone do not reveal the full 

variation in expression of attenuation. In an attempt to 

identify more overt attributes of attenuation, particular 

attention was focused on the physiological and biochemical 

factors, which may account for the attenuation observed. 

978 - Detection of double-stranded RNA from 

Helicobasidium mompa and Rosellinia necatrix 

K. Ikeda 1* , H. Nakamura 1 , M. Arakawa 2 & N. Matsumoto 1 

1 Microbial Ecology Unit, National Institute for Agro- 

Environmental Sciences, 3-1-3 Kan-non dai, Tsukuba, 

Japan. - 2 Faculty of Agriculture, Meijo University, 

Shiogamaguchi, Tenpaku, Nagoya, Japan. - E-mail: 

ikeken@affrc.go.jp 

Helicobasidium mompa and R. necatrix cause root diseases 

of many woody plants including fruit trees. Diseased plants 

show stunting of shoots and yellowing of leaves but are 

seldom killed unless intensive cultural practices are used. 

As the first step to exploit double-stranded (ds) RNA for 

biological control of the diseases, fungal strains were 

examined for the presence of dsRNA. In H. mompa, 72.1% 

(158/219) of strains had dsRNA whereas 24.4% (66/270) in 

R. necatrix. Electrophoresis revealed the presence of 

diverse dsRNAs in both fungi with 1 to 12 bands, ranging 

from ca. 2 to 15 kbp. Hyphal tip isolation was mostly 

unsuccessful to remove dsRNAs except that found in R. 

necatrix with 12 dsRNA segments, which was considered 

to be a member of the Reovididae. Single spore isolates of 

H. mompa from basidiospores were all dsRNA-free. Single 

basidiospore isolates were not pathogenic. Single ascospore 

isolates of R. necatrix did not contain dsRNA and were as 

virulent as those from vegetative hyphae. Sexual 

reproduction functions to remove dsRNA in both fungi. In 

R. necatrix, ascospore offsprings are considered capable of 

survival in nature since they are pathogenic. This is, 

however, not the case with H. mompa; single basidiospore 

isolates may not survive in nature. The difference in the 

detection frequency of dsRNA between H. mompa and R. 

necatrix is, at least partly, ascribed to the difference in the 

efficacy of sexual reproduction to establish dsRNA-free 

offsprings in the field. 

979 - Ecological characteristics and contemporized 

dynamics of ectomycorrhizal ammonia fungi in warm 

temperate forests in Japan 

A. Imamura 1* & Y. Yumoto 2 

1 Graduate School of Human & Environmental Studies, 

Kyoto Univ., Yoshida-nihonmatsu-cho, Sakyo-ku, Kyoto, 

Japan. - 2 Center for Ecological Research, Kyoto Univ., 

Kamitanokami-hiranocho, Otsu, Japan. - E-mail: 

l50580@sakura.kudpc.kyoto-u.ac.jp 

We studied successional speed and dry weight of 

fruitbodies to clarify the correspondence between the 

successional phases of ammonia fungi: the early phase 

(EP) and the late phase (LP) and nutritional types: 

saprophytic and ectomycorrhizal (ECM). Urea treatments 

were conducted to induce fruiting of them in two 

ectomycorrhizal forests in warm temperate Japan. Ten EP 

and six LP species developed their fruitbodies. LP species 

fruited more in biomass than EP, and each fruitbody of LP 

species was larger than that of EP. ANOVA proved that 

fruiting of EP species was affected by treatment season or 

season-vegetation, that of LP by season-vegetation or 

vegetation. EP had short and one fruiting phase, while LP 

had long and multiple phases. Those ecological 

characteristics of EP and LP are corresponding to 

saprophytic and ECM species, respectively. Consequently, 

we measured the contemporized dynamics between ECM 

formation and fruitbody development. We found 1) the 

damaging effect of ammonia released from urea on ECM; 

2) the ECM of two ammonia fungi, Alnicola lactariolens 

and Hebeloma vinosophyllum, by PCR-RFLP analysis; 3) 

the increase and subsequent decrease of the ECM of the 

two fungi preceded the processes made by their fruitbodies; 

and 4) the existence of ECM fungi which do not develop 

fruitbodies in urea-treated soil, which resulted in the 

extension of the definition of ammonia fungi from referring 

solely to fruiting fungi to including proliferating and nonfruiting 

ones. 

980 - Evaluation of the accessibility of ribosomal RNA 

regions to fluorescently labelled probes for in situ yeast 

identification 

J. Inácio 1* , A. Fonseca 1 , R. Amann 2 & I. Spencer-Martins 1 

1 CREM, SABT, Universidade Nova de Lisboa, Quinta da 

Torre, 2829-516 CAPARICA, Portugal. - 2 Max Planck 

Institute for Marine Microbiology, Celsiusstrabe 1, 28359 

BREMEN, Germany. - E-mail: jjis@mail.fct.unl.pt 

Fluorescence in situ hybridisation (FISH) with probes 

targeted at the ribosomal RNA has been successfully 

applied to the rapid and specific identification of 

microorganisms in their natural habitats. However, the 

target site may not be easily reachable by the probe due to 

structural constraints, leading to weak fluorescence 

intensities even when using pure cultures under appropriate 

assay conditions. To circumvent this problem and optimise 

probe design, it is advisable to determine the accessibility 

of the different rRNA regions to FISH. In this work we 

studied the accessibility of the D1/D2 domains of the 26S 

rRNA of Saccharomyces cerevisiae, with a set of ca. 30 

Cy3-labelled oligonucleotide probes covering the full 

length of this region, using flow cytometry. To evaluate 

whether the information obtained for S. cerevisiae could be 

extrapolated to other yeast species, a subset of probes 

belonging to different fluorescence intensity classes were 

hybridised to selected yeasts that had a full complementary 

target site for those probes. The results indicated that the S. 

cerevisiae rRNA accessibility model provides a useful 



guidance tool for the design of species-specific yeast 

probes. To test this hypothesis and illustrate target site 

selection, we designed a DNA probe that specifically binds 

to the rRNA of basidiomycetous fungi, including yeasts, 

but not to ascomycetes. 

981 - Habitat diversity in the Aphyllophorales of 

tropical forests, Albertine Rift, western Uganda 

P. Ipulet 

Makerere University, Institute of Environment and Natural 

Resources, P.O. Box .., Kampala, Uganda. - E-mail: 

ipulet@yahoo.com 

The fungiflora of East Africa is extremely rich in a range of 

genera and the number of species though large areas 

remain totally unexplored (Pegler, 1977). Results on 

habitat diversity of the Aphyllophorales from an ongoing 

Ph. D. research will be presented. The taxonomy follows 

Alexopoulos & Mims (1996), and, Ryvarden & Johansen 

(1980). The study area, the Albertine Rift, with regard to 

non-fungal research,high habitat/species diversity with 

many endemic taxa. Its northern section lies within the 

postulated movement of forest species eastwards from a 

major refugium in eastern Democratic Republic of Congo, 

and, Cameroon/Gabon in western Africa during the 

pleistocene period. The fungi are systematically collected 

from different forest types taking seasonality, 

substrate/host and altitude into account. 

982 - Influence of vegetation types on the distribution of 

tropical forest poroid fungi, Uganda. (Preliminary 

results) 

P. Ipulet 

Institute of Environment, Makerere University, PO Box 

7298, Kampala, Uganda. - E-mail: ipulet@yahoo.com 

Kibale forest covers an area of 560 km 2 with an altitudinal 

range 1,110 - 1590 m. Its annual rainfall is 1,100 - 1600 

mm, a tropical climate with rainfall peaks in March - May 

and September - November. The vegetation can be 

classified as Medium Altitude Evergreen forest, Medium 

Altitude Semi-deciduous forest, Mixed Forest 

Communities, Grassland and Swamp. Though there are 85 

species of polypores known from Uganda, systematic 

collecting has not been done and most of the country 

remains unexplored. In Kibale forest, three forest sites with 

distinct vegetation types, are at different altitudes and have 

different annual rainfall totals, are systematically sampled 

in the last month of each rainy season. Ecological studies 

conducted in temperate forests have indicated no 

differences in fungi species distribution within a forest. 

However due to high diversity of both habitats and species 

in tropical forests, the results could be different. Data 

collection and determination of collected material are going 

on. 

296 


983 - Intraspecific diversity of Thysanophora 

penicillioides and genetic structure in the species 

population 

S. Iwamoto 1* , S. Tokumasu 1 , Y. Ogawa 2 , Y. Suyama 3 & 

M. Kakishima 4 

1 Sugadaira Montane Research Center, University of 

Tsukuba, Sugadaira, Sanada, Chiisagata, Nagano 386- 

2201, Japan. - 2 College of Pharmacy, Nihon University, 7- 

7-1 Narashinodai, Funabashi, Chiba 274-8555, Japan. - 

3 Graduate School of Agricultural Science, Tohoku 

University, Kawatabi, Naruko, Miyagi 989-6711, Japan. - 

4 Institute of Agriculture and Forestry, University of 

Tsukuba, Tsukuba, Ibaraki 305-8572, Japan. - E-mail: 

susumu.iwamoto@nifty.ne.jp 

We investigated the intraspecific variation of 

Thysanophora penicillioides, a widely distributed 

saprophytic hyphomycete analyzing the sequence data of 

rDNA ITS regions to address a possibility of geographic 

isolation between regional or local populations. We 

collected 325 isolates from decaying Abies needles in 20 

local populations in a wide range of Japan. From regional 

populations in Europe and North America, 30 and 5 strains 

isolated were also collected, respectively. As a result of 

analysis of the sequence for all isolates, base changes or 

indels were found in 30 sites in ITS regions, and 18 

haplotypes distributed in seven lineages were detected. In 

Japan, five lineages were detected and major lineages were 

found widely in local populations. One major lineage 

which was not detected from Europe and North America 

dominated in most Japanese local populations and 

accounted for about 65% of all Japanese isolates. From 

European population two lineages were detected and also 

two lineage from North American population. Only one 

lineage was common between Japan and Europe and no 

lineage between Japan and North America, while one 

lineage was common between Europe and North America. 

These results suggest that the genetic structure of Japanese 

population is different from European and North American 

populations. Probably, the distribution of individual lineage 

found in this study is limited to a relatively area, though the 

fungus produces small, dry, wind-dispersed conidia. 

984 - Belowground ectomycorrhizal community 

structure along a local nutrient gradient in a boreal 

forest in Northern Sweden 

J.F. Johansson * & A.F.S. Taylor 


University of Agricultural Sciences (SLU), Uppsala, 

Sweden. - E-mail: Jonas.Johansson@mykopat.slu.se 

The belowground ectomycorrhizal (ECM) community 

structure was investigated along a local nutrient gradient in 

a boreal forest located in Betsele in Northern Sweden. The 

ECM taxa colonising root tips in the O horizon were 

identified using morphological characterisation and PCR-


RFLP of the ITS region of the fungal rDNA. The ECM 

community changed along the investigated transect. A 

Russula sp., Tylospora spp., a Tomentella sp., and 

Cenococcum geophilum were the most commonly found 

ECM taxa along the transect. Species not producing 

conspicuous sporocarps constituted a high proportion of the 

belowground ECM community. Tylospora asterophora, 

Amphinema byssoides and all taxa identified as Cortinarius 

spp. were associated with the nutrient-rich end of the 

transect, whereas the Russula sp. and Piloderma spp. were 

associated with the nutrient-poor section of the transect. 

Nitrogen availability, pH and supply of base cations 

appeared to be the most important soil factors controlling 

the ECM community structure in the O horizon of the 

Betsele site. 

985 - The impact of ash fertilization on the biomass of 

roots and ectomycorrhiza of coniferous trees 

H. Jokinen 1* , R. Strömmer 1 , A. Holma 1 & H.M. Potila 2 

1 University of Helsinki, Department of Ecological and 

Environmental Sciences, Niemenkatu 73 FIN 15140 Lahti, 

Finland. - 2 Forest Research Institute, Parkano Research 

Station, FIN 39700 Parkano, Finland. - E-mail: 

hanna.k.jokinen@helsinki.fi 

Wood ash from the forest industry constitutes a 

considerable waste problem, and recycling it back to the 

forest is of great interest. Ash could be seen as a valuable 

resource since it contains most of the elements needed for 

plant nutrition and growth. The environmental issue 

concerning ash is that it stimulates nitrogen mineralization, 

increases soil pH and also contains heavy metals. Several 

aspects concerning ash fertilization have been studied for 

these reasons, especially the processes in forest soil, like 

microbial activities, nutrient mineralization and responses 

of various soil organisms. We studied the biomasses of the 

roots and ectomycorrhiza in a dry, nutrient poor Scots pine 

(Pinus sylvestris) forest in Muhos, northern Finland before 

and after fertilization with wood ash. The results after the 

first season were unexpected, as both these biomasses 

slightly increased due to strong ash application, 9000 kg 

ha -1 . For getting more thorough picture and long term view, 

the sampling repeated three years later. Another field 

experiment started in Evo, in Southern Finland in a more 

nutrient rich Norway spruce (Picea abies) forest. We also 

established a microcosm experiment for estimating the 

capacity of mycorrhizal root tips and extramatrical 

mycelium in storing N and thus alleviating the high spatial 

and temporal variation in nitrogen mineralization in forest 

soil after ash fertilization. 

986 - Effect of grassland plant species on soil fungal 

communities: a microcosm study 

N. Kennedy * , E. Brodie & N. Clipson 

Dept. Ind. Microbiology, UCD, Belfield, Dublin 4, Ireland. 

- E-mail: nabla.kennedy@ucd.ie 

Grasslands are the main vegetational class in Ireland and 

are of major significance in agriculture. Although 

intensification has been central to increased productivity in 

Ireland, there is concern globally and within the EU about 

the sustainability of such practices and a need to 

understand at a basic level how grasslands function. It is 

known that fungi play a major role in grassland ecosystem 

processes such as carbon, nitrogen and other nutrient 

cycles. In this study, microcosms were planted with seven 

grassland species representing the most commonly found 

plants in both unimproved, species-rich grasslands, and in 

improved, fertilised grasslands, which are typically 

dominated by only a few plant species. After harvest, shifts 

in the fungal community structure of the rhizosphere soil of 

each plant species were assessed. The ergosterol content of 

each soil sample was measured in order to get an indication 

of fungal biomass. In addition, samples were analysed 

using TRFLP (terminal restriction fragment length 

polymorphism). The resulting profiles, which indicated the 

number of fungal species per sample, were compared 

between plant species. In general, fungal communities were 

not significantly altered by plant species. The similarity of 

fungal profiles was even more striking when compared to 

bacterial profiles for the same soils, which differed 

significantly according to plant species type. 

987 - Dynamics of VA mycorrhizal fungi in burned 

Japanese red pine forests in the Setouchi Destrict in 

Western Japan 

S. Kinugasa 1 , M. Fujiyoshi 1 , T. Satomura 1 & T. 

Horikoshi 2* 

1 Graduate School of Biosphere Sciences, Hiroshima 

University, Higashi-hiroshima 739-8521, Japan. - 2 Faculty 

of Integrated Arts and Sciences, Hiroshima University, 

Higashi-hiroshima 739-8521, Japan. - E-mail: 

takao@hiroshima-u.ac.jp 

Many large forest fires have occurred in the Japanese red 

pine forests of the Setouchi District in Western Japan. We 

investigated the seasonal changes of infection rate, 

numbers of spores and sporangiocarps, and composition of 

spore types of VA mycorrhizal fungi at the burned and the 

vegetation naturally recovered sites (BNR sites,7 and 22 

yrs after the fires) and the burned and planted site (BP site, 

7 yrs after). 1. Infection rate is about 90% irrespective of 

the sampling months at the BNR-22 yrs site. Infection rates 

at the BNR-7 and BP-7 yrs sites showed the similar trends, 

70-90% in July-September, and then gradually decreased to 

50-60% in January. 2. Number of spores at the BNR-22 yrs 

site is always higher (400-900/10 g dry soil) than those at 



the BNR-7 and BP-7 yrs sites, and rapidly increased from 

November to January. At the BNR-7 yrs site the number 

gradually decreased from 400 in July to 200 in January. At 

the BP-7 yrs site, on the other hand, the number gradually 

increased from below 100 in July to 200/10 g dry soil in 

January. 3. Number of sporangiocarps at the BNR-22 yrs 

site was always higher than those at the BNR-7 and BP-7 

yrs sites, in the range of 70-110/10 g dry soil, and showed 

the higher level in January. At the BNR-7 and BP-7 yrs 

sites the number is always very low (below 10/10g dry 

soil). 4. At the BNR-7 and BP-7 yrs sites Gigaspora and 

Scutellospora types, at the BNR-22 yrs site Glomus and 

Sclerocystis types were typically observed. 

988 - Seasonal changes in an ectomycorrhizal 

community associated with beech (Fagus sylvatica) 

R. Kjøller 

Department of Mycology, Botanical Institute, University of 

Copenhagen, Øster Farimagsgade 2D, DK-1353, 

Denmark. - E-mail: rasmusk@bot.ku.dk 

The seasonal fluctuations of species abundances in an 

ectomycorrhizal (EM) community associated with beech 

(Fagus sylvatica) are being studied. Throughout 2001 

intensive sampling was performed in a 15 x 15 m plot in 

Lille Bøgeskov, located in mid Zealand. Soil cores were 

taken at eight harvest times covering the growing season 

from March to November. The sampling in this plot will be 

continued in 2002-2003. Fungal species forming EM on the 

beech roots are sorted by morphotyping and ITS-RFLP and 

then identified by sequencing. In the fall 2001, 

basidiocarps of EM species were sampled in the plot 

(collected and identified by Morten Christensen, KVL, 

Denmark). The basidiocarps are being sequenced in order 

to create a reference sequence database to aid in the 

identification of the root tips. As other EM communities 

the beech community turns out to be species rich. So far, 

the two harvests from March and May 2001, which has 

been most thoroughly worked through, has yielded approx. 

40 species. Some of the more abundant species found are 

Cortinarius species, Russulaceae spp., chanterelloid spp. 

including Clavulina cristata, Cenococcum geophilum, 

thelophoroid spp. and Piloderma spp. While some species 

e.g. Lactarius subdulcis are present throughout the growing 

season others e.g. a yet unidentified chanterelloid sp. are 

absent in the spring harvests but are then becoming 

dominant in the fall. Possible explanations for the observed 

seasonal trends will be discussed. 

298 


989 - Rhizopogon spore bank communities: Species 

distribution within and among Californian pine forests 

R. Kjøller 1* & T.D. Bruns 2 

1 Department of Mycology, Botanical Institute, University 

of Copenhagen, Øster Farimagsgade 2D, DK-1353, 

Denmark. - 2 Department of Plant and Microbial Biology, 

University of California Berkeley, 321C Koshland Hall, CA 

94720-3102, U.S.A. - E-mail: rasmusk@bot.ku.dk 

In this study we examine the distribution of Rhizopogon 

species in spore banks from five Californian pine forests. 

Four of the forest sites were discontinuous populations of 

Pinus muricata while the last was a Pinus ponderosa stand 

in the Sierra National Forest. Rhizopogon species were 

isolated from a soil-bioassay using Pinus muricata as host 

followed by isolation from individual root tips with typical 

Rhizopogon ectomycorrhizal morphology. The cultures 

were screened by ITS-RFLP and from each site unique 

patterns were sequenced. These sequences were then 

compared with sequences derived from identified 

sporocarp material. Bioassaying proved to be an efficient 

way to bring Rhizopogon species into culture as 50% of the 

pots contained ectomycorrhizal tips with Rhizopogon like 

morphology and half of these pots then resulted in axenic 

Rhizopogon cultures. Members of all the major pineassociated 

sections were represented within the isolates. 

Our results showed that Rhizopogon spores are 

homogeneously distributed within local forest areas while 

there is significant structuring at the regional scale. High 

spore longevity and homogenization by soil and water 

movement may explain their distribution within local forest 

areas while the latter observation fits with an expectation of 

limited dispersal because of the hypogeous fruiting of 

Rhizopogon species. 

990 - Monitoring of white-rot fungus during 

bioremediation of dioxin-contaminated soils 

R. Kondo * , H. Suhara & C. Daikoku 

Fac. of Agriculture, Kyushu University, Hakozaki, 

Fukuoka, 812-8581, Japan. - E-mail: kondo@brs.kyushuu.ac.jp 

Bioremediation is a low-cost treatment alternative for the 

cleanup of dioxin-contaminated soils and -fly ashes, when 

pollution spread broad-ranging. An interesting fungus was 

isolated from white rotten wood of a broadleaf tree from 

Kyushu Island in Japan, named strain MZ-340. This isolate 

has high ability to degrade dioxin. We have tried to use the 

strain MZ-340 for bioremediation of dioxin-contaminated 

soil on site. On the other hand, we have to consider that 

this trial has potential problems to bring biohazard on 

natural ecosystem, when this organism has been 

naturalized. Therefore, we have to develop a monitoring 

system for the introduced fungus with the examination of 

bioremediation. The bioremediation of the dioxincontaminated 

soils by the fungus MZ-340 has been


examined. We have developed a PCR-based assay to detect 

strain MZ-340 reliably on bioremediation site. DNA, 

isolated by series of CTAB and organic solvent extractions 

followed by silica column purification from the site, was 

amplified by the PCR using specific primer derived from 

ITS sequences of the strain MZ-340. We successfully 

monitored the transition of the population of the strain MZ- 

340 on the bioremediation site. The relationship between 

the population of the fungus and the effect of 

bioremediation by this fungus will be discussed. 

991 - Vegetative compatibility among Botrytis cinerea 

strains in Israel 

N. Korolev * , Y. Elad & T. Katan 

Dept. Plant Pathol. ARO, the Volcani Center, P.O.Box 6, 

Bet Dagan 50250, Israel. - E-mail: vpptlg@agri.gov.il 

A total of 21 B. cinerea strains originating from five host 

plants at 10 sites throughout Israel, were tested for mycelial 

interaction. Complementation between auxotrophic 

selenate-resistant mutants (sul) mutants, and antagonism 

(barrage) between wild type strains served as criteria for 

compatibility: Complementation indicates that the parent 

strains are compatible, whereas the barrage indicates that 

the strains are incompatible. sul mutants resistant or 

sensitive to chromate were paired on MM. Wild type 

growth of complementary heterokaryons was always 

observed between different mutant phenotypes derived 

from the same parent, as well as in some inter-strain 

pairings. Most isolates were placed into the same group 

based either on positive complementation reaction between 

their mutants, or, more often, on positive reaction with 

mutants of bridging strains, compatible with two or more 

isolates that were not compatible directly. Barrage between 

wild type isolates, in the form of a zone of sparse 

mycelium with or without dark pigmentation of the agar 

along the line of confrontation, was observed in about 35% 

of pairings. Although isolates were often incompatible with 

each other, all of them were eventually placed into one 

group based on their compatibility with a common third 

strain (bridging), confirming the data of heterokaryon 

compatibility by sul mutants. sul mutants were pathogenic 

to bean and strawberry and could be used as marked strains 

in epidemiological studies. 

992 - Effect of the Pinus strobus invasion on the 

mycoflora of litter needles 

O. Koukol 

Department of Botany, Faculty of Science, Charles 

University, Benatska 2, Praha 2, 128 43, Czech Republic. - 

E-mail: o.koukol@seznam.cz 

Mycoflora of pine litter and trash (needles, twigs, cones) 

was studied in the National Park Czech Switzerland (Czech 

Republic). A comparison of a native pine species (Pinus 

sylvestris) and an invasive pine species (Pinus strobus) was 

done at two localities in a well preserved P. sylvestris 

forest and in a growth of P. strobus. An experiment using 

litter bags with needles was arranged for a succession 

study. Between years 1999 and 2002, the litter bags were 

exposed in litter of both the pine species. Needles from the 

litter bags were cultivated in damp chambers. Frequencies 

of occurrence of recorded fungi were analysed using 

multivariate analysis and analysis of variance. Sixty-three 

species of ascomycetes in both teleomorph and anamorph 

stage were recorded. Two species: Tympanis neopythia 

Ouell. & Piroz. and Pseudocercospora deightonii Minter 

were found for the first time in the Czech Republic. Most 

of the recorded fungi colonised litter of both the pine 

species but reached different frequencies. Results of the 

analysis proved significant differences of the frequencies 

between the pine species, seasons (spring and autumn) and 

the localities. The following species were recorded in high 

frequencies: Selenosporella curvispora G.Arnaud, 

Chloridium state of Chaetosphaeria preusii W.Gams & 

Hol.-Jech. and Anavirga laxa B.Sutton. However, 

according to the literature they are uncommon at pine litter. 

993 - Spatial distribution of fruitbodies of macrofungi 

in pine forests with Vaccinium myrtillus in northern 

Latvia 

I. Krastina 

Institute of Biology, University of Latvia, Miera str. 3, LV- 

2169, Salaspils, Latvia. - E-mail: 

inguna@email.lubi.edu.lv 

Investigation on spatial distribution of fruitbodies of 

agaricoid and boletoid fungi was carried out during 1993- 

1997 in three Pinus sylvestris stands in the northern Latvia. 

Vaccinium myrtillus and mosses dominated the ground 

cover of forests. 25 permanent subplots, each of 2m x 20m, 

were established at each forest site. Each subplot was 

divided into 40 lesser microplots with size of 2m x 0,5m. 

Counting of fruitbodies was performed 5-6 times per year 

during August-October. A total of 92 species were found in 

the plots during the 1993-1997. The annual number of 

species and their productivity considerably varied from 

year to year. The fruitbodies of some species appeared in 

the same or in the adjoining microplots year after year. The 

distribution of fungal species was patchy. Neither more 

than 3-4 (usually 0-2) species were found at the same 

microplot simultaneously, though a total number of 

recorded species per microplot varied from 0 to 9. A 

negative correlation among species was found. Fruitbodies 

of some species, such as Cortinarius croceus and C. 

semisanguineus, were spread rather evenly through all 

subplots. Fruitbodies of some other species were localized 

in a few microplots only, and their appearance often were 

related to the discriminative features of these microplots- 

soil disturbance, presence of decaying wood or other 

substrates, as well as the type of vegetation. However, no 

spatial patterns of fruitbodies distribution were found for 

the major part of species. 



994 - Leaf decomposition by aquatic fungi in heavy 

metal polluted habitats 

G. Krauss 1* , J. Ehrman 2 & F. Bärlocher 3 

1 Microbiology of Subterrestrial Aquatic Systems Group, 

UFZ Centre for Environmental Research, Leipzig-Halle, 

Theodor Lieser Str. 4; D-06120 Halle/Saale, Germany. - 

2 Department of Biology, Mount Allison University, 63B 

York Street, Sackville, New Brunswick E4L 1G7, Canada. - 

3 Digital Microscopy Facility, Mount Allison University, 

63B York Street, Sackville, New Brunswick E4L 1G7, 

Canada. - E-mail: krauss@halle.ufz.de 

The long history of copper mining and smelting in the 

district of Mansfeld (Central Germany) resulted in high 

loads of heavy metals in streams, lakes and even 

groundwater [1]. We followed decomposition of Alnus 

glutinosa leaves in two streams, one with a high (H4) and 

one with a moderate (H9) load of these metals. In H9, mass 

loss closely followed an exponential decay curve; in H4, 

leaf mass remained constant after a very rapid initial decay 

during the first 4 weeks. Fungal biomass, estimated by 

ergosterol measurements, rose more quickly and to higher 

levels on H9 than on H4 leaves. Conidium production by 

aquatic hyphomycetes was reduced in site H4. Despite the 

huge differences in conidium production between the 2 

streams, fungal species numbers during decomposition 

were similar. After 4 weeks of stream exposure, leaves 

showed greatly increased levels of heavy metals [2]. 

Leaves exposed in site H4 layered with an inconsistent 

assemblage of biofilm material, fungal hyphae, extraneous 

particles of sediment and metallic precipitate. The plant 

material was remarkably well preserved. A relatively high 

leaf decay rate may indicate a fungal community that has 

successfully adapted to the ambient heavy metal levels. 

[1]KRAUSS, G. et al. (2001) Aquatic hyphomycetes occur 

in hyperpolluted waters in Central Germany. Nova 

Hedwigia 72: 419-428 [2]SRIDHAR, K.R. et al. (2001) 

Decomposition of alder leaves in two heavy metal polluted 

streams in Central Germany. Aquat. Microb. Ecol., 26: 73- 

80. 

995 - Fungi and insects - an ecologically important 

relationship in a mountaineous rain forest in South- 

Ecuador 

R. Krettek * & E. Langer 


Str. 40, Germany. - E-mail: roman.krettek@uni-kassel.de 

During two research periods at the Estacion Scientifica San 

Francisco near Loja in South-Ecuador, we discovered that 

specific ecological relationships between fungi and insects 

are an important factor influencing coevolution between 

the two groups. Especially beetles from the Staphylinidae 

have been observed on Agarics. Beetles from the Ptiliidae 

have been discovered to be very well adapted to the 

hymenial morphology of Polyporaceae s.l. 

300 


996 - Microscopic fungi associated with oak bark beetle 

(Scolytus intricatus) in the Czech Republic 

A. Kubátová 1 , D. Novotný 2* & K. Prásil 1 

1 Culture Collection of Fungi (CCF), Department of 

Botany, Faculty of Science, Charles University, Benátská 

2, 128 01 Praha 2, Czech Republic. - 2 Czech Collection of 

Micro-organisms, Faculty of Science, Masaryk University, 

Tvrdého 14, 602 00 Brno, Czech Republic. - E-mail: 

novotdad@natur.cuni.cz 

During 1997-99, surface mycoflora of Scolytus intricatus 

was studied with respect to its role in transmission of 

microscopic fungi. Samples were collected from five 

localities in central Bohemia. Investigation was focused on 

all different stages of life cycle of the beetle: eggs, larvae, 

adults before emergence, adults in generation and 

maturation feeding (altogether 500 samples) and also on 

galleries (400 samples). Insect samples were washed by 

sterile water in ultrasonic cleaner. Suspension and insect 

were separately inoculated onto Petri dishes with maltextract 

agar. The most frequent fungi associated with S. 

intricatus appeared to be Candida rhagii, Geosmithia spp. 

and Penicillium spp. A great attention was paid to 

occurrence of ophiostomatoid fungi. Two species were 

determined: Ophiostoma piceae s.l. and O. cf. prolifera, 

both considered to be saprotrophs or weak pathogens. 

Frequency of all these fungi varied depending on the 

locality and the stage of life cycle of the beetle. During this 

study other interesting micromycetes were isolated, 

belonging to phytopathogenic fungi (e.g. Diplodia mutila), 

wood colonizing fungi (e.g. Acrodontium crateriforme, 

Cytospora sp., Libertella sp., Phomopsis sp., 

Ramichloridium anceps, Phaeoacremonium spp.), 

nematophagous fungi (Esteya vermicola), 

entomopathogenic fungi (e.g. Paecilomyces farinosus) and 

many litter and soil fungi (e.g. Acremonium spp.). The 

study was supported by the Grant Agency of the Czech 

Republic (No. 203/97/0037). 

997 - Study on fungal communities in mediterranean 

pine woods: First results 

A. Laganà 

Dip. Scienze Ambientali 'G. Sarfatti' - University of Siena, 

Via P.A. Mattioli, 4 - I 53100 Siena, Italy. - E-mail: 

lagana@unisi.it 

Fungi can be regarded as useful bioindicators of forest 

ectotrophic stability; mycological studies can be useful for 

predicting the fate of forests subject to different types of 

stresses. Integrated researches on fungal communities and 

their environment provide useful informations about the 

ecology and distribution of fungi in space and time; such 

informations are preliminary for studies on the 

conservation of fungal flora and for the statement of global 

change effects. This type of informations are available for 

many taxa in central and northern Europe, but is largely


lacking for those in the Mediterranean area. The European 

Confederation of Mediterranean Mycology (CEMM.ae) 

recognized the need to fill these gaps and in 1999 research 

was begun on fungal communities of Pinus sp. pl. woods 

by mycologists of the three member countries (France, 

Italy and Spain). Here, the first results of this work are 

reported. Various species linked to pines or to the 

Mediterranean environment were found. Some taxa present 

in the Red Lists of central and northern European countries 

were also present in the studied areas. The percentage of 

mycorrhizal species was calculated for each permanent 

plot. This percentage was found to reflect forest health in 

northern and central Europe. Low values of this index for 

some of the studied pine woods suggests that they are in a 

state of decline. Classification and ordering of the relevés 

showed affinities between the fungal communities. 

998 - Interaction of arbuscular mycorrhizal fungi with 

plant pathogens on Coffea arabica L. 

L. Lebrón 1* , D.J. Lodge 2 & R. Rodríguez 3 

1 Institute for Tropical Ecosystems Studies, University of 

Puerto Rico-Rio Piedras, PO Box 23341, San Juan, PR 

00931-3341, U.S.A. - 2 Center for Forest Mycology 

Research, USDA Forest Service, Forest Products Lab, Box 

1377,Luquillo, PR 00773, U.S.A. - 3 Crop Protection 

Department, University of Puerto Rico-Mayagüez, 

University Station,Mayagüez, PR. 00708, U.S.A. - E-mail: 

llebron@upracd.upr.clu.edu 

Interactions between AM fungi and a pathogen of coffee 

leaves were investigated. First, we evaluated three coffee 

cultivars for differences in the amount of AM extraradical 

hyphae. Extraradical hyphae were significantly less in 

Caturra than in Pacas and Borbón cultivars in all locations. 

In another study, we compared the percentage of 

mycorrhizal root length among paired coffee seedlings with 

and without Cercospora coffeicola lesions. We found that 

seedlings with pathogen lesions on the leaves had 

significantly greater mycorrhizal colonization than 

seedlings without lesions. In a third study, we evaluated 

effects of mycorrhizae and soil nutrient levels on plant 

growth and development of leaf lesions caused by C. 

coffeicola. This experiment used the cultivars Caturra and 

Borbón with four nutrient treatments: a control, 25 ppm 

added N or P, and 25 ppm each of added N and P. Growth 

in Caturra only responded to fertilization while growth in 

Borbon only responded to mycorrhizae. The results also 

suggested that there was a higher cost of mycorrhizal 

association in Borbón than in Caturra. There were no 

significant differences in lesion development among 

treatments in two replicate experiments. 

999 - Community structure of ectomycorrhizal fungi 

beneath fruitbodies of Xerocomus subtomentosus, 

Suillus luteus, Lactarius rufus and Scleroderma citrinum 

T. Leski * & M.L. Rudawska 

Institute of Dendrology, Polish Academy of Sciences, 

Parkowa 5, 62-035 Kornik, Poland. - E-mail: 

tleski@man.poznan.pl 

The goal of our studies was to determine the 

correspondence between above- and below-ground views 

of species composition of ectomycorrhizal fungi in Scots 

pine (Pinus sylvestris) stand in Poland. We sampled 

mycorrhizas from soil cores taken directly below 

fruitbodies of some ectomycorrhizal fungi, which appeared 

with differentiated abundance on tested plots. Suillus 

luteus, Lactarius rufus and Scleroderma citrinum were 

among the most abundant fruitbodies and Xerocomus 

subtomentosus belonged to rather rare specimen. All viable 

ectomycorrhizas removed from cores taken directly below 

fruitbodies were classified into morphotypes based on 

color, size and type of ramification. On average 5-6 

different ectomycorrhizal morphotypes were selected under 

each species and mostly one or two morphotypes 

dominated. RFLP analysis of the ITS region to identify 

species in ectomycorrhizal communities confirmed 

morphological typing. A few ectomycorrhizal tips 

belonging to X. subtomentosus and S. citrinum were 

detected in cores taken under these fruitbodies. In samples 

taken beneath S. luteus and L. rufus root tips bearing 

mycorrhizas of these species were lacking. The results 

suggest that root tips of several ECM species coexist within 

fruitbody of each particular species. 

1000 - Switching partners: ecto- and arbuscular 

mycorrhizal fungi on seedlings of three oak species 

A. Lindahl 1* , L. Egerton-Warburton 2 , C. Blesdoe 3 , D. 

Southworth 4 & M.F. Allen 1 

1 Center for Conservation Biology, UC Riverside, 

University Lab Building, Room 208, Riverside, CA 92521, 

U.S.A. - 2 Chicago Botanic Garden, 1000 Lake Cook Road, 

Glencoe, IL 60022, U.S.A. - 3 Universide of California, 

Davis, 225 Hoagland Hall, One Shields Ave., Davis, CA 

95616, U.S.A. - 4 Southern Oregon University, Science 159, 

1250 Siskiyou Blvd., Ashland, OR 97520, U.S.A. - E-mail: 

AmyL@citrus.ucr.edu 

Interestingly, an increasing number of researchers have 

shown that under field conditions, some Quercus species 

may host both ecto- and arbuscular mycorrhizal fungi. This 

ability to host arbuscular mycorrhizal (AM) fungi may 

allow oaks to successfully establish beyond the 

ectomycorrhizal (EM) root zone of mature oaks. In this 

study, we considered how established EM and AM 

communities might affect colonizing oak seedlings. Three 

oak species (Quercus agrifolia, Q. garryana, and Q. 

douglasii) were transplanted into a Southern Californian 



oak (Q. agrifolia) ecosystem. After a year in the field, the 

seedlings, accompanied by soil cores from their immediate 

environments, were harvested and analyzed. We found that 

the position of planting strongly determined the EM or AM 

status of the seedling; seedlings transplanted under a 

mature oak canopy become colonized with ectomycorrhizal 

(EM) fungi, while those planted into surrounding grassland 

areas typically showed arbuscular mycorrhizal (AM) 

colonization. Morphotyping and RFLP data showed that 

the presence and position of EM species on mature oaks 

strongly predicted EM diversity on seedlings. Several 

species of EM fungi dominated the site and were present 

on both seedlings and mature trees, although many rare 

types were also found. The diversity and identity of 

mycorrhizae associated with transplanted oak seedlings 

appear to be largely regulated by position of planting, and 

not by any mycorrhizal preferences of the three oaks 

species. 

1001 - Population structure of Heterobasidion annosum 

in birch plantations, replanted on infested sites 

V. Lygis * , R. Vasiliauskas & J. Stenlid 


University of Agricultural Sciences, P.O.Box 7026, SE-750 

07 UPPSALA, Sweden. - E-mail: irklas2000@yahoo.com 

The persistence of Heterobasidion annosum on infested 

areas and its transfer to a new forest generation was studied 

in three forest sites with similar histories. The sites 

represented H. annosum disease centres (each 0.2 ha in 

size) in 50-60 year-old Pinus sylvestris stands, which were 

clear-felled and replanted with Betula pendula 25 years 

previously. Fungal isolation from trees and stumps on each 

site encompassed both replanted B. pendula and 

surrounding P. sylvestris from the previous generation. A 

total of 121 isolates of H. annosum was obtained from 

spatially separated resource units. All of them belonged to 

the P intersterility group. Each isolation locality was put on 

a local map. Vegetative compatibility tests with the isolates 

revealed extensive territorial clonality: from the first site, 

23 isolates clustered into 5 clones (4.6 isolate per clone), 

from the second - 63 into 6 (10.5), and from the third - 35 

into 5 (7.0). Three largest clones consisted of 30, 18 and 10 

isolates and covered the respective areas up to 25, 32 and 

48 m across. It was not uncommon for the territorial clone 

to cover areas that encompassed both P. sylvestris and B. 

pendula forest generations. The results of this study show 

that H. annosum is able to persist on infested sites for 

decades and readily attack replanted trees of the new forest 

generation. This is the first extensive study dealing with the 

population structure of H. annosum P group in birch stands. 

302 


1002 - Chytridium lagenula Scheffel is a parasite of 

Tribonema gayanum Pash. (Xanthophyceae) 

M.A. Mamkaeva * , A.V. Pljushc & K.A. Mamkaeva 

Biological Research Institute of St.Petersburg University, 

Oranienbaumskoye sch.2, Stary Petergof, 198504, 

St.Petersburg, Russia. - E-mail: mamkasha@yandex.ru 

Development of Chytridium lagenula was observed in the 

cultures inoculated by water samples. This parasite 

develops only on the alive cells of alga. We observed 

development of C. lagenula on Tribonema gayanum and T. 

vulgare Pasch. We did not observed development of this 

parasite on dead cells and there was not growth on the 

pine's pollen and on the free medium (without alga), that is 

why we think, that C. lagenula is obligate parasite of algae. 

C. lagenula has a sessile upright or slightly titled 

sporangium with a thin smooth wall. Growing sporangium 

is globular, ripening pear-shaped, 8-13 µm length, 4-6 µm 

breadth. Zoospores of the parasite are globular with one big 

oil droplet. The diameter of zoospore is 3-4 µm. C. 

lagenula is able to form globular resting spores, 6 µm in 

diameter. Endobiotic system of C. lagenula consist of a 

rhizoid. Also we have a facts about quantity circulation of 

C. lagenula in nature. The number of infectious units of C. 

lagenula was evaluated in Ladoga lake and adjacent water 

bodies. It was 10 stations examinated. C. lagenula was 

detected in 4 stations examined. Quantity of infective units 

(sporangium, zoospores) was varied from 0.1 to 0.36. (It is 

need to take into account, that concentrations were low, 

because water was filtered). Reservoir it is possible to 

speak, that C. lagenula is widespread parasite of alga in 

nature, which mainly inoculate genus Tribonema. 

1003 - Single sequence repeats markers reflect 

population diversity and geographic barriers of 

Ceratocystis polonica in Eurasia 

M. Marin 1* , O. Preisig 1 , B.D. Wingfield 2 , T. Kirisits 3 & 

M.J. Wingfield 1 



University of Pretoria, Pretoria 0002, South Africa. - 



Pretoria 0002, South Africa. - 3 Institute of Forest 

Entomology, Forest Pathology and Forest Protection, 

Universität für Bodenkultur Wien, Hasenauerstrasse 38, 


Mauricio.Marin@fabi.up.ac.za 

The blue stain fungus Ceratocystis polonica and its 

associated bark beetle vectors, in particular Ips 

typographus, cause huge losses to spruce (Picea abies) in 

Eurasia. The aim of this study was to develop Single 

sequence repeats markers (SSRs) in order to determine the 

population structure and genetic diversity of C. polonica in 

Europe and Japan. RAMS primers were used to target


SSRs regions. The amplified PCR products were cloned, 

sequenced and analyzed for SSRs. Primers were designed, 

flanking the SSRs found in these amplicons. Seventy-five 

isolates from four populations were tested with ten SSR 

markers. A high gene diversity was found (h=0,56). 

Analysis of the G statistics showed a low degree of 

population structure in Central Europe and a high level of 

gene flow between populations (Gst=0,02, Nm=17,17). In 

contrast, a high population subdivision was found between 

the Japanese and the European populations, indicating 

strong geographic isolation. These data based on C. 

polonica closely resemble those recently produced for its 

insect vector, I. typographus (Stauffer et al., Mol. Ecol. 8, 

763-773). The SSR primer pairs could also amplify PCR 

products from six other Ceratocystis spp. from conifers in 

the C. coerulescens complex. Results from this study 

suggest that quarantine strategies in Europe should 

consider the entire range of genetic diversity on the 

continent and that movement of C. polonica and its vectors 

between Europe and Japan should be avoided. 

1004 - Differentiation of the macrofungal communities 

in high diversity forest ecosystems of central Balkan 

peninsula 

Z. Marjanovic 1* & B. Karadzic 2 

1 Eberhard Karls Universität Tuebingen, Physiol. Ökologie 

der Pflanzen, Auf der Morgenstelle 1, 72076 Tuebingen, 

Germany. - 2 Institut za bioloska istrazivanja 'Sinisa 

Stankovic', 29 Novembra 142, 11 000, Beograd, 

Yugoslavia. - E-mail: zaklina.marjanovic@unituebingen.de 

To study macrofungal communities in forest ecosystems 

we chose a canyon in Western Serbia where the vegetation 

showed very high biodiversity and differentiation due to 

the climatic conditions. Three ecological groups of 

macrofungi (wood and litter decayers and mycorrhizal) 

were detected by observing sporocarps during 6 years. 

Plant and fungal community differentiation were examined 

comparing three techniques of multivariate analyses. To 

determine the influence of abiotic factors on differentiation 

of plant and macrofungal communities we used a 

calibration method for humidity, temperature, soil acidity, 

available nitrogen and light intensity. We used canonical 

correspondence analysis for analysing influence of abiotic 

parameters, as well as for analysing influence of dominant 

phanerophytae species on differentiation of macrofungal 

communities. Four vegetation complexes were described: 

xerophyllous forests of black hornbeam, forests of 

xerophyllous oaks, mixed mesophyllous forests of black 

hornbeam and mesophyllous beech forests. The highest 

macrofungal diversity, as well as the best community 

structure and differentiation were detected in oak and beech 

forests. The main factors that influenced differentiation of 

plant communities were temperature, available nitrogen 

and light intensity. Abiotic factors showed much lower 

influence on differentiation of macrofungal communities. 

The main factor for this process was presence of specific 

phanerophytae species. 

1005 - Fungal associations of mycorrhizal roots of 

conifer seedlings in Lithuanian forest nurseries 

A. Menkis * , R. Vasiliauskas, A.F.S. Taylor & R.D. Finlay 

Department of Forest Mycology & Pathology, Swedish 

University of Agricultural Sciences, P.O. Box 7026, SE-750 

07 Uppsala, Sweden. - E-mail: 

Audrius.Menkis@mykopat.slu.se 

Morphological and molecular identification of fungi 

associated with mycorrhizal roots was conducted on 60 

pine and spruce seedlings in 5 forest nurseries in Lithuania. 

The plants were either grown in nursery beds as bare root 

seedlings, in plastic trays or in wrapped polyethylene (WP) 

containers. Bare root greenhouse seedlings were also 

examined. In total, 5364 & 12441 root tips were 

investigated for pine and spruce, respectively. Of the pine 

roots, 2131 (39%) were mycorrhizal, and 6527 (52%) of 

the spruce roots. The cultivation system had a profound 

impact on mycorrhizal colonization for both tree species. In 

pine, it was most abundant in the nursery bed bare root 

system (49%), while in spruce - in the WP containers 72% 

of the roots were mycorrhizal. In total 8658 mycorrhizal 

roots were morphotyped, and 7 species were found on pine 

and 8 on spruce roots. On pine an unidentified ascomycete 

was most common (58% of mycorrhizal tips), while on 

spruce a basidiomycete Amphinema byssoides colonized 

54% of the mycorrhizal tips. Isolation of the fungi into pure 

culture was attempted from 8163 mycorrhizal root tips, 

yielding 625 isolates, which were divided into 144 groups 

according to morphological characteristics of the 

mycelium. One representative of each mycelial group was 

subjected to sequencing of the ITS region of the rDNA and 

43 fungal species have been determined using the NCBI 

BLAST. The most common species were Phialocephala 

fortinii (29%) and Phialophora finlandia (37%). 

1006 - Rhizomorph Foraging Patterns of Armillaria 

gallica and A. mellea from the Ozark Mountains of the 

central U.S.A. 

J.D. Mihail * & J.N. Bruhn 

Dept. of Plant Microbiology & Pathology; University of 

Missouri, 108 Waters Hall, Columbia, Missouri, 65211, 

U.S.A. - E-mail: mihailj@missouri.edu 

Our research is driven by the need to understand inter- and 

intra-specific interactions of co-occurring Armillaria genets 

with respect to oak deline. Foraging patterns of A. gallica 

and A. mellea rhizomorphs were measured in three 

experimental series: 1) temporal pattern changes measured 

in a uniform environment (agar); 2) pattern responses to 

woody baits measured in sand; and 3) pattern responses to 

the presence of neighboring genets of the same or different 

species, measured in sand. Four response metrics were 

used: fractal dimension, number of foraging tips, total 

rhizomorph length, and total rhizomorph surface area. 

Preliminary analysis indicates that rhizomorph foraging by 



A. gallica was stochastic where target woody food bases 

were placed laterally with respect to the colonized source; 

however rhizomorph foraging was significantly directed 

when the target woody food base was placed above or 

below the source. Presence of con-specific neighbor genets 

altered the allocation of rhizomorph length and the number 

of foraging tips without affecting fractal dimension for 

both A. gallica and A. mellea. Similarly, neighboring 

genets of different species did not influence the fractal 

dimension of A. gallica or A. mellea rhizomorph systems. 

Thus, Armillaria rhizomorph system pattern is influenced 

in abundance and physical location in the landscape by 

adjacent genets of the same or different species, while the 

inherent foraging pattern remains unresponsive to such 

influences. 

1007 - Micro-evolution in the ectomycorrhizal fungus 

Suillus luteus 

L.A.H. Muller, K. Adriaensen, J. Vangronsveld & J.V. 

Colpaert * 

Limburgs Universitair centrum, Laboratory of 

Environmental Biology, Universitaire Campus, 3590 

Diepenbeek, Belgium. - E-mail: jan.colpaert@luc.ac.be 

Previous research has shown that populations of the 

ectomycorrhizal fungus Suillus luteus found in areas 

polluted with high levels of heavy metals (Zn, Cd) have 

acquired adaptive tolerance to elevated levels of these 

metals. Furthermore, these populations showed a reduced 

genetic variability, as measured by ISSR markers, 

compared to control populations from unpolluted sites. In 

this study we aim to analyse the microevolution of 

populations of Suillus luteus in areas polluted with heavy 

metals. Therefore, the genetic structure of these 

populations, as well as the structure of populations from 

unpolluted areas, the amount of gene flow between these 

populations and the occurrence of bottlenecks will be 

described using microsatellite and AFLP markers. In a later 

stage, mtDNA markers will also be used, after having 

resolved the inheritance mode of the mitochondria in S. 

luteus. After identification of genes related with the heavy 

metal tolerance, natural selection will be investigated on a 

molecular level using these genes and the selectively 

neutral molecular markers. 

1008 - Genetic diversity within French populations of 

Tuber melanosporum Vittad. 

C. Murat 1* , P. Luis 2 , J. Diez 2 , C. Delaruelle 2 , C. Dupré 3 , G. 

Chevalier 3 & F. Martin 2 

1 University of Torino, Viale Mattioli, 25, 10125 Torino, 

Italy. - 2 UMR 'Interaction Tree-Microorganisms' INRA of 

Nancy, 54280 Champenoux, France. - 3 INRA Unity of 

Mycology, Crouelle, 63039 Clermont-Ferrand cedex 2, 

France. - E-mail: claude.murat@unito.it 

304 


Truffles are ectomycorrhizal ascomycetes and edible fungi 

of economic value. They have been the focus of numerous 

studies of genetic diversity. Certain species show a high 

genetic diversity (e.g. Tuber uncinatum), whereas others 

have low polymorphism (e.g. T. melanosporum). T. 

melanosporum was regarded as a clonal species without 

any geographic genetic structure. We have analyzed the 

genetic diversity of T. melanosporum populations from 

different geographic regions in France by looking for 

polymorphic loci. We have used RAPD (Random 

Amplified Polymorphism DNA) profiles, genomic 

sequences previously identified as SCAR (Sequence 

Characterized Amplified Random) and sequences from the 

internal transcribed spacers (ITS) of nrDNA. The RAPD 

phenotypic analysis and the sequence analysis of genomic 

tags have confirmed the genetic homogeneity of this 

species. RAPD phenotypes can not be correlated to their 

origin region. However, we have found some polymorphic 

populations and, according to the Shannon's index, isolates 

from 'Bourgogne' were the most variable. Sequencing of 

the ITS allowed us to identify four haplotypes (I, II, III and 

IV) in 47 ascocarps. Haplotypes I and II were the most 

frequent and they were found in every populations except 

for haplotype I in 'Lorraine'. On the other hand, haplotypes 

III and IV were more scarce and restricted to particular 

French regions (e.g. Haplotype IV was found only in 

'Lorraine') suggesting a genetic structure in T. 

melanosporum. 

1009 - Ceratocystis wilt of Acacia mearnsii in Uganda 

G. Nakabonge * , I. Barnes, J. Roux & M.J. Wingfield 


University of Pretoria, 0002, Pretoria, South Africa. - Email: 

grace.nakabonge@fabi.up.ac.za. 

Ceratocystis wilt caused by C. albofundus is a severe 

disease of Acacia mearnsii, in Africa. In a recent survey, 

this disease was found on wounded A. mearnsii in South 

Western Uganda. Diseased trees showed extensive 

discoloration of the wood, dieback, gummosis, wilting and 

death. Previous studies of the gene diversity of a South 

African population using CAT5 marker and 8 microsatellite 

markers revealed a high gene diversity. The high gene 

diversity and presence of C. albofundus on native Protea 

species has led to a hypothesis that it is native to South 

Africa. The aim of this study was to determine the gene 

diversity of a Ugandan population and compare it to that of 

the South African population. 36 isolates, were obtained 

from A. mearnsii trees in South Western Uganda. Eight 

microsatellite primers pairs previously designed for C. 

fimbriata were used to amplify the microsatellite rich 

regions of the genome. Analysis of the microsatellite data 

revealed 7 loci to be polymorphic. Nei's gene diversity (H) 

showed that the South African population is slightly more 

diverse than the Ugandan population with a value of 

0.4320 compared to 0.4082. Results further revealed that 

very few alleles are shared, with little gene flow between 

the two populations. The high gene diversity in both 

populations strongly suggests that C. albofundus is native 

to the African continent and not necessarily only to South 

Africa, as previously suggested.


1010 - Roles of ascospores in the white root rot fungus, 

Rosellinia necatrix 

H. Nakamura 1* , K. Ikeda 1 , M. Arakawa 2 & N. Matsumoto 1 

1 National Institute for Agro-Environmental Sciences, 3-1-3 

Kan-non dai, Tsukuba 305-8604, Japan. - 2 Faculty of 

Agriculture, Meijo University, 1-501 Shiogamaguchi, 

Tempaku-ku, Nagoya 468-8502, Japan. - E-mail: 

nakamh@affrc.go.jp 

Rosellinia necatrix Prilleux, the ascomycetous white root 

rot pathogen, causes destructive damage to numerous 

woody and herbaceous plants, especially to fruit trees, 

throughout the world. The fungus produces teleomorph 

rarely on diseased plants in nature, and the roles of 

ascospores in the life cycle remains unclear due 

presumably to the scarcity of teleomorph production. In 

this study, stromata produced on root fragments were 

studied to elucidate the roles of ascospores. Stromata were 

produced on 23 out of 47 diseased root samples from 

Japanese pear, grapevine and Chloranthus glaber, which 

had been placed in a hollow on the ground surface in the 

shade of trees and covered with rice straw. Also, stromata 

were obtained on mulberry twigs inoculated with two out 

of six isolates. Readiness of stroma production was not 

related to host plant species, dates of treatment, or the 

presence of double-stranded (ds) RNA. In 18 single 

ascospore isolates from six stroma samples, mycelial 

interactions between isolates from the same samples were 

incompatible, and the isolates were pathogenic to Lupinus 

luteus with a few exceptions. dsRNAs in vegetative hyphae 

of 10 stroma samples were not transferred to single 

ascospore isolates. Thus, ascospores in R. necatrix are 

effective in extending genetic variation, producing 

pathogenic offspring and eliminating infectious factors 

such as dsRNA. 

1011 - Population dynamic of Pyrenochaeta terrestris on 

the basis of cropping patherns and infection percentage 

in onion fields 

M. Nasr-Esfahani 

Plant pests and diseases research Dep., Agricultural 

research center, P.o. Box 81785-199, Esfahan, Iran. - Email: 

M_nasresfahni@yahoo.com 

Studies on population dynamic of the fungus on the basis 

of infection, indicated that the cropping patherns having a 

significant effects in infection of the fungus. 441 onion 

fields were taken into considerations during 1997-98, 

where there was five manin cropping patherns including 

alfa-alfa (Medicago sativa L. usually for 4-5 years), cereals 

(wheat, barley, rice and seldom millet), vegetables (tomato, 

bringal, mask-melon, cauliflower, cabbage, etc), orchard 

(where the fruit trees were up rooted and became under 

cultivation) and onion frequently for the last four years 

before to onion. A hundred onion plants were up rooted 

near maturity stage and their roots were examined for the 

presence of the infection based on infection percentage of 

the roots in three scoring scales (>10%, >25% and more 

than 25% infection). The higher infection was observed in 

frequent onion growing fields, fallowed by alfa-alfa with 

36.16 and 27 percent infection respectively, where the 

orchard soil had the least infection with 2.83 percent 

infection on onion roots. Out of which 12.58, 9.58 and 0.50 

percent were in the ranges of more than 25 percent 

respectively with a significant effects, where cereals and 

vegetables were in a separate group. These results revealed 

that the build up of population inoculum is an improtant 

factor in natural enivronments, but the more inportant one 

is the population potentiality, which can measured 

infectionally and/or differentially with a distinct scoring 

scales. 

1012 - Coprinaceae in wood and straw degradation: 

implications for ecology and biotechnological use 

M. Navarro-Gonzalez 1 , M. Hoffmann 1 , O. Holdenrieder 2 & 

U. Kües 1* 

1 Molecular Wood Biotechnology, Institute for Forest 

Botany, Georg-August University Goettingen, Buesgenweg 

2, D-37077 Goettingen, Germany. - 2 Section of Forest 

Pathology & Dendrology. Department of Forest Sciences. 

Federal Institute of Technology, Raemistr. 101, CH-8092 

Zurich, Switzerland. - E-mail: ukuees@gwdg.de 

Coprinaceae have been defined as a family of about 800 

species, following morphological and physiological 

characteristics such as lamella and spore colour and their 

abilities to self-digest upon fruit body maturation. Recent 

molecular analysis suggests Coprinaceae are not 

monophyletic but comprise four different genera (1). 

Coprini are usually saprotrophic on soil or dung of 

herbivores. Species growing on decaying wood or plant 

debris in soil and a few plant pathogens have also been 

described. Here we present an overview on species relating 

to wood and straw degradation. Enzymes implicated in 

wood and straw degradation (phenol oxidases including 

laccases; peroxidases; xylanases) have been described in a 

few species. Biotechnological use have been made of C. 

cinereus peroxidase (2) and one laccase (3) - genes for at 

least six different laccases are known (3,4). It is thought 

that at least some of these enzymes link to developmental 

processes, for example fruiting body development (C. 

congregatus; 5) and spore formation (C. cinereus; 6). Our 

experimental work aims at elucidating wood degrading 

abilities of various Coprinaceae, at making use of enzymes 

in biotechnology and at determining functions of enzymes 

in fungal development. (1) Redhead et al. 2001 Taxon 

50:203; (2) Kauffmann et al. 1999 J Biotechnol 73:71; (3) 

Yaver et al. 1999 AEM 65:4943; (4) Bottoli et al. 1999 J 

Microbiol Meth 35:129; (5) Choi et al. 1987 Mycologia 

79:166; (6) Vnenchak, Schwalb 1989 Mycol Res 93:546. 



1013 - Physiologic races of stem rust of wheat in Egypt 

during 1994 - 1996 seasons 

M. Nazim 1 , M. Awad 1 , S. Abu El-Naga 2* & I. Youssef 2 

1 Agricultural Botany Dept., Faculty of Agriculture, 

Minufiya Univ., Shebin El-Kom, Egypt. - 2 Plant Pathology 

Inst., Agriculture Research, Center, Giza, Egypt. 

The problem of stem rust incidence has been solved since 

the release of resistant wheat varieties in Egypt. Also, the 

early sowing of wheat varieties made the problem less 

serious. However, the appearance of new races is expected. 

Therefore, continuous identification of physiological races 

of the causal organism is required. In the course of this 

study, identification of stem rust physiologic races was 

carried out during two successive seasons (1994-1995 & 

1995-1996) using the North American wheat stem rust 

monogenic differentials. The results obtained revealed the 

presence of different physiologic races in Egypt during 

both seasons of the study. Races RT-, RR-, TR-, TT- 

comprised 92.02% of the total isolates. While, races RT-, 

RR-, TR- and TT-comprised 7.98% of the isolated races in 

1994/1995 season. On the other hand, in the second season 

(1995-1996) races TT-, RT-, RR-, MT- and QF- comprised 

47.04% of the total isolates. While, the rest were 29 races 

comprised 52.96% of the isolated races. The great 

variability of the physiologic races showed the dynamic 

state of the pathogen. Also, showed that wheat plants are 

liable to be attacked by different races from different 

sources. 

1014 - Frequency of virulence of Puccinia triticina on 

wheat in Egypt during 1998-2001 

M. Nazim 1* , S. Sharif 2 , I. Shinoda 2 , O. Bolot 2 & I. Imbabi 2 

1 Botany Dept., Faculty of Agriculture, Menofiya 

University, Shebin El-Kom, Egypt. - 2 Plant Pathology 

Institute, Agriculture Research Center, Giza, Egypt. - Email: 

abfahmi@hotmail.com 

Leaf rust of wheat is common under the Egyptian 

conditions due to the favorable environmental climate for 

disease incidence and development. Many wheat varieties 

were discarded because of their susceptibilty to the disease 

due to the appearance of new virulences and the dynamic 

state of the causal organism. The frequency of virulence 

was studied against 32 local genotypes and 45 monogenic 

lines for leaf rust resistance in addition to the highly 

susceptible cv. Thatcher. The results revealed that more 

than 75% of the obtained races were virulent to six 

genotypes i.e cvs. Giza 139, Giza 155, Giza 157, Giza 160 

and Giza 164. Whereas 50-75% of the total races were 

virulent to 15 genotypes i.e. cvs. Giza 162, Giza 163, Giza 

165, Giza 167, Sakha 8, Sakha 92, Sids I, Sids 2, Sids 3, 

Sids 4, Sids 5, Sids 9, Gemmieza 1, Gemmieza 3, and 

Sakha line 202. Less than 50% of the total races were 

virulent to 11 genotypes i.e. cvs. Giza 168, Sakha 61, 

Sakha 69, Sakha 93, Sids 6, Sids 7, Sids 8, Gemmieza 5, 

306 


Gemmieza 7, Gemmieza 9 and Sakha line 206. On the 

other hand, no virulence was detected against Lr 19. Low 

frequecies were found to Lrs; 15, 21, 24, 28, 29, 36, 38, 39, 

40, 41, 42, and 43. Therefore, these genes should be 

considered in breeding programme for leaf rust resistance. 

High frequencies (more than 80%) were found against Lrs; 

1, 2c, 3, 12, 13, 14 a, 22b, 23, 33 and Lr B whereas, 

frequencies from 51 - 80% were found on the rest of genes. 

1015 - Contribution to the knowledge of the endophytic 

mycoflora of roots of sessile oak (Quercus petraea) 

D. Novotný 

Czech Collection of Microorganisms, Faculty of Science, 

Masaryk University, Tvrdého 14, 602 00 Brno, Czech 

Republic. - E-mail: novotdad@natur.cuni.cz 

The composition of endophytic mycoflora of roots of 

sessile oak (Quercus petraea) was studied. This research 

was conducted in six oak stands in Krivoklátsko region in 

Central Bohemia (Czech Republic). The fungi were 

isolated from fine roots (0.1-0.3 cm diam) and from 

peridemal bark, subperidermal bark and wood of medium 

roots (2-5 cm diam) and coarse roots (0.5-1 cm diam). 

After surface sterilisation the roots were cut into segments 

and placed onto malt extract agar. More than 60 species of 

microscopic fungi were identified from the samples. Fungi 

occurred most frequently in fine roots and in peridermal 

bark of medium and coarse roots. Wood of coarse roots 

was colonised at least. The most frequent fungi were 

Phialophora cf. fastigiata, Cryptosporiopsis radicicola, 

Cystodendron-like hyaline anamorph, dark septate 

endophyte, Umbelopsis nana, Trichoderma viride, 

Oidiodendron griseum, Chloridium preusii and 

Sporidesmium cf. anglica. Species of genera 

Sporidesmium, Chloridium, Chaetosphaeria and 

Menispora were mainly isolated from the fine roots. 

Rhizomorphs of Armillaria and different mycelia of 

Basidiomycetes were recorded in some roots. Bacteria 

were common in roots. The study was supported by the 

Grant Agency of the Charles University (No. 243/1997/B 

BIO/PøF). 

1016 - Biodegradation of drying oil claddings by 

micromycetes 

N. Nugaeva 1* , E. Lebedeva 1 , E. Vedenyapina 1 , I. Smirnov 2 

& G. Markov 2 

1 Komarov Botanical Insitute of the Russian Academy of 

Sciences, 2 Prof. Popov Str., St. Petersburg 197376, 

Russia. - 2 RPA V. G. Khlopin Radium Institute, 28 2nd 

Murinski pr., St. Petersburg 194021, Russia. - E-mail: 

fungi@mail.ru 

In the drying-related industry the oil components are of 

great utility for covering wood, metal and other materials 

and constructions. And there is the problem of its complete


degradation after some period of service. It is known that 

natural and synthetic polymeric materials undergo a 

process of decomposition after a period of service and the 

main biological agents of it are filamentous microscopic 

fungi-micromycetes (MM). The goal of the present study 

was to find out the community of MM for a maximal 

biodegradational activity against the drying oil cladding on 

the base of the natural component (linseed oil). Active 

strains of MM which were isolated from different 

polymeric materials were used to make the inoculation 

communities: I - including Aspergillus niger, A. versicolor, 

Penicillium aurantiogriseum, P. funiculosum,Trichoderma 

koningii; II - Aspergillus fumigatus, A. flavus, 

Cladosporium cladosporioides, Penicillium frequentans, P. 

notatum; III - Aspergillus glaucus, A. terreus, A. ustus, 

Alternaria alternata, Penicillium brevicompactum. It was 

found that the different communities of MM colonized oil 

cladding to different extent. The areas of colonization 

varied from 60% to 100% for II and III communities and 

from 40% to 50% for the I community. The inoculation by 

the I community resulted in maximal loss of oil cladding 

(up to 13.3% of the starting weight). So, the first 

community of MM has the maximal biodegradational 

activity against the drying oil cladding. 

1017 - Formation of communities of micromycetescolonizers 

of municipal waste during the composting 

N. Nugaeva * , E. Vedenyapina & E. Lebedeva 

Komarov Botanical Insitute of the Russian Academy of 

Sciences, 2 Prof. Popov Str., St. Petersburg 197376, 

Russia. - E-mail: fungi@mail.ru 

Micromycetes (MM) are the best known decomposers of 

different polymeric products, which comprise the main part 

of municipal waste (MW). The research has been focused 

on mycological monitoring of MW composting and was 

first carried out in Russia. The results obtained 

demonstrated that the spontaneous communities of 

micromycetes-colonizers of MW were undergoing seral 

succession and mechanisms of species replacement in 

fungal communities were different. It was established that 

species and biochemical diversity of MM increased to the 

the end of the composting process. Successions of MM 

tended to formation of a stable heterogenic and 

polyfunctional community. This heterogenic climax 

community can withstand the environmental changes and 

can provide the production of a high-quality compost. 

Biologically high-quality compost should serve not only as 

a source of organic matter at soil applications but also as a 

source of soil suppressivness inhancement and biological 

control. Compost should be free from phytotoxic and 

pathogenic species of MM, the microbe community of the 

final product should be stable and include beneficial for 

plants species. However, it was found that final compost 

communities contained species with negative effect on 

plant and human populations. Certain complexes of MM 

possesing positive properties for composting and 

enviroment were suggested for the introduction at the 

definite stages of succession during composting. 

1018 - Habitat differences of coprophilous organisms on 

moose dung 

Å. Nyberg 1* & I.-L. Persson 2 

1 Department of Ecology and Environmental Science, Umeå 

University, SE-901 87, Umeå, Sweden. - 2 Department of 

Animal Ecology, Swedish University of Agricultural 

Sciences, SE-901 83, Umeå, Sweden. - E-mail: 

asa.nyberg@eg.umu.se 

Dung from cervids supports a species-rich community, but 

few studies have been done on forest-living species. In 

order to test whether differences in species composition 

may be explained by habitat-specific differences, we 

studied coprophilous fungi developing on moose dung of 

homogenous origin, composition and age in an area in 

northeastern Sweden. Of the 26 species of fungi found, 12 

species were new to the region, 17 had never been 

observed on moose dung and 2 species were not previously 

described. We found a significant difference in species 

composition between the three habitats studied, with low 

number of species in the spruce forest and about a three 

fold increase in the pine forest and the open mire. Species 

diversity was negatively associated with degree of insect 

attack. This suggests that insects feeding either on the dung 

or the fungi (spores, mycelium) may be an important factor 

explaining the observed pattern. In order to test this 

hypothesis we need experiments including insects 

exclusion. 

1019 - Comparative enumeration of anaerobic fungi 

from different herbivores 

E. Ozkose 1* , G.W. Griffith 2 & D.R. Davies 3 

1 

Sutcu Imam University, Animal Science Department, 

2 

Kahramanmaras, Turkey. - University of Wales 

Aberystwyth, Biological Sciences, Aberystwyth, SY23 3BY, 

3 

U.K. - Institute of Grassland and Environmental 

Research, Animal Science department, Aberystwyth. SY23 

3EB, U.K. - E-mail: eozkose@ksu.edu.tr 

Determination and comparison of anaerobic fungal 

populations from the faeces of cattle, sheep and horses 

were carried out. Similarities and differences, in terms of 

population density between the faeces and rumen digesta of 

same individual were also assessed. The effects of using 

different substrates in the most probable number (MPN) 

dilution series on the population density were also studied. 

The MPN technique was used to enumerate fungal 

populations and counts were expressed as thallus forming 

units (Log 10 tfu g DM -1 ). Mean value of MPN counts 

obtained from fresh faeces were as follow; cattle = 5.61; 

sheep = 4.41 and horse = 4.76. Fungal counts obtained 

from horse and sheep were similar (P > 0.05), whilst cattle 

contained significantly more anaerobic fungal tfu in their 

faeces than either sheep or horse (P < 0.05). The mean 

MPN values of faeces and ruminal digesta of 3 cattle were 

as follows; fresh faeces = 5.22; frozen faeces = 4.92 and 



fresh ruminal digesta = 5.31. MPN values obtained from 

fresh faeces, frozen faeces and fresh ruminal digesta were 

similar (P > 0.05). The mean values from faecal and 

ruminal samples of 3 cattle for different substrates were as 

follow; glucose = 5.11; cellobiose = 5.23; xylan = 4.75; 

wheat straw = 5.53 and cellulose = 5.11. Wheat straw gave 

significantly (P < 0.05) higher viable counts than glucose, 

xylan and cellulose. Population densities, obtained when 

glucose, cellulose and xylan used, were similar (P> 0.05). 

1020 - Ectomycorrhizal fungal dynamics of one- and 

two-year-old Norway spruce seedlings after planting to 

mounded forest clear-cut 

T. Pennanen 1* & J. Heiskanen 2 

1 Finnish Forest Research Institute, Vantaa Research 

Centre, P.O.B. 18, 01301 Vantaa, Finland. - 2 Suonenjoki 

Research Station, 77600 Suonenjoki, Finland. - E-mail: 

taina.pennanen@metla.fi 

Container Norway spruce seedlings were planted on two 

clearcut and mounded Myrtillus type sites in Central 

Finland. Seedlings were planted on mounds and on 

untreated spots between mounds. Seedlings were sampled 

before and one and two growing seasons after planting for 

detailed measurement of root and shoot attributes and for 

morphological and molecular characterization of 

ectomycorrhiza (ECM) in roots grown out into the soil. 

Different morphological groups were subsampled for PCR- 

DGGE analysis of 18S rRNA gene region in order to 

separate ECM in detail. During the first growing season, 

one-year old seedlings grew better in shoot height than the 

two-year-old. However, root egress from containers into 

the surrounding soil was greater in the two-year-old 

seedlings during both first and second summer after 

planting. The outgrown root length and the number of root 

tips per length were greater on mounds than on untreated 

spots. Totally 17 different morphotypes were observed and 

6 of them were present in all treatments. Neither age of the 

seedling nor duration in the field did affect the richness of 

ECM morphotypes but there was a clear temporal shift in 

the proportions of 3 major morphotypes. The smooth white 

type was the most dominant morphotype before outplanting 

but almost disappeared in two years in the field. In 

contrast, the proportion of the smooth brown type increased 

up to 58-78% after two years. The smooth dark brown type 

increased especially on mounds. 

1021 - Pleurotus spp. characterization by microsatellitebased 

DNA fingerprinting 

P. Rapanà 1* , A. Rubini 2 & F. Di Mario 1 

1 CNR IBAF - IBESM, Area della Ricerca di Roma, Via 

Salaria km 29.300; 00016 Monterotondo Sc. (Roma), Italy. 

- 2 CNR IRMGPF, Via della Madonna Alta n.130, 06128 

Perugia, Italy. - E-mail: pompilio.rapana@mlib.cnr.it 

308 


Characterizing fungal species and varieties through 

microsatellite is a powerful approach to evaluate 

biodiversity and genetic relationships among populations. 

Four ISSR (inter simple sequence repeats) primers have 

been utilized to characterize fungal species and varieties 

among Pleurotus genus. Five strains of each 

species/varieties (P. ostreatus, P. eryngii, P. eryngii var. 

ferulae, P. eryngii var. nebrodensis) have been analyzed 

and ISSR data were subject to cluster analysis performed 

with UPGMA method using Dice and Jaccard coefficients. 

This analysis clearly showed two distinctive cluster, one 

related to P. ostreatus and one to P. eryngii var. ferulae, 

whereas less clear relationships appeared between P. 

eryngii and P. eryngii var. nebrodensis. Further 

investigation are in progress to analyze additional 

Pleurotus strains and to test different ISSR primers as well 

as to identify microsatellite-containing amplicons. 

1022 - The polymorphic nature of the rRNA LSU gene 

of arbuscular mycorrhizal fungi (AMF) spores 

A. Rodriguez 1* , J.C. Dodd 2 & J.P. Clapp 3 

1 University of Hohenheim, Universität Hohenheim (340), 

D-70593 Stuttgart, Germany. - 2 Sittingbourne Enterprise 

Hub, Sittingbourne Research Centre, Sittingbourne, Kent 

ME9 8AG, U.K. - 3 University of Kent at Canterbury, 

Canterbury, Kent CT2 7NJ, U.K. - E-mail: aliarodr@unihohenheim.de 

Sequencing of ribosomal RNA genes has been extensively 

exploited in attempts to identify and determine 

phylogenetic relationships and genetic heterogeneity of 

AMF. The heterogeneity of rRNA genes within single 

spores and isolates of AMF has been recognised for several 

years, however the magnitude of this variation has been 

ignored in most taxonomic and phylogenetic analyses. 

Sequence diversity has crucial implications for the 

interpretation of AMF biodiversity and the species concept 

in these fungi. We report here the first investigation of 

genetic diversity in AMF where variation has been 

assessed using sample sizes of several hundred sequences. 

The study concentrated on a well characterised species of 

AMF, Glomus coronatum, but also included three other 

Glomus species with similar spore morphologies. In total 

435 clones containing the LSU D2 regions from 7 isolates 

of G. coronatum (80% of isolates currently available) as 

well as isolates of Glomus mosseae, Glomus geosporum 

and Glomus constrictum, were screened for sequence 

variation using a PCR-single strand conformational 

polymorphism (PCR-SSCP) technique. The variation 

encountered was higher than expected, 138 representative 

sequences were obtained and analysis indicated that most 

were unique. This variation could not be attributed to DNA 

polymerase or cloning artefacts. Implications for the 

species concept in AMF, the use of rRNA sequences to 

estimate biodiversity and in situ detection in field ecology 

are discussed.


1023 - Temporal distribution of ectomycorhizae in 

Quercus rubra L. plantations of the Basque Country 

and evaluation of their infective potential 

N. Rodriguez, I. Salcedo * , E. Sarrionandia & M.K. 

Duñabeitia 

Dpt. Plant Biology and ecology. University of the Basque 

Country, Apdo. 644. 48080 Bilbao, Spain. - E-mail: 

gvpsalai@lg.ehu.es 

Due to its fast growth and the commercial value of the 

wood, Q. rubra has been used in many Basque Country 

plantations which have been located in Q. robur L. 

habitats. The evaluation of infection ability and density of 

the mycorhizal propagules is one of the main factors to 

take into account in reforestation and restoration 

programmes (Kendrick 1988). The principal aim of this 

study is to explore the temporal variation throughout a year 

(2000-01) of the species richness and diversity of 

ectomycorhizae in six localities. Likewise, greenhouse's 

experiments were carried out in order to determine the 

infective potential of the forest soils. Maximum values of 

diversity were recorded during autumn and winter; whereas 

the minimum was in summer. The ubiquitous Cenococcum 

geophilum is one of the few morphotypes that appears 

throughout the year. According to the mycorhization rate 

biometric and physiological variables of the seedlings 

grown in soil of different localities of Q. rubra forests were 

determined. A positive correlation has been found between 

mycorhization rate and measured variables. 

1024 - Growth promoting effects of endophytic 

colonisation of the roots of larch seedlings (Larix 

decidua) with Cryptosporiopsis sp. and Phialophora sp. 

A.-K. Römmert 1* , M. Oros-Sichler 2 , T. Lange 3 , H.J. Aust 1 

& B. Schulz 1 

1 

Institut für Mikrobiologie der Technischen Universität 

Braunschweig, Spielmannstr. 7, 38106 Braunschweig, 

Germany. - 2 Biologische Bundesanstalt für Land- und 

Forstwirtschaft, Messeweg 11-12, 38104 Braunschweig, 

Germany. - 

3 Botanisches Institut der Technischen 

Universität Braunschweig, Mendelssohnstr. 4, 38106 

Braunschweig, Germany. - E-mail: ak.roemmert@tu-bs.de 

Symbiotic associations of endophytic fungi with their hosts 

vary from mutualistic to latently pathogenic. In order to 

discern the nature of endophytic infections of the roots, 

those of axenically cultured larch seedlings were inoculated 

either with one of two endophytes, Cryptosporiopsis sp. or 

Phialophora sp., previously isolated from roots of larch, or 

with the pathogen Heterobasidion annosum. All three 

isolates colonised the roots systemically and both inter- and 

intracellularly. Whereas colonisation with each of the fungi 

significantly improved growth of the seedlings in 

comparison to the non-inoculated control, the greatest 

improvement resulted from endophytic colonisation. 

However, in contrast to colonisation with the endophytes, 

infection with H. annosum also led to development of 

disease symptoms. Application of a mycelium extract of 

Phialophora sp. to roots of the host also improved growth 

of the roots and seedlings. The capability of all three fungi 

to produce IAA may explain the growth enhancing effects 

of the mycelium extract and of colonisation. Additional 

virulence factors may be responsible for pathogenicity of 

H. annosum. In conclusion, endophytic colonisations of the 

roots of larch with Cryptosporiopsis sp. or Phialophora sp. 

are mutualistic symbioses and we hypothesise that 

endophytic colonisation of the roots, in contrast to that of 

the shoots, is generally systemic and mutualistic1. 1Boyle, 

Götz, Dammann-Tugend & Schulz (2001). Symbiosis 31: 

259-281. 

1025 - Spatial and temporal genetic variation among 

arbuscular mycorrhizal fungi from undisturbed 

vegetation 

S. Rosendahl 

Department of Mycology, University of Copenhagen, Oster 

Farimagsgade 2D, DK-1353 Copenhagen K, Denmark. - 

E-mail: soerenr@bot.ku.dk 

Mycorrhizal roots from Hieracium pilosella in a Danish 

coastal grassland were sampled in May, August, October 

and November along a 50 m transect. Arbuscular 

mycorrhizal fungi in the roots were characterised from 

partial LSU rDNA sequences obtained by nested PCR. In 

the specific PCR, the primers were designed to amplify a 

clade within Glomus including species as G. mosseae, G. 

caledonium, and G. intraradices. Three sequence types 

were found to be common. One type showed high 

similarity with Glomus intraradices, whereas the other 

could not be grouped with known AM species. The 

sequences showed some variation, but the same sequence 

could be sampled from several plants along the transect. 

Most of the variation in sequence types could be attributed 

to the sampling time. The results showed that the high 

degree of sequence variation reported from spores, could 

not be found in roots from undisturbed vegetation systems. 

1026 - A novel cotton swab sampling method for fungal 

spores 

C. Roux 1 , P.C. van der Merwe 1 , M. Smith 1 & I.H. Rong 2* 

1 Mycology Unit, Biosystematics Division, ARC - Plant 

Protection Research Institute, Private Bag X134, Pretoria 

0001, South Africa. - 2 ARC - Biometry, Private Bag X519, 

Silverton 0127, South Africa. 

A sampling method for continual monitoring of fungal 

spores, from various surfaces, such as bins in silos, 

harvesters and other farm implements has been developed 

with the use of standard cotton swabs (Johnson & Johnson 

Cotton balls TM ). The swabs are used to lift all spores from 

the surfaces sampled. The swabs are then washed with a 



surfactant and, if necessary, washed through a series of 

sieves to eliminate particles of irrelevant sizes. The residue, 

in the form of a pellet, is obtained by centrifugation. The 

top surface of the pellet is mounted on microscope slides 

(usually 2) to which a drop or two of lactophenol has ben 

added. The slides are sealed with nail varnish and studied. 

With this method one can monitor the presence of 

particular spores and diseases in bulk grain can 

consequently be monitored successfully. Spores from 

diseases that are dificult to detect in the field, as well as 

other fungi present in the grain and particularily smuts and 

rusts, can be confirmed using this technique. This method 

could be extremely cost effective as the samples can be 

taken by a lay parson, sent by post and analysed. The 

overhead expenses are kept to a minimum. The only 

drawback is that a surface can only be sampled once, 

giving a retrieval rate of about 90%, which is ideal for 

monitoring compared with other grain washing methods. 

1027 - Ceratocystis albofundus, an African fungus 

causing a wilt disease on exotic Acacia mearnsii 

J. Roux 1* , G. Nakabonge 1 , I. Barnes 1 , B.D. Wingfield 2 & 

M.J. Wingfield 1 







jolanda.roux@fabi.up.ac.za 

Ceratocystis albofundus is a relatively newly discovered 

fungus that causes a rapid wilt disease of economically 

important Acacia mearnsii trees. Until recently it was 

known only from South Africa, on exotic Australian 

Acacia spp. and on native Protea spp. In 1999, the fungus 

was discovered in Uganda and a year later, it was also 

found in Kenya. In both cases it was associated with 

vascular wilt of A. mearnsii trees. Although based on only 

three records, C. albofundus has been found on native 

Protea spp. in South Africa, leading to the hypothesis that 

it is native to this country. Recent studies on the population 

diversity of South African isolates using a range of 

microsatellite markers, have shown high levels of genetic 

diversity in C. albofundus, similar to that found in other 

native Ceratocystis spp. The genetic diversity of a Ugandan 

population has also been studied, showing that the 

Ugandan population has a similar diversity to that in South 

Africa. These studies lead us to believe that C. albofundus 

is native to the African continent. It appears to have 

acquired the ability to infect exotic Acacia spp., causing 

serious losses to the forestry industry. Surveys of native A. 

mearnsii in Australia have failed to yield C. albofundus 

and we do not believe that it is present in that country. Our 

studies suggest that the fungus poses a serious threat to A. 

mearnsii in its native range and that quarantine measures 

should be imposed to prevent its spread to Australia. 

310 


1028 - Endophytic fungi from Cassia fistula L. 

N. Ruchikachorn 1* , A.J.S. Whalley 1 & P. Sihanonth 2 


University, Byrom Street. Liverpool, L3 3AF, U.K. - 


Bangkok, 10330, Thailand. - E-mail: 

bmsnruch@livjm.ac.uk 

Cassia fistula, known as Khoon (Thai) or Golden Shower, 

is a deciduous tree which is found all over Thailand. It is 

often planted as an ornamental, but is also an important 

medicinal plant. Historically all parts of the plant have 

been used in native remedies. The leaves of the plant are 

used for curing boils, coughs and skin diseases such as 

ringworm while the roots can be used to treat snake bites 

and dysentery. During the current study it has been found 

to contain a wide range of fungal endophytes. Isolates from 

three separate geographical locations in Thailand are 

compared and the data discussed in relation to specific 

habitat. A significant number of isolates have been 

recognised as members of the Xylariaceae and 

identification of a number of these has been confirmed 

following teliomorph induction. Testing of the biological 

activity of a selection of the isolates indicates antibacterial 

ability and, in a few cases, antifungal. Herbicidal and 

insecticidal activity has also been demonstrated in a 

number of cases. We have recently identified 

fluoroglucinol as a major metabolite from one of the 

isolates and this has been shown to exhibit antifungal 

activity. 

1029 - Ectendomycorrhizal communities of Pinus 

sylvestris seedlings from forest nurseries in Poland 

M.L. Rudawska * & T. Leski 

Institute of Dendrology, Polish Academy of Sciences, 

Parkowa 5, 62-035 Kornik, Poland. - E-mail: 

mariarud@man.poznan.pl 

Scots pine is the most common tree produced in bare-root 

nurseries in Poland. Mycorrhizas present on 1-0 and 2-0 

Pinus sylvestris seedlings produced in bare-root nurseries 

situated in north-west part of Poland were estimated. Most 

seedlings were spontaneously colonized by naturally 

present ecto- and ectendomycorrhizal symbionts. However 

ectendomycorrhizal colonization very often exceeded the 

occurrence of ectomycorrhizas. Predominance of 

ectendomycorrhizas was significantly correlated with 

nitrogen fertilization. Fungal communities associated with 

ectendomycorrhizas of tested seedlings were assessed by a 

combination of morphological and molecular techniques. 

Morphological typing of mycorrhizas based on the 

structure and colors of their external hyphae, fungal mantle 

and anatomical analysis distinguished several 

ectendomycorrhizal morphotypes. To identify these 

mycorrhizas, fungal rDNA was amplified using polymerase 

chain reaction and digested with endonucleases (HinfI,


MboI, TaqI). Morphologically variable complex of 

ectendomycorrhizal morphotypes consisted from at lest 

three symbionts giving different RFLP pattern. The fungal 

taxa involved as dominating ectendomycorrhizal symbiont 

were ascomycetes Wilcoxina sp. Investigation is under way 

to clarify the systematical position of all 

ectendomycorrhizas found. 

1030 - Diversity and function of ectomycorrhiza during 

transformation of forests 

M. Rumberger 1* , F. Ehrig 2 , P. Lentzsch 3 , B. 

Münzenberger 3 & R.F. Hüttl 1 

1 Brandenburg University of Technology, Chair of Soil 

Protection and Recultivation, P.O. Box 101344, 03013 

Cottbus, Germany. - 2 Federal Centre fro Breeding 

Research on Cultivated Plants (BAZ), Institute of 

Resistence Research and Pathogen Diagnostics (IRP), 

Theodor-Roemer-Weg 4, 06449 Aschersleben, Germany. - 

3 Centre for Agricultural Landscape and Land Use 

Research (ZALF) e.V., Institute of Primary Production and 

Microbial Ecology, Eberswalder Strasse 84, 15374 

Müncheberg, Germany. - E-mail: rumberg@zalf.de 

During the transformation of forests biological niches are 

created that are thought to result in enhanced biological 

variety. In the present study diversity and function of 

mycorrhizas were investigated in a pure stand of Scots pine 

(Pinus sylvestris L.), in a chronosequence of Scots pine 

stands that were underplanted with beech (Fagus sylvatica 

L.) and in a pure beech forest. Mycorrhizal diversity was 

higher in the pure stand of beech compared to the pure 

stand of Scots pine. Beech trees in the mixed forests had 

dominating morphotypes that were very similar to the ones 

dominating in Scots pine. Therefore, it seems plausible that 

mycorrhiza coenosis of Scots pine is influencing the 

coenosis of beech in Scots pine stands being transformed 

into mixed stands of Scots pine and beech. However, trees 

in the pure beech stand were mycorrhized with other types. 

These types were distributed in even dominances. Along 

the chronosequence, frequency per soil volume and 

nutrient amount of beech mycorrhizas increased. Nutrient 

uptake was higher for mycorrhizas of older beech than for 

mycorrhizas of older pine. This indicates that older beech 

trees are able to accumulate specific nutrients in the upper 

mineral soil and the forest floor layer. This observation 

supports the hypothesis of the so-called 'base-pump effect' 

of the older beech trees. 

1031 - PCR analysis reveals diverse fungal community 

in smut sori of Anthracoidea spp. on host plants, Carex 

spp. 

V. Salo 1* & R. Sen 2 

1 University of Helsinki, Department of Applied 

Biology/Botany, P.O.Box 27 (Latokartanonkaari 5), FIN- 

00014 University of Helsinki, Finland. - 2 University of 

Helsinki, Department of Biosciences, Division of General 

Microbiology, P.O.Box 56 (Viikinkaari 9), FIN-00014 


vanamo.salo@helsinki.fi 

Twelve Anthracoidea populations representing six species 

were collected from natural populations of Carex spp. in 

Finland. The aim of the study was to use RFLP markers 

from the rDNA ITS region to complement a previous study 

on isozyme variation of these species. Twelve smut sori 

from different inflorescences in each population were 

examined under a light microscope before DNA extraction 

from teliospores and PCR amplification. Six PCR samples 

per population were analysed using three restriction 

enzymes. The results revealed considerable variation 

between sori in size and number of PCR products and 

RFLP patterns. Also the size of RFLP bands often 

exceeded the size of the original PCR product. All samples 

were analysed in high resolution gels which showed that 

many single bands were actually two bands almost of 

similar size. A set of samples producing one and multiple 

bands was chosen for further analysis. Single bands were 

isolated, cloned and sequenced. ITS sequences were 

compared to sequences in the GenBank database. Most 

bands pointed to totally different organisms than smuts, 

viz. Cladosporium spp. together with some other fungi. 

None of these fungi has previously been found in 

association with smuts. True Anthracoidea sequences 

turned out to be the longest ones detected and are among 

the longest fungal ITS sequences reported. The nature of 

the previously unknown fungal community in 

Anthracoidea sori, and possible relation to the smut fungus, 

will be discussed. 

1032 - Homothallism in Cordyceps militaris 

H. Sato * & M. Shimazu 

Forestry and Forest Products Research Institute, Kukizaki, 

Ibaraki 305-8687, Japan. - E-mail: 

hirokis@ffpri.affrc.go.jp 

Strains from mono-ascospore, mono-part-spore and monoconidium 

of Cordyceps militaris produced stromata from a 

species of lepidopteran pupae. Hyphal bodies (yeast-like 

vegetative stage) of Cordyceps militaris were injected into 

the pupal haemocoel of Spodoptera litura and stromata 

formation was observed. C. militaris were isolated in July 

1995, the season of the stromata production of this fungus, 

from the air by placing agar plates in a beech forest. 

Originally, four isolates were used for experiments. Hyphal 

bodies of the four isolates were injected to the host pupae 

independently, and four mono-ascospore-strains were 

established from their stromata. Three out of the four 

mono-ascospore strains produced stromata. By re-isolation 

from a stroma-produced-strain in the experiment above, 

five mono-ascospore-strains and three mono-part-sporestrains 

were established. The two out of the five (monoascospore) 

and all the three strains (mono-part-spore) 

produced stromata. Seven mono-conidium strains were 

established from the two among the original four isolates. 

Four out of the seven strains produced stromata. Both a 

part spore and a conidium contained a nucleus by 

ultrastructural observation. From these findings, we 



considered that C. militaris has homothallism. For this 

entomopathogenic fungus, homothallism is thought to be 

effective character for producing stromata against rare 

chance to encounter host insects. 

1033 - Some saprobic fungi from Typha and their 

competitive interactions on leaves 

M.J. Schulz 1* , M.N. Thormann 1 , E.A. Smreciu 2 & R.S. 

Currah 1 

1 University of Alberta, Dept. of Biological Sciences, 

Edmonton, AB T6G 2E9, Canada. - 2 Wild Rose Consulting, 

Inc., 15109 77th Ave, Edmonton AB T5R 3B5, Canada. - Email: 

mkschulz@hotmail.com 

Typha latifolia is a major detritus producer in many 

wetlands. A diverse group of fungi decompose this matter, 

but how they work to partition the substrate spatially and 

chemically is unknown. Psathyrella typhae, Talaromyces 

ucrainicus, Hymenoscyphus repandus, H. scutula, 

Hymenopsis typhae, Phoma sp., Cladosporium herbarum 

and Sclerotium cf. hydrophilum were isolated from Typha 

leaves at various stages of decomposition, and their 

enzymatic abilities tested using a suite of in vitro tests. 

Corresponding mass loss determinations using natural 

substrate showed little correlation for many species, 

indicating in vitro tests may not accurately represent 

decompositional abilities in vivo. We also looked at 

competitive interactions between two common saprobes 

from T. latifolia, P. typhae and Sclerotium cf. 

hydrophylum, by using differential morphology 

(presence/absence of clamps, hyphal diameter) and 

enzymatic abilities (pH tolerance and ability to acidify 

casamino acid medium) to disinguish their relative 

development in co-inoculated Typha leaf fragments. Even 

though Sclerotium grew faster, P. typhae was the superior 

competitor, able to inhibit and invade substrate already 

colonized by Sclerotium. 

1034 - From orchids (Neottia nidus-avis) to forest tree 

roots: the mycorrhizal web of the Sebacinaceae 

(heterobasidiomycetous 'Rhizoctonia') 

M.-A. Selosse 1* , M. Weiß 2 , R. Bauer 2 & A. Tillier 1 

1 Institut de Systématique (IFR CNRS 1541) du Muséum, 43 

rue Cuvier, F-75005 Paris, France. - 2 Botanisches Institut, 

Universität Tübingen, Auf der Morgenstelle 1, D-72076 

Tübingen, Germany. 

Mycoheterotrophic (MH) plants are achlorophyllous 

organisms fed by fungi. Up to now, only 

homobasidiomycetous ectomycorrhizal fungi have been 

found in MH plants, while several MH orchids are 

probably associated to Rhizoctonias, 

heterobasidiomycetous fungi that are often parasitic or 

saprobic. We investigated the MH Neottia nidus-avis, a 

European orchid supposedly associated to a fungus of the 

312 


Rhizoctonia complex. 61 Neottia root systems were 

analysed by amplification and sequencing of the fungal 

rDNA, showing a specific association to a subclade of 

Sebacinaceae. Ectomycorrhizae (ECM) growing near 

Neottia roots were shown, through rDNA sequencing, to be 

also colonized by the sebacinoid associates of the orchid 

(83% of the typed ECM). Additional data suggests that the 

same genets colonize the Neottia and neighbouring tree 

roots. Neottia therefore confirms the emerging features of 

MH plants, specifically associated with tree-colonizing 

fungi. Up to now, available data did not exclude that 

sebacinoids are saprobic or mycoparasites on ECM. Our 

TEM analysis shows that they form typical ECM, with 

dolipore ultrastructure supporting the molecular 

identification of the fungus. Sequencing of the rDNA ITS 

of the host plant showed that sebacinoids are not treespecific, 

colonizing Carpinus, Fagus, Quercus, Tilia, etc. 

This, together with some GenBank data, suggests that 

Sebacinaceae is an overlooked mycorrhizal family, where 

many species await morphological description. 

1035 - Rare mechanism of genetic recombination in 

Pleurotus ostreatus natural populations 

A.V. Shnyreva 

Department of Mycology & Algology, Faculty of Biology, 

Moscow State University, Vorob'yevy Gory, Moscow 

119992, Russia. - E-mail: shnyreva@herba.msu.ru 

The higher basidiomycete fungi form panmictic 

populations resulting from haploid progeny hybridization. 

However, a dikaryon is also capable of contributing 

fertilizing nuclei to a haploid monokaryon resulting in a 

new dikaryon. Little is known about occurring diploidhaploid 

mating in nature due to rather difficulties in 

observing such phenomenon. Having analysed Pleurotus 

ostreatus population structure, we have revealed rather rare 

mechanism of genetic recombination via di-mon-mating in 

a local population. We have analysed a sample of 

heterokaryotic isolates collected within a log. All the 

heterokaryotic isolates within an individual substrate were 

somatically compatible and identical at 12 polymorphic 

isozyme loci. To examine distribution of mating 

incompatibility factors among basidiocarps within a single 

fruit body cluster on a log, matings between their 

monospore offspring were carried out. Some of the 

dikaryons were identically constituted with respect to A 

and B factors resulted in 25% of compatible pairings, while 

the others were distinguished at one of the two mating loci 

(75% of compatible pairings). This can be explained by 

occurring diploid-haploid hybridization. Dikaryotic 

individual occupying the substrate fuses with alien 

monospore germling that leads to random combination of 

three types of nuclei in cells of different basidiocarps 

within a fruit body cluster. Data on population 

structure,reproductive strategy will be also presented. 

RFBR grant 01-04-49447.


1036 - Evaluating colonization of ECM fungi at Quercus 

petraea seeds, seminal roots and at the roots of 

greenhouse seedlings 

A. Silberstein 1* , S. Kotter 2 , J. Wöllecke 1 , B. 

Münzenberger 3 & R.F. Hüttl 1 

1 

Brandenburg University of Technology Cottbus, Chair of 

Soil Protection and Recultivation, Universitätsplatz 3-4, 

03044 Cottbus, Germany. - 2 Tinplant Biotechnics and 

Plant Propagation Inc., Magdeburger Str. 33, 39164 Klein 

Wanzleben, Germany. - 

3 Center for Agricultural 

Landscape and Land Use Research (ZALF) e.V., Institute 

of Primary Production and Microbial Ecology, 

Eberswalder Str. 84, 15374 Müncheberg, Germany. - Email: 

silber@tu-cottbus.de 

In order to recultivate postmining landscapes, to diversify 

existing coniferous forests and to re-afforest uneconomic 

agricultural areas millions of young oak tree plants 

(Quercus spec.) are needed. But we are faced with many 

problems when cultivating the young oak plants. This is, 

above all, true as regards the germinating process of the 

acorns, as their seminal roots are often infected by 

pathogenic fungi. The plant raising enterprises are, 

therefore, facing huge costs. The project is accomplished 

with the Tinplant company that is specialized in the raising 

of containerized greenhouse seedlings of Q. petraea and 

aims at evolving and testing simple and economical 

methods of mycorrhization of oaks. To utilize the positive 

effects of ECM fungi at a very early stage of plant 

development, the hyphae should be established at the 

stored or germinating seeds already. Plant raising 

enterprises could look forward to better results during the 

seed storage and germinating stages, as compared to 

existing procedures. Recultivating companies could expect 

a lower plant mortality rate after oak tree afforestation, by 

making use of ectomycorrhizal fungi, suitable to the 

respective location. 

1037 - Population structure and pathogenicity of 

Fusarium oxysporum isolated from soil and root 

necrosis of pea 

K. Skovgaard 1* , L. Bødker 2 & S. Rosendahl 1 

1 

Department of Mycology, University of Copenhagen, 

Oester Farimagsgade 2D, 1353 Copenhagen K, Denmark. 

2 

- Danish Institute of Agricultural 

Sciences, Research 

Centre Flakkebjerg, Dept. of Crop Protection, 4200 

Slagelse, Denmark. - E-mail: kerstins@bot.ku.dk 

Forty-nine strains of Fusarium oxysporum were isolated 

from five different sample locations within two 

neighbouring pea fields. Of these, thirty-nine strains were 

isolated from soil and ten from pea plants showing 

symptoms of root rot. Twenty-eight of the isolates were 

tested for pathogenicity towards pea. Based on percentage 

discoloration of the roots and the stem base the isolates 

were divided into three groups. Seven strains were 

pathogenic, fourteen strains were weakly pathogenic, and 

seven strains were non-pathogenic towards pea. To assess 

the genetic relatedness of all forty-nine strains, gene 

genealogies were constructed from aligned DNA sequences 

from part of translation elongation factor, nitrate reductase, 

beta tubulin, and mitochondrial small subunit rDNA. 

Maximum parsimony analysis of the combined dataset 

yielded a single most-parsimonious tree containing three 

strongly supported clades which may represent cryptic 

species. No correlation was observed between the 

multigene phylogeny and pathogenicity toward pea, strain 

geographic origin and substrate (soil or plant) from which 

the strains were isolated. Strains of F. oxysporum that were 

either non-pathogenic, weakly pathogenic or pathogenic 

sometimes shared the same multilocus genotype. These 

results suggest that strains pathgenic and putatively nonpathogenic 

to pea are very closely related genetically. 

1038 - Increased below-ground diversity of 

ectomycorrhizal fungi after removal of litter and humus 

determined from fungal hyphae in bulk soil 

E. Smit 1 , C. Veenman 1 & J. Baar 2* 

1 National Institute of Public Health and the Environment, 

Microbiological Laboratory for Health Protection, P.O. 

Box 1, 3720 BA Bilthoven, The Netherlands. - 2 Applied 

Plant Research, Wageningen University and Research 

Center, P.O. Box 6042, 5960 AA Horst, The Netherlands. - 

E-mail: j.baar@ppo.dlo.nl 

Ectomycorrhizal fungi have declined in coniferous stands 

in The Netherlands over the last decades due to nitrogen 

deposition originating from air pollution. The effects of 

restoration practices on the ectomycorrhizal communities 

in nitrogen-enriched Pinus sylvestris (Scots pine) stands in 

The Netherlands were studied by Baar (1995). Litter and 

humus were removed to reduce nitrogen availability in the 

soils. This resulted in increased diversity of 

ectomycorrhizal fungi within three years. However, all data 

were obtained by counting sporocaps. The effects of litter 

and humus removal on the below-ground ectomycorrhizal 

communities remained unknown. In the present study the 

composition of the below-ground ectomycorrhizal 

communities in a P. sylvestris stand in The Netherlands 

was determined five years after litter and humus removal. 

Fungal DNA was extracted directly from soil samples and 

PCR was applied with basidiomycete-specific ITS primers. 

Samples were analyzed by DGGE to visualize diversity. To 

assess species composition, ITS regions of the amplified 

fragments were cloned and sequenced. Sequences were 

compared with known sequences and were analyzed 

phylogenetically. The analyses showed that the diversity of 

the ectomycorrhizal fungi below ground in the treated plots 

was higher than in the untreated plots. Rhizopogon luteolus 

was the most abundant species after litter and humus 

removal, while Lactarius and Russula species were the 

most common species in the untreated plots. 



1039 - Molecular ecology of Bistorta vivipara 

ectomycorrhiza in alpine tundra communities 

J.H. Sønstebø * , K. Høiland & T. Schumacher 

University of Oslo, Department of Biology, Division of 

Botany and Plant Physiology, P.O. Box 1045 Blindern, 

0316 Oslo, Norway. - E-mail: jornso@bio.uio.no 

We have studied the diversity and phylogenetic affinities of 

mycobionts of Bistorta vivipara ectomycorrhizas in four 

alpine tundra plant communities in Norway, using ITS1-5.8 

sequences as a diversity marker. ITS1-5.8 sequences 

obtained by direct sequencing of 114 ectomycorrhizal root 

tips compared with sequences from known fungal taxa 

showed a high diversity of fungal mycobionts, including 

mycobionts from a number of distantly related taxonomic 

groups. The composition and taxonomic specificity of the 

ectomycorrhizal fungi varied considerably among the study 

sites. This suggests that B. vivipara is capable to interact 

with a number of ectomycorrhizal fungal partners in its 

many divergent habitats of the lowlands as well as in the 

alpine zone. Frequent members of the B. vivipara 

ectomycorrhizal symbiosis include Tomentella, Russula, 

Inocybe, Cortinarius, Laccaria, Hebeloma, Cenococcum 

and some unidentified members of Sebacinaceae. The 

observed high abundance and diversity of Tomentella 

mycobionts add to the earlier observations of importance of 

this genus as ectomycorrhizal former also in boreal forest 

ecosystems. The sebacinaceous ectomycorrhizal fungal 

partners showed affinity (95% similarity) to orchid 

mycorrhizal isolates from root tips of Neottia nidus-avis 

retrieved from GenBank. 

1040 - Analysing the genetic structure of Botrytis spp. in 

lily and tulip fields using molecular markers 

M. Staats * , P. van Baarlen & J.A.L. van Kan 

Wageningen University, Laboratory of Phytopathology, PO 

Box 8025, 6700 EE Wageningen, The Netherlands. - Email: 

Martijn.Staats@fyto.dpw.wau.nl 

Botrytis spp. that are pathogenic on flower bulb crops are 

generally assumed to be host-specific. On lily, B. elliptica 

produces primary necrotic lesions which potentially expand 

into healthy tissue. On the non-host tulip, however, B. 

elliptica can only cause primary necrotic lesions. The 

reciprocal holds true for B. tulipae, which uses tulip as its 

host. The mechanistic and genetic basis of host specificity 

of the two Botrytis species mentioned is unclear. 

Distinction between the species thus far relies on 

morphological characteristics. At this moment it is 

unknown to what extent sexual reproduction contributes to 

genetic variation in local populations of Botrytis spp. in lily 

and tulip fields. A sexual stage has so far been detected in 

B. elliptica but not in B. tulipae. The purpose of this study 

is to develop DNA markers to characterise the level of 

inter- and intraspecific heterogeneity and the amount of 

genetic recombination in B. elliptica and B. tulipae field 

314 


isolates. To reach these objectives we will use a 

combination of sequence analysis and novel DNA 

fingerprinting methods. - This research is funded by the 

Dutch Technology Foundation STW. 

1041 - Molecular markers associated with virulent and 

avirulent phenotypes in populations of Uromyces 

appendiculatus from bean fields in the Central USA 

J.R. Steadman * , A.T. Alleyne, J.G. Fenton & K.M. 

Eskridge 

University of Nebraska, 406 Plant Sciences, Lincoln, NE 

68583-0722, U.S.A. - E-mail: jsteadman1@unl.edu 

Uromyces appendiculatus, cause of bean rust, is 

characterized by highly diverse virulence phenotypes. The 

use of race-specific rust resistance genes has been 

complicated by the 'boom and bust' cycle. Pinto Olathe was 

released as a rust resistant bean cultivar containing the Ur- 

6 gene in 1979. The resistance only lasted 5 years. 

Resistance gene pyramiding is a strategy to strengthen 

race-specific resistance. Phenotyping the rust population is 

time-consuming and costly. Pathogen variation can also be 

monitored by genotyping. Previous work on molecular 

variation in rust pathotypes have examined geographic 

significance of rust races and the association between 

resistance and virulence patterns. In the present study we 

monitored the changing pattern of virulence on pinto 

'Olathe' over the past 20 years using both molecular and 

phenotypic markers. A population of over 130 isolates 

from the Central USA representing both avirulent and 

virulent pathotypes on pinto Olathe were analyzed using 

molecular markers such as rep-PCR and RAPD-PCR. 

These markers were found to discriminate between rust 

pathogen genotypes with virulent or avirulent interactions 

with the Ur-6 gene. In addition, cluster analysis of the 

molecular data using UPGMA methods showed agreement 

between statistical analysis and cluster associations of these 

pathotypes with selected markers. 

1042 - Annual production of leaf-decomposing fungi in 

streams 

K. Suberkropp 1* , B.R. Methvin 1 , M.D. Carter 2 & H.S. 

Weyers 3 

1 University of Alabama, Tuscaloosa, AL, U.S.A. - 2 Noth 

Carolina State University, Raleigh, NC, U.S.A. - 3 U. S. 

Geological Survey, Dover, DE, U.S.A. - E-mail: 

ksuberkp@bama.ua.edu 

We have determined the annual production of fungi 

associated with decomposing leaves in seven streams in the 

southeastern United States using rates of acetate 

incorporation into ergosterol to calculate in situ growth 

rates and ergosterol concentrations to estimate fungal 

biomass. Annual fungal production ranged from 8-551 

g/m 2 in these streams. Fungal production was significantly


correlated with the mean amount of leaf litter present in the 

streams throughout the year (r=0.72, p


1046 - Structure of mycobiota and organic matter 

decomposition in undisturbed biogeocenoses of the 

southern taiga 

V.A. Terekhova 

Institute of Soil Science, Lomonosov Moscow State 

University and Russian Academy of Sciences, Vorob'evy 

Gory, Moscow, 119899, Russia. - E-mail: vat@soil.msu.ru 

The structural and functional peculiarities of soil 

microscopic fungi in typical biogeocenoses of the southern 

taiga have been characterised. Mycological analysis has 

been carried out in the Central Forest State Biosphere 

Reserve (CFSBR, Tver oblast, Russia). The information on 

taxonomic diversity of mycobiota has been obtained using 

the plating method and description of microscopic fungi 

species from different plant material, layers of litter and 

soil profile horizons in sphagnum spruce forest, bilberry 

spruce forest, wood sorrel-nemoral spruce forest, nemoral 

spruce forest, black alder grassy-swampy spruce forest. 

The soils of biogeocenoses differ significantly in total 

reserves of the organic carbon, rate of organic matter 

decomposition. The data obtained show high fungal 

diversity (Shannon index) in all biogeocenoses, but 

seasonal changes in this index are much greater than those 

related to the substrate type. The increasing of the rate of 

organic matter mineralization and fungal diversity have 

been observed only under optimal temperature and water 

conditions. We have tried to estimate the role of organic 

matter composition, and interspecies relations on the 

destructive activity of fungi in model laboratory 

experiments. Using cross introduction it has been shown 

quite high adaptation of fungi community to the 'native' 

decomposed substrate. The research has been supported by 

Russian Foundation for Basic Research grant 02-04-48870. 

1047 - Nematode-trapping fungi in Victoria Land 

(Continental Antarctica) 

S. Tosi * , G. Caretta & G. Del Frate 

Dipartimento di Ecologia del Territorio e degli Ambienti 

Terrestri, Sezione di Micologia, Università degli Studi di 

Pavia, via S. Epifanio, 14, 27100, Pavia, Italy. - E-mail: 

stosi@et.unipv.it 

Nematode-trapping fungi have a world wide distribution. 

They were also found in Antarctica. In the present work 

nematode-trapping fungi isolated in Victoria Land 

(Continental Antarctica) are reported. Fortythree samples 

of soil, mosses and ornithogenic materials were collected 

during 1987-1999. The investigated area ranges from 74°S 

to 76°S and 162°E to 165°E. Nematode-trapping fungi 

were isolated by means of sprinkled-plate technique. A 

suspension of the nematode Caenorhabditis elegans was 

added as a bait. Predaceous hyphomycetes were isolated 

from twelve samples. Thirty strains of Arthrobotrys spp. 

and one strain of Nematoctonus were isolated and 

identified. All Arthrobotrys strains were morphologically 

316 


similar; differences were noted in the general features of 

the colony (colours, zonations). They are able to capture 

namatodes by means of adhesive nets. The strain of 

Nematoctonus is the first record for Maritime and 

Continental Antarctica. It is characterized by the presence 

of clamp connections on its hyphae, typical of many 

heterocaryotic Basidiomycota. It produces adhesive 

trapping devices formed by a secretory cell with a 

characteristic hour-glass shape. The results here reported, 

show that nemade-trapping fungi are abundant in 

Continental Antarctica, above all in the moss where there 

are nematodes and other small organisms. On the base of 

their massive presence and high grade of specialization, 

they can be considered indigenous fungi of the Antarctica. 

1048 - In vitro mycorrhization of Cistus incanus 

seedlings by Tuber melanosporum mycelium 

Y. Ventura 1* , V. Kagan Zur 1 , A. Bustan 1 , D. Mills 1 & N. 

Roth-Bejerano 2 

1 Ben Gurion University of the Negev, Institutes for Applied 

Research, P.O. Box 653, Beer Sheva 84105, Israel. - 2 Ben 

Gurion University of the Negev, Department of Life 

Sciences, P.O. Box 653, Beer Sheva 84105, Israel. - Email: 

yventura@bgumail.bgu.ac.il 

Establishment of mycorrhizal associations in vitro between 

Cistus incanus and Tuber melanosporum strain 1015 was 

studied. Cistus incanus, a Mediterranean flowering plant 

(Cistaceae), is a known symbiont of the black truffle. It can 

easily be germinated and rooted in vitro. Three growth 

media were evaluated for plant development, fungal 

growth, and mycorrhization: (i) N5, an MS medium 

containing one fifth the nitrogen content of the standard 

version, (ii) M, a minimal medium developed for in vitro 

mycorrhization of VAM, and (iii) half-strength Hoagland 

solution. All media were solidified with 0.8% agar. No 

significant differences were found between the media in 

respect to plant development (height, number of nodes and 

dry weight). The tuber mycelium grew vigorously on 

medium M, while on half-strength Hoagland and N5 its 

growth was very significantly reduced (by 62.5% and 

84.6%, respectively). The incidence of Hartig net 

establishment was close to 100% on medium M, versus 

10% and zero on half-strength Hoagland and medium N5, 

respectively. Direct irradiation with light as low as 25 µmol 

m -2 sec -1 strongly inhibited T. melanosporum mycelium 

growth and mycorrhization failed to occur, whereas diffuse 

light showed no effect on fungal development and enabled 

mycorrhiza formation. It appears that in vitro growth of 

Tuber mycelium is affected by the nature of the medium 

and by light intensity and correlates with mycorrhization 

degree.


1049 - Survival of insect target and biocontrol fungus in 

a greenery plantation 

S. Vestergaard * , C. Nielsen, S. Harding & J. Eilenberg 

Royal Veterinary and Agricultural University, 

Thorvaldsensvej 40, 1871 Frb. C, Denmark. - E-mail: 

sve@kvl.dk 

The insect family Curculionidae contains some of the 

economically most important pest insect species in 

greenery production for Christmas decorations in Denmark. 

In particular, the two species Strophosoma 

melanogrammum and S. capitatum are important pest 

species. At present no chemical treatment of these weevils 

is allowed in Denmark. Biological control including the use 

of insect pathogenic fungi from Hyphomycetes may thus 

provide a potential to minimise the damage caused by these 

weevils. Isolates of the insect pathogenic fungi 

Metarhizium anisopliae Beauveria bassiana and 

Paecilomyces spp. were studied in the laboratory and field. 

In the laboratory, all tested isolates were able to infect and 

cause mycosis in both adults and larvae of S. 

melanogrammum and S. capitatum. Based on the results 

from the laboratory bioassays, isolates were chosen for 

release under field conditions in an Abies procera 

plantation, where different application strategies were used. 

Spatial-temporal distribution of the released fungus in the 

soil was monitored by cfu per g soil. Weekly sampling 

from emergence traps monitored survival of adult weevils. 

Adults were collected, counted and determined to species 

level and incubated individually in plastic cups to estimate 

the prevalence of the applied fungus as well as the 

prevalence of natural infections. Preliminary results proved 

that populations of weevils in the treated plots were 

reduced significantly. 

1050 - Do Piceirhiza bicolorata mycobionts form both 

ecto- and ericoid mycorrhizas? 

L. Villarreal-Ruiz 1* , I. Anderson 2 & I.J. Alexander 1 

1 Department of Plant and Soil Science, University of 

Aberdeen, Cruickshank Building St Machar Drive, 

Aberdeen, AB24 3UU, Scotland, U.K. - 2 Soil Quality & 

Protection, The Macaulay Institute, Craigiebuckler, 

Aberdeen, AB15 8QH, Scotland, U.K. - E-mail: 

l.virl@abdn.ac.uk 

Ectomycorrhizas of the Piceirhiza bicolorata morphotype 

were found commonly associated with Pinus sylvestris 

seedlings and mature trees in Glen Tanar Native Pinewood, 

Northeast Scotland. Fungal isolates from P. bicolorata 

mycorrhizas are thought to be part of the Hymenoscyphus 

ericae aggregate, and this raises the possibility that they 

form mycorrhizas with both the pine trees and the 

understorey ericoid vegetation. In this study, strains from 

P. bicolorata were isolated from soil cores collected in a 

160 year-old natural stand. The mycobiont was cultured 

and identified by extracting genomic DNA and sequencing 

the PCR- amplified ITS regions. Phylogenetic analysis 

showed that our isolate grouped in clade 4 of Vrålstad et al. 

(New Phytol., 2002) along with many other isolates from P. 

bicolorata-type ectomycorrhizas, including Phialophora 

finlandia. Mycorrhizal synthesis were performed to test the 

ability of our isolate to form ECM and ERM on P. 

sylvestris seedlings and ericoid plants respectively, and to 

cross infect between P. sylvestris + P. bicolorata 

(previously synthesized) and Vaccinium myrtillus. The 

synthesis with Scots Pine seedlings produced black 

morphotypes with or without hyaline tips and with 

emanating hyphae. Pure culture synthesis with V. myrtillus 

seedlings produced typical ERM: these were also produced 

in the cross infection experiments. In both cases the growth 

of Vaccinium plants forming ERM was stimulated relative 

to the growth of uninfected plants. 

1051 - Strategies for sustainable land management: 

quarry recovered through truffle-culture 

M. Vinay * , L. Baciarelli Falini & M. Bencivenga 

Dip. Biologia vegetale e biotecnologie agroambientali- 

Università di Perugia, Borgo xx giugno 74 06121 Perugia, 

Italy. - E-mail: mavinay@tin.it 

Building materials come from quarries. The opening of a 

quarry is a geological damage and it causes the complete 

destruction of vegetation. Italian laws provide for each 

open quarry project an environmental recovery and 

steadying plan through soil-modelling and vegetationplanting 

in order to repair the damage and avoid 

subsequent trouble. A concrete case of quarry recovery 

through truffle-cultivation is analysed. 

1052 - The identification and frequency of occurrence 

of higher filamentous marine fungi on mangroves in a 

south Florida mangle 

C. Vogel 1* , S. Schatz 2 , H. Laubach 3 & A. Rogerson 1 

1 Nova Southeastern University, College of Oceanography, 

8000 North Ocean Drive, Dania FL 330044, U.S.A. - 

2 Nova Southeastern University, Colleges of Optometry and 

Oceanography, Davie FL 33314, U.S.A. - 

3 Nova 

Southeastern University, College of Medical Sciences, 

Davie FL 33314, U.S.A. - E-mail: vogelc@nova.edu 

Little is known regarding the occurrence and distribution of 

the higher filamentous fungi on mangroves in South 

Florida, USA. Previous studies have demonstrated that 

marine fungi are an important degradative component and 

assume an important role in nutrient recycling systems in 

estuarine and near-shore ecosystems. The purpose of this 

study was to collect, identify and prepare a key to the 

higher filamentous fungi occurring in the mangle of J. U. 

Lloyd State Park on the southeast coast of Florida, USA. 

Over a period of ten months 33 species were identified 

including 1 Basidiomycete, 8 Deuteromycetes, and 24 



Ascomycetes. The driftwood collected was mainly 

comprised of pieces of Rhizophora mangle and 

Conocarpus erectus. The predominant species, by 

frequency of occurrence, include the Ascomycetes 

Hypoxylon oceanicum (8.3%), Leptosphaeria australiensis 

(15.2%), Lulworthia grandispora (6.8%), and Nais glitra 

(8.3%) as well as the Deuteromycete Humicola 

alopallonella (7.6%). In addition, two new species of 

Ascomycetes were collected and will be presented. New 

records for Florida include the Ascomycetes Lineolata 

rhizophorae and Massarina velatospora. A new host 

record for Phaeosphaeria gessneri occurring on R. mangle 

is reported. Overall, the marine mycota of South Florida 

appears to be very similar to that reported for other tropical 

and subtropical regions. 

1053 - An ecological study of woody and leafy 

endophytes in the tropical mangrove tree, Kandelia 

candel 

L.L.P. Vrijmoed * , N. Plaingam & E.B.G. Jones 

Department of Biology and Chemistry, City University of 

Hong Kong, 83 Tat Chee Avenue, Kowloon, Hong Kong 

S.A.R., China. - E-mail: bhlilian@cityu.edu.hk 

Kandelia candel is the only tree member of the 

Rhizophoraceae which occurs in the subtropical Asian 

Pacific mangroves. We have undertaken a preliminary 

survey of endophytes in woody and leafy tissues of K. 

candel in Hong Kong. Samplings were carried out in two 

distinct climatic period so as to evaluate possible effects on 

their occurrence. Four fully grown trees were selected. 

Wood sections were cut from branches of different 

diameters and separated into bark and xylem tissue 

segments. Leafy tissue segments were also removed from 

mature leaves of branches sampled. All segments were 

incubated in malt extract agar after surface sterilisation. A 

total of 987 strains of fungi was isolated from 1280 bark 

tissues, 1280 xylem tissues and 320 leafy tissues segments, 

with infection rates of 60, 12 and 25% respectively. This is 

represented by 22 identified genera and 28 morpho-types. 

The dominant species in bark included Pestalotiopsis spp., 

Tryblidiopycnis sp., Xylaria sp., morpho-types 3 and 5. 

Frequent xylem endophytes were Phoma sp., some 

unidentified pycnidial coelomycetes, Geniculosporium sp., 

and morpho-type 13. Guignardia sp. and morpho-type 26 

were abundant on leaves. These initial results confirm 

endophytic infection in trees is not vertical and each tissue 

type is colonised by a distinct group of endophytes. 

1054 - Lichen genotyping with fungus specific 

microsatellites 

J.-C. Walser 1* , C. Sperisen 1 , M. Soliva 2 & C. Scheidegger 1 

1 Swiss Federal Research Institute WSL, Zuercherstrasse 

111, CH-8903 Birmensdorf, Switzerland. - 2 Geobotanical 

Institute ETH Zurich, Zollikerstr. 107, CH-8008 Zürich, 

Switzerland. - E-mail: jean-claude.walser@wsl.ch 

318 


Little is known about the genetic variation of lichenforming 

fungi. Yet, corresponding knowledge would be 

required for the accurate delimitation of individuals, 

populations, or species and for studies of reproductive 

systems and dispersal strategies. This will also help to 

develop and evaluate measures for conserving lichen 

biodiversity. Lacking genetic variation and, hence, low 

adaptability to environmental changes such as habitat 

fragmentation may explain why many epiphytic lichens are 

declining over much of Europe. We suppose that 

population genetic studies on lichen-forming fungi are 

mainly limited by the lack of suitable molecular markers. 

An ideal genetic marker type for population studies of 

lichen-forming fungi would combine the detection of high 

levels of genetic variation with the fungal specific 

amplification of DNA. Both requirements would be 

fulfilled by microsatellite markers. Microsatellites consist 

of short tandemly repeated sequences and proved to be 

highly informative markers in evolutionary, population 

genetic and conservation biological studies. We show that 

highly polymorphic, fungus-specific microsatellite loci also 

exist in the epiphytic lichen species Lobaria pulmonaria. 

Their fungus specific nature, their potential to identify 

multiple alleles and their transferability make these STMS 

markers a powerful and reliable new tool for genetic 

mapping, large and small scale genetic diversity analyses 

of L. pulmonaria and related species. 

1055 - Nutrients are released during mycelial 

interactions in soil 

J.M. Wells * , L. Boddy & H.K. West 

Cardiff School of Biosciences, Cardiff University Po Box 

915, Cardiff Cf10 3TL, U.K. - E-mail: john@grade-sixsupplies.co.uk. 

Cord-forming saprotrophic basidiomycetes represent a 

major nutrient resevoir within forest ecosystems and appear 

to be highly conservative of acquired nutrients. At some 

stage they must release nutrients to soil, but information on 

this is lacking. We have investigated nutrient release in the 

wood decay fungi Hypholoma fasciculare and 

Phanerochaete velutina. These were grown in soil 

microcosms in the laboratory in unpaired, self-paired and 

competing combinations. Inocula were prelabelled 32P 

orthophosphate, and losses from the soil systems were 

quantified by analysing water which had percolated 

through different areas of the systems following simulated 

rainfall events. 32P was released in unpaired systems and, 

though amounting to to less than 1% of total recovery, is 

direct demonstration of nutrient release to soil. Interaction 

with self resulted in significant mobilization of 

accumulated P, and significant increases in losses to soil. 

Interaction with non-self mycelia usually resulted in greater 

P mobilization than in interactions with self, even in the 

case of interaction between different isolates of the same 

species. P. velutina and one isolate of H. fasciculare 

preferentially translocated mobilized P to the interaction 

zone, but a another isolate of the latter preferentially 

translocated P away from the interaction zone.


1056 - Does human impact influence the species 

composition of epiphytic macrolichen communities? 

S. Werth 1* , H. Tømmervik 2 & A. Elvebakk 3 

1 

Swiss Federal Research Institute WSL, Zürcherstrasse 

2 

111, 8903 Birmensdorf/ZH, Switzerland. - NINA, 

Department for Arctic Ecology, Polarmiljøsenteret, N-9296 

Tromsø, Norway. - 3 Ecological Botany, Institute of 

Biology, University of Tromsø, N-9027 Tromsø, Norway. - 

E-mail: silke.werth@wsl.ch 

Epiphytic macrolichen communities were sampled on 

lower stems (≤ 2 m) of deciduous trees in plots of size 400 

m 2 along a regional macroclimatic gradient in Troms 

county, northern Norway. Canonical Correspondence 

Analysis (CCA) with variation partitioning revealed the 

following key factors for macrolichen communities: 1. 

Macroclimate was the primary factor controlling epiphytic 

macrolichen communities, with strictly macroclimatic 

variation accounting for 35% of total variation explained 

(TVE). 2. Two other key factors were spatial variation and 

tree substrate, amounting to 25.5% and 17% of TVE, 

respectively. 3. Human impact explained little (5.2%), 

forest fragmentation and natural patchiness none of the 

variation in species composition. The largest amount of 

shared variation was pooled between the sets of spatial and 

climatic variables, while the set of human impact variables 

shared no variation with other sets of explanatory variables. 

The reliability of species groupings was confirmed by 

DCA (Detrended Correspondence Analysis), showing an 

ecologically sound aggregation of the species optima. DCA 

revealed two main groups of species, characterised by 

differing photobionts. Key factors controlling epiphytic 

macrolichen communities are discussed in relation to 

lichen ecology, physiology and distribution patterns. 

1057 - Modelling colonies of fungi - a generic process 

based approach 

N.A. White * , R.E Falconer, J.L Bown & J.W Crawford 

SIMBIOS, University of Abertay, Kydd Building, Dundee, 

DD1 1HG, Scotland, U.K. - E-mail: 

mltnaw@abertay.ac.uk 

A fundamental challenge in ecology is to derive a 

mechanistic understanding of biodiversity of natural 

communites. This challenge is particularly difficult in 

fungal communities since these systems are indeterminate 

and demonstrate a high degree of plasticity in response to 

both biotic and abiotic stimuli. The approach described 

considers fungal communities in terms of constituent 

colonies, and begins by identifying a generic, process 

based description of individual colonies. An individualbased 

model of fungal colony development is presented. 

The model considers the spatio-temporal growth of mycelia 

over heterogeneous substrates. The model is spatially 

explicit where an individual colony is represented as a 

number of interconnected cells. Communication among 

cells is facilitated via fundamental ecological processes. 

The processes of resource uptake, diffusion of internal 

nutrients within the colony and conversion of resource 

taken up into biomass are represented. In addition, the 

directional translocation of nutrients over large distances is 

supported. Colonies are characterised in terms of 

physiological traits: different colony forms have different 

traits; traits parameterise the model processes. Importantly, 

processes within the model are amenable to quantitative 

parameterisation from experiment. This model of 

development of individual colonies serves as a platform for 

investigations into fungal community dynamics through the 

inclusion of interspecific interaction terms. 

1058 - The widespread occurrence of dark septate 

endophyte fungi in grassland communities 

E.M. Wilberforce 1* , G.W. Griffith 1 , L. Boddy 2 & R. 

Griffiths 3 



3DA, Wales, U.K. - 2 School of Biosciences, Cardiff 

University, PO Box 915, Cardiff CF10 3TL, Wales, U.K. - 

3 National Botanic Garden of Wales, Middleton Hall, 

Llanarthne, Carmarthenshire, SA32 8HG, Wales, U.K. - Email: 

emw96@aber.ac.uk 

This study describes unexpectedly high levels of isolation 

of dark septate endophytes (also known as mycelium 

radicis atrovirens [MRA]) from surface-sterilised plant 

roots from both undisturbed and agricultural mesotrophic 

grasslands in Wales. We have also examined spatial and 

temporal variables in their distribution. Most reports of 

these fungi have previously related to subalpine and boreal 

habitats. Their failure to sporulate in culture has hindered 

further studies. Analysis of sequence and restriction 

fragment polymorphism has been used to assess diversity 

of these fungi. Preliminary data suggesting a potential 

ecological role in these habitats is presented, based on 

microcosm studies of their interactions with other fungi 

isolated from the same site. 

1059 - Cyanobiont specifity of lichens on Livingston 

Island, maritime Antarctica 

N. Wirtz 1* , H.T. Lumbsch 1 , B. Schroeter 2 , R. Türk 3 & L.G. 

Sancho 4 

1 

Universitaet Essen, Fachbereich 9, Universitätsstr. 5, 

2 

45117 Essen, Germany. - Botanisches Institut, 

Universitaet Kiel, Olshausenstr. 40, 24098 Kiel, Germany. 

- 3 Universitaet Salzburg, Inst. f. Pflanzenphysiologie, 

Hellbrunnerstraße 34, 5020 Salzburg, Austria. - 

4 

University Complutense, Depto.Biologia Vegetal II, 

Fac.Farmacia, 28040 Madrid, Spain. - E-mail: 

nora.wirtz@uni-essen.de 



Nitrogen is a major factor of the lichen biodiversity in 

maritime Antarctica. At coastal habitats, there is a luxuriant 

lichen flora because of nutrition by the sea and animals, 

while the lichen flora in inland habitats is rather poor. The 

percentage of taxa containing nitrogen-fixing cyanobacteria 

either as primary or secondary photobiont is increased in 

inland communities. We have investigated the cyanobionts 

of lichens occurring at two ice-free locations on Livingston 

Island: inland habitats of South Bay and Byers Peninsula. 

The cyanobionts of two lichens with primary cyanobiont 

(Leptogium puberulum, Massalongia carnosa) and three 

species containing Nostoc in cephalodia (Placopsis 

contortuplicata, P. parellina, Psoroma cinnamomeum) 

were examined using molecular techniques and compared 

with free-living Nostoc colonies found in the area. 

Nucleotide sequences of the cyanobacterial tRNALeu 

intron were employed as genetic markers. Preliminary 

results suggest that different species at the same location 

share a common cyanobiont. 

1060 - Small scale distribution pattern of 

ectomycorrhizal types in a red oak stand 

J. Wöllecke * , S. Gebhardt & R.F. Hüttl 

Brandenburg University of Technology Cottbus, Box 

101344; D-03013 Cottbus, Germany. - E-mail: 

jenswoellecke@ektomykorrhiza.de 

Forest soils, especially the organic layers are very 

heterogenious substrates. Parameters like water content, pH 

value or the organic content could vary in the range of 

millimetres. Fungal mycelia or ectomycorrhizae are 

dispersed dependent of the heterogeneity of micro-habitat 

conditions. Beyond, the spatial distribution of 

ectomycorrhizae is influenced by interspecific interactions. 

As a result of all these influences, the ectomycorrhizal 

coenosis form a mosaic structure in the soil. At present, 

very few data on micro-scaled distribution of 

ectomycorrhizal fungi are available. First investigations 

started in recent years. To find characteristic structures of 

ectomycorrhizal coenosis in different scales, is one of our 

goals. Therefor we map the ectomycorrhizae in a coherent 

soil volume. This project is established in red oak stands of 

former mining sites and compare these afforestations with 

stands growing on undisturbed soil. Soil units of 6 cm 

width, 100 cm length and 3 cm depth taken from the top 

organic soil layer are divided in subunits of two to two cm 

and one cm depth. These subunits are solved in water and 

the ectomycorrhizae are qualitatively recorded. Maps will 

visualize the distribution of the ectomycorrhizae. We know 

from investigations in Scots pine stands, that the 

assemblage of ectomycorrhizal types can vary between 

sides and that nearly the half of all combination of 

ectomycorrhizal types never grow together in the same 

micro-habitat. 

320 


1061 - The longevity of Cordyceps humberti on Icaria 

wasps (Hymenoptera: Vespidae) 

A. Wongkaeo 1 , N.L. Hywel-Jones 2* , S. Sirimungkararat 1 & 

W. Saksirirat 1 

1 Khon Kaen University, Facultu of Agriculture, Khon 

Kaen, 40002, Thailand. - 2 BIOTEC-Mycology, National 

Center for Genetic Engineering and Biotechnology, 

National Science and Technology Development Agency, 

113 Pahonyothin RD., Khlong 1, Khlong Luang, Pathum 

Thani 12120, Thailand. - E-mail: nigelhj@biotec.or.th 

Cordyceps stromas persist for some time but little work has 

been done on longevity of individual stromas. 

Identification of a Cordyceps humberti epizootic in Nam 

Nao National Park, Thailand provided an opportunity to 

examine longevity of individual stromas. Cordyceps 

humberti and Hirsutella saussurei infected Icaria sp. The 

nest (ca. 80 cm long, 50 cm across) was in a tree at ca. 10 

m. Infected wasps were found up to 30 m radius from the 

nest. Wasps were attached by their mouth-parts to the leaf 

margin. Bamboo was a preferred substrate for settling but 

dicotyledonous plants were also used. The first wasps were 

seen in August (1999). Thirty wasps were tagged and 

observations made every 3-4 dy. Five of 27 specimens 

were Cordyceps while the rest were the Hirsutella form. 

However, after 1 month, of 23 specimens remaining 13 

were the Cordyceps form. Of 27 specimens recorded, 17 

were lost within 100 dy while 4 survived 100-200 dy and 5 

specimens were documented from 200-300 dy. One 

specimen was still present at the end of the work (513 dy). 

Significantly, of persistent specimens, 2 produced 2 crops 

of perithecia. One specimen remained in the Hirsutella 

form for 169 dy before synnemata were lost and it entered 

a dormant state. After ca. 15 days of dormancy new 

synnemata developed and ca. 30-40 dy later the Cordyceps 

developed. This work demonstrates that in Cordyceps 

humberti perithecia can be produced over an extended 

period with hosts providing material over more than one 

season. 

1062 - The structure of symbiotic communities: 

population-level patterns of association between lichen 

fungi and their algal photobionts 

R. Yahr 1* , P.T. DePriest 2 & R. Vilgalys 1 

1 Duke University, Department of Biology, Box 90338, 

Durham, NC 27708, U.S.A. - 2 Smithsonian Institution, 

National Museum of Natural History, Washington, D.C. 

20560, U.S.A. - E-mail: ry2@duke.edu 

Morphological and molecular studies have largely rejected 

broad coevolutionary patterns between lichen fungi and 

their photobionts, but theory predicts that populations of 

intimate symbionts become locally coadapted to each other 

and to their environment. In the place of broad-scale 

phylogenetic congruence, we test this alternative 

hypothesis of fungal-algal association on a population-level


geographic scale. Over 150 samples representing eight 

Cladonia species with their green algae Trebouxia were 

collected from five populations and sequenced for ITS. 

Three distinct well-supported clades of algae were 

identified, and each clade was found in each sampled 

population. Of the eight sampled fungi, six are restricted to 

a single clade of algae, and the other two fungi can be 

found with either of two clades of algae, even in the same 

population. Patterns of association between fungi and their 

algae differ among lichens, but in all cases, fungi appear 

selective, consistently associating with only a subset of all 

algal clades available in the local environment, regardless 

of where the lichens are collected. Furthermore, at the 

genotype level, some evidence is found for even stronger 

patterns of association, with four cases of single algal 

genotypes restricted to one fungal species in a given 

population. These patterns of association of fungi and algae 

vary but are consistent with strong fungal preference and in 

some cases support the model of local geographic patterns 

of coevolution. 

1063 - Ophiostoma species associated with bark beetles 

infesting three Abies species in Okunikko, Japan 

Y. Yamaoka 1* , H. Masuya 2 , N. Ohtaka 1 , H. Goto 3 & S. 

Kaneko 4 

1 Inst. Agr. For., Univ. of Tsukuba, Tennodai, Tsukuba 

Ibaraki 305-8572, Japan. - 2 Tohoku Res. Ctr., For. Forest 

Prod. Res. Inst., Shimokuriyagawa,Morioka, Iwate 020- 

0123, Japan. - 3 For. Forest Prod. Res. Inst., Kukisaki, 

Inashiki, Ibaraki 305-8687, Japan. - 4 Kansai Res. Ctr., 

For. Forest Prod. Res. Inst., Momoyama, Fushimi, Kyoto 

612-0855, Japan. - E-mail: 

yyamaoka@sakura.cc.tsukuba.ac.jp 

Ophiostoma species were isolated from bark beetles and 

Abies mariesii, A. veitchii and A. homolepis infested with 

the beetles in Okunikko, Tochigi, Central Honshu, Japan. 

One to two Ophiostoma species were frequently isolated 

from each species of bark beetle. Ophiostoma subalpinum 

was the most common associate of Cryphalus montanus. 

This fungus is known to be the most dominant associate of 

C. montatus and C. piceae in wave regeneration forests of 

Abies species in central Japan. Ophiostoma sp. B as well as 

O. subalpinum were common fungi associated with 

Polygraphus proximus. Ophiostoma europhioides was 

isolated from Dryocoetes hectographus and D. 

autographus as one of the common associates. Ophiostoma 

sp. J and sp. S were frequently isolated from D. 

autographus and O. striatus, respectively. These fungi 

seem to have specific relationships with particular bark 

beetles. Ophiostoma sp. B, sp. J and sp. S have unique 

morphological characteristics and appear to be new 

species. 

1064 - Investigation of phylloplane yeasts in Taiwan 

S.-H. Yang & P.-H. Wang * 

Depatment of Microbiology, Soochow University, 70, Lin- 

Hsi Rd, Shin-Lin, Taipei 11102, Taiwan. - E-mail: 

phwang@mail.scu.edu.tw 

Seventy nine yeast strains were isolated from phylloplane 

of eight plants species in the campus of Tunghai University 

in Taiwan. Eleven species were identified and five of them 

presented on one plant only: Kodamaea kakaduensis and 

Sporidiobolus johnsonii on Ipomoea cairica; Cryptococcus 

magnus, Rhodotorula glutinis and Rhodotorula sp. on 

Hibiscus rosa-sinensis. Six yeast species distributed on 

different plants. Aureobasidium pullulans presented on 

Euphorbia tirucalli L., Oxalis violacea Linn. and Hibiscus 

rosa-sinensis; Discosphaerina fagi and Sporidiobolus 

pararoseus presented on Calocedrus formosana and Oxalis 

violacea Linn.; Pseudozyma aphidis presented on Oxalis 

violacea Linn., Mussaenda parviflora and Ageratum 

houstonianum; Cryptococcus laurentii presented on 

Calocedrus formosana, Oxalis violacea Linn., Hibiscus 

rosa-sinensis, and Ageratum houstonianum; Cryptococcus 

sp. presented on Acer serrulatum Hay. and Ageratum 

houstonianum. Black yeast found from all plants we 

investigated except Ipomoea cairica. Distributions of some 

phylloplane yeasts seem to associate with the species of 

plants, and some yeasts were widespread on different 

plants. 

1065 - Identification of fungi associated with algae using 

PCR-amplified sequences of the 18S and 28S rDNA 

fragments and DGGE 

A. Zuccaro 1* , B. Schulz 1 & J.I. Mitchell 2 

1 Institut für Mikrobiologie Thechnische Universität 

Braunschweig, D-38106, Germany. - 2 School of Biological 

Sciences King Henry Building Portsmouth, Po1 2DY, 

England, U.K. - E-mail: alga.zuccaro@t-online.de 

Since the introduction of molecular techniques, such as 

PCR and DGGE, in environmental microbiology, great 

steps forward in the comprehension of microbial diversity 

have been made. This method is now routinely used to 

analyse bacterial community diversity and dynamics. The 

application of these techniques to estimate fungal diversity 

and follow community structure, however, is poorly 

developed. Consequently, fungi are often omitted from 

ecological community analyses. In this study, PCR primers 

and DGGE conditions were optimised for use in 

environmental studies of marine fungi associated with 

macroalgae. Current fungal community analysis usually 

involves only PCR-amplified sequences of the 18S rRNA 

gene. PCR primers used for this gene show little fungal 

specificity, which is required to detect fungal sequences 

that might be rare in the environment. Five different PCR 

primer pairs were tested to amplify rRNA genes from 20 

fungi isolated from Fucus serratus. Two primer pairs 



(AZ24-AZ709; AZ107-AZ709) were designed for the PCR 

amplification of fungal 28S rDNA (covering D1, D2 and 

D3 regions) from marine fungi and environmental algal 

samples. Two other pairs for 18S (NS1-FR1; FF390-FR1) 

were chosen from previously published work for their 

fungal specificity and their ability to achieve a separation 

for most of the amplification products obtained from a 

reference collection. The remaining primer pair (FF390- 

NS24) was fungal specific for the 3-prime end of the 18S 

rRNA gene. 

322 


IMC7 Main Congress Theme V: CELL BIOLOGY AND PHYSIOLOGY Posters 

1066 - Exploring the transcriptome of the nematodetrapping 

fungus Monacrosporium haptotylum 

D.G. Ahrén, C. Fekete * , E. Friman, T. Johansson & A. 

Tunlid 

Department of Microbial Ecology, Ecology Building, SE 


csaba.fekete@mbioekol.lu.se 

Monacrosporium haptotylum infects nematodes using 

adhesive knobs, which are developed on branches of 

vegetative mycelium. To identify genes and metabolic 

pathways that are expressed during development of traps 

and infection of nematodes, we have analyzed 4767 

expressed sequence tags from three different cDNA 

libraries, vegetative mycelium, knobs, and knobs infecting 

the nematode Caenorhabditis elegans. The ESTs were 

clustered into 2006 contigs representing 2006 putatively 

unique transcripts. Only 7.6% of the contigs contained 

ESTs from more than one cDNA library, thus the patterns 

of genes expressed in the three libraries were significantly 

different. Approximately 26-37% of all ESTs displayed 

significant similarities to sequences found in the GenBank 

database. Most markedly, the frequency of transcripts 

related to protein synthesis was up-regulated in the knobs. 

On the other hand the knob library contained in comparison 

with the mycelium cells and the fungus infected the 

nematode lower frequencies of ESTs involved in cell 

growth, cell division and DNA synthesis, transcription, 

protein destination and cellular communication/ signal 

transduction. Among the most prevalent transcripts 

expressed during the infection of nematodes were ESTs 

displaying sequence similarities to extracellular proteases, 

transportes and toxins. 

1067 - Identification of an HMG-domain protein 

implicated in nuclear migration and mating in Coprinus 

cinereus 

M.C. Aime * & L.A. Casselton 

University of Oxford, Department of Plant Sciences, South 

Parks Road, Oxford OX1 3RB, U.K. - E-mail: 

cathie.aime@plants.ox.ac.uk 

Mating between compatible homokaryons in Coprinus 

cinereus leads to the formation of a mycelium known as a 

dikaryon. In the dikaryon nuclei of both mates remain 

paired in each cell and are partitioned each time the cell 

divides via a structure known as the clamp connection. The 

formation and maintenance of the dikaryon is under the 

control of two sets of genes, the A mating type genes that 

encode a transcription factor that activates clamp cell 

formation and the B mating type genes that encode peptide 

pheromones and their cognate receptors, and which 

activate nuclear migration following compatible cell fusion 

and clamp cell fusion once the dikaryon is established. We 

are trying to understand how pheromone signalling 

regulates these quite distinct stages in dikaryon 

development. As a first step towards our goal we cloned a 

gene, hmg1, which encodes a High Mobility Group (HMG) 

protein. Homologous replacement of hmg1 implicates 

HMG1 in nuclear migration, but not in clamp cell fusion. 

∆hmg1 strains are capable of donating nuclei to a wild type 

partner, but unable to receive nuclei. Homozygous ∆hmg1 

dikaryons display unusually vigorous growth, but nuclear 

distribution is irregular and sporulation is defective. 

Current studies explore the role of Hmg1 in pheromone 

signalling and the role it may have in regulating nuclear 

migration at other stages in sexual development. 

1068 - Heterologous diploid nuclei in the violet root rot 

fungus, Helicobasidium mompa 

* 

T. Aimi , Y. Iwasaki, S. Kano, Y. Yotsutani & T. 

Morinaga 

School of Bioresources,Hiroshima Prefectural University, 

Nanatsuka 562,Shobara-city,Hiroshima Prefecture 727- 

0023, Japan. - E-mail: taimi@bio.hiroshima-pu.ac.jp 

Allelic genes hga1-1 and hga1-2, which encode G protein 

alpha subunit from violet root rot fungus, Helicobasidium 

mompa, were sequenced and characterized. Restriction 

fragment polymorphism (RFLP) analysis determined that 

the gene is present as a single locus in the single 

basidiospore isolates, while strain V169 possessed both 

alleles of this gene. Therefore, although basidiospore 

isolates are dikaryon, they are homokaryon. Field-isolated 

strain V169, the putative parent strain, is dikaryotic 

heterokaryon. hga1-1 and hga1-2 segregated to almost 1:3 

among single basidiospore isolates from the same fruiting 

body. These results suggest that one of the nuclei in the 

V169 parent strain is homologous diploid and the other 

heterologous diploid. This parent strain produced four 

homokaryotic and dikaryotic basidiospores on each 

basidium. The basidiospore germinated to homokaryotic 

and dikaryotic mycelium, and the homokaryon strains 

generated secondary mycelium by mating between 

compatible homokaryon. 

1069 - Water relations, photosynthesis and growth of 

mycorrhizal bean plants under water stress 

1* 2 3 

M.R. Amerian , W.S Stewart & H. Griffiths 

1 

Shahrood University, 7th Tir Square, Shahrood, Iran. - 

2 

Agricultural and Environmental Sciences Dept., 

Newcastle University, Newcastle upon Tyne, U.K. - 

3 

Department of Plant Sciences, Cambridge University, 

Cambridge, U.K. - E-mail: M.R.Amerian@ncl.ac.uk 

Arbuscular mycorrhizae symbiosis often altered water 

relations with consequent effects on gas exchange rate and 

photosynthesis. The effect of arbuscular mycorrhizal (AM) 

colonisation on photosynthesis, stomatal conductance, 

water relations, leaf area and dry weight of droughted and 

well-watered Phaseolus vulgaris L. was assessed after 3 



months growth in sandy loam pot culture in greenhouse 

condition inoculated at the seedling stage with the fungi 

Glomus mosseae (Nicol. & Gerd.) Gerd. and Trappe or 

Glomus interaradices Schenck & Smith, or uninoculated. 

During the drought period and after rewatering, leaf water 

potential, CO2 assimilation rate, stomatal conductance and 

transpiration of plants were measured. Leaf water potential, 

CO2 assimilation rate and stomatal conductance in 

mycorrhizal plants during the unwatered period were, in 

most cases, significantly greater than in the nonmycorrhizal 

plants, particularly in those infected by G. 

interaradices. There was no significant mycorrhizal effect 

on transpiration rate. Mycorrhizal plants were able to 

postpone the onset of wilting. During recovery from 

drought, there were significant differences in speed of 

recovery of leaf water potential, transpiration rate and 

stomatal conductance in mycorrhizal plants, but not in 

assimilation rate. Leaf areas and dry weight of mycorrhizal 

plants were generally greater than non-mycorrhizal plants 

after drought and recovery from drought. 

1070 - Lignocellulose degradation by Pleurotus ostreatus 

in the presence of heavy metals 

P. Baldrian * & J. Gabriel 

Institute of Microbiology AS CR, Videnska 1083, CZ- 

14220, Prague 4, Czech Republic. - E-mail: 

baldrian@biomed.cas.cz 

The ability of white-rot fungi to decompose lignin, 

cellulose, hemicelluloses and a range of structurally related 

compound including several xenobiotics has found several 

technological applications. The contents of heavy metals is 

among the most important factors that affect the 

lignocellulose-decomposing system of these fungi. In 

Pleurotus ostreatus, Cu and Cd increase laccase activity, 

whereas other metals (Hg, Pb, Ag, Zn) have negative effect 

on enzyme production. Mn-peroxidase is negatively 

affected by the presence of Cd. Heavy metals also affect 

the carbon and energy-supplying system of 

polysaccharides-decomposing enzymes. Addition of Cd to 

P. ostreatus cultures increased the activities of 

endocellulase, exocellulase, and β-1,4-glucosidase, 

whereas β-1,4-xylosidase and β-1,4-mannosidase activity 

was negatively affected. During cultivation of wheat straw, 

the presence of Cd also decreased the substrate utilisation 

(weight loss). Furthermore, some heavy metals, e.g. Cd and 

Hg negatively affect fungal colonisation of both 

lignocellulose substrate and nonsterile soil at low 

concentrations, that do not affect the activity of ligninolytic 

enzymes. This has to be taken into account when 

applications of white-rot fungi for degradation of 

xenobiotic contaminants in soil are considered, since these 

processes are dependent upon external substrate input and 

effective spread of mycelia in the substrate. This work was 

supported by the Grant Agency of the Czech Republic 

(204/02/P100). 

324 


1071 - High rates of extracellular superoxide 

production by lichens in the suborder Peltigerineae 

R.P. Beckett 1* , F.V. Minibayeva 2 & T. Tolpysheva 3 

1 School of Botany and Zoology, University of Natal, PBag 

X01, Scottsville 3209, South Africa. - 2 Institute of 

Biochemistry and Biophysics, Russian Academy of Science, 

PO Box 30, Kazan 420503, Russia. - Department of 

Mycology, Moscow State University, Vorobjevi Gory, 

Moscow 119899, Russia. - E-mail: beckett@nu.ac.za 

Rates of extracellular superoxide radical formation were 

measured in 34 species of lichens from different taxonomic 

groupings and contrasting habitats before and after 

desiccation stress. All 20 species tested from the suborder 

Peltigerineae produce superoxide extracellularly at high 

rates, even when they are not stressed. In addition, some 

species show a burst of superoxide production during 

rehydration following desiccation. In general, production 

of high levels of superoxide and the existence of an 

inducible oxidative burst were best developed in species 

growing in wet microhabitats. Extracellular production of 

superoxide was almost absent from the 14 species from 

other lichen groups. Preliminary studies on the identity of 

the superoxide producing enzymes suggested that they do 

not possess the classical characteristics of those suggested 

to produce reactive oxygen species in higher plants. 

Patterns of superoxide production are discussed in terms of 

their possible role as a defence against pathogenic fungi 

and bacteria. 

1072 - Cross talk between cAMP and calcium signalling 

in the filamentous fungus Aspergillus niger 

M. Bencina 1 , O. Kozlova-Zwinderman 2 , M. Legisa 1 & 

N.D. Read 2* 

1 Laboratory for Biotechnology and Industrial Mycology, 

National Insitute of Chemistry, Hajdrihova 19, SI-1000 

Ljubljana, Slovenia. - 2 Fungal Cell Biology Group, 

Institute of Cell and Molecular Biology, University of 

Edinburgh, Rutherford Building, Edinburgh EH9 3JH, 

U.K. - E-mail: Nick@fungalcell.org 

Very little is known about cross-talk between cAMP and 

calcium signalling in filamentous fungi. The aim of this 

study was to analyse the influence of cAMP and protein 

kinase A (PKA)-dependent phosphorylation on calcium 

signalling in Aspergillus niger. For this purpose, cytosolic 

free calcium was measured in living hyphae expressing 

codon-optimised aequorin whilst applying pharmacological 

agents which increased cAMP levels or inhibited 

phosphorylation by cAMP-dependent PKA. Calcium was 

also measured in mutant strains in which phosphorylation 

by PKA was increased or lacking. Our results indicate that 

cAMP-dependent phosphorylation regulates calcium 

homeostasis in A. niger, probably by activating calcium 

channels. Further evidence for cross-talk between cAMP 

and calcium signalling came from the analysis of a mutant 

in which the catalytic subunit of PKA was under the 

control of an inducible promoter. The consequence of PKA 

3


induction was a transient increase in cytosolic free calcium 

which correlated with a polar?apolar transition in hyphal 

morphology. A transient increase in calcium was not 

observed in this mutant when the morphological shift was 

in the opposite direction. These results indicate that PKAdependent 

phosphorylation may be important for 

increasing cytosolic free calcium to induce a polar to apolar 

shift in hyphal morphology. 

1073 - Biological decolorization of textile dyes by 

Funalia trogii 

H. Bermek 1* , T. Deveci 2 , M. Erdogan 3 , O. Pekcan 3 & C. 

Tamerler 1 

1 Istanbul Technical University, Dept. of Molecular Biology 

and Genetics, Maslak, Istanbul, 80626, Turkey. - 2 Istanbul 

Technical University, Dept. of Environmental Engineering, 

Maslak, Istanbul, 80626, Turkey. - 3 Istanbul Technical 

University, Physics Department, Maslak, Istanbul, 80626, 

Turkey. - E-mail: bermek@itu.edu.tr 

Synthetic azo and disperse dyes are extensively used in 

textile industries. A significant proportion of these dyes is 

eluted in wastewaters. Biodegradation is an alternative to 

physical and chemical methods used for waste dye 

removal. In this study, the decolorization capability of a 

white-rot fungus Funalia trogii for eleven synthetic azo or 

disperse textile dyes were tested. Preliminary results 

showed that the fungus was capable of removing four dyes, 

Remazol blue RR, Dianix blau BRL, Dianix blau ACE and 

Telon schwarz by 38.6%, 50.9%, 32.8% and 54.5% 

respectively during a period of 5 days. For various 

dyestuffs, mechanism of decolorization appeared to be 

different. The effect of inoculum age and agitation on dye 

removal was investigated for obtaining maximum 

decolorization. Degree and type of dye removal was found 

to be dependent on the time of dye addition to the fungal 

cultures. Consequently, morphological differences in 

mycelial growth were also observed depending on addition 

time of dyestuffs. 

1074 - Effect of oxygen in the production of manganese 

peroxidase and laccase by a newly isolated white-rot 

fungus Trichophyton rubrum LSK-27 

H. Bermek 1* , I. Gulseren 1 , H. Jung 2 , K. Li 3 & C. Tamerler 1 

1 Istanbul Technical University, Molecular Biology and 

Genetics, Maslak - Istanbul 80626, Turkey. - 2 Sunchon 

National University, Department of Forest Resources, 

Sunchon 540-742, Korea. - 3 Oregon State University, 

Department of Wood Science and Engineering, 102 

Richardson Hall, Corvallis, OR97331-5751, U.S.A. - Email: 

bermek@itu.edu.tr 

Fungal mycelium morphology is a key concept in the 

production of enzymes. Oxygen transfer is one of the most 

important parameters that affect the morphology, 

consequently the enzyme production. Our studies on the 

morphology and ligninolytic enzyme production of a newly 

isolated white rot-fungus Trichophyton rubrum LSK-27 

constitute a very good example to this fact. Three 

ligninolytic enzymes, manganese peroxidase (MnP), lignin 

peroxidase (LiP) and laccase productions exhibited 

variations accordingly. In shake flasks, 3 different working 

volumes (wv: 10%, 25% and 50%) were studied, in which 

the organism exhibited significantly different pellet size 

and shape formation. Oxygen limited conditions (50% wv) 

led to the formation of large puffy balls (radii ^20 mm), 

however in 10% and 25% wv, cultures appeared similar in 

terms of pellet size and shape with a slightly flattened 

spheres (radii ^1 mm). Moreover, 10% cultures, exposed to 

least oxygen limitation, contained some hyphal growth in 

addition to the pellets. Our results clearly demonstrated that 

this microorganism preferred oxygen rich conditions for 

enzyme production. MnP and laccase productions were the 

highest at 10% and 25% wv respectively No MnP and 

laccase produced at 50% wv. LiP was not detected at all 

cases. 

1075 - Elemental analysis of edible mushroom brown 

birch scaber stalk Leccinum scabrum (Bull. ex Fr.) S. F. 

Gray growing in various sites in Poland 

L. Bielawski * & J. Falandysz 

Department of Environmental Chemistry and 

Ecotoxicology, University of Gdansk, 18 Sobieskiego St., 

PL 80-952 Gdansk, Poland. - E-mail: 

leszek29@chemik.chem.univ.gda.pl 

The concentrations of 13 elements such as Al, Ba, Ca, Cu, 

Cd, Fe, K, Mn, Mg, Na, Sr, Zn, and Hg were determined in 

fruiting bodies of Brown Birch Scaber Stalk Leccinum 

scabrum (Bull. ex Fr.) S. F. Gray collected in summer and 

autumn 2000 from different sites in Poland. Fresh 

mushrooms after cleanup from plants and soil were dried 

for several days in room temperature and further dried in 

electric oven at 40 degree C for 24 hours. The samples 

(0.2-0.5 g) of dried and powdered pooled samples of 

mushrooms (separately caps and stalks) were wet digested 

with 6 ml of concentrated nitric acid under pressure in 

closed TFM vessels in a microwave oven (CEM, Mars 5). 

The PerkinElmer Optima 2000 DV ICP-OES instrument 

was used for the analysis of mushrooms samples. In the 

case of mercury CV-AAS technique was used. The method 

was validated by analysing certified reference materials: 

CTA-OTL-1 (Oriental Tobacco Leaves, Institute of 

Nuclear Chemistry and Technology) and IAEA-359 

(Cabbage Leaves, International Atomic Energy Agency). 

There were some variations between trace elements 

concentration and collecting places. Magnesium, zinc and 

manganese were determined in quite similar concentrations 

regardless of sampling site while in the case of other 

elements some variations were noted. The essential 

elements such sodium, potassium, magnesium and calcium 

were found in higher concentrations than toxic mercury 

and cadmium. Potassium is the most abundant amongst of 

the elements quantified. 



1076 - Weathering in a saxicolous lichen community: A 

geobiological research project 

T. Bjelland 

Department of Botany, University of Bergen, Allégaten 41, 

N-5007 Bergen, Norway. - E-mail: 

torbjorg.bjelland@bot.uib.no 

The weathering in a saxicolous lichen community growing 

on sandstone has been studied. Lichen and rock samples 

have been analysed by high performance thin-layer 

chromatography/thin-layer chromatography, scanning 

electron microscopy, and X-ray diffraction analysis, and 

rock samples also by X-ray fluorescence spectroscopy 

analysis. The study clearly indicates that lichen mediated 

weathering leads to fragmentation and chemical dissolution 

of the rock and its component minerals. Despite the 

heterogeneity of the rock, and the fact that the studied 

weathering rinds are the result of complex interactions of 

physical, chemical, and biological weathering processes 

during the postglacial period, the results show that some 

crustose lichen taxa are clearly more aggressive in aiding 

weathering than other taxa in a lichen community. There is 

a positive correlation between the degree of weathering and 

species with high amounts of hyphae within the rock. 

Differences in content of lichen compounds seem to be a 

more likely explanation for the variations in chemical 

weathering, than differences in oxalic acid production. The 

results further imply that there is an indirect relationship 

between weathering effect and ecology in saxicolous 

lichens, but whether this is a specific strategy for nutrient 

requirements remains unclear. It is also suggested that 

lichens generally increase the weathering processes, except 

at locations with extremely high abrasion, where they may 

protect the surface. 

1077 - Biochemical pathway studies on the loline 

alkaloids of the grass endophyte, Neotyphodium 

uncinatum 

J.D. Blankenship 1 , M.J. Spiering 1 , L.P. Bush 2 , R.B. 

Grossman 3 & C.L. Schardl 1* 

1 University of Kentucky, Dept. of Plant Pathology, 

Lexington, KY 40546, U.S.A. - 2 University of Kentucky, 

Dept. of Agronomy, Lexington, KY 40546, U.S.A. - 

3 University of Kentucky, Dept. of Chemistry, Lexington, KY 


The insecticidal loline alkaloids (1-aminopyrrolizidines) 

are produced in mutualistic associations of Festuca and 

Lolium species with their respective fungal endophytes. 

Reliable production of N -formylloline (NFL) and N - 

acetylnorloline (up to a total of 1000 µg/ml) in cultures of 

the meadow fescue endophyte, Neotyphodium uncinatum, 

has facilitated studies on the loline alkaloid biosynthetic 

pathway. Based on loline alkaloid structures, it was 

previously suggested that lolines are polyamine products. 

However, our precursor feeding experiments, employing 

GC-MS and NMR analyses, indicated that lolines share 

common precursors with, but are not derived from, 

326 


polyamines. Labels from 5-[ 13 C]-ornithine and 1,2-[ 13 C]ornithine 

were incorporated into specific positions of the 

B-ring in the pyrrolizidine-ring structure of NFL, but the 

pattern of incorporation differed from what would be 

expected if a polyamine intermediate was involved. 

Feeding studies with 4-[ 13 C] and [ 15 N]-aspartic acid gave 

enrichment in the expected position of the pyrrolizidine Aring 

(at C-3) and the 1-amine, respectively. Universally 

[ 13 C]-labeled methionine did not incorporate in the 

pyrrolizidine ring system, but labeled the 1-amino methyl 

and 1-amino formyl groups. These studies demonstrated 

the origins of the carbon atoms in NFL: the B ring is 

derived from ornithine, the A ring is from aspartic acid, 

and the N-methyl and N-formyl carbons are from the Smethyl 

carbon of methionine. 

1078 - Cell state dynamics and pattern formation in 

polymorphic fungi 

E.V. Bogomolova 

St.Petersburg State University, Dept. of Botany, 

Universitetskaya emb. 7/9, 199034 St.Petersburg, Russia. - 

E-mail: evgenia@EB6711.spb.edu 

One of the most topical problems of cell and complex 

systems biophysics is elucidation of driving forces, 

mechanisms and dynamics of pattern formation and 

growth. Modern molecular genetics do not allow to 

understand fully fundamental principles of morphological 

plasticity. The main idea of our work is to reveal and 

describe the generic principles of regulation of cell states 

dynamics in polymorphic fungi, of morphogenesis and 

differentiation from the point of view of self-organization 

theory. Experimental study of cell state dynamics in some 

strains of polymorphic fungi (black yeasts) has shown cell 

state transitions (spontaneous or induced by various culture 

conditions). The main morphogenetic factors (governing 

parameters) were revealed on the basis of experimental 

analysis of the data obtained for several black yeast strains. 

In preliminary surveys we have found that regulation of 

cell state switch exhibits some dynamic properties. Such 

properties include: 1) all-or-none transitions between 

discrete states (with a stochastic component); 2) 

reversibility vs. irreversibility of such state transitions 

(asymmetry of transition probabilities); 3) hysteresis (in 

response to transition-triggering agents); 4) autocatalysis or 

autoinhibition of cell state transitions; 5) symmetry 

breaking events. Cell state transitions (in particular, 

mycelial/yeast) result in changes of colony form, and 

consequently, in diverse development strategies and 

substrate colonisation patterns of fungi.


1079 - Modelling fungal growth and function in 

heterogeneous environments 

G.P. Boswell 1 , H. Jacobs 2 , F.A. Davidson 1 , G.M. Gadd 2 & 

K. Ritz 3* 

1 Department of Mathematics, University of Dundee, 

Dundee, DD1 4HN, Scotland, U.K. - 2 Division of 

Environmental and Applied Biology, Biological Sciences 

Institute, School of Life Sciences, University of Dundee, 

Dundee, DD1 4HN, Scotland, U.K. - 3 Soil-Plant Dynamics 

Group, Scottish Crop Research Institute, Invergowrie, 

Dundee, DD2 5DA, Scotland, U.K. - E-mail: 

k.ritz@cranfield.ac.uk 

Modelling mycelial growth in nutritionally or spatially 

heterogeneous environments is particularly challenging 

since such growth is a consequence of small-scale 

processes (e.g. hyphal tip extension) that are manifest at 

considerably larger spatial scales (i.e. the mycelium) and 

each of which has important consequences for the spatiotemporal 

development of the fungus and its environment. 

We describe a mathematical model that accommodates 

these factors and explicitly simulates mycelial development 

in heterogeneous environments at both scales. The model is 

derived through the discretization of a previously 

formulated partial differential equation model. The 

modelled mycelial network is defined on a lattice allowing 

branching and anastomosis to be explicitly observed. 

Specifically, the model involves the number, location and 

orientation of hyphal tips; active and inactive hyphae (i.e. 

involved/not involved in translocation); internal and 

external substrate concentration; and spatial organization of 

substrate and habitable space. On calibrating the model 

using known properties for the species Rhizoctonia solani, 

information is provided on growth behaviour that is shown 

to be in very good qualitative and quantitative agreement 

with experimental data. Development of such an accurate 

model enables important predictions to be made on the 

functional consequences of fungal growth in heterogeneous 

environments and the key role of translocation within 

mycelia in enabling such responses. 

1080 - Elements in ectomycorrhizal fungus Paxillus 

involutus (Batsch ex Fr.) Fr. from various sites in 

Poland 

A. Brzostowski 1* , R. Kubota 2 , T. Kunito 2 , J. Falandysz 1 & 

S. Tanabe 2 

1 Department of Environmental Chemistry and 

Ecotoxicology, University of Gdansk, 18 Sobieskiego St. 

PL 80-952 Gdansk, Poland. - 2 Department of Environment 

Conservation, Center for Marine Environmental Studies, 

Ehime University, 3-5-7 Tarumi, Matsuyama 790-8566, 

Japan. - E-mail: abrzost@chemik.chem.univ.gda.pl 

Data on metallic elements accumulation of wild growing 

ectomycorrhizal fungus Poison Pax Paxillus involutus 

(Batsch ex Fr.) Fr. collected from different (contaminated 

and uncontaminated) ecosystems in Poland are presented. 

Concentrations of essential metals (e.g. K, Na, Mg, Ca, 

Mn, Fe, Cu, Zn, Ni) and other which have no apparent 

essential function (e.g. Hg, Pb, Cd, Al, Ag) were 

determined in caps, stalks or whole fruiting bodies. The 

mushrooms were collected between 1999 and 2001 from 

sites located in northern, central and southern Poland. 

Dried samples were wet digested with concentrated nitric 

acid in TFM vessels XP-1500 under pressure in microwave 

oven (Mars 5, CEM, USA). The elements were detected 

and quantified using atomic emission spectrometer with 

induced coupled plasma (Optima 2000 DV, Perkin-Elmer, 

USA) for some elements and with an inductively coupled 

plasma-mass spectrometer (HP-4500, Hewlett-Packard, 

USA). Accuracy of the method was assessed with standard 

reference materials: CTA-OTL-1, IAEA-359. Poison Pax is 

very common fungus and is well known to grow in 

symbiosis with various trees (e.g. Pinus sylvestris) this 

kind of complex research could be very useful in 

understanding interactions in fungus-metal-environmenttree 

system. On the other hand it may be one step further in 

investigation this specimen and its metal accumulation 

abilities. All these elements could be absorbed and 

accumulated by different physico-chemical mechanisms 

and transport systems. 

1081 - Exchange processes across the ectomycorrhizal 

interface: interactions between phosphate and 

carbohydrate flow 

H. Bücking 

Centre for Environmental Research and Technology 

(UFT), Applied Botany, Physiological Plant Anatomy, 

Leobener Str., D-28359 Bremen, Germany. - E-mail: 

heibueck@uft.uni-bremen.de 

Basis of the compatible interaction between plant and fungi 

in an ectomycorrhizal symbiosis is a bidirectional transfer 

of nutrients across the ectomycorrhizal interface. However, 

the normal flows of P and carbohydrates through the 

plasma membranes into the interfacial apoplast are 

calculated to be insufficient to maintain the symbiosis. 

Therefore, conditions in the interface, which cause an 

enhanced efflux or a decrease in the level of competing 

uptake systems has been proposed by several authors (e.g. 

Smith et al. 1994). Presently, we have only rare 

information about a regulation of these transfer processes 

occuring across this specialized interface and the 

mechanisms involved in polarizing the transfer. Here, new 

results are presented, which show, that the carbohydrate 

supply by the photosynthetic host plant have an influence 

on the P absorption by the mycobiont and the P transfer 

across the ectomycorrhizal interface. By the carbohydrate 

flow across the interface the P allocation between different 

P pools in the mycobiont and the P efflux across the fungal 

plasma membrane into the interfacial apoplast is affected. 

A model system is presented, which show, in which way 

the carbohydrate and P transfer might be linked and how 

the exchange processes between both symbiotic partners 

might be regulated. Smith SE, Gianinazzi-Pearson V, 

Koide R, Cairney JWG (1994): Plant Soil 159:103-113. 



1082 - The localization of nutrient transfer in the 

arbuscular mycorrhizal symbiosis 

H. Bücking * & W. Heyser 

University of Bremen, Center for Environmental Research 

and Technology, Leobener Str., D-28359 Bremen, 

Germany. - E-mail: heibueck@uft.uni-bremen.de 

The absorption of 33 P-inorganic phosphate and the element 

distribution in mycorrhizal roots of cucumber inoculated 

with the arbuscular mycorrhizal fungus Glomus mosseae 

was analysed by microautoradiography and energy 

dispersive X-ray spectroscopy (EDXS). Arbuscules with 

their extensive contact area to the plasma membrane of 

inner cortical cells are the main site for P transfer from an 

arbuscular mycorrhizal fungus to its host plant. The X-ray 

microanalytical results indicate, that across this 

mycorrhizal interface possibly Na and Mg can also be 

transferred. However, there are no indications, that the 

arbuscules are the only site for P transfer. Both methods 

showed that P can also be released from intercellular 

hyphae into the interfacial apoplast. The results are 

discussed with respect to the effect of an arbuscular 

mycorrhizal infection on the nutrient supply of its host 

plant and the interface regions which might be involved in 

nutrient transfer. 

1083 - Rock and mould: transformation of carbonate 

minerals by fungi 

E.P. Burford 1* , S. Hillier 2 & G.M. Gadd 1 

1 Division of Environmental and Applied Biology, 

Biological Sciences Institute, School of Life Sciences, 

University of Dundee, Dundee, DD1 4HN, Scotland, U.K. - 

2 Macaulay Land Use Research Institute, Craigiebuckler, 

Aberdeen, AB15 8QH, Scotland, U.K. - E-mail: 

e.p.burford@dundee.ac.uk 

In terrestrial environments, fungi serve as important 

geochemical agents in rock and soil. Fungi promote rock 

weathering and contribute to the dissolution of mineral 

aggregates in soil through the excretion of H + , organic 

acids and other ligands, or through redox attack of mineral 

constituents such as Mn and Fe. Fungi can also play an 

active or passive role in mineral formation through the 

precipitation of secondary minerals, e.g. oxalates, and 

through the nucleation of crystalline material onto cell 

walls, resulting in the formation of biogenic micro-fabrics 

in mineral substrates. Such interactions between fungi and 

minerals are of fundamental importance to biogeochemical 

cycles including those of C, N, S and P which are 

important nutrients required for plant, fungal and other 

microbial growth. Our research has demonstrated that fungi 

may play an important role in the transformation of microfabrics 

in limestone and dolomite. Scanning electron 

microscopy and energy-dispersive X-ray micro-analysis of 

rehydrated limestone samples has provided direct evidence 

of mineralized fungal filaments. The crystal coatings on 

328 


fungal hyphae exhibited different crystalline lattices and 

elemental composition from the initial limestone rock. 

Powder X-ray diffraction analysis indicated that these were 

probably secondary (re-distribution) carbonates, providing 

a novel demonstration of biogenic mineral formation 

mediated by fungal activity. 

1084 - Toxic metals in edible mushrooms: Is 

hyperaccumulation caused by synthesis of metalbinding 

proteins? 

C. Collin-Hansen 1* , E. Steinnes 1 & R.A. Andersen 2 

1 

Department of Chemistry, NTNU, 7491 Trondheim, 

Norway. - 

2 

Department of Zoology, NTNU, 7491 

Trondheim, Norway. - E-mail: Christian.Collin- 

Hansen@chembio.ntnu.no 

A common feature of many of the most popular edible 

mushrooms is the capability of concentrating certain 

potentially toxic metals to high levels in the fruiting body, 

thus introducing the metals to terrestric food chains, 

including humans. Throughout the years, several reports 

have revealed high concentrations of toxic metals such as 

Cd and Hg in wild-growing, edible mushroom species. 

Firstly, these reports may suggest negative effects on 

persons consuming large quantities of mushrooms, but they 

also call for a better understanding of the mechanisms 

involved in metal uptake, transfer and storage. Metalbinding 

proteins of several classes have been isolated from 

macrofungi and characterized previously. Some of these 

proteins share several characteristics with metallothioneins 

(MT). We here present data on a new Cd-binding protein 

from the popular edible mushroom Boletus edulis, which is 

a hyperaccumulator of both Cd and Hg. Radiotracer studies 

clearly suggest this new protein as an important binding 

site for Cd in this species. The presence of aromatic amino 

acids and the virtual absence of cystein shows that this 

protein does not belong to the MT family. When the Cd 

radiotracer was added to cytosol of B. edulis sampled from 

soils rich in Cd, more than 90% of the radiotracer was 

bound by the new protein, suggesting a role for the protein 

in Cd transport and storage. These findings are of 

importance in view of toxicology and food chemistry, but 

also environmental protection. 

1085 - Metal effect on fungal toxin, genetic expression, 

and growth regulation: Use of RT-PCR, microarray 

and electrospray ionization 

R. Cuero 1* , T. Ouellet 2 , J. Yu 3 & N. Mogongwa 4 

1 Prairie View A&M University, CARC, PO BOX 685, 

Prairie View, Texas 77446, U.S.A. - 2 Eastern and Oilseed 

Research Center, Agriculture-Food, Ottawa, Ontario K1A 

0C6, Canada. - 3 Southern Regional Research Center, 

USDA, ARS, New Orleans, LA 70179, U.S.A. - 4 Prairie 

View A&M University, Chemistry Dept., PO BOX 685, 

Prairie View, Texas 77446, U.S.A. - E-mail: 

olimpa@aol.com


The effects of the metal ions (Zn+2, Cu+2 and Fe+2 ), as 

single or in mixed treatments were observed through 

changes in A. flavus RNA, RT-PCR levels, and by 

differential genetic expression using microarray analysis, 

and also assessed by fungal dry weight accumulation, 

aflatoxin and OMST production. The interaction between 

the metal ions and the aflatoxin and/or its precursor OMST 

was determined by electrospray ionization mass 

spectrometer. The direct effects of metal ions on fungal 

growth, aflatoxin, and OMST synthesis, varied according 

the type of metal ion and expressed to either single ion or 

amendment of ion mixture. All ion treatments induced 

changes of fungal total RNA and mRNA levels and 

associate fungal growth, biosynthesis of aflatoxin and its 

precursor OMST. All mixed metal ions treatments boosted 

total RNA synthesis and enhanced expression of fungal 

RNA and the RT-PCR, as well as fungal biomass and 

synthesis of aflatoxin and OMST. These treatments thus 

induce changes in pattern of gene expression of A. flavus 

related to fungal growth and synthesis of aflatoxin and its 

precursor OMST. The differential genetic effects of these 

treatments were also clearly expressed by the microarray 

results. The electrospray ionization mass spectrometer 

analysis showed distinct differential binding of the metal 

ions to OMST. 

1086 - Carbon and nitrogen allocation patterns in the 

two green algal lichens Hypogymnia physodes and 

Platismatia glauca in relation to nutrient supply 

L. Dahlman 1* , J. Persson 2 , T. Näsholm 2 & K. Palmqvist 1 

1 Ecology and Environmental Science, Umeå University SE- 

901 87 Umeå, Sweden. - 2 Forest Genetics and Plant 

Physiology, Umeå Plant Science Center SE-901 83 Umeå, 

Sweden. - E-mail: Lena.Dahlman@eg.umu.se 

We investigated resource investments in lichens, by 

quantifying major carbon and nitrogen pools in two green 

algal lichens in nutrient enriched and poor habitats. 

Investments between the symbionts were determined by 

quantifying ribitol and chlorophyll a for the photobiont, 

and mannitol, ergosterol and chitin for the mycobiont. Two 

soluble carbohydrates, arabitol and glucose, as well as 

amino acids and proteins were also quantified. The soluble 

carbohydrate concentrations were twice as high, c. 4% of 

thallus dry weight, and thallus N concentrations were 2-4 

times higher in thalli from the nutrient enriched compared 

to the nutrient poor habitat. The proportion of thallus N 

invested in proteins was similar in the two habitats, while 

the amino acid pool was doubled in the fertilized thalli. 

This increase could be attributed to the N- rich amino acid 

arginine. Both lichens displayed an increased proportion of 

photobiont cells in relation to mycobiont hyphae in the 

nutrient enriched habitat. Resulting in a significantly higher 

ribitol to mannitol ratio, and a higher Chl a to ergosterol 

ratio, in thalli from the nutrient enriched habitat. 

Suggesting that the photobiont had benefited more than the 

mycobiont from the fertilization, and the increased arginine 

concentration suggests that the mycobiont may suffer from 

a relative carbon shortage. Both lichens were still healthy 

and alive, so the altered balance between the bionts might 

be an adaptation to handle the fertilization stress. 

1087 - Proteinases involved in skin degradation by the 

oomycete pathogen Pythium insidiosum 

D.J. Davis 1* , C. Bonati 1 , K. Lanter 1 , S.D. Makselan 1 & 

N.P. Money 2 

1 College of Mount St. Joseph, Department of Chemistry, 

Cincinnati, Ohio, 45233, U.S.A. - 2 Miami University, 

Department of Botany, Oxford, Ohio, 45056, U.S.A. - Email: 

diana_davis@mail.msj.edu 

The oomycete Pythium insidiosum is the only stramenopile 

pathogen of humans and other mammals. In essence, the 

mechanism of tissue invasion utilized by this 

microorganism is identical to the process employed by 

every other filamentous fungus that colonizes animal or 

plant tissues: invasive growth depends upon the exertion of 

force by the hyphal apex, and the degree to which the 

substrate is degraded by secreted enzymes. Biomechanical 

experiments prove that Pythium insidiosum achieves a 100fold 

reduction in the strength of cutaneous and subcutaneous 

tissue through proteinase secretion. In an attempt 

to identify the enzymes that play a primary role in tissue 

degradation, we have isolated and characterized a number 

of unique serine- and metallo-proteinases. By analyzing the 

effectiveness of each protein at facilitating hyphal 

penetration of collagen-rich membranes it is possible to 

distinguish between their nutritional and 'barrier-dissolving' 

roles. 

1088 - Lichens as a model-system for symbiotic 

organisms under most extreme conditions 

J.-P. de Vera & S. Ott * 

Botanical Institute, University of Duesseldorf, 

Universitätsstr.1, D-40225 Duesseldorf, Germany. - Email: 

otts@uniduesseldorf.de 

As a consequence of the symbiotic state of lichens both the 

bionts are able to colonize habitats where the separate 

bionts would not be able to survive. The symbiosis of 

lichens reflects a high degree of complexity and plasticity. 

The combination of both enables these organisms to 

colonize most extreme habitats. Besides the already well 

investigated microorganisms lichens are good modelsystems 

to examine adaptation strategies to most extreme 

environments. Because of the symbiotic nature of the 

lichens a 3-component-system can be used for 

investigations: the mycobiont, the photobiont and the 

lichen itself. Our investigations are based on such a system. 

The influence of different doses of UV A, B, C on the 

vitality of fungal (mycobiont) spores and the germination 

process has been investigated. The spores are cultivated on 

a variety of different subtrates to testing the influence of 

the UV radiation. The influence of vacuum conditions has 

been investigated. The aim of this research is to test the 

reaction of a symbiotic organism and its respective bionts 

to highly extreme conditions. For interpretation of results 



the method of modern confocal lasermicroscopy (CLSM) - 

a novel method in lichenology - is presented. 

1089 - Phosphate transporter expression in the soybean 

AM symbiosis 

J.D.W. Dearnaley 1 , B.A. Learmonth 1 & D.S. Bougoure 2* 

1 The University of Southern Queensland, West Street, 

Toowoomba, 4350, Australia. - 2 The University of Western 

Sydney, Locked Bag 1797, Penrith, Australia. 

We are currently studying genes that are potentially 

involved in the process of phosphate uptake in the 

arbuscular mycorrhizal (AM) symbiosis of soybean. We 

have extracted mRNA from AM colonised and uncolonised 

soybean roots using biotinylated poly-T probes and 

streptavidin-coated magnetic particles. Total mRNA 

extracted by this method was reverse-transcribed into 

cDNA with poly-T primer and reverse transcriptase. cDNA 

was next PCR-amplified with primers for a previously 

published soybean root phosphate transporter gene and 

amplified sequences were visualized on silver-stained 

acrylamide gels. In situ hybridization studies were carried 

out to localise the phosphate transporter in root tissues. 

Root cryosections were probed with a fluorosceinconjugated 

antisense probe and bound targets were 

visualized either via alkaline phosphatase labeling or with 

an antifluoroscein texas-red antibody. Results of these 

investigations are presented here. 

1090 - Profiling differences in metabolic activity 

between isolates of Penicillium camemberti using 

dielectric spectroscopy (impedimetric measurements) 

M. Decker * & P.V. Nielsen 

Technical University of Denmark, BioCentrum-DTU, 

Building 221, Technical Univeristy of Denmark, DK-2800, 

Kgs. Lyngby, Denmark. - E-mail: mad@biocentrum.dtu.dk 

In this work we characterise Penicillium camemberti starter 

cultures in order to predict the quality of white mould 

cheeses. Impedimetric changes in the first 100 hours after 

inoculation are detected using a Bactometer at a 1 kHz 

frequency. At this low frequency, the capacitance and 

conductance changes in the medium are measured, thereby 

indirectly measuring the metabolic activity of the fungi 

growing in the medium. Traditional impedimetric methods 

are used for detecting lag phase and growth rate. In the 

present work we try to use all impedimetric changes in the 

first 100 hours as a profile of the each isolate and 

subsequently treat the data using chemometric methods. As 

the metabolic activity of fungi depends strongly on the 

environmental conditions, this study is done using a cheese 

medium developed in our laboratory. 

330 


1091 - Intracellular enzymatic activity of the mycelium 

of Pleurotus spp. grown on media containing 2-deoxy- 

D-glucose 

G. Díaz-Godínez * & C. Sánchez 

Universidad Autónoma de Tlaxcala, Apartado Postal 129, 

Tlaxcala, Tlax. C.P. 90000, Mexico. - E-mail: 

gdg@cci.uatx.mx 

The strain selection of edible mushrooms has often been 

done by trial and error because there appear to be no clear 

correlation between mycelial growth rates evaluated on 

Petri dishes at laboratory level and productivity under 

production plant conditions. It has recently been found that 

the tolerance of some P. ostreatus strains to 2-deoxy-Dglucose 

(DG) at laboratory level, is correlated with 

improved yield of these strains at mushroom production 

level. In this work, the effect of DG on the intracellular 

activity of laccases (L), proteases (P), &beta-1,3glucanases 

(G) and endoglucanases (E) of the mycelium of 

strains of Pleurotus was evaluated. Six strains of P. 

ostreatus; 201216, 32783, 58052, 38537, 201218 from 

ATCC, and 3526 from NRRL, and two strains of P. 

pulmonarius; PPL27 and PPL34 from the Chinese 

University of Hong Kong Collection were studied. The 

strains were grown on a starch-based media with 0.01% 

(SDG) and without addition of DG (S) at 25 °C for 7 days. 

On S, all the strains presented L, P, G and E activity. 

However, on SDG, the strains 201216, 32783, 58052, 

38537 (previously reported as sensitive to DG) did not 

present any enzymatic activity. On SDG, the strains 

201218, 3526, PPL27 and PPL34 (previously reported as 

tolerant to DG) presented higher P, G and E activity than 

those presented on S. It suggests that the DG tolerant 

strains are derepressed for producing L, P, G and E, which 

could be importantly related to the mushroom productivity. 

1092 - Twin arbuscules in Linum usitatissimum L. 

S. Dickson 1* , P. Schweiger 2 , F.A Smith 1 , B. Söderström 3 & 

S.E. Smith 1 

1 The University of Adelaide, Waite Campus, PMB 1, Glen 

Osmond, South Australia 5064, Australia. - 2 Institute of 

Ecology and Conservation Biology, Vienna University, A- 

1090 Vienna, Austria. - 3 Department of Microbial Ecology, 

Ecology Building, Lund University, S-223 62 Lund, 

Sweden. - E-mail: sandy.dickson@adelaide.edu.au 

The 'twin' arbuscule development of Arum-type 

mycorrhizal colonization in Linum usitatissimum L. were 

investigated using a time-course study to show formation 

and senescence of the structures. Roots were freezesectioned 

longitudinally, mycorrhizal structures were 

visualized using 1) nitroblue tetrazolium as a vital stain to 

indicate metabolic activity of arbuscules and intercellular 

hyphae and 2) acid fuchsin counterstaining. Arbuscules 

occur in pairs in adjacent cortical cells arising from a 

single, radial intercellular hypha. These 'twin' structures 

often appeared to be at different stages of metabolic 

development. Arbuscules and their relationship to


intercellular hyphae and plant cortical cells were imaged 

using laser scanning confocal microscopy (LSCM). Using 

the optical xy confocal slices, a 3D reconstruction of a twin 

structure was produced and the surface area of each 

arbuscule was measured. The measurement of surface area 

together with logistic regression calculated over the time 

course study indicated that there was a delay in 

development of the second arbuscule. Intercellular spaces 

within the root cortex appear responsible for determining 

the structural type of colonization. Crooks (1933) described 

the unusual cell divisons (and hence intercellular spaces) in 

L. usitatissimum roots which is used to produce a 

conceptual model of this Arum-type mycorrhiza. Crooks 

DM. 1933. Botanical Gazette 95: 209-239. 

1093 - In vitro retention of 137 Cs and potassium by the 

mycelium of mycorrhizal and saprotrophic 

basidiomycete fungi 

I. Druzhinina 1* , V. Karg 2 & M. Berreck 3 

1 Institute of Botany, University of Vienna, Rennweg 14, A- 

1090, Vienna, Austria. - 2 Institute of Microbiology, 

University of Innsbruck, Innsbruck, Austria. - 3 Federal 

Institute for Agricultural Research, Vienna, Austria. - Email: 

druzhini@mail.zserv.tuwien.ac.at 

After Chernobyl disaster it has become apparent that 

natural losses of 137 Cs from the European forest soils are 

proceeding extremely slowly due to the ability of various 

fungi to accumulate this radionuclide. Large differences 

were observed in 137 Cs levels in fruit bodies of various 

edible fungal species often independent from the site and 

time of sampling. This phenomenon has led a number of 

radioecological modelers to conclude that radionuclides 

contamination in fungi is very difficult to predict. This 

paper presents results of the experimental investigation of 

137 Cs uptake by fungal mycelium versus undisturbed 

potassium metabolism. A special methodology was 

employed to study the ability of mycelium of mycorrhizal 

species - Suillus variegatus, S. grevillei - and saprotrophic - 

Pleurotus ostreatus, Stropharia rugosoannulata - to retain 

137 Cs and potassium dependent on: (i) genomic factor 

(strain and species), (ii) cultivation conditions (composition 

of the medium and the growth rate), (iii) metabolism 

activity and (iv) in a stream of time. Since high 

radiocaesium/potassium ratios were demonstrated for 

ectomycorrhizal fungi only, we postulate the existence of a 

high-affinity potassium efflux system in cells of 

mycorrhizal fungi; it is likely that this system promotes the 

retention of accumulated radiocaesium inside the cell, 

while potassium can be released back to the medium. 

1094 - Mapping of psychro- and/or halotolerance of 

Penicillium spp. and Aspergillus spp. and correlations of 

enzyme profile and habitat 

M.J. Due * , K. Karlshøj & J.C. Frisvad 

BioCentrum-DTU, Technical University of Denmark, 

Søltofts Plads, Building 221, DK-2800, Kgs. Lyngby, 

Denmark. - E-mail: c958218@student.dtu.dk 

The extracellular enzymatic activity of psychro- and/or 

halotolerant Penicillium spp. and Aspergillus spp. were 

studied and the correlations to habitat investigated. 170 

isolates were screened. As expected, the results showed 

that the Penicillia tend to be more psychrotolerant and the 

Aspergilli have better halotolerance. Except for the 

halophiles, it was observed that the higher the incubation 

temp., the better the halotolerance. Only mild halophiles 

were found. Of these many tended to stay halophile at 

lower temp. Among the Aspergilli only psychrotolerant 

isolates were found. The Penicillia yielded some 

psychrofilic isolates. The fastest growing isolates were 

selected for semi-quantitative enzyme screening. These 

were inoculated on triglyceride, casein and lignocellulosic 

liquid media with various amounts of NaCl added. The 

enzyme activities in the media were tested after 10 to 14 

days of growth. Assays were performed for proteinases, 

hemi- and cellulases and lipases on assay plates. It is 

expected that the isolates will exhibit habitat specific 

enzyme profiles, with the enzyme activity dependant on 

temperature and A w of the habitat. This hypothesis 

originates from the fact, that most fungi are highly habitat 

specific and not metropolitan. It is also expected, that while 

all (or most of) the isolates will have activity for the 

substrates screened against, there will be a difference in 

activity correlated to their habitat. 

1095 - A MAT-2 mating-type gene in the homothallic 

fungus Aspergillus nidulans 

P.S. Dyer 

School of Life and Environmental Sciences, University of 

Nottingham, University Park, Nottingham, NG7 2RD, U.K. 

- E-mail: Paul.Dyer@Nottingham.ac.uk 

Mating-type (MAT) genes have been identified from the 

pyrenomycete, loculoascomycete and discomycete classes 

of ascomycete fungi. By using hot-start PCR with 

degenerate primers, together with thermal asymmetric 

interlaced (TAIL)-PCR, it has been possible to identify a 

characteristic MAT-2gene from the plectomycete fungus 

Aspergillus nidulans. It includes a conserved high mobility 

group (HMG)-domain. RACE-PCR analysis has confirmed 

transcription of the gene during sexual reproduction and 

the presence of an intron in a conserved position within 

MAT-2 genes. Further analysis of the flanking regions of 

the MAT-2 gene revealed the presence of a 5S ribosomal- 

DNA sequence, a putative gene with homology to an 

element of the anaphase-promoting complex (APC) of 

Schizosaccharomyces pombe, and a putative gene with 

homology to the transmembrane receptor SYG1 from 

Saccharomyces cerevisiae. The presence of an APC 

homologue provided evidence of microsyteny around the 

MAT locus. However, no MAT-1 alpha-domain gene could 

be detected. This suggests either that a MAT-1 homologue 

may be present elsewhere in the genome, not directly 

adjacent to the MAT-2 locus, or that A. nidulans may 

contain only a MAT-2 mating-type gene. This would be a 



unique situation among homothallic euascomycetes so far 

analysed, which contain either a MAT-1 gene or both MAT- 

1 /MAT-2 genes. 

1096 - Field evaluation of yield and N2-fixation of 

chickpea (Cicer arietimum L.) as affected by 

phosphorus and biofertilizers 

I. El-Ghandour * , S. Soliman & Y. Gaal 

Soil & Water Dept., Nuclear Research Center, Atomic 

Energy Authority, 13759, Abu-Zabal, Egypt. - E-mail: 

elghandour_1@yahoo.com 

As a result of our previous work held under green-house 

conditions, we have decided to develop our experiment 

under field trial to generalize our results. So, a field 

experiment was carried out to evaluate the influence of 

biofertilizers, Mycorrhizae (AM) and / or Rhizobium (Rh), 

combined with super or rock - on the growth and N2fixation 

in chickpea grown in sandy soil. A minimal dose 

of super and rock-P were added at two doses (15 and 30 kg 

p/h) as well as 15 kg N /h as ordinary and labeled (15 NH4) 

2 SO4. Dry matter yield (DYM) of inoculated plants were 

higher than control treatments. Combined inoculation (AM 

+ Rh) in case of super or rock-P increased DYM than 

single inoculation. Also, the results showed that, single 

inoculation with Rhizobium increased yield in case of 30 

kg P/ h than 15 kg P / h application on the other- hand 

mycorrhizal infection increased chickpea production with 

15 kg P / h rather than with 30 kg P / h addition. AM 

infection increased nodule number, so nitrogen uptake 

increased in dual inoculation than the single one, at the 

same time N2 fixation quantities were so much in 

combined inoculation in case of super-P than with rock-P 

application. We can conclude that, from an economical and 

environmental point of view biofertilizers (AM or Rh) ) 

can save a great amounts of P and N fertilizers to increase 

crop production and this cleared the importance of 

biofertilizers and rock-P in clean sustainable agriculture 

systems. 

1097 - Identification of essential groups at the active site 

of inulinase from Penicillium cyclopium 

H.M. El-Shora 

Botany Department, Faculty of Science, Mansoura 

University, Mansoura, Egypt. - E-mail: 

shora@mans.edu.eg 

Amino acid residues involved at the active site of inulinase 

from Penicillium cyclopium were determined. DEPC 

(diethylpyrocarbonate) totally inactivates the enzyme but 

not in the presence of substrate. The inactivation reaction 

follows pseudo-first order kinetics with a second-order rate 

constant of 0.05 mM -1 min -1 . Reversal of inactivation in the 

presence of hydroxylamine leads to the inference that 

histidyl residues are essential for catalysis. EEDQ (2- 

332 


ethoxy-1-ethoxy-1,2-dihydroquinoline) inactivates the 

enzyme, but not in the presence of inulin, following 

pseudo-first order kinetics with a second-order rate 

constant of 0.02 mM -1 min -1 . This is indicative of the 

involvement of residues with a carboxyl group in the 

catalytic activity. Further kinetic analysis of the 

inactivation caused by EEDQ, strongly implies that 

modification of a single residue of aspartate or glutamate 

inactivates the enzyme. Chemical modification of inulinase 

with tryptophan specific reagent NBS (Nbromosuccinimide) 

inactivated the enzyme but the enzyme 

was protected with inulin. The inactivation with NBS 

indicated that tryptophan residues are essential for the 

enzyme catalysis. Treatment of the enzyme with pHMB (phydroxy-mercuribenzoate) 

and PMSF (phenylmethylsulphonyl 

fluoride) did not influence activity, thus 

eliminating the possibility that cysteine or serine participate 

in catalysis. Chemical modification of inulinase with (NAI) 

N-acetylimidazole and TNM (Tetranitromethane) caused 

inactivation of the enzyme but in the absence of inulin. 

These results suggest the necessity of tyrosyl group in the 

enzyme catalysis. 

1098 - In vitro element accumulation by macrofungi 

G. Farron * , D. Delavy, M. Aragno & D. Job 

Université de Neuchâtel, Lab. de Microbiologie, Case 

Postale 2, 2007 Neuchâtel, Switzerland. - E-mail: 

gilles.farron@unine.ch 

Element accumulation by macrofungi in natural conditions 

(especially at the level of carpopohores) has been studied 

since thirty years. Results often showed mycorrhizal 

species to be better heavy metals accumulators than 

saprophytes. In vitro studies can be used to find 

explanations on the phenomenon of element accumulation 

occuring in natural conditions (e.g. absorption, localization 

and resistance). Two kinds of research are usually 

performed: short term experiment without any growth or 

growth in element enriched medium or substrates. Our in 

vitro study integrate and compare both of these research 

possibilities, using a selection of five mycorrhizal and ten 

saprophytic fungi, three heavy metals (copper, iron and 

zinc) and one semi metal (selenium). Firstly, we analysed 

under growth condition the absorption (AAS, 

spectrophotometry) as a function of pH and element 

concentration. Secondly, we analysed the absorption of 

those species in short term experiments under the same 

conditions but without any growth. A comparison of both 

experiments showed the absorption depending on many 

factors such as pH, initial element concentration, species 

and kind of method chosen. As perspectives, we would like 

to optimize the absorption and localisation of copper, iron 

and selenium into different parts of mycelium and 

carpophore. Possible applications, e.g. in the cosmetic 

industry, where selenium is used as antioxydant or in the 

alimentary industry (functional food).


1099 - Demonstration of DHN-melanin pathway and 

genetic analysis of the gene encoding scytalone 

dehydratase in sapstain fungi 

C. Fleet 1 , C. Breuil 1 & A. Uzunovic 2* 

1 University of British Columbia, Dept. of Wood Science, 

Vancouver, B.C., Canada. - 2 Forintek Canada Corp., 2665 

East Mall, Vancouver, B.C., Canada. - E-mail: 

adnan@van.forintek.ca 

The presence of the 1,8-dihydroxynaphthalene (DHN) 

melanin biosynthesis pathway was demonstrated in several 

sapstain fungi using both chemical inhibitors and molecular 

techniques. The inhibitor compounds tricyclazole and 

carpropamid effectively reduced pigmentation at low 

concentrations in all tested fungal species, but also lead to 

growth inhibition at higher concentrations. The inhibitor 

cerulenin prevented fungal growth in all tested fungi at all 

tested concentrations, likely due to its inhibitory effect on 

another enzyme, the metabolically critical fatty acid 

synthase. Partial DNA sequences for the gene encoding 

scytalone dehydratase (SD) were obtained from species of 

Ceratocystis and Ophiostoma and found to have homology 

with known respective DHN-SD gene sequences. Sequence 

analysis of the partial SD amino acid sequences showed 

greater than 80% similarity among the sapstain species, and 

corresponded well with known phylogenies of sapstain 

fungi based on rDNA sequences. Aside from the work 

carried out on the isolate O. floccosum 387N, this is the 

first known documentation of the melanin pigmentation 

pathway used by species of the sapstain fungi from 

Ceratocystis, Leptographium and Ophiostoma (Eagen et. 

al. 2001, Wang et al. 2001, Wang and Breuil submitted). 

Furthermore, since no fungus has ever been found, to our 

knowledge, to have more than one melanin synthesis 

pathway, we can state that these species likely only use the 

DHN pathway for melanin production. 

1100 - Clay effects on the fungal growth morphology 

and copper sorption ability 

M. Fomina * & G.M. Gadd 

Division of Environmental and Applied Biology, Biological 

Sciences Institute, School of Life Sciences, University of 

Dundee, Dundee DD1 4HN, Scotland, U.K. - E-mail: 

m.fomina@dundee.ac.uk 

As a prelude to the development of biomineral sorbents for 

toxic metals, this study assesses the influence of clay 

minerals (bentonite, kaolinite and palygorskite) on the 

morphology of mycelial pellets produced by melaninforming 

microfungi and on the copper sorption properties 

of fungal/clay mixtures. In general, a reduction of pellet 

size, an increase in the length of surface hyphae of the 

pellets, and a reduction in exopolymer production were 

observed with increasing clay mineral concentrations up to 

5%(w/v). It was found that the clay particles were involved 

in the development of pellet structure of Cladosporium 

cladosporioides at all stages of growth. A general model of 

the structure of a fungal pellet grown in clay-containing 

medium is proposed. The pellets consist of three main 

layers: a central core with densely packed mycelium 

aggregated with solid clay minerals or a matrix of 

clay/polysaccharides; a middle layer with looser mycelium 

mixed with clay mineral flakes; and an outer, or 'hairy' 

zone, with loose hyphae surrounded by clay mineral flakes. 

A study of equilibrium Cu uptake from pH-buffered 

solutions showed that bentonite addition to the medium 

increased the sorption of Cu by Aureobasidium pullulans 

and C. cladosporioides grown in this medium. The 

mechanism of changed sorption capacity of the combined 

biomineral sorbents is suggested to occur by blocking or 

modification of binding sites on biotic and abiotic 

components of the 'biomineral' association. 

1101 - Effect of nutrient resources on growth and 

morphology of fungal mycelia penetrating toxic metal 

domains 

M. Fomina 1* , K. Ritz 2 & G.M. Gadd 1 

1 Department of Environmental and Applied Biology, 



2 National Soil Resources Institute, Cranfield University, 

Silsoe, MK45 4DT, England, U.K. - E-mail: 

m.fomina@dundee.ac.uk 

Natural environments contain a heterogeneous distribution 

of metal concentrations, and the ability of fungi to colonise 

such metal-contaminated domains will be influenced by the 

resources available to fungi. The aim of this study was to 

investigate the growth responses of some common soil 

fungi (Trichoderma viride and Gliocladium roseum) 

towards copper and cadmium under different nutritional 

conditions and using a system of tessellated agar tiles. The 

growth parameters recorded in this study demonstrated a 

decrease in metal toxicity with increasing concentration of 

available carbon source. It was shown that maximum 

extension rates and efficacy of carbon substrate utilization 

of both cultures decreased with increasing concentration of 

toxic metals. It was observed that in the gap between 

metal-free tiles and metal-containing tiles, the presence of 

the toxic metals led to negative chemotropic reactions and 

stopping of growth, swelling and lysis of some hyphal tips. 

Penetration of the hyphae into the metal-containing domain 

was often followed by the formation of very dense 

mycelium or mycelial 'bushes'. After fungi entered the 

toxic metal-containing domain, they often produced long 

sparsely-branched or branchless explorative hyphae. Our 

data have demonstrated that fungi efficiently use both 

'phalanx' and 'guerrilla' states of the mycelial system as a 

response to toxic metal stress combined with nutritionallypoor 

conditions. 



1102 - Ruderal and combatative strategies in insect 

pathogenic fungi examined by EST analysis 

F.M. Freimoser * , G. Hu & R.J. St. Leger 

Department of Entomology, University of Maryland, 4112 

Plant Sciences Building, College Park, MD, 20742, U.S.A. 

- E-mail: ff34@umail.umd.edu 

Metarhizium anisopliae (Ascomycota) and Conidiobolus 

coronatus (Zygomycota) are facultative saprophytes that 

are pathogenic to many insect species. About 2000 EST 

cDNA clones from each species were sequenced to analyze 

gene expression during growth on host insect cuticle and/or 

nutrient rich media. Proteases were the commonest 

transcripts with both species producing multiple subtilisins, 

as well as trypsin, metalloprotease and aspartyl protease 

activities capable of degrading host tissues and disabling 

anti-microbial peptides. However, compared to M. 

anisopliae, C. coronatus produced many fewer 

extracellular hydrolases (e.g., no phospholipases), 

antimicrobial agents, toxic secondary metabolites and no 

ESTs with putative roles in the generation of antibiotics. 

Instead, C. coronatus produced a much higher proportion 

of ESTs encoding ribosomal proteins and enzymes of 

intermediate metabolism consistent with the rapid growth 

characteristics of C. coronatus. These results are consistent 

with M. anisopliae using a combatative strategy to defend 

captured resources, while C. coronatus has modified the 

saprophytic ruderal-selected strategy using rapid growth to 

overwhelm the host and exploit the cadaver before 

competitors overrun it. Both strategies are consistent with 

specialization to pathogenicity. Thus, induction of 

proteases by host cuticles (mucoralean fungi do not 

produce proteases under these conditions) demonstrates 

that C. coronatus is adapted to entomopathogenicity. 

1103 - Induction of defense-related enzyme activities in 

poplar roots inoculated with compatible and noncompatible 

isolates of Paxillus involutus 

A. Gafur 1* , A. Schützendübel 2 & A. Polle 2 

1 Department of Plant Protection, University of Lampung, 

Jalan S. Brojonegoror 1, Bandar Lampung 35145, 

Indonesia. - 2 Institute for Forest Botany, University of 

Göttingen, Büsgenweg 2, D-37077 Göttingen, Germany. - 

E-mail: gafur@uwalumni.com 

Defense-related enzyme activities, especially those of 

peroxidases, were examined in short roots of control, noncompatible 

and compatible interactions between hybrid 

poplar (Populus tremula L. x P. tremuloides Michx.) and 

Paxillus involutus (Batsch) Fr. Peroxidase activities 

increased significantly in poplar inoculated with noncompatible 

isolate of the fungus compared to control, an 

indication that the plant recognized invasion of the noncompatible 

isolate and subsequently responded with 

activation of the defense responses. Elevated levels of the 

enzymatic activity were maintained after seven weeks of 

334 


inoculation. On the other hand, accumulation of the same 

enzyme was not detected in compatible associations. The 

failure of induction of peroxidase activity in mycorrhizal 

root tips may suggest low-scale or even lack of plant 

defense responses in the compatible association between 

poplar and P. involutus. In other experiments, an 

appreciable amount of hydrogen peroxide (H 2O 2) was 

surprisingly detected only in compatible tissues, leading to 

speculation on the presence of stress conditions in the 

tissues. It has to be noted, however, that the accumulated 

H2O 2 in the tissues may have been released by fungal 

mycelia forming mantle and Hartig net in compatible 

interactions. It has been recognized that mycelia of some 

ectomycorrhizal fungi, including P. involutus, produce 

H2O 2. 

1104 - Overflow of organic acids in Penicillium 

simplicissimum 

M. Gallmetzer & W. Burgstaller * 

Institute of Microbiology, Technikerstrasse 25, 6020 

Innsbruck, Austria. - E-mail: 

wolfgang.burgstaller@uibk.ac.at 

The efficiency of carbon source utilization by filamentous 

fungi is strongly influenced by environmental conditions. 

This is of advantage in biotechnological processes, because 

it is easier to vary environmental conditions than to change 

the metabolic properties of a fungus. Understanding how 

environmental conditions influence a fungal metabolism is 

therefore important. We used continuous cultivation to 

study the influence of the specific growth rate and of 

environmental conditions (glucose-, ammonium-, nitrate-, 

phosphate- and potassium-limited growth; various pH 

values, osmolarities and inhibitors) on the excretion of 

organic acids by Penicillium simplicissimum. P. 

simplicissimum is used in heterotrophic metal leaching 

processes. Citrate was under all conditions the main 

excreted intermediate. This excretion can be termed an 

energy spilling process, because it avoids the formation of 

NADH. The sum of the excreted organic acids was 

inversely correlated with the biomass yield. Glucose uptake 

was strongly influenced by the specicfic growth rate, but 

only weakly by the tested environmental conditions. The 

highest carbon overflow was observed with phosphate 

limitation (12% of the total consumed carbon, compared to 

1% during glucose limitation). Stress factors (alkaline pH, 

high osmolarity, addition of benzoic acid) in general 

increased both glucose uptake and carbon overflow. 

1105 - Calcium oxalate and gypsum on the thallial 

surface of the lichen Ramalina lacera exposed to 

polluted air 

J. Garty 1* , P. Kunin 1 , J. Delarea 2 & S. Weiner 3 

1 Tel Aviv University, Dept. of Plant Sciences, Tel Aviv 

69978, Israel. - 2 Tel Aviv University, Lab. Electron 

Microscopy, Tel Aviv 69978, Israel. - 3 Weizmann Inst. of


Science, Dept. of Structural Biology, Rehovot 76100, 

Israel. - E-mail: garty@post.tau.ac.il 

The formation of calcium-containing structures on the 

thallial surface of the lichen Ramalina lacera (With.) J.R. 

Laund. in response to air pollution and to simulated acid 

rain, was studied in in situ and transplanted thalli. In situ 

thalli were collected from an unpolluted site and 

transplanted to heavily polluted and to less polluted sites 

for a ten-months period. Additional thalli were treated 

either with double distilled water or with simulated acid 

rain. SEM and infrared spectrometry revealed that thallial 

surfaces of in situ R. lacera samples collected in unpolluted 

sites were covered with two kinds of calcium oxalate 

crystals. These aggregates of calcium oxalate crystals 

appear to disintegrate and provide a crystal layer on the 

thallial surface. Infrared spectroscopy of powder scrapped 

from thallial surfaces of transplants, retrieved from nonpolluted 

sites, showed the presence of whewellite and 

weddelite whereas powders obtained from thalli retrieved 

from polluted sites contained whewellite, weddelite and 

gypsum. It is suggested that a certain fraction of the 

gypsum detected in crater-like structures in transplants 

from polluted sites and in thalli treated with simulated acid 

rain is endogenous and should be considered a biomineral. 

1106 - Biotechnological aspects of basidiomycetes study 

V.P. Gavrilova 1* , N.S. Yakovleva 1 , O.V. Koroleva 2 , E.V. 

Stepanova 2 & E.O. Landesman 2 

1 V.L.Komarov Institute of Botany Russian Academy of 

Sciences, Prof. Popov Str., 2, St. Petersburg, 198022, 

Russia. - 2 A.N.Bach Institute of Biochemistry Russian 

Academy of Sciences, Leninskii pr.33, Moscow, 117071, 

Russia. - E-mail: Valeria@VG2438.spb.edu 

The study of physiological and biochemical peculiarities of 

Basidiomycetes opens a wide range of possibilities for their 

using in biotechnology. The first step of creation of 

biotechnological process starts with an investigation of 

physiology of fungal producer. The Collection of LE(BIN), 

containing a great number of Basidiomycetes provides the 

opportunity to carry out the study on biology of 

macromycetes development in culture including the 

investigation of enzymatic activities of macromycetes. The 

kinetic regularities (duration of lag-period, specific rate of 

growth, the velocity of carbohydrate consumption, 

economical coefficient, the influence of pH on the growth 

and metabolism) and peculiarities of ligninolytic enzymes 

biosynthesis under the submerged cultivation of 

Basidiomycetes belonging to families Polyporaceae, 

Ganodermataceae and Strophariaceae have been studied. It 

has been shown that rate of culture growth and enzyme 

synthesis can be regulated by cultivation conditions and 

desirable level of enzymes produced can be achieved by 

screening of appropriated producers. The combination of 

fast method developed for fungi screening on agar plates 

and the study of biosynthetic peculiarities of macromycetes 

under the conditions of submerged cultivation allows to 

chose the most prominent strains for different 

biotechnological applications, in particular for obtaining of 

ligninolytic enzymes, biologically active substances as well 

as biomass with high protein concentration. 

1107 - Identification of transposon-like sequences in the 

genome of fungi belonging to the Glomales 

A. Gollotte * , O. Chatagnier, C. Arnould, D. van Tuinen, S. 

Gianinazzi & V. Gianinazzi-Pearson 

UMR 1088 BBCE-IPM, INRA, BP86510, 21065 Dijon 

Cedex, France. - E-mail: a.gollotte@epoisses.inra.fr 

Arbuscular mycorrhizal fungi (Glomales) which are 

important for plant health and survival are ancient 

organisms which appeared with the first land plants more 

than 400 million years ago. Their genome is large (108 to 

109 bp) and is rich is repeated sequences. A high level of 

genetic polymorphism exists within their multinucleate 

spores. The aim of this work was to characterise repetitive 

DNA elements and provide a greater insight into the 

structure of the genome in the Glomales. Repeated 

sequences were isolated by screening lambda ZAPII 

genomic libraries from Glomus mosseae BEG12, 

Gigaspora rosea BEG9 and Gigaspora margarita BEG 34 

or by genomic self-priming PCR (GSP-PCR). Insert length 

ranged from 900 to 6000 bp and the copy number of each 

repeated sequence in the genome was between 70 and 

8000. Of 25 sequences that were isolated, all were rich in 

AT. Most of the sequences had no open reading frame and 

showed no homology to known sequences. However, short 

regions in these sequences were identical to elements 

present in transposons in other organisms. Two sequences 

showed similarity to reverse transcriptases. The distribution 

of these sequences in the genome is being determined by 

Southern blot and FISH. The presence of transposon-like 

sequences in the genome of the Glomales may have 

contributed to the appearance of high genetic 

polymorphism in the large genome of these fungi during 

evolution. This work is part of the european project 

GENOMYCA (QLRT-2000-01319). 

1108 - Black fungal colonies as units of survival: hyphal 

mycosporines synthesized by rock dwelling fungi 

A. Gorbushina 1* , M. Volkmann 1 , Th. Dornieden 1 , H. 

Ruetters 2 & J. Rullkoetter 2 

1 Geomicrobiology, ICBM, P.O.Box 2503, D-26111 

Oldenburg, Germany. - 2 Organic Geochemistry, ICBM, 

P.O.Box 2503, D-26111 Oldenburg, Germany. - E-mail: 

anna.gorbushina@uni-oldenburg.de 

Microcolonial fungi (MCF), long living vegetative mycelia 

frequently occurring on desert rock surfaces, are exposed to 

strong ultraviolet radiation, desiccation and nutrient 

scarcity. Fungal rock dwelling microcolonies lack 

ascospores and all their cells possess a thick melanised cell 

wall. These simply organised mycelial structures grow by 

mitotic cell divisions, but frequently are more stress- 



tolerant than lichens. Colonies of several randomly selected 

MCF strains were analyzed by cryo-SEM and TEM 

electron microscopy and for the presence of mycosporines. 

This class of UV-absorbing compounds, common in spores 

and other survival structures, is unknown in vegetative 

hyphae and was thought to be limited to reproductive 

morphogenesis. Mycosporine were analysed by LC/MS 

and LC/MS/MS (using a HPLC with a UV-VIS Detector 

and a ELSD interfaced with an ion trap MS). Intracellular 

intrahyphal growth and recolonization of old cells by new 

ones were observed in all MCF strains investigated. 

Mycosporines ubiquitous presence in the vegetative MCF 

cells help explain the high survival potential and longevity 

of these fungi. Territorial expansion of MCF is limited and 

mycosporines -known as inhibitors of conidia germination- 

may also serve as regulators of non-expansive restricted 

growth inherent for MCF. Also multiple cell functions such 

as UV-protection and the intracellular signaling for cell 

division and/or restriction of hyphal branching may be 

mediated by light-absorbing mycosporines. 

1109 - Mutational dissection of morphogenesis in 

Neurospora 

A. Griffiths, A. Virag, O. Gavric * & E. Jovel 

Botany, UBC, 6270 University Blvd. Vancouver V6T 1Z4, 

Canada. 

Morphogenesis in filamentous fungi is driven by 

programmes of tip growth and branching. The main 

components of current cellular models are the actin 

cytoskeleton, calcium homeostasis and morphogenes. We 

are investigating these components using the Neurospora 

model. First, we are characterizing many of the available 

morphological mutants by examining their responses to 

calcium and manganese at the macroscopic and cellular 

levels. This analysis has generated new groupings that 

could reflect function. Second, we are examining the 

functional interactions between mutations using epistasis 

analysis. The results require redefinition of the modes of 

interaction into full epistasis, partial epistasis, co-epistasis, 

novelty and cumulative or multiplicative action. This work 

also generates functional groups that might represent 

morphogenetic networks. Third, a mutation of the 

structural gene for actin has been isolated, cloned and 

sequenced. This clone has permitted more mutations to be 

isolated by repeat-induced point mutation (RIP). The 

properties of the mutant strains are being studied at the cell 

and molecular levels. They show a variety of types of 

aberrant growth, branching, aerial growth and sexual 

structures. Attempts at actin staining are in progress. 

[Supported by a grant from the Natural Sciences and 

Engineering Research Council of Canada.] 

336 


1110 - Natural protease inhibitors from fruit bodies of 

mushrooms 

K. Grzywnowicz * & K. Sobczyk 

University of Maria Curie-Sklodowska, Dept. of 

Biochemistry, Pl. M.C. Sklodowskiej 3, 20-031 Lublin, 

Poland. - E-mail: grzyw@hermes.umcs.lublin.pl 

Proteolytic enzymes have been firmly established as main 

regulatory components in a number of cellular, tissue and 

physiological processes. The most important factors 

influencing the proteolytic enzymes and pathways of 

proteolysis are natural protease inhibitors which form 

complexes with target proteases to inactivate and/or to 

regulate their activity. There is growing interest in 

inhibitors of proteases, not only synthetic but also naturally 

occurring (mushrooms' ones among them), due to their role 

in various human diseases. Searching for new, bioactive 

metabolites of basidiomycetous fungi we isolated and 

characterized some low and high molecular natural 

protease inhibitors of serine, asparagine, cysteine and 

metallo-proteases. They were isolated from fruit bodies and 

mycelia of edible, cultivable and potentially medicinal 

mushrooms (Pleurotus ostreatus, Lentinus edodes, 

Agaricus bisporus, Trametes versicolor). Isolation of 

inhibitors was achieved by ion exchange and size exclusion 

chromatography. Characterization of their inhibitory 

activity, pH and temperature optima, and molecular mass 

were analysed. It appeared that low molecular inhibitors of 

serine proteases were typical for both mycelium and fruit 

bodies, and high molecular inhibitors ones were 

characteristic for fruit bodies only. Inhibitors of asparagine 

proteases were typical for mycelium mainly. And inhibitors 

of cysteine proteases and metalloproteaes were found 

mainly in fruit bodies. 

1111 - Deliquescing of fruit bodies of Coprinus comatus: 

new insight into proteolysis of the process 

K. Grzywnowicz * & T. Typinski 

University of Maria Curie-Sklodowska, Dept. of 

Biochemistry, Pl. M.C. Sklodowskiej 3, 20-031 Lublin, 


Species of Coprinus are well recognized by their 

deliquescing of fruit bodies. Genus is also mentioned from 

the point of view of C. cinereus, which model organism to 

study development processes in the Homobasidiomycetes. 

The species C. comatus is characterized by melanized 

spores and inaequi-hymeniferous gills that tend to 

deliquesce into black liquid that drips from the 

disintegrating cap. The autolysis is evidently part of the 

developmental program of fruit body. The autolysis of the 

mature cap is caused mainly by chitinases. Process effects 

cap tissues, but not the outer veil layers and the stipe. Apart 

chitinases, proteases and glucanases also have been 

identified. In our work we try to evaluate proteolysis events 

during deliquescing process. We have been using 'wild' C.


comatus model instead of 'laboratory' C. cinereus one. 

During analysis of enzymology of stages of development 

of fruit body, we were investigating levels of enzymes 

connected with melanization proces and proteolysis events. 

We have been searching of also the levels of natural 

protease inhibitors of various groups of proteases. 

Comparison of changes in levels of protease activities and 

of their inhibitors may give us new, interesting insight into 

relations of various types of proteases and their natural 

inhibitors, and into a role of proteolysis during 

development and deliquescing of basidiocarps of genus 

Coprinus mushrooms. 

1112 - Characteristics of secondary metabolites from 

lichen mycobionts 

N. Hamada 1* , T. Tanahashi 2 & H. Miyagawa 3 

1 Osaka City Institute of Public Health & Environmental 

Sciences, 8-34 Tojo-cho, Tennoji, Osaka 543-0026, Japan. 

- 2 Kobe Pharmaceutical University, 4-19-1 Motoyamakitamachi, 

Higashinada, Kobe 658-8558, Japan. - 3 Div. of 

Applied Life Science, Graduate School of Agriculture, 

Kyoto University, Sakyo, Kyoto 606-8502, Japan. - E-mail: 

Nobuo.Hamada@iphes.city.osaka.jp 

Secondary metabolites were often obtained from colonies 

of spore-derived lichen mycobionts cultured on 

conventional malt-yeast extract media with 10% added 

sucrose. Some substances not detected in the lichens, were 

often new, and crystallized on the surface of mediumcultured 

mycobiont. These substances, which differed 

among the various lichen mycobionts, included 

graphenone, graphisquinone, dibenzofurans, xanthones, 

hybocarpone, isocoumarins and other substances. These 

substances appear to be often bioactive or toxic to 

photobionts. In addition to previous work identifying new 

mycobiont substances, further studies are performed in 

order to elucidate the biological significance of substances 

produced by the lichen mycobionts and the origin of lichen 

symbiosis. In the present study, we have examined the 

distribution and variation in dibenzofurans, which are 

common in lichen mycobionts, and distributed over many 

families, e.g. Stereocaulon, Evernia, Usnea, Lecanora. On 

the other hand, dibenzofurans were not found in lichens. 

This biosynthetic pathway was thought to dormant in 

lichenized condition, and was induced in cultured lichen 

mycobiont. The structure of dibenzofurans varies with 

species of lichens, and some dibenzofurans are chlorinated 

derivatives. These substances appear to be toxic to an algal 

partner. We tried to confirm this hypothesis in 

experimental procedure. The biological significance of 

these metabolites is discussed from the viewpoint of lichen 

symbiosis. 

1113 - T-DNA based genetic transformation of the 

ectomycorrhizal fungi 

M. Hanif 1* , M. Raudaskoski 1 , A.G. Pardo 2 & M. Gorfer 3 

1 

University of Helsinki, PL 56 viikinakari 9 00014-FIN HY, 

Finland. - 2 Programa de Investigación en Interacciones 

Biológicas (PIIB), Roque Saénz Peña 180, (B1876BXD) 

Bernal, Argentina. - 

3 Eco work lab, Vienna, 

Längenfeldgasse 27, 1120, Austria. - E-mail: 

hanif@mappi.helsinki.fi 

The T-DNA of Agrobacterium tumefaciens can be 

transferred to plants, yeasts, fungi and human cells. Using 

this system, a technique was developed for transforming 

ectomycorrhiza forming basidiomycetes Suillus bovinus, 

Hebeloma cylindrosporum, and Paxillus involutus. The 

selection marker employed was the Shble gene conferring 

resistance to phleomycin under control of the 

Schizophyllum commune GPD promoter and terminator. 

Putative transformants were shown by PCR to contain the 

GPDScP-Shble-GPDScT construct, although the fate of the 

foreign DNA could not be determined. In order to improve 

the system, dikaryotic mycelia of S. bovinus were 

transformed with recombinant hygromycin B 

phsphotransferase (hph) and enhanced green fluorescent 

protein (EGFP) genes fused with a heterologous fungal 

promoter and CaMV 35S terminator. Transformation 

resulted in hygromycin B resistant clones, which were 

mitotically stable. Putative transformants were analysed for 

the presence of hph and EGFP genes by PCR and Southern 

analysis which proved both multiple and single copy 

integrations of the genes. Several genes encoding small 

GTPases have been charcterized from S. bovinus, including 

SbCdc42 SbRac1, SbRas1 and SbRas2. The introduction of 

in vitro mutagenized dominant forms of these genes into S. 

bovinus genome by the transformation system described 

here will greatly advance our understanding of the function 

of the actin cytoskeleton and small GTPases in vegetative 

and symbiotic hyphae of S. bovinus. 

1114 - Characterization of copper-philic mycobiont 

from lichen Tremolecia atrata 

K. Hara * , H. Fujii, M. Komine, H. Hattori & Y. Yamamoto 

Akita Pref. Univ., 241-7 Shimoshinjo-nakano, Akita 010- 

0195, Japan. - E-mail: kojiro_h@akita-pu.ac.jp 

It is well known that lichens are able to accumulate high 

amount of heavy metals in their thalli. To clarify the 

possibility of application of lichens for bio-remediation, we 

had screened lichen mycobiont cultures for coppertolerance. 

As a result of the screening, a mycobiont derived 

from Tremolecia atrata found to be copper-philic rather 

than copper-tolerant. The copper ion level for optimum 

growth in liquid medium was between 25 to 30 ppm. The 

upper limit adequate for growth was around 125 ppm. 

Analyses by an atomic absorption spectrophotometer 

revealed an accumulation of copper ion within mycobiont 



aggregates. To cast further light on molecular mechanisms 

of this copper-philicity, differential protein expression 

analyses were performed, resulting in different protein 

profiles between mycobiont aggregates cultured with and 

without copper ion. 

1115 - Determining the relative role of ligand versus 

proton-promoted dissolution of metal phosphates by 

fungi 

F.A. Harper * & G.M. Gadd 

Department of Environmental and Applied Biology, 

University of Dundee, Biological Sciences Institute, 

Dundee, DD1 4HN, Scotland, U.K. - E-mail: 

f.a.harper@dundee.ac.uk 

Many soil fungi can solubilize sparingly soluble metal 

phosphates, and are thus important agents in nutrient 

cycling of phosphate and metal ions. Despite rapid 

screening methods to identify isolates with proficient 

phosphate solubilizing activity, there lacks quantitative 

data on the mechanisms by which dissolution occurs. Two 

non-mutually exclusive mechanisms involving efflux of (i) 

protons and/ or (ii) metal complexing ligands (such as 

organic acid anions) are frequently acknowledged in this 

role, although their relative importance in the dissolution 

process is unclear. In this work we used Aspergillus niger, 

Fusarium sp. and Coriolus versicolor, to develop 

contrasting models to provide a better mechanistic 

understanding of phosphate dissolution. We show that the 

relative role of proton versus ligand-promoted dissolution 

is influenced by many factors, including nitrogen-source, 

with marked differences occurring between fungal species. 

Many fungi subsequently immobilize metals, such as 

recrystallization as oxalates, while all fungi exhibit surface 

sorption and/or intracellular accumulation. Similar to the 

mechanisms of mobilization, these lack proper 

quantification. A sequential extraction technique has been 

used to fractionate metal in fungal biomass and agar into 

soluble, phosphate and oxalate phases, in order to quantify 

the fate of metal and achieve a mass balance. The 

functional significance of our findings for fungi in 

biogeochemical processes will be discussed. 

1116 - Production of neutral and alkaline proteases by 

the thermophilic fungus, Scytalidium thermophilum, 

grown on microcrystalline cellulose 

I. Hasbay Ifrij * & Z.B. Ogel 

Middle East Technical University, Department of Food 

Engineering, 06531 Ankara, Turkey. - E-mail: 

incinur@metu.edu.tr 

Scytalidium thermophilum protease production in Avicelcontaining 

media was analysed. Proteases produced by 

Scytalidium thermophilum were found to be most active at 

neutral and alkaline pH and at a temperature range of 37-45 

338 


°C. Highest protease activity was observed at early 

exponential phase where endoglucanase activity was low. 

The production of proteases and endoglucanases on 

different days suggested a possible negative effect of 

proteases on endoglucanase activity. The effect of protease 

inhibitors were examined to determine the type of 

proteases. Those were pCMB, antipain, PMSF, E-64, 

EDTA and pepstatin A. Antipain, pCMB and PMSF were 

observed to be effective on proteases produced by the 

fungus. Two major protease inhibition peaks at day 3 and 5 

suggest production of thiol-containing serine protease and 

serine protease, respectively. The presence of protease 

inhibitors in culture medium or cell-free supernatants failed 

to show a significant effect on endoglucanase activity and 

the ability of endoglucanases to adsorb onto Avicel. 

1117 - Development of an Aspergillus sojae expression 

system 

M. Heerikhuisen * , A. Drint-Kuyvenhoven, A.R. Albers, 

C.A.M. van den Hondel & P.J. Punt 

TNO Nutrition and Food Research Institute, Utrechtseweg 

48, Zeist 3704 HE, The Netherlands. - E-mail: 

Heerikhuisen@voeding.tno.nl 

Aspergillus species are known as high-level expression 

hosts for the production of enzymes or metabolites. In the 

last two decades for the industrially used species for 

Aspergillus niger, A. foetidus, A. tubigensis, and A. oryzae 

expression systems have been developed. We describe the 

development of an expression system based on the koji 

mold Aspergillus sojae for the production of homologous 

and heterologous proteins. Transformation based on 

auxotrophic (pyrG, niaD) and/or dominant markers 

(amdS), was used to introduce the genes of interest. To 

improve the yield of produced protein, Aspergillus sojae 

mutants with lower protease activity, either by gene 

disruption or by UV mutagenesis, were isolated. To 

improve the fermentation yields of Aspergillus sojae also 

morphological and so-called fermentor adapted mutants 

with lower viscosity, were isolated. As one of the examples 

for heterologous protein production, the production of 

human interleukin 6 was used. References: Heerikhuisen, 

M. et al. (2001) World Patent Application WO 01/09352. 

1118 - Isolation, structure elucidation and biological 

activities of novel secondary metabolites from Pochonia 

chlamydosporia (Goddard) Zare & W. Gams 

V. Hellwig 1 , A. Mayer-Bartschmid 1 , W. Zitzmann 1 , G. 

Greif 2 , H.V. Tichy 3 , G. Kleymann 1 & M. Stadler 1* 

1 Bayer AG, Pharma Research, Bayer Pharma Research 

Center, P.O.B. 10 17 09, D-42115 Wuppertal, Germany. - 

2 Bayer AG, Animal Health Research & Development, 

Bayer AG, Research Center Monheim, Bldg. 6700, D- 

51368 Leverkusen, Germany. - 3 TUEV ISB, Engesserstr. 

4b, C-79108 Freiburg, Germany. - E-mail: 

marc.stadler@t-online.de


Extracts from the fungus strain P0297 showed strong 

activities in two concurrent screenings of natural extracts 

for new antiviral compounds active against Herpes Simplex 

Virus 1 (HSV1) and for novel antiparasitic agents active 

against the protozoan pest Eimeria tenella, respectively. 

Strain P0297 was identified as Pochonia chlamydosporia 

var. catenulata sensu Zare et al. [Nova Hedwigia 73 (1-2) 

51-86, 2001] by morphological studies and comparison of 

DNA sequences with data from type strains. Fermentation 

of the fungus in 10 litre scale and bioassay-guided 

preparative HPLC of the crude extracts yielded several 

biologically active metabolites, which were identified by 

NMR spectroscopy and mass spectrometry. Besides 

Monorden (Radicicol), several structurally related 

resorcylic acid lactones, for which the trivial names 

Pochonins are proposed, were obtained as congeners. In 

addition, the spiro-alkaloid Pseurotin A was isolated. Upon 

modification of fermentation conditions, a drastic shift in 

the secondary metabolite profiles of strain P0297 occurred 

when bromide salts were added to the medium. 

Interestingly, the fungus now produced the monocillins, 

which constitute non-halogenated analogues of monorden 

and pochonins. All compounds were studied on their 

antiviral and anticoccidial properties in cellular replication 

assays. Furthermore, their activities against estrogenic 

receptors were evaluated. Their isolation, structure 

elucidation and biological activities are discussed. 

1119 - Heterologous expression of Phanerochaete 

manganese peroxidases in Pleurotus ostreatus 

Y. Honda 1* , C. Imamura 2 , H. Takahashi 2 , T. Watanabe 1 & 

M. Kuwahara 3 

1 Wood Res Inst., Kyoto Univ., Gokasho, Uji 611-0011, 

Japan. - 2 Toyota Central R&D labs., Yokomichi, Nagakute, 

Aichi 480-1192, Japan. - 3 Inst. Wood Technol., Akita Prif. 

Univ., kaieizaka, Noshiro, Akita 016-0876, Japan. - Email: 

yhonda@kuwri.kyoto-u.ac.jp 

Manganese peroxidases catalyze oxidation of Mn(II) to 

Mn(III) in the presence of hydrogen peroxide and form a 

class of extracellular peroxidases of white rot 

basidiomycetes. The enzymes have been focus of research 

interests because of their importance in lignin 

biodegradation and potential for utilization in many 

industrial processes. Genomic and cDNA sequences 

encoding manganese peroxidase have been cloned and 

characterized from variety of white rot basidiomycetes, 

including Ceriporiopsis subvermispora, Phanerochaete 

chrysosporium, Pleurotus ostreatus and Pleurotus eringii. 

However, overexpression of active ligninolytic peroxidases 

is difficult with non-basidiomycetous host systems such as 

Escherichia coli, Saccharomyces cerevisiae and 

Aspergillus spp. We have developed a recombinant gene 

expression system in P. ostreatus, using promoter and 

terminator sequences of sdi1 which encodes iron-sulfur 

subunit of succinate dehydrogenase. The system was 

successfully used to overexpress one of its manganese 

peroxidase genes, mnp3 under the control of homologous 

sdi1 expression signals. Here we report heterologous 

expression of wild-type and artificially mutagenized 

manganese peroxidases from P. chrysosporium using the 

gene expression system in P. ostreatus. 

1120 - Antifreeze proteins from snow mold fungi 

T. Hoshino * , M. Kiriaki, I. Yumoto & S. Tsuda 

National Institiute of Advanced Industrial Science and 

Technology (AIST), 2-17-2-1, Tsukisamu-Higashi, 

Toyohira-ku, Sapporo 062-8517, Japan. - E-mail: 

tamotsu.hoshino@aist.go.jp 

Many living organisms have biochemical strategies to 

protect themselves against freezing. Antifreeze protein 

(AFP) are one of biochemical defense mechanisms by 

which intracellualr ice formation is inhibited. Typhula 

ishikariensis, one of snow molds secreted a 22kDa protein 

that only accumulated in culture at a subzero temperature. 

Reported fish AFPs bound to prism faces of ice crystals 

that were formed hexagonal bipyramid. The short axes (aaxes) 

of ice crystals of fish AFPs were limited by the size 

of the initial ice. However, fungal AFP-bound ice could 

grow not only in the c-axis direction but also in the a-axis 

direction. Therefore, fungal AFP formed ice that was 10fold 

larger than that of fish AFP-bound ice and became a 

distorted hexagonal bipyramid shape resembling Stone Age 

knives. Fish AFPs can cover all ice-growth sites of prism 

faces and those AFP-bound ice crystals can grow only in 

the direction of basal faces (c-axis). Our results showed 

that fungal AFP-bound ice could grow not only in the caxis 

direction but also in the a-axis direction. Therefore, it 

is assumed fungal AFP-bound ice has extra space for 

crystal growth in prism faces. It is thought that the number 

of binding fungal AFPs per area of prism face is less than 

those of fish or plant AFPs. However, the highest 

measurable value of antifreeze activity of fungal AFPs is 

higher than average values reported for fish AFPs. It is 

therefore thought that fungal AFPs inhibit ice growth by 

another mechanism. 

1121 - Repression of chsB expression reveals the 

functional importance of class IV chitin synthase gene 

chsD in Aspergillus nidulans 

M. Ichinomiya * , H. Horiuchi & A. Ohta 

Department of Biotechnology, The University of Tokyo, 1- 

1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. - E-mail: 

aa07117@mail.ecc.u-tokyo.ac.jp 

Chitin is one of the major constituents of the cell wall of 

Aspergillus nidulans. To date, five chitin synthase genes 

(chsA-D, and csmA) in this fungus have been isolated and 

characterized in our laboratory. We have reported 

previously that a chsB single disruptant exhibits severe 

growth defects, whereas chsA, chsC, and chsD single 

mutants do not show any obvious growth defects. Here, we 

constructed conditional chsB mutants in which chsB 

expression was controlled under the repressible alcA 



promoter (alcA(p)-chsB). Under repressing condition, the 

mutant exhibited similar phenotypes to those of the chsB 

disruptant. To examine the functional relationships of other 

chitin synthase genes with chsB, we constructed three types 

of double mutants in which the chsA, chsC, or chsD 

deletion mutation was combined with alcA(p)-chsB. The 

chsD chsB double mutants grew more slowly than the chsB 

single mutant under high osmolarity conditions. The 

hyphae of the double mutants appeared to be more 

disorganized than those of the chsB single mutant. These 

results indicate the importance of the ChsD function in the 

absence of chsB expression. The chsA chsB and chsC chsB 

double mutants also showed different phenotypes from the 

chsB single mutant under alcA(p)-repressing conditions; 

the chsA chsB double mutant produced less aerial hyphae, 

and the chsC chsB double mutant showed reduced cell 

mass. These observations suggest that chsA and chsC each 

play some different roles in hyphal morphogenesis. 

1122 - Effect of the wood degrading basidiomycete 

Coniophora puteana on the chemical and physical 

properties of pinewood 

I. Irbe * , B. Andersons & J. Chirkova 

Latvian State Institute of Wood Chemistry, 27 Dzerbenes 

St., Riga, LV 1006, Latvia. - E-mail: ilzeirbe@edi.lv 

Coniophora puteana is a wood destroying basidiomycete, 

widespread both in nature and construction wood. The 

fungus produces a brown-rot in soft- and hardwood, 

primarily degrading its polysaccharides, while lignin is 

only modified. The fungus is often encountered in the 

buildings in Latvia, causing considerable damage to wood 

in service. The aim of this study was to investigate the 

chemical, physical and ultrastructural changes in the 

pinewood degraded by C. puteana (Schum.:Fr.) Karst. 

(BAM Ebw.15) isolate. For this, the Scots pine sapwood 

blocks were subjected to the attack of fungus for 4 months 

in laboratory conditions. Decayed wood was analyzed by 

ion exchange chromatography, water vapor sorption and 

scanning electron microscopy (SEM) methods. At the 

wood weight loss of 40%, the losses of hemicelluloses 

forming monosaccharides mannose and galactose were 

78% and 56%, while xylose and arabinose were lost by 

47% and 39%, respectively. The losses of cellulose 

forming monosaccharide, glucose, comprised 32%. Based 

on the water sorption isotherms, value of the accessible 

specific surface A, and constant am, characterizing the 

mass hydrophilicity, were determined. Correlation between 

the value A and wood weight losses was obtained. The 

hydrophilicity am tended to decrease as the share of lignin 

in decayed wood increased. SEM micrographs illustrated 

the pattern of thinning and cracking of the wood cell wall, 

confirming the considerable loss of wood carbohydrates. 

1123 - Solubilization of metal phosphates by fungi as a 

consequence of carbon translocation 

340 


H. Jacobs 1* , G.P. Boswell 2 , K. Ritz 3 , F.A. Davidson 2 & 

G.M. Gadd 1 

1 Division of Environmental and Applied Biology, 



2 Department of Mathematics, University of Dundee, 

Dundee, DD1 4HN, Scotland, U.K. - 3 Soil-Plant Dynamics 

Group, Scottish Crop Research Institute, Invergowrie, 

Dundee, DD2 5DA, Scotland, U.K. - E-mail: 

h.jacobs@dundee.ac.uk 

The spatial distribution of nutrients in most soils is 

heterogeneous due to factors that include soil type and 

structure. Pollutants, such as toxic metal-contaminated 

wastes, and soil amendments such as fertilizers, also 

increase the heterogeneity of the edaphic environment. The 

ability of fungi to solubilize insoluble metal phosphates, by 

the production of organic acids, is important for both plant 

and microbial nutrition. Whilst filamentous fungi are well 

adapted for growth in heterogeneous conditions, 

experimental information in relation to the functional 

consequences of such phenomena is sparse. A model 

system consisting of concentric rings of discrete agar 

droplets was developed which allowed study of fungal 

growth in vitro in nutritionally-heterogeneous conditions. 

Droplets containing different combinations of glucose and 

calcium phosphate were used to study the consequences of 

spatially separating these components in relation to metal 

phosphate solubilization by Rhizoctonia solani. In the 

presence of the fungus, solubilization of calcium phosphate 

only occurred when glucose was present in the underlying 

medium. However, solubilization occurred in droplets 

containing calcium phosphate, but no glucose, when 

glucose was present in other droplets within the tessellation 

and where fungal hyphae spanned the droplets. This 

demonstrates that substrate was transported via mycelia 

from glucose-containing domains, with the functional 

consequence of metal phosphate solubilization. 

1124 - Modulation of phospholipid metabolism in 

cAMP-dependent protein kinase mutants of Aspergillus 

niger 

K. Jernejc * & M. Bencina 

National Institute of Chemistry, Hajdrihova 19, SI-1000 

Ljubljana, Slovenia. - E-mail: katarina.jernejc@ki.si 

Cyclic AMP-dependent protein kinase (PKA) plays a 

crucial role in the regulation of metabolic pathways by 

means of enzyme phosphorylation and therefore in changes 

of the activities of key cellular enzymes. Genes encoding 

PKA catalytic and regulatory subunits in Aspergillus niger 

were cloned, characterized, overexpressed and/or deleted. 

The correlation between cAMP-dependent protein kinase 

activity and phospholipid biosynthesis in wild type A. niger 

strain and strains with overexpressed and deleted genes for 

regulatory pkaR and/or catalytic pkaC subunit was 

followed. Isolated mutants displayed different lipid, 

especially phospholipid metabolism. In mutant strains with 

abolished PKA activity amount of total lipids was 1.6 times 

higher, with much more neutral lipids than in other mutant


strains. Relative as well as absolute amounts of 

phospholipids and glycolipids were 1.3 and 4.9 times 

higher in mutants with regulatory subunit deleted than in 

the control strain. Differences in individual phospholipids 

were most distinctive. Mutant strains with increased pkaC 

and disrupted pkaR genes showed several times lower 

amount of phosphatidylcholine with concomitant increase 

in phosphatidylethanolamine and 

lysophosphatidylethanolamine, as well as differences in 

phospatidylinositol in relation to other followed strains. 

Therefore consequences of mutations of PKA on 

phospholipid synthesis could be interpreted through cAMP 

signaling. 

1125 - Copper ions might have a significant role in 

redirecting metabolic fluxes in Aspergillus niger 

K. Jernejc & M. Legisa * 

National Institute of Chemistry, Hajdrihova 19, SI-1001, 

Ljubljana, Slovenia. - E-mail: matic.legisa@ki.si 

In commercially important micro-organisms, such as 

Aspergillus niger, lipid synthesis was often regarded as an 

unwanted process since it diverts metabolic fluxes from 

final bio-product synthesis - citric acid. The presence of 

copper ions was found to induce the citric acid overflow, 

while concomitantly lower levels of total lipids were 

detected in the cells. Its effect was more obvious in the 

medium with magnesium ions as a sole divalent metal ions, 

while in the substrate with magnesium and manganese the 

addition of copper had less pronounced effect. Since the 

malic enzyme was recognised as a supplier of reducing 

power in the form of NADPH for lipid bio-synthesis, its 

kinetic parameters in regard to different concentrations of 

copper ions were investigated and a strong competitive 

inhibition of enzyme by Cu2+ ions proposed. It seemed 

that copper ions compete with Mg2+ and Mn2+ ions for 

the same binding site on the protein, since the deactivation 

of malic enzyme after the addition of metal ions was a time 

dependent process. Sesamol, another substance reported to 

reduce the lipid content in some fungi, had no significant 

effect in A. niger cells. The data illustrated that copper ions 

were capable of inhibiting the A. niger malic enzyme, 

which might lead to redirection of metabolic fluxes from 

lipid synthesis toward citric acid overflow. 

1126 - Ammonium assimilating GS/GOGAT genes of 

the ectomycorrhizal basidiomycete Suillus bovinus 

S. Jokela 1* , L. Paulin 2 , R. Sen 1 & J.T. Juuti 1 

1 

Department of Biosciences, PL 56 (Viikinkaari 9), 00014 

2 

University of Helsinki, Finland. - Institute of 

Biotechnology, PL 56 (Viikinkaari 9), 00014 University of 

Helsinki, Finland. - E-mail: sanna.jokela@helsinki.fi 

Nitrogen is a major plant growth limiting nutrient in boreal 

forests where the bulk of soil nitrogen is bound as organic 

complexes inaccessible to uptake by plants. Root symbiotic 

ectomycorrhizal fungi are able to mineralize organic soil 

nitrogen to ammonium and subsequently translocate 

nitrogen as amino acids to mycorrhizal roots. Glutamine 

synthetase (GS) and gluamate synthase (GOGAT) are key 

enzymes responsible for ammonium incorporation into host 

plant and fungal cellular amino acid pools. Genomic and 

cDNA libraries were constructed for the ectomycorrhizal 

basidiomycete Suillus bovinus, which is commonly found 

in Scots pine forest soils. Full genomic GS and GOGAT 

sequences, several GS cDNAs and one GOGAT cDNA 

were isolated and sequenced. The complete GOGAT 

cDNA sequence was determined using RT-PCR. 

Transcription initiation sites were identified using primer 

extension and confirmed by utilising 5'-RACE methods. 

Fungal transcription levels in the Scots pine 

mycorrhizosphere were assessed by Northern analysis. 

Although present as single copy genes, the amount of GS 

transcripts was much higher than that of GOGAT. The 

majority of GS transcripts seemed to be non-translatable, 

due to loss of mRNA 5'-ends, making the actual amounts of 

functional GS and GOGAT transcripts comparable. Total 

fungal GS transcript levels seem to be high throughout the 

mycorrhizosphere. 

1127 - Surface structure and composition of Aspergillus 

nidulans walls 

S. Kaminskyj 1* , K. Gough 2 & T. Dahms 3 

1 Biology Dept. Univ Saskatchewan, Saskatoon SK S7N 

5E2, Canada. - 2 Dept. Chemistry, Univ Manitoba, 

Winnipeg MB R3T 2N2, Canada. - 3 Dept. Chemistry, Univ 

Regina, Regina SK S4S 0A2, Canada. - E-mail: 

Susan.Kaminskyj@usask.ca 

Wildtype Aspergillus nidulans has highly polarized growth 

producing narrow tubular hyphae, but at 42 °C the 

conditional non-lethal hypA1 mutation leads to nonpolarized 

growth producing wide, tapered cells. TEM 

shows that 42 °C hypA1 tip walls are four-fold thicker than 

those produced at 28 °C, with wall thickness increasing 

subapically. Subapical walls of wildtype hyphae are 

inextensible, however, the 42 °C; hypA1 growth pattern 

implies they can remain plastic, consistent with Bartnicki- 

Garcia's model that wall softening is an integral part of 

deposition. Can differences between 28 °C and 42 °C walls 

of hypA1 strains be detected at the surface? hypA1 colonies 

were prepared for cryo-SEM after growth at 28 °C, 42 °C 

or a shift from 42 °C to 28 °C. Surprisingly, the wall 

surface was similar for all growth regimes: a finely pebbled 

texture, and no evidence of stretching or insertion of 

material even at tips. Atomic force microscopy (AFM) can 

create a detailed surface image, and in some modes can 

also provide chemical information, potentially monitoring 

changes as a hypha grows past a fixed point. Here we show 

that AFM images reveal the same wall textures in fixed 

chemically hypA1 cells as are seen with cryo-SEM. 

Nevertheless, despite the similarity between these surface 

images, infrared spectroscopy of wildtype hyphae showed 

that tips were relatively enriched for protein compared to 

subapical regions, which were enriched for carbohydrate. 



1128 - Nuclear NAD metabolism in fungi and yeast 

A.M. Kerbalaeva * , S.M. Nasmetova, D.M. Ruzieva & T.G. 

Gulyamova 

Institute of Microbiology AS RU, A.Kadiry 7 B, Tashkent- 

700128, Uzbekistan. - E-mail: imbasru@uzsci.net 

Activity of NAD metabolism enzymes in nuclei at 

stimulation of NAD production by F. sambucinum and S. 

cerevisiae selected earlier as NAD-producers has been 

studied. Cultivation of these fungi and yeast in the medium 

containing 1 mM nicotinamide was accompanied by up to 

3-fold increasing of intracellular NAD concentration in 

steady state growth phase. The increasing of intracellular 

NAD concentration accompanied by falling of its 

intranuclear pool caused by acceleration of coenzyme 

turnover due by growing up of nuclear NADpyrophosphorylase 

and NAD-ase activites. Also we found 

that the decreasing of nuclear concentration of NAD during 

stimulation of its synthesis in cells by nicotinamide 

correlates with its utilization at ADP-riboslylation of 

nuclear proteins. It was detected that increasing of basal 

and total ADP-ribosyl transferase activity attendant with 

appropriate changes in the level of modification of nuclear 

proreins as determined by the incorporation of 14 C -adenine 

from NAD into fungi and yeast nuclei. Moreover, as it was 

established by fractionation of nuclear proteins, although 

80% radiolabel is localized in the fraction of histones, the 

NAD content fluctuations mostly influences on the level of 

ADP-ribosylation of nonhistone proteins. The conclusion 

was made that ADP-rybosylation of nuclear proteins might 

be one of the possible mechanisms of NAD oversynthesis 

regulation in fungi and yeast. 

1129 - Alkaline-tolerant fungi from Thailand: a source 

for alkaline enzymes? 

W. Kladwang 1* , A. Bhumiratana 2 , N.L. Hywel-Jones 1 & L. 

Lange 3 

1 BIOTEC, 73/1 Rama VI Rd. Radjhevee, Bangkok, 

Thailand. - 2 Mahidol University, Rama VI Rd, Rajdhevee, 

Bangkok, Thailand. - 3 Novozymes, Bagsvaerd 2880, 

Denmark. - E-mail: wipapat@biotec.or.th 

A collection of 490 isolates of alkaline-tolerant fungi was 

established using PDA buffered at pH11. This was from 

sand, soil, tree-holes and assorted plant material collected 

from southern, central and northern Thailand. If possible 

the sample pH was recorded and was between pH4 and 9. 

Although alkaline habitats were good sources of alkalinetolerant 

fungi it was significant that alkaline-tolerant fungi 

could be isolated from samples that were pH4. Strains were 

screened for their ability to hydrolyze arabinan, amylose, 

potato-galactan and skimmed milk at pH10. Eleven genera 

had activity against at least one of these substrates. Of 

these 11, 7 were members of the Hypocreales. 

Furthermore, of 57 positives, 51 belonged to the 

Hypocreales and 38 were Acremonium/Stilbella spp. 

342 


Acremonium was therefore selected for further study. 

Alkaline enzyme production was tested using 9 substrates. 

Four different groups of Acremonium were separated based 

on their enzyme production profiles. From these, isolate 

WK276 (neutral rock-hole, south of Thailand) was selected 

for further study. Of the nine enzymes, the α-amylase 

properties of isolate WK276 were characterised. The Nterminal 

of WK276 α-amylase was studied for developing 

a specific primer. This was used to determine the complete 

sequence coding for the α-amylase. 

1130 - A time course study of functional and structural 

differences between two Glomus mosseae isolates 

L.M. Knudsen 1* , S. Dickson 2 , I. Jakobsen 1 & S.E. Smith 2 

1 Risoe National Laboratory, PO BOX 49, 4000 Roskilde, 

Denmark. - 2 Dept. of Soil and Water, University of 

Adelaide, PMB 1, Glen Osmond, SA 5064, Australia. - Email: 

lisa.munkvold@risoe.dk 

A time course study was conducted with symbioses 

between Cucumis sativus and two Glomus mosseae isolates 

BEG161 and BEG29 previously shown to differ in hyphal 

growth pattern. Four consecutive harvests were employed 

to investigate plant growth and P uptake through time as 

well as the development of intra- and extraradical 

mycelium of the two fungi. Intraradical fungal structures 

were visualised by light microscopy and laser scanning 

confocal microscopy of NBT- and acid fuchsin-stained 

sections. Mycorrhizal cucumber plants grew better and 

reached higher P contents than non-mycorrhizal plants 

within 29 days. However, the magnitude of the mycorrhizal 

growth response depended on the fungal isolate. The rate 

of intraradical development differed between the two 

isolates, but reached the same final level. The most rapid 

colonisation occurred in the time intervals 14-23 days and 

23-29 days for BEG161 and BEG29, respectively. The 

structure of the intraradical mycelium was similar, but 

arbuscular branching patterns generally seemed very 

variable. Finally, the present data show that cross walls, as 

reported by Dickson & Smith (2001), are also formed in 

arbuscular trunk hyphae of G. mosseae colonising C. 

sativus. 

1131 - The pecularieties of laccases biosynthesis by cocultivated 

white-rot fungi Cerrena maxima and Coriolus 

hirsutus 

O.V. Koroleva 1 , V.P. Gavrilova 2* , E.V. Stepanova 1 , I. 

Yavmetdinov 1 & A. Antipov 1 

1 A.N.Bach Institute of Biochemistry Russian Academy of 

Sciences, Leninskii pr.33, Moscow, 117071, Russia. - 

2 V.L.Komarov Institute of Botany Russian Academy of 

Sciences, Prof. Popova 2, St. Petersburg, 198022, Russia. - 

E-mail: Valeria@VG2438.spb.edu


Laccase (EC 1.10.3.2) belongs to a group of enzymes 

called blue copper oxidases which uses oxygen as electron 

acceptor to remove hydrogen from phenolic hydroxyl 

groups. This reaction leads to the formation of free radicals 

that can undergo rearrangements leading to alkyl-aryl 

cleavage, oxidation of benzyl alcohols, and cleavage of 

side chains and aromatic rings. Presently of great interest is 

the examination of laccase in relation to the possibility of 

enzyme applications in biotechnology in particular for pulp 

biobleaching. The main limitation of laccase-mediator 

system application is very high dosage of laccase and redox 

mediator needed for pulp beableaching or detoxification 

processes. In order to overcome this factor it is necessary to 

investigate the mechanism of lignin degradation, the 

interaction between lignolytic enzymes and especially their 

synergism during lignin degradation. The two strains 

Cerrena maxima and Coriolus hirsutus producing high 

yield of extracellular Mn-peroxidase and laccase under cocultivation 

have been found. It was shown that both C. 

maxima and C. hirsutus produced three isoenzymes pI 

3.53, 3.91, 4.25 and 3.6, 4.0, 4.5, respectively. However, 

during co-cultivation of these strains, only one laccase 

isoenzyme pI 4.0 has been produced. The physico-chemical 

properties and N-terminal amino acid sequence of this 

isoenzyme have been studied in comparison with 

isoenymes produced by pure fungal cultures of C. maxima 

and C. hirsutus. 

1132 - Aquatic hyphomycetes living under heavy metal 

stress 

G.-J. Krauss 1* & G. Krauss 2 

1 Martin-Luther University, Dept. 

Biochemistry/Biotechnology, Div. Ecology and Plant 

Biochemistry, K. Mothes Str.3, D-06099 Halle/Saale, 

Germany. - 2 Microbiology of Subterrestrial Aquatic 

Systems Group, UFZ Centre for Environmental Research, 

Leipzig-Halle, Theodor -Lieser -Str. 4, D-06120 

Halle/Saale, Germany. - E-mail: krauss@biochemtech.unihalle.de 

In a former copper shale mining district in Central 

Germany, weathering of slag heaps and dumps resulted in 

groundwater, lakes and streams with extremely high heavy 

metal and metalloid concentrations. Communities of 

aquatic hyphomycetes (AQH) in such streams are clearly 

impoverished but still surprisingly diverse, and leaf 

decomposition and conditioning proceeds even [1, 2]. 

Crystalline components form a crust over the developing 

fungal biofilms. Two strains of Heliscus lugdunensis 

isolated from a harsh and a moderately polluted habitat 

respectively, had clearly adapted to these conditions. 

Conidia of both strains are significantly distinct in shape 

and size. Their capacity for biosorption and accumulation 

of Cd(II) and Cu(II) in liquide culture was quite different. 

Under Cd stress both strains showed specific response in 

their production and turnover of glutathione (GSH) and 

sulfur-rich peptides. Both strains synthesized significantly 

more GSH in the presence of Cd ranging from 25 µM to 

100 µM, but quantitiy and time course were different. It 

seems likely that resistant AQH have undergone 

considerable physiological and genetic changes. [1] 

KRAUSS, G. et al. (2001) Aquatic hyphomycetes occur in 

hyperpolluted waters in Central Germany. Nova Hedwigia 

72: 419-428 [2] SRIDHAR, K.R. et al. (2001) 

Decomposition of alder leaves in two heavy metal polluted 

streams in Central Germany. Aquatic Microbial Ecology, 

26: 73-80. 

1133 - Diversity of genome size in zygotic meiosis of 

Pleurotus studied by flow cytometry 

B. Kullman 

Institute of Zoology and Botany, Estonian Agricultural 

University, 181 Riia Str., 51014 Tartu, Estonia. - E-mail: 

bellis@zbi.ee 

For quantitative evaluation of nuclear DNA and protein 

contents of spores, the flow cytometer (PAS) with staining 

DAPI SR101 was employed. The spore print of P. 

ostreatus (TAA 142824) is applicable as a standard 

(24Mb). The spore nuclei of P. ostreatus are unreplicated 

(Kullman, 2000). Two spore prints collected from wild (PV 

and PU) fruitbodes and one from a commercial (PM) 

fruitbody were studied. Two subpopulations were 

discovered in the spore print of PM and in PU and one in 

PV. The diversity of a spore print reflect the fate of hybrid 

genomes during meiosis. The spores of PM are divided in 

to two equally represented groups. One of them has the 

smallest DNA and protein content per spore compared with 

wild mushrooms. At the same time, these characters, 

especially protein content, are more variable. It can be 

suggested that in meiosis two hybrid genomes be divided. 

They differ 1.5 times in DNA content and 4.1 times in 

protein content, which corresponds to the loss of many 

chromosomes. Different chromosome numbers and 

genome sizes reported by several authors for one and the 

same species may also refer to the existence of a true 

aneuploidy indicating the plasticity of the fungal genome. 

Meiosis can occur even in the case of low density of 

homology between chromosomes (CLP and aneuploidy) 

and may ensure distribution of highly different strains. If in 

the zygote two nuclei are only a little conjugated than their 

mitotic haploidisation may results in dimorphism of spores. 

1134 - Isolation and characterization of thermophilous 

fungi from temperate soil of north India 

R. Kumar 

Guru Nanak Dev University, Department of Botanical and 

Environmental sciences, Guru Nanak Dev University, 

amritsar - 143 005, India. - E-mail: 

rajsalar@rediffmail.com 

Thermophilic fungi are now a well-recognised group for 

their temperature relationships, taxonomic characterization, 

industrial application and for wide occurrence. In the 

present study 19 species of thermophilic and 

thermotolerant fungi belonging to 14 genera were isolated 



and identified from temperate soils of varying altitudes of 

North India. The isolation of Chaetomium senegalense 

belonging to Ascomycetes, Myceliophthora fergusii and 

Synnmukerjiomyces thermophile belonging to mitosporic 

fungi are reported for the first time from India. The later 

species is a new synnematous hyphomycetous fungus. 

Detailed studies on importance value index (IVI) and 

numerical values for population number were conducted. 

Soil pH, moisture and altitude of their habitat were recored. 

In the present work, an attempt was also made to evaluate 

the thermostable enzymes especially, amylase, cellulase, 

lipase and xylanase employing Chaetomium thermophile, 

Thermomyces lanuginosus, Malbranchea sulfurea and 

Torula thermophila species. In the course of present 

investigation, it was further realised that the exploration of 

the biodiversity of thermophilic fungi has immense 

potential for the future. 

1135 - Gene expression in a whitish truffle during the 

phase transition from mycelium to fruitbodies 

I. Lacourt 1 , S. Duplessis 2 , S. Abba' 1 , F. Martin 2 & P. 

Bonfante 1* 

1 Dipartimento di Biologia Vegetale - Università di Torino 

and Istituto per la Protezione delle Piante del CNR-sezione 

di Torino, V.le Mattioli, 25 - 10125, Torino, Italy. - 2 UMR 

INRA/UHP, Centre de Recherches de Nancy, 54280 

Champenoux, France. - E-mail: p.bonfante@csmt.to.cnr.it 

The transition from vegetative mycelium to the fruit body 

in truffles requires differentiation processes which lead to 

edible ascomata consisting of different cell and tissue 

types. The identification of genes differentially expressed 

during these developmental processes can contribute 

greatly to a better understanding of truffle morphogenesis. 

A cDNA library was constructed from vegetative 

mycelium RNAs of the white truffle Tuber borchii and 214 

cDNAs were sequenced. Up to 58% of the ESTs were 

coding for known genes. The majority of the identified 

sequences represented 'housekeeping' proteins, i.e., 

proteins involved in gene/protein expression, cell wall 

formation, primary and secondary metabolism and 

components of signaling pathways. We screened 171 

arrayed cDNAs by using cDNA probes constructed from 

mRNAs of vegetative mycelium and ascomata to identify 

fruit body-regulated genes. Comparisons of signals from 

vegetative mycelium and fruit body, bearing 15% or 70% 

mature spores, revealed significant differences in the 

expression levels for up to 33% of the investigated genes. 

The expression level of six highly regulated genes was 

confirmed by RNA blot analysis. Expression of glutamine 

synthetase, 5-aminolevulinic acid synthetase, isocitrate 

lyase, thioredoxin, glucan 1,3-ß-glucosidase and UDPglucurosyl 

transferase were highly up-regulated suggesting 

that amino acid biosynthesis, glyoxylate cycle pathway and 

cell wall synthesis are strikingly altered during 

morphogenesis. 

344 


1136 - Methyl radical derived from non-enzymatic 

coordinated-copper system and its potential relation to 

the selective delignification by white-rot fungi 

P. Lamaipis 1* , K. Fackler 2 , T. Watanabe 3 , F. Pohleven 4 , M. 

Sentjurc 5 & K. Messner 2 

1 

Fermentation and Biochemical Engineering Laboratory, 

National Center for Genetic Engineering and 

2 

Biotechnology, Bangkok, Thailand. - Institute for 

Biochemical Technology and Microbiology, University of 

Technology Vienna, Vienna, Austria. - 3 Wood Research 

Institute, Kyoto University, Kyoto, Japan. - 4 Biotechnology 

Faculty, Department of Wood Science and Technology, 

University of Ljubljana, Ljubljana, Slovenia. - 5 Jozef Stefan 

Institute, Ljubljana, Slovenia. - E-mail: 

plamaipis@biotec.or.th 

White-rot fungi are able to degrade lignin more or less 

selectively. A non-enzymatic catalytic system consisting of 

a Cu-coordinating compound and hydroperoxides was 

proposed to play a role in the selective delignification of 

wood. White-rot fungi produce hydroperoxide containing a 

pyridine nucleus. Cu is present in wood. Hydroperoxide 

could be either obtained from lipid contained in resin or 

derived by fungi. Due to this hypothesis the radicals 

produced oxidatively by coordinated-Cu system could be 

the compound mediating the depolymerization of lignin. 

EPR measurement is the method of choice to elucidate the 

radicals produced from organic peroxides by coordinated 

transition metals. Methyl-, hydroxyl-, methoxyl-, and acyl 

radicals were generated and trapped with DEPMPO. The 

strongest signal was found for the methyl radical when Cu 

was coordinated with 4-AP. In uncoordinated reactions as 

well as when Fe or Mn was used, the hydroxyl radical was 

predominated. When these systems were applied on wood 

sections, fast delignification without major visual damage 

of the cellulose part of wood cell wall was only found with 

the coordinated Cu system. Faster delignification occurred 

on hardwood than softwood. These correlated to the 

preference of white-rot fungi on native lignin. Therefore, 

we propose that the methyl radical formation from 

coordinated-Cu and organic peroxide is the key step 

leading to selective delignification in native wood probably 

involve in selective delignify fungi. 

1137 - Genome characterization of the ectomycorrhizal 

fungus Paxillus involutus 

A. Le Quéré, T. Johansson, D. Wright, P. Samson, B. 

Söderström * & A. Tunlid 

Department of Microbial Ecology, Ecology Building, Lund 

University, Sölvegatan, 37 SE-223 62 Lund, Sweden. - Email: 

bengt.soderstrom@mbioekol.lu.se 

The basidiomycete Paxillus involutus is forming 

ectomycorrhizal (ECM) symbiosis with a broad range of 

forest trees. Reassociation kinetics and genomic 

reconstruction analyses on nuclear DNA indicated that P. 

involutus has a haploid genome size of 21.5 Mb including 

11% of repetitive DNA. Analyses of a 33-kb genomic


sequence indicated the presence of twelve potential ORFs, 

and by extrapolation the haploid genome of P. involutus 

was estimated to contain approximately 7,700 genes. In 

order to obtain information on genes specifically expressed 

during the symbiotic interactions with forest trees, 3,555 

Expressed Sequence Tags (ESTs) have been analyzed in a 

cDNA library constructed from an ECM formed between 

P. involutus and birch (Betula pendula). In parallel, cDNA 

libraries from saprophytically growing fungus (3,964 

ESTs) and from axenic plants (2,532 ESTs), respectively, 

have been analyzed. By assembly of all ESTs, 2,284 

unique transcripts have been identified either of fungal or 

plant origin. Approximately, 51-75% of the ESTs displayed 

significant homology to sequence information found in the 

GenBank (nr) protein database and these have been 

annotated to various functional and metabolic groups. The 

expression levels of the identified transcripts are examined 

using RNA blots and microarray analyses. 

1138 - Pathogenicity tests on Heterobasidion annosum 

hybrids from a S-type/P-type crossing 

M. Lind, Å. Olson & J. Stenlid * 

Dep. Forest mycology and pathology, SLU, Box 7026, S- 

750 07 Uppsala, Sweden. - E-mail: 

jan.stenlid@mykopat.slu.se 

Little is known about the pathogenicity of Heterobasidion 

annosum on a molecular level. It is however crucial to 

develop an understanding of the genes involved in order to 

fully understand the infection process. Knowledge of this 

process would be drastically enhanced if the genes of 

pathogenicity could be cloned and analysed, e.g. what 

proteins are involved, what singal pathways are used etc. 

The aim of this project is to study the pathogenicity of 

Heterobasidion annosum on pine at a molecular level. This 

goal will be obtained by the construction of a genomic 

map, using AFLP markers, and defining regions of the 

genome which contain genes of interest for the 

pathogenicity. These regions will be fished out of a 

genomic library and transformed into Heterobasidion 

annosum, using Agrobacterium tumefaciens or particle 

bombardment technique. In this first step, a S- and a P-type 

of the fungus has mated in laboratory conditions. The 

dikaryotic hybrid has been isolated and allowed to form 

fruitbodies. From these, 100 single spore isolates has been 

isolated. The pathogenicity of these isolates were tested on 

pine saplings, 2-3 weeks old. The results were measured in 

percentage of dead pines every two days for a 25 days 

period. These results will be correlated to the genomic 

map, in order to find regions of interest common for all the 

highly virulent isolates and not present in those of lower 

virulence. 

1139 - Efficient production by Aspergillus awamori of a 

Llama antibody fragment fused to a peroxidase 

B.C. Lokman 1 , V. Joosten 1* , R.J. Gouka 2 , C.T. Verrips 3 & 

C.A.M. van den Hondel 1 

1 TNO Nutrition and Food Research, P.O.box 360, 3700 AJ 

Zeist, The Netherlands. - 2 Unilever Research Vlaardingen, 

P.O.box 114, 3130 AC Vlaardingen, The Netherlands. - 

3 Department of Molecular Celbiology, Utrecht University, 

Padualaan 8, 3584 CH Utrecht, The Netherlands. - E-mail: 

joosten@voeding.tno.nl 

The development of fusion proteins consisting of antibody 

fragments and enzymes is of great medical and industrial 

importance. Previously, our group has demonstrated that 

single-chain Fv antibody fragments (scFv) could be 

efficiently produced by A. awamori (Frenken et al. 1998). 

Recently we have studied the production of Llama variable 

heavy-chain antibody fragments (VHH) by A. awamori. The 

advantage of V HH over scFv fragments is that V HHs are 

devoid of light chains. Furthermore, VHHs lack the 

hydrophobic regions that are normally facing the variable 

domain of the light chain and are therefore suggested to be 

better secreted than scFv fragments. A suitable enzyme for 

industrial applications is Arthromyces ramosus peroxidase 

(ARP). This 41 kDa monomeric glycoprotein has a broad 

specificity for phenolic and anilinic hydrogen donors. In 

previous studies we have demonstrated that there is no 

heme limitation during overproduction of ARP in A. 

awamori. Under control of the endoxylanase promoter 

secretion of active ARP was achieved up to 0.8 g/L in 

shake flask cultures (Lokman et al. submitted). Fusions 

between enzymes and VHHs permit interesting applications 

due to the fact that V HHs can direct enzymes to the place 

where they should act. This study shows the production of 

N-terminal and C-terminal fusions of ARP with a V HH 

fragment against the azo-dye RR6. RNA and protein 

analyses were performed to investigate possible limitations 

in production of the fusion proteins by A. awamori. 

1140 - Water stress effects on water/turgor potentials of 

mycelium of Agaricus bisporus and relationship with 

polyol accumulation/translocation of nutrients 

N. Magan 1* , T. Beecher 1 , S. Gray 2 & K. Burton 3 

1 Applied Mycology Group, Cranfield University, Silsoe, 

Bedford MK45 4DT, U.K. - 2 Department of Biology and 

Health Science, University of Luton, Park Square, Luton, 

U.K. - 3 HRI-Wellesbourne, Warwickshire, CV35 9EF, U.K. 

- E-mail: N.Magan@cranfield.ac.uk 

The mechanisms by which A. bisporus is able to 

translocate water and nutrients under dynamic and 

changing environmental conditions is little understood. We 

have studied the growth of A. bisporus strains from wet to 

dry conditions and vice versa to examine the changes in 

internal mycelial osmotic and turgor potentials and 

correlated this with accumulation of sugars and sugar 



alcohols for the first time. These studies demonstrated that 

matric and solute potential had makedly different effects on 

this relationship. Studies using radio-labelled nutrients 

showed that accumulation of phospohate varied with regard 

to water potential, growth rate of strain and with water 

potential. 

1141 - Effect of concentration of Tris-HCl buffer 

(hydroxymethyl aminomethane) on sporulation of 

arbuscular mycorrhizal fungi (AMF) 

L.C. Maia 1* , F.S.B. Silva 1 & A.M. Yano-Melo 2 

1 Universidade Federal de Pernambuco, Depto. de 

Micologia/CCB/UFPE, 50670-420 Recife, PE, Brazil. - 

2 Embrapa Semi-Árido, BR 428, km 152, Zona Rural. Caixa 

Postal 23, 56300-000 Petrolina, PE, Brazil. - E-mail: 

leonorcmaia@hotmail.com 

Organic buffers can stimulate growth of arbuscular 

mycorrhizal fungi through maintenance of stable hydrogen 

ion concentration, and by the stimulatory capability of the 

molecule, being an alternative for increasing inoculum 

production of AMF. In order to determine the effect of 

concentrations of Tris-HCl buffer on sporulation of AMF, 

fifth spores of: Glomus etunicatum Becker & Gerdemann, 

Gigaspora albida Schenck & Smith, Scutellospora 

heterogama (Nicol. & Gerdemann) Walker & Sanders, and 

Acaulospora longula Spain & Schenck were inoculated, 

separately, in Panicum miliaceum L. seedlings and five 

replicates of each treatment were maintained for 85 days at 

the greenhouse (27°C ±4°C and 45 - 81% relative 

humidity). The pots were irrigated every other day with 

nutrient solution supplemented with 0, 10, 25, 50, and 75 

mM de Tris-HCl and, once a week, the plants were 

irrigated with distilled water. Different results on spore 

production were related with the buffer concentration. 

Growth and sporulation of S. heterogama were not 

benefited by addition of buffer in the nutrient solution. 

Conversely, comparing with the control, 10 mM and 25 

mM of Tris-HCl were enough to increase significantly the 

sporulation, respectively, of G. etunicatum and G. albida. 

For A. longula, production of spores was significantly 

stimulated when the nutrient solution was supplemented 

with 50 or 75 mM of buffer. The results indicate that Tris- 

HCl can be used for improving sporulation of some AMF. 

1142 - Physiological and molecular variability in strains 

of a white rot fungus Physisporinus rivulosus 

P. Maijala, V. Salo * , C. Olsson, A. Dresler-Nurmi & A. 

Hatakka 

University of Helsinki, Department of Applied Chemistry 

and Microbiology, P.O. Box 56, FIN-00014 University of 

Helsinki, Finland. - E-mail: vanamo.salo@helsinki.fi 

An isolate of Physisporinus rivulosus has been shown to 

have excellent properties for the use as a pretreatment step 

346 


for mechanical pulping. In this study, several strains from 

type culture collections and the wild strain T241i were 

compared for the cultural morphology, growth 

characteristics and ability to decolorise Poly R dye and 

oxidise ABTS. Fungal growth in spruce wood was studied 

by light and confocal laser scanning microscopy using 

conventional anatomical stains, chitin binding lectin Alexa 

Fluor 660 conjugate, lipid staining Nile red as well as 

ethidium bromide and DAPI. Another basidiomycete 

Ceriporiopsis subvermispora was used in comparison. 

Extracellular enzyme activities of lignin degrading 

manganese peroxidase (MnP) and laccase were recorded. 

Strain variation at molecular level was investigated using 

sequence data from MnP encoding genes. Growth habit and 

speed on malt agar, in and on wood blocks revealed 

differences among Physisporinus strains. Variation was 

noted in their ability to oxidise the dyes used in the study. 

All Physisporinus strains produced MnP, although 

heterogeneity was found in the MnP isoenzymes secreted 

during fungal growth in wood chips. The strain T241i was 

the only one capable to lower the pH value of the 

surrounding growth medium during the growth. 

Morphological, physiological and molecular data is 

pointing to the conclusion that P. rivulosus is heterogenous 

and may be in need of further systematic study. 

1143 - Impact of ectomycorrhiza formation on the 

expression of water channel proteins (aquaporins) in 

fine roots of Norway spruce 

Z. Marjanovic 1* , N. Uehlein 2 , R. Kaldenhoff 2 , R. Hampp 1 

& U. Nehls 1 

1 

Eberhard-Karls-Universität, Physiologische Ökologie der 

Pflanzen, Auf der Morgenstelle 1, D-72076 Tübingen, 

Germany. - 

2 

Julius-von-Sachs Institut für 

Biowissenschaften, Lehrstuhl für molekulare 

Pflanzenphysiologie und Biophysik, Julius-von-Sachs Platz 

2, D-97084 Würzburg, Germany. - E-mail: 

zaklina.marjanovic@uni-tuebingen.de 

Root fungus symbiosis (mycorrhiza) improves the nutrient, 

and also the water supply of the host plant. Water flow 

across membranes is facilitated by water channel proteins, 

aquaporins. In order to detect a possible effect of 

mycorrhiza formation on root aquaporins, we used a sterile 

Petri dish system to obtain ectomycorrhiza between spruce 

(Picea abies) seedlings and fly agaric (Amanita muscaria). 

Two transcripts were detected with sequence homologies to 

plant plasma membrane aquaporins (PaPip1-1 and PaPip2- 

1). Translation of full length cRNA in Xenopus oocytes 

identified the respective proteins as functional aquaporins. 

Oocyte swelling studies showed that one of them, PaPip2- 

1, supported very high rates of water flow (Pf values about 

200), up to now only known for aquaporins from motor 

cells of leaf pulvini. An organ-specific Northern blot 

analysis showed that the expression of PaPip2-1 was 

mainly confined to fine roots, and that the amount of the 

transcript was decreased to about 25% of the control upon 

mycorrhization. In contrast, PaPip1-1 dominated in 

hypocotyl tissue, facilitated only low rates of swelling in 

oocytes (Pf values around 25), and its expression was not 

affected by mycorrhization. We suggest, that PaPip2-1 is


mainly responsible for facilitating water uptake via fine 

roots of spruce, and that the down-regulation of gene 

expression is an adaptation to the improved water supply 

mediated by the symbiotic fungus. 

1144 - Characterisation of a phosphate transporter in 

Candida albicans 

P. Martineau * , T.-L. Tremblay & L. Giasson 

Laval University, School of Dentistry, GREB, Quebec, Qc, 

G1K 7P4, Canada. - E-mail: 

philippemartineau@videotron.ca 

The human pathogen Candida albicans is a dimorphic 

fungus that can reversibly alter its mode of growth from 

yeast-like cells to mycelial cells, depending upon its 

environment. As the mycelial form is usually associated 

with candidiasis, genes involved in the control of 

dimorphism represent potential targets for the development 

of antifungal drugs. Using the differential display 

technique, we have identified a gene preferentially 

expressed in the mycelial cells. We have cloned and 

sequenced that gene. Sequence analysis revealed that this 

gene displays 70% homology with the Saccharomyces 

cerevisiae Pho89 gene, encoding a phosphate transporter. 

The C. albicans CaPho89 gene was cloned into a S. 

cerevisiae expression vector and transformed into a S. 

cerevisiae pho89 null mutant strain. Phosphate transport in 

the transformants was evaluated using [ 32 P]orthophosphate. 

The results obtained showed that CaPho89 

could complement the S. cerevisiae pho89 mutation, 

suggesting that CaPho89 encodes a functionnal phosphate 

transporter. 

1145 - Mitochondrial group-I intron in a higher 

basidiomycete, Pleurotus ostreatus, is mobile in sexual 

crosses 

T. Matsumoto * & Y. Fukumasa-Nakai 

Tottori Mycological Institute, Kokoge-211, Tottori-shi, 

Tottori, Japan. - E-mail: kin-matu@infosakyu.ne.jp 

Inter-parental recombination between mitochondrial (mt) 

genomes of Pleurotus ostreatus occurred at high frequency 

in sexual crosses. To reveal the process leading to this 

recombination, we estimated a certain recombination site in 

each parental mitochondrial genome contributing to the 

generation of the new restriction fragments which 

characterized a RFLP pattern of recombinant mtDNA, and 

determined their nucleotide sequences. It was found that 

the regions of recombination within parental mtDNAs 

possessed consensus sequences in common, which were 

divided by some regions of non-consensus sequences. 

Meanwhile, the nucleotide sequences in the new restriction 

fragments and their surrounding regions in recombinant 

mtDNA were harbored all consensus and non-consensus 

sequences recognized in both parental mtDNA, suggesting 

that this structure was caused by the integration of regions 

with non-consensus sequences that exist in only one 

parental mtDNA to the other. All non-consensus sequences 

were homologous to the sequences of the group I intron 

reported in fungi. They also included the ORF sequences 

that encode the LAGLIDADG motif, a characteristic of 

endonuclease involved in intron mobility. These results 

indicate that the manner of recombination for generating 

the new restriction fragment is elucidated by gene 

conversion, achieved by the mobility of the introns. This 

suggests that, in P. ostreatus, intron mobility is playing a 

large role for the increase of diversity of mtDNA. 

1146 - Effect of carminic acid and other mediators on 

lignosulfonates degradation by extracellular enzymes of 

Trametes versicolor 

A. Matuszewska * , G. Nowak & A. Leonowicz 

M. Curie-Sklodowska University, Departament of 

Biochemistry, Pl. M. Curie-Sklodowskiej 3, 20-031 Lublin, 

Poland. - E-mail: amatusze@hermes.umcs.lublin.pl 

Many low-molecular mass compounds have been 

suggested as possibly mediating mobile factors during 

biodegradation of lignin. Degradation of lignosulfonates by 

ligninolytic enzymes produced by the white-rot fungus 

Trametes versicolor was studied. The 29,6 kDa fraction of 

Peritan Na obtained by a gel filtration method using a 

Sephadex G-100 column was used as a substrate. The 

substrate was incubated with extracellular idiophasic 

enzymes of Trametes versicolor and with mediators. HBT 

(1-hydroxybenzotriazole), ABTS (2,2'azinobis-(3ethylbenzenthiazoline-6-sulfonic 

acid) and carminic acid 

were used as mediators. Following the incubation products 

were separated using gel filtration on a Sephadex G-50 

column and capillary electrophoresis. When the mediators 

were added to the incubation mixture, depolymerization 

was more extensive than in the experiment with no 

mediators. In the presence of carminic acid the 

depolymerization was the most extensive and as it's result a 

fraction of about 2 kDa appeared. Similar results were 

observed when HBT or ABTS were used. However, 

products of higher molecular weight besides the 2 kDa 

fraction were also obtained. Experiments with the reaction 

mixtures supplemented with H2O 2 were also performed. 

The above experiments revealed that only in the case of 

HBT used as a mediator H 2O 2 addition resulted in the 

increased depolymerization of lignosulfonates. Supported 

by the EC Contract ICA 2-CT-2000-10050, and by KBN 

grant 139/E-SPUB-M-5PR-UE/DZ 280/2000. 



1147 - Comparative biochemical analysis of enzyme 

electrophoretic spectra from mycorrhizal roots 

R.C. Maximilian 1* , E.M. Carasan 1 , A. Brezeanu 1 & A. 

Rosu 2 

1 

Institute of Biology, Spl.Independentei 296, 79651, P.O. 

Box 56-53, Bucharest, Rumenia. - 

2 

Faculty of 

Biotehnology, University of Agricultural Sciences, 

Str.Marasti 59, 71331, Bucharest, Rumenia. - E-mail: 

cmaxi@ibiol.ro 

Metabolic changes induced by mycorrhizal colonization 

can be evaluated by modifications appear in enzyme 

activities. We have investigated the potential use of 

electrophoretic analysis for detection the specific isozyme 

that presents differences in metabolic pathway in maize 

colonized roots. Staining protocols for esterases, alkaline 

phosphatases (ALP) and malate dehydrogenases (MDH) 

provided the enzymes for the mycorrhizas. We found 

differences between spectra of these enzymes at various 

time of cultivation. We used a starter inoculum of Glomus 

mossae (SI) and a current granular inoculum of Glomus sp 

(CGI). For esterases, the electrophoretic spectrum revealed 

5 bands for mycorrhizal roots comparative with 3 bands of 

non-mycorrhizal roots using SI. After 42 days of 

colonization on the gel appeared 2 electrophoretic bands. 

For MDH the electrophoretic spectrum showed 6 bands at 

12, 34 days and 2 bands at 42 days of symbiosis. We found 

no differences between electrophoretic spectra of SI and 

CGI. We used also various concentrations of inocula. The 

electrophoresis spectra emphasized the following: 9 bands 

for SI 5% and 10 bands for SI 10% comparative with 8 

bands for control and 9 bands for both concentrations 5% 

and 10% CGI for esterases. 8 electrophoretic bands using 

5% and 10% concentrations of SI and CGI, and also 8 

bands at control for detection isozymes of MDH. 3 

electrophoretic bands of isozymes of ALP were revealed 

comparative 2 bands for control. 

1148 - Early molecular communication among plant 

and fungus: detection of specifically expressed genes 

during pre-symbiotic interaction 

M. Menotta 1* , A. Amicucci 1 , A.M. Gioacchini 2 , D. Sisti 3 & 

V. Stocchi 1 

1 Istituto di Chimica Biologica Giorgio Fornaini, Università 

degli Studi di Urbino, Via Saffi 2 I 61029 Urbino (PU), 

Italy. - 2 Istituto di Ricerca Attività Motoria Università degli 

Studi di Urbino, Via Sasso I 61029 Urbino (PU), Italy. - 

3 Istituto e Orto Botanico Università degli Studi di Urbino, 

Via Bramante 28 I 61029 Urbino (PU), Italy. - E-mail: 

mmenotta@uniurb.it 

Tuber borchii Vittad. is an ascomycetous fungus which 

forms ectomycorrhizae on the roots of angiosperms and 

gymnosperms. After the achievement of this symbiotic 

association, the mycelium can form the hypogeous 

fruitbody, commonly known as truffle. Ectomycorrhizae 

348 


formation is a highly regulated process that is accompanied 

by molecular reorganization of both partners during 

symbiosis. An analogous molecular mechanism also takes 

place during the pre-symbiotic phase, when the partners 

exchange molecular signals, in order to take their stand and 

prepare both organisms for symbiosis instauration. We 

studied this latter genetic reorganization in T. borchii 

during the interaction with its symbiotic plant Tilia 

americana. For this purpose we set up a culture system 

where the mycelium interacts with the plant, but avoiding 

physical contact between the two organisms. By 

suppressive subtractive hybridization, we identified several 

specifically expressed genes. All selected clones were 

further analysed by Northern blot hybridization comparing 

transcript levels of control RNA (mycelium grown alone) 

and tester RNA (mycelium grown in presence of T. 

americana), both obtained from a tested in vitro 

ectomycorrhizae synthesis system. The same analysis was 

conducted on RNA of T. borchii extracted at different stage 

of its life cycle. Some differentially expressed genes are 

involved in nuclear rearrangement, while some others seem 

to be involved in extra cellular signal transduction. 

1149 - Molecular karyotype of Thraustochytrium 

striatum Schneider 

J.I. Mitchell & J. Slatter * 

School of Biological Sciences, University of Portsmouth, 

King Henry Building, King Henry I Street, Portsmouth 

PO1 2DY, England, U.K. 

The thraustochytrids comprise a group of marine protists 

that are important ecologically and commercially. Strains 

of thraustochytrids can produce commercially significant 

quantities of docosahexaenoic acid (DHA), a medically 

important long chain polyunsaturated fatty acid. 

Ecologically, thraustochytrids have been identified as 

important degradative organisms, breaking-down naturally 

occurring recalcitrant carbon-containing polymers, and an 

important food source for picoplankton. Despite the rising 

importance of thraustochytrids, little is known about their 

life cycle or their molecular biology. This study presents 

preliminary findings on the genome of Thraustochytrium 

striatum Schneider, a member of the marine protistan 

group the thraustochytrids. Pulsed field gel electrophoresis 

studies identified 18 presumptive chromosomes from the 

zoospores and thalli of T. striatum. The sizes of these 

molecules varied from 0.4 to 2 Mbp and gave a genome 

size of 18.77 Mbp. The chromosome locations of the 

D15/n-3 desaturase gene, 18S rRNA genes and thymidine 

kinase gene were determined. Results indicated that there 

were at least four copies of the desaturase gene on separate 

chromosomes; multiple copies of the 18S rRNA gene 

present on all chromosomes and two copies of the 

thymidine kinase gene. The unusual gene distribution and 

unequal distribution of chromosome bands suggests that 

these organisms might be processing their chromosomes.


1150 - Ecophysiological manipulation of fermentation 

improves viability of the biocontrol yeast Pichia 

anomala 

S. Mokiou * & N. Magan 


University, Silsoe, Bedford MK45 4DT, U.K. - E-mail: 

s.mokiou.s00@cranfield.ac.uk 

For effective biocontrol to be achieved it is important that 

cheap and economic substrates can be used to produce 

ecologically competent inocula. To this end we have 

examined and compared the production of the biocontrol 

yeast P. anomala on rich defined media (NYDB) and on 

molasses. Manipulation of the physiology of the yeast by 

modification of water stress [water activity (aw) of 0.98 and 

0.96] using different compatible solutes/sugars and NaCl 

markedly affected yield and quality of the cells. 

Endogenous water potential of cells, and sugar/sugar 

alcohol contents were significantly modified.In general, 

accumulation/synthesis of trehalose or sugar alcohols was 

affected by the solute used in media. For example, use of 

proline, glucose and sorbitol in molasses-based media 

resulted in accumulation of the desiccation protectant 

trehalose, while proline, NaCl, glucose, sorbitol and 

glycerol in media resulted in an accumulation/synthesis of 

glycerol and varying amounts of arabitol. Ecological 

competence of the yeast treatments was examined by 

plating on non-stressed (0.995 aw) and water stressed media 

(0.96 a w). Viability was significantly improved by the use 

of some solutes in molasses-based media. Such studies 

have implications for improving shelf-life and perhaps the 

production of ecologically stable biocontrol agents. 

1151 - Identification of mating type sequences in 

toxigenic Fusarium species known as asexual fungi 

A. Moretti 1* , Z. Kerényi 2 , G. Mulè 1 , C. Waalwijk 3 & L. 

Hornok 2 

1 Institute of Science of Food Production (CNR), Viale 

Einaudi, 51, Bari, Italy. - 2 Agricultural Biotechnology 

Center, P.O. box 16, Szebet-Gyorgy 4, 2100 Godollo, 

Hungary. - 3 Plant Research International, Business Unit 

Biointeractions and Plant Health, P.O. box 16, 6700 AA, 


moretti@area.ba.cnr.it 

Fusarium species currently known as asexual fungi showed 

to possess mating type (MT) sequences similar to those of 

species in which sexual recombination regularly occurs. 

Two pairs of degenerated oligonucleotide primers based on 

known fungal MT sequences were designed. Each of the 

two conserved MT sequences, ALPHA- and HMG-boxes, 

could be detected by PCR in F. camptoceras, F. cerealis, 

F. culmorum, F. poae, F. langsethiae, F. semitectum, F. 

sporotrichioides, F. redolens and F. oxysporum. Sizes of 

MAT-1 and MAT-2 amplicons were 200 bp, and 260 bp 

long. Both MAT-1 and MAT-2 sequences were identified 

for all species, but F. camptoceras (MAT-2) and F. 

langsethiae (MAT-1) for which just one MT was detected. 

As the sexual behaviour of F. avenaceum is not clear, this 

fungus was also studied. No strain of F. avenaceum was 

found to harbour both MT sequences, although it has been 

reported as weak homothallic species. On the other hand 

both MAT-1 and MAT-2 sequences occurred in single 

strain of F. graminearum, a typical homothallic species. 

RT-PCR experiment was set up to find the transcription of 

MAT genes in F. avenaceum, F. culmorum, F. poae and F. 

sporotrichioides. By using the primers pairs, 

fusALPHAfor/fusALPHArev and fusHMGfor/fusHMGrev, 

one characteristic PCR amplicon was produced in all 

samples, indicating that MT genes are transcribed in all 

asexual Fusarium strains included in these experiments. 

Support of EU Fifth Framework (Detox fungi QLK1-CT- 

1999-001380). 

1152 - Analysis of CHS8 - a fourth chitin synthase 

isoenzyme of Candida albicans 

C.A. Munro, R.K. Adam, H.B. Hughes * , S. Selvaggini, M. 

Reilla & N.A.R. Gow 

Department of Molecular and Cell Biology, Institute of 

Medical Sciences, Foresterhill, Aberdeen, AB25 2TP, 

Scotland, U.K. - E-mail: b.hughes@abdn.ac.uk 

All fungi studied to date have multigene families of chitin 

synthase enzymes. In the dimorphic pathogen Candida 

albicans, three genes encoding chitin synthases have 

previously been identified. CHS1 encodes a chitin synthase 

involved in primary septum formation and is essential for 

viability. CHS2 is not essential for viability, but is 

responsible for the majority of chitin synthase activity 

detectable in vitro. CHS3 is responsible for producing most 

of the chitin present in both yeast and hyphal cell walls, 

and deletion of the gene results in attenuation of virulence. 

Recently a fourth chitin synthase gene CHS8 encoding a 

second class I enzyme was identified in the genome 

sequence. The chs8 null mutant was more sensitive to 

certain agents disruptive to cell wall integrity, had a 

significant reduction in the in vitro hyphal chitin synthase 

activity and a slight reduction in hyphal chitin content. 

Microsomal preparations of chs2/chs8 double mutant had 

very low chitin synthase activity but the organism had 

normal morphology. Class I enzymes are therefore 

dispensable for normal growth of this organism. 

1153 - Cloning of chitin deacetylase gene from 

Phycomyces blakesleeanus and its expression in 

Escherichia coli 

T. Murayama 1* , A. Miyazaki 2 & K. Imamura 1 

1 College of Engineering, Kanto-Gakuin Univ., 1-50-1 

Mutsu-ura Higashi, Kanazawa-ku, Yokohama 236-8501, 

Japan. - 2 Graduate School of Life Sciences, Tohoku 

University, Katahira, Aoba-ku, Sendai 980-8577, Japan. - 

E-mail: murayama@kanto-gakuin.ac.jp 



Chitosan is a biopolymer with unique properties favorable 

for a broad variety of industrial, biomedical, and 

agricultural application. The chitosan is produced from 

crab shell through the chemical procedure which leaves a 

lot of industrial waste at present. Chitin is converted to 

chitosan by chitin deacetylase. To develop an enzymatic 

process for chitosan production that reduces the waste, we 

have planned to isolate a clone of chitin deacetylase gene 

from Phycomyces blakesleeanus cDNA library which we 

made in the _ZAP vector. The 218bp-DNA fragment 

amplified from genomic DNA of P. blakesleeanus using 

PCR (polymerase chain reaction) primers to two highly 

conserved sequences within chitin deacetylase gene of 

Mucor rouxii was the probe for isolation the clone from the 

library. A positive cDNA clone which is composed of 

1,613bp was isolated. The deduced protein encoded by this 

clone is composed of 459 amino acids. The protein 

sequence comparison revealed that the protein is 

significantly similar to chitin deacetylase of M. rouxii. 

After ligation of the clone to the expression vector pET32 

Ek/LIC, chitin deacetylase activity was detected in the 

purified His-tagged protein fraction of the crude extract 

from a E. coli transformant. These results show that the 

chitin deacetylase gene has been successfully cloned from 

P. blakesleeanus. The cloned gene was named PbCD and 

the nucleotide sequence of PbCD was registered to 

GenBank (accession #AB046690). 

1154 - Genome analyses of Micromucor ramanniana and 

Micromucor isabellina 

Á. Nagy 1* , M. Pesti 1 & C. Vágvölgyi 2 

1 

University of Pécs, Department of General and 

Environmental Microbiology, Pécs, Ifjúság u. 6, H-7624, 

Hungary. - 

2 

University of Szeged, Department of 

Microbiology, Szeged, P.O. Box 533, H-6701, Hungary. - 

E-mail: nagi@freemail.hu 

The Micromucor species (genus Mortierella) belong 

among the most common saprophytic soil fungi. Some 

members of the genus (Mortierella species) are potent 

producers of the long-chain polyunsaturated fatty acids 

(e.g. arachidonic acid). These compounds are important 

both nutritionally and pharmacologically. These fungi are 

biotechnologically important, but the organisation of their 

nuclear genome has not been described so far. 

Electrophoretic karyotype analysis using different 

approaches of pulsed field gel electrophoresis (PFGE) has 

led to significant progress in fungal genetics: the physical 

karyotypes of numerous previously genetically 

uncharacterised fungal species have been established. In 

the present study, orthogonal field alternation gel 

electrophoresis (OFAGE) and the contour clamped 

homogeneous electric field (CHEF) technique were applied 

to obtain preliminary information on the organisation and 

intrageneric variability of the nuclear genome in three 

Mortierella (Micromucor) strains of 2 different species (M. 

isabellina and M. ramanniana). Conditions for the 

preparation of highly-intact chromosome-size DNA 

molecules and for the separation of DNA molecules were 

established. Furthermore, through the use of homologous 

and heterologous gene probes (e.g. 3-hydroxy-3- 

350 


methylglutatyl coenzyme A reductase) gene assignment 

experiments have been performed. This research was 

supported financially by the Hungarian Scientific Research 

Fund (OTKA) D29113 and T 032738. 

1155 - Is the interfacial pH a measure of the efficiency 

of nutrient transfer in ectomycorrhizas? 

A. Nenninger 1* , G.W. Griffith 1 & W. Heyser 2 



3DA, Wales, U.K. - 2 Centre of Environmental Research 

and Technology, University of Bremen, D-28359 Bremen, 

Germany. - E-mail: ann@aber.ac.uk 

Interfacial pH-conditions in Pinus nigra ectomycorrhizas 

were determined using the pH-sensitive ratio-dye Cl-NERF 

and a confocal laser scanning microscope (CLSM). A 

pseudo-ratiometric method is commonly used for 

determining apoplastic pH-values. In the present study, a 

non-invasive ratiometric method relying on a single 

fluorescence dye was devised and tested. Results show 

homogeneous pH-values near 3 in non-mycorrhizal roots 

from different plants and heterogeneous interfacial pHconditions 

between 3.1 and 4 in various pine 

ectomycorrhizas. In mycorrhizas, reduced sucrose and 

phosphate leads to decreasing interfacial pH-conditions, 

whereas external applied glucose induced an increase of 

the interhyphal pH-values in the mantle. Based on the 

results we developed a model for the interfacial transfer 

processes in ectomycorrhizas. Transfer processes on the 

ectomycorrhizal interface are not pH-regulated. The 

processes generate an interfacial pH-value to maintain the 

symbiotic balance. In balance you find optimal pHconditions 

for the nutrient transfer. The lack of only one of 

the transfer goods induces a decrease of the interfacial pHconditions. 

A decreasing pH-value in the interface of 

ectomycorrhizae leads to reduce transfer processes and the 

mutualistic balance in the symbiosis can be maintained. 

1156 - Effect of ferulic acid on guaiacylglycerol-βguaiacyl 

ether degradation by extracellular enzymes of 

Trametes versicolor 

G. Nowak, A. Matuszewska * & M. Nowak 

Departament of Biochemistry M. Curie-Sklodowska 

University, Pl. M. Curie-Sklodowskiej 3, 20-031 Lublin, 

Poland. - E-mail: amatusze@hermes.umcs.lublin.pl 

Trametes versicolor, the effective lignin decomposer, 

produces a set of enzymes acting on bonds in lignin 

polymer, especially on most abundant in lignin ether β-O-4 

bond. In our studies the most active enzymes are secreted 

into the medium in idiophasic cultures of Trametes as a 

result of carbon starvation. During experiments destaining 

of Remazol Brilliant Blue and/or carminic acid were used 

as an indirect method and degradation of guaiacylglycerol-


β-guaiacyl ether as a specific method for the measurement 

of ligninolytic activity. Secreted enzymes were prepared 

from media of carbon-starved, ferulic acid supplemented 

cultures of Trametes. Capability to degradation of β-O-4 

bond was measured in reaction mixtures containing 

enzymes, the dimer substrate, hydrogen peroxide and 

ferulic or p-coumaric acid. Reaction products were 

analysed by capillary electrophoresis. Presence either 

ferulic or p-coumaric acid in reaction mixture resulted in 

significant decrease of dimer degradation. Both 

hydroxycinnamates are parts of lignocellulosic polymer, 

forming bridges between hemicelluloses and lignin, and 

both are known as natural antioxidants. When released by 

relevant esterases during lignocellulosics degradation, they 

stimulates production of ligninolytic enzymes and 

simultaneosly diminish enzymatic activities splitting the 

ether bond β-O-4. This paradoxical effect deserves further 

reasearch. Supported by the EC Contract ICA 2-CT-2000- 

10050 and by KBN grant 139/E-SPUB-M-5PR-UE/DZ 

280/2000. 

1157 - Can wooddecay fungi use chitosan as a N-source 

at extreme C/N-ratios? 

M.S.C. Nyborg * & S. Rosendahl 

University of Copenhagen, Botanical Institute, Dept. of 

Mycology, Øster farimagsgade 2 D, 1353 Kbh. K, 

Denmark. - E-mail: mathilden@bot.ku.dk 

Chitosan has been proposed as an alternative, 

environmentally safe wood preservative. In vitro tests of 

wood preservatives are often carried out on media with 

moderate C/N-ratios in contrast to the high C/N-ratio in 

wood. Chitosan contains nitrogen and fungi could therefore 

be suspected to use this as an N-source under extreme C/Nratios. 

In this study, we investigated the relationship 

between the inhibiting effect of chitosan and the C/N-ratio 

of the media. The fungi used were Trametes versicolor, 

Heterobasidion annosum, Gloeophyllum trabeum and 

Oligoporus placenta. The experimental set-up was a 

factorial design with 5 concentrations of chitosan and 6 

C/N-ratios in a solid caboxymethylcellulose medium. The 

growth was measured as colony diameter. In general, the 

growth of the fungi was much more affected by chitosan 

than by the C/N-ratio. The C/N-ratio had only limited 

effect on Trametes versicolor, Heterobasidion annosum 

and Gloeophyllum trabeum, whereas Oligoporus placenta 

was clearly stressed by high C/N-ratios. However 

Oligoporus placenta was also inhibited by the chitosan. 

The effect of chitosan was independent of the C/N-ratios, 

indicating that wooddecay fungi are not able to use 

chitosan as an alternative N-source at extreme C/N-ratios. 

1158 - Environmental signals associated to the 

transcriptional activation of the mycoparasitism related 

gene prb1 in Trichoderma atroviride 

V. Olmedo-Monfil * , A. Mendoza-Mendoza, I. Gómez, C. 

Cortés & A. Herrera-Estrella 

Department of Plant Genetic Engineering, Centro de 

Investigación y Estudios Avanzados. Unidad Irapuato, 

Apartado Postal 629. 36500, Irapuaro, Gto, Mexico. - Email: 

volmedo@ira.cinvestav.mx 

Trichoderma atroviride parasites a large variety of 

phytopathogenic fungi. This characteristic has allowed its 

use as a biological control agent. The production of 

hydrolytic enzymes appears to be a key element in the 

parasitic process. Among the enzymes released by 

Trichoderma, the proteinase Prb1 plays a major role. 

Herewith we show that the corresponding gene (prb1) is 

subjected to Nitrogen Catabolic Repression. Accordingly, 

induction of prb1 transcription by Rhizoctonia solani cell 

walls and by osmotic stress requires the release from a 

represed condition which is determined by nitrogen 

availability. Furthermore, the transcription pattern of the 

prb1 gene was not affected when a p38-Hog1 inhibitor was 

used. In contrast, a MEK1/2 inhibitor blocked prb1 

transcription in response to nitrogen limitation, indicating 

that the pathway employed in the nitrogen response 

involves proteins similar to p42-p44. Fusions of the prb1 

promoter with the gfp reporter gene allowed the detection 

of a novel regulatory element and represent the first insight 

into relevant sites that control prb1 expression. 

1159 - Heavy metal accumulation and distribution in 

mycorrhizal and nonmycorrhizal roots of Plantago 

lanceolata, AAS and micro-PIXE analysis 

E. Orlowska 1 , Sz. Zubek 1 , J. Mesjasz-Przybylowicz 2 , W. 

Przybylowicz 2 , A. Jurkiewicz 1 & K. Turnau 1* 

1 Institute of Botany, Jagiellonian University, ul. Lubicz 46, 

31-512 Krakow, Poland. - 2 Materials Research Group, 

iThemba LABS, PO Box 722, Somerset West 7129, South 

Africa. - E-mail: ubturnau@kinga.cyf-kr.edu.pl 

Mycorrhizal and nonmycorrhizal Plantago lanceolata were 

grown for six weeks under greenhouse conditions in rhizoboxes 

filled with industrial waste. According to atomic 

absorption spectroscopy (AAS) the shoots of plants 

inoculated with Glomus spp. contained less Zn and Pb than 

nonmycorrhizal ones; their roots did not differ statistically. 

Selected nonmycorrhizal and mycorrhizal roots were 

studied using proton microscopy accompanied by PIXE 

microanalysis. Significant differences in element 

distribution were found between investigated roots. 

Especially a strong accumulation of Zn within the cortex of 

mycorrhizal roots was visible. The present results do not 

allow judgements on whether the metals are located 

exclusively in fungal structures, or both by the fungus and 

the colonised plant cells. In mycorrhizal roots more Zn and 

Pb were found within the vascular tissues than in 

nonmycorrhizal ones. Therefore, mycorrhiza is not 

effective as a filtering mechanism; still, its role in the 

transformation of heavy metals into non-toxic compounds 

cannot be excluded. According to AAS these compounds 

are retained in roots, as a decrease of these elements was 

observed in shoots. This could also suggest the presence of 

additional mechanisms depending on the plant, which 

enable it to cope with metals transferred by the fungus. 



These findings confirm our previous observations of 

mycorrhizas under heavy metal stress. 

1160 - Metabolism in ectomycorrhizal preinfection: 

Expression of malate synthase and acetylCoA 

acetyltransferase genes from Laccaria bicolor 

G.K. Podila 1* , S. Balasubramanian 2 & S.T. Hiremath 3 

1 University of Alabama, Dept. of Biological Sciences, 

Huntsville, AL 35899, U.S.A. - 2 Michigan Tech University, 

Dept. of Biological Sciences, Houghton, MI 49931, U.S.A. - 

3 USDA-NEFES, 359 Main Road, Delaware, OH 43015, 

U.S.A. - E-mail: podilag@email.uah.edu 

The ectomycorrhizas are beneficial fungal associations 

with temperate tree roots. Ectomycorrhizal fungi improve 

survivability, pathogen resistance, drought tolerance and 

provide mobilized mineral nutrient in exchange for fixed 

carbon from the plant. Molecular level analysis suggests 

that, in response to the perception of the plant root, there is 

a directional shift in fungal cell physiology as early as four 

hours into the interaction. During the interaction between 

Laccaria bicolor and Pinus resinosa, we have observed the 

upregulation of metabolic genes such as Lb-MS coding for 

a malate synthase and Lb-AAT that codes for an acetylCoA 

acetyltransferase. The former, one of the two unique 

enzymes of the glyoxylate pathway and the latter the 

terminal thiolase of beta oxidation of fatty acids. The 

expression of these two genes in conjunction with others 

(LB-AUT7) suggests that recycling of cell contents and 

breakdown of reserve food and channeling towards growth 

occur early in the preinfection stage of the symbiosis. 

Regulation of Lb-MS and Lb-AAT under interaction 

conditions and under alternate carbon sources have been 

studied. Lb-MS has been analyzed at the protein level and 

gene structure of Lb-AATg has been elucidated. We 

present our findings on these two metabolic genes and their 

significance in the symbiosis via the agency of the 

peroxisome. 

1161 - Pre-infection and nutrient starvation expression 

profiling of interaction related ESTs from the 

ectomycorrhizal fungus Laccaria bicolor 

G.K. Podila 1* , M.J. Hymes 2 & J.H. Brand 2 

1 University of Alabama, Dept. of Biological Sciences, 

Huntsville, AL 35899, U.S.A. - 2 Michigan Tech University, 

Dept. of Biological Sciences, Houghton, MI 49931, U.S.A. - 

E-mail: podilag@email.uah.edu 

In this study, a 384 member macroarray was created 

consisting of 340 ESTs derived from mRNA of Laccaria 

bicolor during early interaction with Pinus resinosa. The 

macroarray also contained several positive and negative 

controls as internal standards and to confirm 

reproducibility and reduce experiment-to-experiment 

variability. This array was used to study multiple factors of 

352 


interaction, including expression patterns at various time 

points of early interaction (


Tehran, Iran. - 3 Faculty of Medical Sciences, Department 

of Mycology,Tarbiat Modares University, 14115- 

111,Tehran, Iran. - E-mail: mrab@institute.pasteur.ac.ir 

Aflatoxins are secondary fungal metabolites produced by 

toxigenic strains of Aspergillus flavus and A. parasiticus. 

The relationship between aflatoxin synthesis with 

glutathione S-transferase (GST) activity and microscopic 

morphology in A. parasiticus grown under different culture 

conditions has been investigated in order to find out the 

regulatory mechanism(s) of aflatoxin biosynthesis at 

cellular level. Aflatoxin formation was inhibited in A. 

parasiticus grown in the presence of various 

concentrations(1.56-50%, v/v) of aqueous neem leaf 

extract with a maximum about >90% at 50% concentration 

(P


channels have been cloned from filamentous fungi and 

only a few recordings of ion channel activity have been 

made from hyphal cells. In an attempt to gain an insight 

into the role of ion channels in fungal hyphal physiology, 

NcTOKA was cloned from the filamentous fungus, 

Neurospora crassa. Hydropathy analysis showed NcTOKA 

to be a member of the two-pore domain K+ channel family. 

The patch clamp technique was used to investigate the 

biophysical properties of NcTOKA after heterologously 

expression of NcTOKA in yeast. NcTOKA mediated 

mainly time-dependent outward whole cell currents which 

represented K+ efflux. However, expression of NcTOKA 

was able to overcome the K+ auxotrophy of a yeast mutant 

missing the K+ uptake transporters, TRK1 and TRK2, 

suggesting that NcTOKA mediated K+ influx. Consistent 

with this, close inspection of NcTOKA-mediated currents 

revealed small inward K+ currents at potentials negative of 

EK. NcTOKA single channel activity was characterised by 

rapid flicking between the open and closed states with a 

unitary conductance of 14 pS. NcTOKA was effectively 

blocked by extracellular Ca2+, verapamil, quinine and 

TEA but insensitive to Cs+, 4-aminopyridine and 

glibenclamide. The physiological significance of NcTOKA 

is discussed in the context of its biophysical properties. 

1167 - Identification of fungal enzymes in faecal 

droplets from leaf-cutter ants 

S. Rønhede * & S. Rosendahl 

Department of Mycology, Institute of Botany, University of 

Copenhagen, O.Farimagsgade 2D, DK 1353, Copenhagen 

K, Denmark. - E-mail: stigr@bot.ku.dk 

The enzymatic capability of the symbiotic fungus is crucial 

for the ability of leaf-cutter ants to use fresh leaves as a 

substrate. The ants place their faeces as droplets on the 

plant material before it is incorporated into the fungusgarden, 

and these faecal droplets contain enzymes of which 

some are of fungal origin. The aim of the present study was 

to identify specific fungal enzymes in faecal droplets from 

the leaf-cutter ants Acromyrmex echinatior and Atta 

colombica to clarify the role of these enzymes in 

degradation of leaves in the fungus-garden. CMCases, 

laccase, proteases and pectinases were identified in faecal 

droplets by isoelectric focusing and staining in overlays. 

The amount of fungal pectin esterase in faecal droplets was 

comparable to that in pure mycelium indicating that the 

enzymes pass the ant-gut without loosing activity. The 

importance of the fungal enzymes for the evolutionary 

transition of the ants from feeding on dead organic material 

to be able to exploit fresh leaves is discussed. 

1168 - ABC transporters in Trichoderma harzianum 

M. Ruocco 1* , V. Scala 2 , S. Woo 2 , S. Lanzuise 2 , G. Del 

Sorbo 2 , F. Scala 2 & M. Lorito 2 

1 Istituto CNR per la Protezione delle Piante IPP, Via 

Università, 100, 80055 Portici (NA), Italy. - 2 Dept. 

354 


ARBOPAVE- Plant Pathology, University of Naples, Via 

Università, 100, 80055 Portici (NA), Italy. - E-mail: 

miruocco@unina.it 

ATP-binding cassette (ABC) transporters are membrane 

glycoproteins that utilize the energy derived from the 

hydrolysis of ATP to drive transport of endogenous 

metabolites and toxic xenobiotics through biological 

membranes. Overexpression of some ABC's determines 

simultaneous resistance to many chemically unrelated 

compounds whereas disruption of genes encoding ABC 

transporters is associated with increased sensitivity to the 

same compounds. We hypothesize that ABC transporters 

of Trichoderma harzianum have an important role in a 

number of processes such as resistance to environmental 

toxicants produced either by soil microflora or introduced 

by human activity (e.g. fungicides, heavy metal pollutants), 

secretion of factors (mycotoxins and cell wall degrading 

enzymes) necessary for the establishment of a compatible 

interaction with a host fungus. Several oligonucleotides 

were designed from conserved regions of the sequences of 

many ABC transporters from different filamentous fungi, 

and used to clone and partially sequence several genes 

encoding putative ABC transporters from genomic DNA of 

Trichoderma harzianum P1. Functional analysis to 

investigate their role during mycoparasitic events are in 

progress. We are using gene disruption and overexpression 

in Trichoderma to determine the involvement of these 

transporters in biocontrol and fitness. 

1169 - Influences of temperature and light on 

morphological changes during fruit body formation in 

Flammulina velutipes 

Y. Sakamoto 1* & Y. Tamai 2 

1 Ibaraki Prefectural Forestry Research Insutitute, 

Nakamachi To 4692, Nakagun, Ibaraki 311-0122, Japan. - 

2 Division of Environmental Resources, Graduate School of 

Agriculture, Hokkaido University, North 9, West 9, Kitaku, 

Sapporo, 060-8589, Japan. - E-mail: 

yuichisakamoto@hotmail.com 

Influences of light and temperature on morphological 

changes during fruit body formation in Flammulina 

velutipes were investigated. Fruit bodies of F. velutipes 

were induced after temperature reduction in complete 

darkness. However, fruit bodies formed in complete 

darkness had a long stipe with undeveloped pileus on the 

top (pinhead fruit body), thinner and whiter than normal 

fruit bodies formed under light illumination. This suggests 

that F. velutipes can be induced fruit bodies, but cannot 

mature in complete darkness. We subsequently irradiated 

light to pinhead fruit bodies formed in complete darkness. 

It is revealed that pinhead fruit bodies formed in complete 

darkness can be induced pileus after light stimulation. We 

could observe hymenium formation immediately after light 

stimulation. We also observed thickening and pigmentation 

after light stimulation, suggesting that light can directly 

induce fruit body maturation in F. velutipes. Protein 

expression patterns were investigated by two-dimensional 

electrophoresis, and 22 protein spots were newly expressed 

in the mycelia and fruit bodies after temperature reduction


in complete darkness. We also revealed proteins spots that 

were newly expressed after light stimulation and localized 

in pileus. These suggests that many of proteins involved in 

fruit body induction in F. velutipes are expressed after 

temperature reduction, and involved in pileus induction are 

expressed after light stimulation. 

1170 - Ultrastructure and biochemistry of hyphae of 

different zone of growth in a fruiting colony of 

Pleurotus pulmonarius 

C. Sánchez 1* , G. Díaz-Godínez 1 & D. Moore 2 

1 Universidad Autónoma de Tlaxcala, Apartado Postal 129, 

Tlaxcala, Tlax. C.P. 90000, Mexico. - 2 The University of 

Manchester, School of Biological Sciences, Manchester 

9PT, U.K. - E-mail: sanher6@hotmail.com 

In a fungus colony, hyphae from the peripheral zone (i.e. 

young hyphae) are responsible for colony expansion. On 

the other hand, hyphae from the central zone (i.e. mature 

hyphae) may differentiate into reproductive structures. It 

has previously been observed that when P. pulmonarius 

cultures were stained by using conventional histological 

stains the peripheral zone of the colony (PZC) and the 

fruiting structures were stained, but the central zone of the 

colony (CZC) was unstained. In this research, the 

ultrastructure and biochemistry of hyphae from the PZC 

and CZC of a fruiting colony of P. pulmonarius, developed 

on potato dextrose agar were studied. The wall thickness of 

hyphae from the CZC was approx. twice that of hyphae 

from the PZC (0.15 and 0.07 µm, respectively). Hyphae 

from the PZC had about twice glycogen content and 50% 

higher protein content (501 and 18 mg/g dry biomass (dX), 

respectively) than those from the CZC (237 and 11 mg/g 

dX, respectively). The cell wall of hyphae from the CZC 

had about twice soluble and insoluble glucans content (7.7 

and 6.4 mg/g dX, respectively) that of hyphae from the 

PZC (3.8 and 3.4 mg/g dX, respectively). The histological 

differences between young and mature hyphae of a colony 

of P. pulmonarius, particularly in the cell wall and the 

amount of cytoplasmic material present in the hyphae, 

suggest that the wall is used as a temporary nutrient reserve 

during development of Pleurotus. 

1171 - Stability of some commercial mushrooms in 

Thailand after continuous subcultures of mycelia for 

production 

W. Sattayaphisut 1 , N. Boonkerd 1 , N. Teaumroong 1 & S. 

Rodtong 2* 

1 School of Biotechnology, Institute of Agricultural 

Technology, Suranaree University of Technology, Nakhon 

Ratchasima 30000, Thailand. - 2 School of Microbiology, 

Institute of Science, Suranaree University of Technology, 

Nakhon Ratchasima 30000, Thailand. - E-mail: 

sureelak@ccs.sut.ac.th 

The decrease in fruiting body yields commonly occurs in 

the mushroom production in Thailand when mycelia 

obtained from continuous subcultures are used. The 

stability of commercial mushrooms: Auricularia auricula, 

Agrocybe cylindracea, Lentinula edodes, Lentinus 

polychrous, L. squarrossulus, Pleurotus ostreatus, 

Pleurotus species under names P. cystidiosus and P. sajorcaju, 

and Tricholoma crassum, after continuous 

subcultures of mycelia for production twenty times was 

investigated. Mycelial growth rates and fruiting body 

yields were determined. The PCR-RFLP technique was 

also tried to be employed for the preliminary detection of 

fungal genetic stability. After the twentieth subculture, 

growth rates and fruiting body yields of all mushrooms 

were found to be rather consistent except Auricularia 

auricula giving the decrease in both its growth and yield 

after the third subculture then reversing to be stable 

through the subsequent subcultures. For the detection of 

genetic stability, all fungal species gave consistent DNA 

patterns (600-800bp PCR products from ITS4 and ITS5 

primers, and the unique RFLP pattern of each species when 

the DNA was digested with either AluI, TaqI, MboI, or 

HinfI) except A. auricula giving the different pattern of 

HinfI digest after the third subculture which was 

corresponding to its growth and yield reduction. The 

genetic stability of mushrooms should be examined after 

continuous subcultures of mycelia prior to application in 

the large scale production. 

1172 - Long-term exposure of lichens to high 

atmospheric CO 2 under controlled conditions: effects 

on morphology and chemistry 

M. Saunders * & D. Armaleo 

Duke University, Department of Biology, Box 90338, 

Durham, NC 27708, U.S.A. - E-mail: saunders@duke.edu 

The lichens Usnea strigosa and Parmotrema hypotropum 

were collected at the FACE (Free Air CO2 Enrichment) site 

in operation near Duke University. FACE technology is 

able to monitor and maintain open-air sites with 

atmospheric CO2 levels kept either at ambient (^500 ppm) 

or enriched (^700 ppm) values. Flora is grown under 

natural conditions realistically simulating future 

concentrations of atmospheric CO 2. This setup, in 

operation for 15 years, is ideal for monitoring long-term 

effects of CO 2 enrichment throughout the lichens' lifespan, 

with a degree of control not possible in earlier studies. Both 

species of lichens were collected almost exclusively from 

the bark of Sweetgum (Liquidambar styraciflua) trees from 

both CO2-enriched and control sites. Each specimen was 

photographed, sectioned, and observed by standard and 

fluorescence microscopy. The secondary compounds were 

extracted to near completion (^95%) and quantified (in 

reference to thallus dry weight) by digital analysis of Thin 

Layer Chromatography data. Significant differences in 

micro-anatomy and chemistry were found between the 

lichens collected in the high CO2 and control sites. 



1173 - Correlation of parasitism and gradual selectivity 

between lichen bionts - molecular data and culture 

experiments verify morphological observations 

T. Schaper 1 , A. Beck 2 & S. Ott 1* 

1 Botanical Institute, University of Duesseldorf, 

Universitätsstr. 1, D-40225 Duesseldorf, Germany. - 

2 Department Biologie I, Bereich Biodiversitätsforschung, 

L.-M.-Universität Muenchen, Menzinger Str. 67, D-80638 

Muenchen, Germany. - E-mail: otts@uni-duesseldorf.de 

The mutualistic interactions between lichen species range 

from different steps of symbiosis to parasitism. Intra- and 

interspecific interactions characterise the lichen species 

within the close association on the gravel alvar of Gotland, 

Sweden. Concerning these complex interactions the degree 

of selectivity of the lichen bionts is of great importance. 

For lichens only reproducing sexually the availability of 

the suitable photobiont is a crucial prerequisite for 

relichenisation. The ITS rDNA sequences of the 

photobionts demonstrate the diversity of the photobiontal 

pool within the lichen community. Different species of 

Trebouxia belonging to Clade I which is divided in two 

subgroups as well as species of the genus Asterochloris has 

been found. The molecular data verify and support the 

morphological observations of the parasitic contact 

between Fulgensia bracteata and Toninia sedifolia. Both 

lichens share the same photobiont. In culture experiments 

with the isolated photobionts of five lichen species in 

combination with the mycobiont of F. bracteata gradual 

stages of the first steps of the relichenisation process get 

obvious. These observations indicate to a reduced 

selectivity of the mycobiont of F. bracteata in its 

photobiontal choice. The mycobiont of F. bracteata forms 

a temporary aggregate with different ? maybe less - 

compatible bionts. This enables the mycobiont to survive 

until the association with a compatible photobiont is 

established and thallus formation is initiated. 

1174 - Mating reactions, sexual morphogenesis, and 

trisporic acid in zygomycetes 

C. Schimek * , K. Kleppe, A.R. Saleem, A. Petzold & J. 

Wöstemeyer 

Friedrich-Schiller Universität, Institute of General 

Microbiology and Microbial Genetics, Neugasse 24, D- 

07743 Jena, Germany. - E-mail: B9SCCR@nds.rz.unijena.de 

It is understood that mating reactions in zygomycetes are 

mediated by trisporic acid. But this generalization was only 

valid for mucoralean fungi, as no other orders had been 

examined in that respect. We present data on several 

species of the order Mortierellales covering physiological, 

biochemical, and molecular biological approaches. We 

found that in Mortierellales, too, the sexual responses are 

mediated by trisporoids. So in zygomycetes, the same 

signalling system is used not only beyond family limits but 

also crossing order barriers, a feature unique in fungi. 

Trisporoids are derivatives of beta-carotene that are 

356 


synthesized in a cooperative process by the different 

mating types. Some trisporoids act as pheromones, others 

show hormone-like activities. Although specific trisporoid 

isoform patterns are typical for certain species, 

interspecific reactions occur quite frequently and we have 

also observed inter-order reactions. Interspecific reactions 

are especially interesting in parasitism. A few species, e.g. 

Parasitella parasitica are parasites to other zygomycetes in 

a mating type-dependent manner: interactions occur only if 

the partners belong to opposing mating types. The 

susceptibility to parasitism by Parasitella can therefore be 

used as a marker for studying the relationship between 

species. Using in-situ techniques, the involvement of the 

same key enzymes for trisporoid metabolism could be 

shown in sexual as well as in parasitical interactions. 

1175 - Regulation of sex-pheromone biosynthesis and its 

role in mating and parasitic interactions of mucoralean 

fungi 

K. Schultze, J. Wöstemeyer & A. Burmester * 

Friedrich-Schiller-University Jena,Institute of 

Microbiology, Neugasse 24, D-07743 Jena, Germany. - Email: 

Anke.Burmester´@rz.uni-jena.de 

Successful infection of Absidia glauca (Mucorales, 

Mucoraceae) by Parasitella parasitica (Mucorales, 

Mucoraceae) requires that the partners belong to 

complementary mating types. These observations point 

towards a physiological relationship between parasitism 

and the sexual pathway. In all mucoralean fungi analyzed, 

sexual differentiation depends on the synthesis of the sex 

pheromone trisporic acid. Trisporic acid is synthesized via 

the complementary biosynthetic action of both mating 

types. One of the genes for an enzyme of the trisporic acid 

biosynthesis pathway was cloned from host and parasite. 

Expression studies at the transcriptional level show 

transcripts only after stimulation by mating and infection. 

DNA sequence analysis of the genomic surrounding detects 

a second gene in close neighborhood of the first one, with 

an unknown function. Nevertheless, this gene is transcribed 

in P. parasitica. Recently, vectors were constructed 

containing the gene for green fluorescent protein (GFP) for 

transformation of A. glauca strains. Now, also in vivo 

expression can be studies to obtain more insights in the 

interaction of parasite and host. 

1176 - Screening of environmental and clinical 

Cladosporium isolates for cellobiose dehydrogenase 

production 

M. Shams 1* , A. Fazli 1 , A. Lotfi 2 & M. Razzaghi abyaneh 3 

1 Dept.of Mycology, Faculty of Medical Sciences,Tarbiat 

Modares University, P.O.Box:14115-111, Tehran, Iran. - 

2 Dept.of Biochemistry, Tarbiat Modares University, 

P.O.Box:14115-111, Tehran, Iran. - 3 Dept.of Myclogy, 

Pasteur Institute of Iran, P.O.Box:13164, Tehran, Iran. - 

E-mail: shamsm@modares.ac.ir


Cellobiose dehydrogenase(CDH)is an extracellular enzyme 

produced by wood-degrading and also other fungi. In this 

communication, CDH activity of 24 Cladosporium isolates 

was measured using a zymogram procedure for 

determining the possible role of the CDH in the fungal 

pathogenesis. 2,6-Dichlorophenolindophenol (DCPIP) was 

added into czapek agar medium containing Avicel (1% 

w/v) and cellobiose (20mM). The blue coloured agar plates 

were inoculated with fungal spores and incubated for 5 

days at 25C. Production of CDH was assessed by the 

measuring zones of clarification in the agar around the 

growing colonies. This clarification was higher for 10 

clinical Cladosporium isolates (5-6 mm) as compared with 

14 environmental ones (0.5-1.4 mm)(P3)-glucan, the major matrix wall component, 

was immuno-localized in all regions of elongating and nonelongating 

hyphae. A number of cytochemical, 

biochemical and physiological controls were performed to 

assure the reliability of these findings. We suggest that in 

elongating regions, the matrix is synthesized first and 

synthesis of microfibrilar component follows. Another 

explanation for our results is that localized apical cellulose 

hydrolysis by endoglucanase creates plastic wall regions 

consisting mainly of β-(1-3)-glucans, which expand under 

turgor and/or cytoskeleton pressure. Cellulose deposition 

quickly follows. 

1178 - Aspergillus nidulans polarity genes swoA, a 

protein mannosyl transferase, and swoF, an Nmyristoyl 

transferase 

B.D. Shaw * & M. Momany 

University of Georgia, Plant Biology, 2502 Plant Science, 

Athens, GA, 30602, U.S.A. - E-mail: 

bshaw@botany.uga.edu 

Hyphal growth in filamentous fungi requires first 

establishment and then maintenance of polarity and is 

necessary for disease in many fungal pathosystems. 

Germination of Aspergillus nidulans conidia involves 

isotropic (nonpolar) expansion of the cell through the first 

two nuclear divisions. Subsequent development is polar 

with production of a germ tube and establishment of 

vegetative growth. The A. nidulans swo mutants are 

temperature sensitive mutants defective in one or both 

stages of polar growth during the process of spore 

germination. A. nidulans swoAp is a protein mannosyl 

transferase, a protein responsible for the first stage of Oglycosylation. 

A glycosylated substrate of swoAp may be a 

cell wall associated protein important for polar growth. 

swoFp is an N-myristoyl transferase, a protein responsible 

for the co-translational addition of a small fatty acid to the 

N-terminus of its substrate thereby giving it higher affinity 

for association with the membrane. In Saccharomyces 

cerevisiae N-myristoyl transferase substrates are 

commonly signaling proteins. The temperature-sensitive 

allele changes a conserved aspartic acid to a tyrosine. The 

wild type D appears to stabilize a Beta strand bend through 

two hydrogen bonds and an ionic interaction. A 

myristoylated substrate of swoFp may be a signaling 

molecule important for polar growth. A proteomics 

approach is being taken to characterize the substrates of 

these proteins. 

1179 - Effect of bacteria on germination and mycelial 

growth of arbuscular mycorrizal fungi, in vitro 

S.A.G. Soares 1 , L.C. Maia 1 , R.L.R. Mariano 3 , U.M.T. 

Cavalcante 2 & G.A. Silva 1* 

1 Departamento de Micologia - UFPE, 50670-420 Recife - 

PE, Brazil. - 2 Departamento de Biologia - UFRPE, 52171- 

900 Recife - PE, Brazil. - 3 Departamento de Agronomia - 

UFRPE, 52171-900 Recife - PE, Brazil. - E-mail: 

gladstonesilva@yahoo.com 

The effect of plant growth promoted rhizobacteria (PGPR) 

on germination and mycelial growth of Gigaspora albida, 

Glomus etunicatum and Scutellospora heterogama was 

studied. Aqueos suspensions of Bacillus cereus (C210), 

Bacillus sp. (RAB9), B. thuringiensis subvar. kurstakii 

(ENF10) and Pseudomonas chlororaphis (GN1212) were 



adjusted to 2 × 10 7 ufc mL - 1 and 50 mL inoculated into 

Petri dishes with 8% water agar; six superficially 

disinfected (NaOCl) AMF spores/plate were then placed on 

the medium and incubated in the dark, at room temperature 

(28 °C). Experiments combining PGPR and AMF, with 

five replicates, were performed and evaluated at the 21th 

and 28th days. The PGPR promoted different effects on 

germination and mycelial growth of the AMF. Spore 

germination of G. albida was stimulated by RAB9 and 

GN212 and the mycelial growth stimulated by the latter, 

but inhibited by the other bacterial strains. Neither the 

incubation period nor the PGPR affected germination of S. 

heterogama. However, the mycelial growth was stimulated 

since the 21th day by GN1212, and later by the other 

bacteria. Germination of G. etunicatum was stimulated by 

the PGPR, but at the 28th day did not differ from the 

control, while mycelial growth was benefited by ENF10 

and C210. In general, among the AMF, higher germination 

and mycelial formation were obtained with G. albida. 

Financial support: CAPES and CNPq. 

1180 - Effects of photosynthetically active radiation 

(PAR) and carbohydrate source on UV-B induced 

parietin synthesis in Xanthoria parietina 

K.A. Solhaug * & Y. Gauslaa 


Agricultural University of Norway, P.O. Box 5014, NO- 

1432 Ås, Norway. - E-mail: knut.solhaug@ibn.nlh.no 

Lichen secondary compunds can be removed by rinsing in 

100% acetone without affecting thalli viability. Acetonerinsed 

parietin-free Xanthoria parietina thalli were 

cultivated in two growth chamber experiments. Thalli were 

soaked in water or carbohydrate solution once a day during 

both seven days experimental periods. In the first 

experiment a factorial design with ±UV-B (0.75 Wm-2), 

±PAR (200 µmol photons m-2s-1) and 0, 2 or 5 g ribitol 

per liter water was used. No resynthesis of parietin took 

place in the abcence of UV-B. For thalli exposed to UV-B 

both PAR and ribitol contributed to the parietin resynthesis. 

Thalli exposed to PAR and the highest concentration of 

ribitol resynthesized more than 20% of the original content 

during the seven days experimental period. In the second 

experiment thalli were exposed to UV-B in the dark and 

moistened with various carbohydrate solutions. Thalli 

moistened with ribitol or sucrose resynthesized about 14% 

of their original parietin content, those moistened with 

mannitol or glucose resynthesized about 8%, while 

moistening with sorbitol or water resulted in resynthesis of 

2-4% parietin only. The results are discussed in relation to 

the role the different carbohydrates may play in the lichen 

symbiosis e.g. that ribitol is the carbohydrate that the 

photobiont Trebouxia export to the fungal partner in the 

lichen X. parietina. 

358 


1181 - Eucalypt wood pretreatment with Phanerochaete 

crassa and Peniophora lycii decreased pitch content 

after laboratory kraft cooking 

M. Speranza 1* , A. Gutíerrez 2 , J.C. del Río 2 , J. Romero 3 , 

A.T. Martínez 1 & M.J. Martínez 1 

1 Centro de Investigaciones Biológicas, CSIC, Velázquez 

144, E-28006 Madrid, Spain. - 2 Instituto de Recursos 

Naturales y Agrobiología de Sevilla, P.O. Box 1052, E- 

41080 Sevilla, Spain. - 3 Centro de Investigación y 

Tecnología (ENCE), Cra. Campañó s/n, E-36157 

Pontevedra, Spain. - E-mail: marielas@cib.csic.es 

During last years eucalypt wood treatment with 

basidiomycetes is being studied as an alternative to 

traditional methods for pitch control. In this way, 

Phanerochaete crassa and Peniophora lycii, isolated from 

the Uruguay forestry, have been selected by their ability to 

remove both sterols, which are responsible for pitch 

deposition, and lignin from eucalypt wood. Here we 

confirm the effectiveness of Eucalyptus globulus wood 

pretreatment with these two fungi to reduce the amount of 

free and esterified sterols from pulp and black liquors after 

laboratory kraft pulping, and the distribution of lignin and 

free sterols in the pretreated wood is analyzed. P. lycii and 

P. crassa decreased lignin content after two weeks of 

industrial chips treatment (24 and 16% respectively). An 

extensive alteration of lignin was observed by confocal 

laser scanning microscopy of wood sections. Filipin 

staining (a selective method to localize free sterols) showed 

that most of sitosterol was degraded after wood 

pretreatment, although some deposits remained in the 

lumen of parenchymatic cells. Chemical analysis after 

laboratory kraft cooking showed that both fungi decreased 

the content of free sterols (32 and 31%, respectively) and 

sterol esters (39 and 45%, respectively) in brown kraft 

pulps, and the free sterol content in black liquors (87 and 

73%, respectively). The optical and papermaking 

properties of the bio-pulps obtained are being analyzed. 

1182 - A gene cluster associated with production of 

insecticidal loline alkaloids in the fungal endophyte 

Neotyphodium uncinatum 

M.J. Spiering 1* , H.H. Wilkinson 2 , C.D. Moon 1 , J.D. 

Blankenship 1 & C.L. Schardl 1 

1 University of Kentucky, Lexington, Kentucky 40546, 

U.S.A. - 2 Texas A&M University, College Station, Texas 

77843, U.S.A. 

Grass plants can acquire chemical defenses against insect 

and vertebrate herbivores by living in symbiosis with 

fungal endophytes of the genus Epichloë (anamorphs, 

Neotyphodium). In several Epichloë/Neotyphodium - grass 

associations, insecticidal 1-aminopyrrolizidine (loline) 

alkaloids are produced, which hold promise as natural plant 

protectants. The objective of this study was to identify the 

genes involved in loline production. N. uncinatum can


produce lolines under certain culture conditions, and a 

culture system was developed for regulated loline 

expression in this fungus. With this system, genes 

differentially expressed during loline accumulation were 

isolated by subtractive hybridization. Two genes isolated, 

lolA and lolC, were similar to genes encoding aspartate 

kinases and homocysteine synthase, respectively, which are 

enzymes in methionine biosynthesis. lolA and lolC were 

highly expressed during loline production in culture, and 

the two genes were present only in endophytes with lolineproducing 

phenotypes. Close linkage of lolA and lolC in N. 

uncinatum was found by PCR, corroborated by data from a 

genomic library survey of the closely related endophyte E. 

festucae. At least five putative genes were found clustered 

with lolA and lolC. These putative genes have similarity to 

polyamine biosynthesis/interconversion genes, and genes 

for monooxygenases, oxidoreductases, demethylases and 

aminotransferases, which are frequently found in 

secondary-metabolite gene clusters. 

1183 - Vacuolar and non-vacuolar protein degradation 

routes in the white-rot fungus Phlebia radiata: their 

influence on ligninolytic enzymes 

M. Staszczak 1 , I. Glina 2 , A. Leonowicz 1 & K. 

Grzywnowicz 1* 

1 Department of Biochemistry, Maria Curie-Sklodowska 

University, Pl. M.Curie- Sklodowskiej 3, 20-031 Lublin, 

Poland. - 2 Department of Cell Biology, Maria Curie- 

Sklodowska University, ul. Akademicka 19, 20-033 Lublin, 


Continuous protein turnover is involved in basic cellular 

functions such as the modulation of the levels of proteins, 

adjustment to stress, as well as preferential removal of 

defective proteins. Eukaryotic cells contain two major 

systems for protein degradation: lysosomal (vacuolar) and 

non-lysosomal (non-vacuolar). The lysosome was long 

believed to be the only site for protein breakdown in cells, 

but it is now clear that intracellular proteolysis is largely 

accomplished by a highly selective non-lysosomal pathway 

that requires a large multicatalytic proteinase complex 

known as the proteasome. The 26S eukaryotic proteasome 

catalyses the ATP-dependent degradation of 

polyubiquitinated proteins. Lignin-modifying enzymes of 

white-rot fungi are mainly expressed during the secondary 

phase of growth (idiophase), when the limitation of carbon 

and nitrogen occurs. It has been demonstrated for many 

eukaryotic organisms, that both the vacuolar and nonvacuolar 

proteolytic systems are activated by nutrient 

starvation. A major goal of the present study was to 

examine whether the specific agent (CbzLLLal, MG 132) 

that blocks the function of yeast and mammalian 26S 

proteasome and the lysosomotropic agent inhibiting 

intralysosomal degradation of proteins (chloroquine) can 

affect the levels of ligninolytic enzymes of the white-rot 

fungus Phlebia radiata during the shift to idiophase 

triggered by nitrogen or carbon starvation. Supported by 

the EC Contract ICA2-CT-2000-10050, 5PR-UE/DZ 

280/2000. 

1184 - Degradation of natural and artificial humic acids 

by the litter-decomposing basidiomycete Collybia 

dryophila 

K.T. Steffen 1* , M. Hofrichter 1 & A. Hatakka 2 

1 Department of Applied Chemistry and Microbiology, 

P.O.Box 56, Viikinkaari 9, Bicenter 1, FIN-00014 

University of Helsinki, Finland. - 2 Dep. of Environmental 

Biotechnology, International Graduate School Zittau, 

Markt 23, D-02763 Zittau, Germany. - E-mail: 

Kari.Steffen@Helsinki.Fi 

The litter decomposing fungus Collybia dryophila is a 

common basidiomycete colonizing different types of forest 

soils. It decomposed in our experiments a natural humic 

acid isolated from pine-forest litter (LHA) and a synthetic 

14 C-labeled humic acid ( 14 C-HA) prepared from [UL- 

14 C]catechol in liquid culture. Polar, lower-molecular mass 

fulvic acid (FA) and carbon dioxide were formed during 

the degradation. Manganese (200 µn; M Mn 2+ ) enhanced 

considerably the decomposition of HA leading to 75% 

conversion of LHA and 50% mineralization of 14 C-HA 

compared to 60% and 20%, respectively, in the absence of 

Mn 2+ . These findings strongly indicated an involvement of 

the ligninolytic enzyme manganese peroxidase (MnP), the 

production of which was noticeably increased in Mn 2+ - 

supplemented cultures and which was found to be capable 

of converting LHA in vitro. The enzyme was produced as a 

single protein with a pI of 4.7 and a molecular weight of 44 

kDa. In solid-state culture (pine-forest litter), C. dryophila 

mineralized 14 C-HA as well and released substantial 

amounts of water-soluble FA from the insoluble litter 

material. The results indicate that litter-decomposing 

basidiomycetes such as C. dryophila are involved in humus 

turnover by recycling high-molecular mass humic 

substances. 

1185 - Ultrastructural analysis of sterile elements of 

basidiomes and fruitbodies in holobasidiomycetes 

A.A. Stepanova 

National Institute of Desert, Flora and Fauna Ministry of 

Nature Protection Of Turkmenistan, 

Ashgabad,744000,Bitarap Turkmenistana,15, 

Turkmenistan. - E-mail: iccnet@onlinee.tm 

Comparative ultrastructural analysis of morphogenesis of 

sterile elements of basidiomes (B) in 6 species of the 

Aphyllophorales and fruitbodies in 17 of Agaricales and 1 

Gasteromycetes showed that the most specialised are the 

cells of subhymenium, having the features of meristematic 

cells. During the grows of B and FB the cells of sterile 

elements are vacuolised, reserve substances, accumulated 

in many species, disappear, cytosol and number of 

organelles are reduced.For all studied species (expect the 

species of Coprinaceae) cells of sterile elements has 

ultrasructure indicated of low level of its metabolism. It 

was shown, that mass senescence and death of the cells of 



sterile elements occur before the formation of mature 

basidium. The process of cell senescence and death in B 

(Aphyllophorales) occur in the direction from the trama of 

pileus to trama hymenophore and subhymenium. In FB of 

Agaricales and Gasteromycetes this processes occur in 

direction, from the base of stipe to its upper part, later they 

occupied the trama of pileus, trama pf hymenophore and 

subhymenium. It was supposed that during the 

morphogenesis of B and FB in Holobasidiomycetes they 

are early transforming in to the autonomous from 

mycelium way of morphogenesis. 

1186 - Construction of a gene-trap vector, pPTR- 

EGFP1 and pUsCdelPA2, for Aspergillus oryzae 

S. Suzuki * & Y. Kashiwagi 

National Food Research Institute, Kan-nondai 2-1-12 

Tukuba Ibaraki 305-8642, Japan. - E-mail: 

satosuz@nfri.affrc.go.jp 

A promoter-trap vector that uses GFP as a reporter for 

Aspergillus oryzae gene expression and a polyA-trap vector 

using auxotrophic selection marker were constructed for 

the first time. The promoter-trap vector, pPTR-EGFP1, and 

polyA-trap vector, pUsCdelPA2 can be used to investigate 

unknown gene function. These gene-trap vector were 

integrated into a host genome randomly and transformants 

were selected by drug resistans or auxotrophic marker gene 

on the vector. The endogenous promoter activity trapped 

by the pPTR-EGFP1 vector was used to express the 

reporter gene egfp. The endogenous polyA addtion signal 

trapped by the pUsCdelPA2 vector was used to express the 

auxotrophic marker gene sC on the vector. By the 

transformation using pPTR-EGFP, approximately 300 drug 

resistant transformants were obtained, one of which 

showed the same strong fluorescence as GFP. The 

expression of egfp gene was confirmed by Northern blot 

analysis. The flanking genome regions of the integrate site 

was isolated by plasmid rescue method. Our resuls show 

that this newly constructed vector, pPTR-EGFP1, can trap 

endogenous promoter activity. The transformation using 

pUsCdelPA2 is under continuation now. 

1187 - Production of CsmA, a class V chitin synthase 

with a myosin motor-like domain of Aspergillus 

nidulans, is regulated in multiple ways 

N. Takeshita, A. Ohta & H. Horiuchi * 

Department of Biotechnology, The University of Tokyo, 1- 

1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. - E-mail: 

ahhoriu@mail.ecc.u-tokyo.ac.jp 

Chitin, a β-1,4-linked homopolymer of Nacetylglucosamine 

(GlcNAc), is one of the major structural 

components of the fungal cell wall. Chitin synthases are 

membrane-bound proteins and have been classified into 

five groups, classes I to V, on the basis of the structures in 

360 


their conserved region. csmA gene of Aspergillus nidulans 

encodes a class V chitin synthase with a myosin motor-like 

domain at its N-terminus. csmA null mutants showed 

remarkable abnormalities in polarized growth, hyphal wall 

integrity, and conidiophore development. In this study, to 

investigate the intracellular behavior of the csmA product 

(CsmA) and the regulation of its production, we 

constructed strains that produced CsmA tagged with nine 

repeats of hemagglutinin A (HA) epitope at its C-terminus, 

CsmA-HA, instead of CsmA. Western blot analysis with 

anti-HA antibody proved that the entire coding region of 

csmA was translated as a single polypeptide with an 

approximate molecular mass of 210 kDa. CsmA-HA was 

found in the membrane fraction of the CsmA-HA 

producing strain. CsmA-HA was produced during 

vegetative growth, and its amount increased under low 

osmotic conditions. A discrepancy in the amounts of 

CsmA-HA protein and csmA transcript as determined by 

northern analysis under those conditions suggests that the 

production of CsmA is regulated at both the transcriptional 

and post-transcriptional levels. 

1188 - Media optimization for manganese peroxidase 

production by response surface methodology in a newly 

isolated white-rot fungus Trichophyton rubrum LSK-27 

C. Tamerler 1* , U.O.S. Seker 1 , K. Li 3 , H. Jung 2 & H. 

Bermek 1 

1 Istanbul Technical University, Molecular Biology and 

Genetics, Maslak - Istanbul 80626, Turkey. - 2 Sunchon 

National University, Department of Forest Resources, 

Sunchon 540-742, Korea. - 3 Oregon State University, 

Department of Wood Science and Engineering, 102 

Richardson Hall, Corvallis, OR97331-5751, U.S.A. - Email: 

tamerler@itu.edu.tr 

Response surface methodology (RSM) is a useful 

technique for media optimization, which is a crucial step 

for maximizing enzyme production in fungal cultures. In 

current study, RSM was applied for media optimization in 

a newly isolated white rot-fungus T. rubrum LSK-27, for 

manganese peroxidase (MnP) production. Carbon, nitrogen 

and also phosphorus are known to have significant impact 

in the production of fungal enzymes. Manganese stimulates 

MnP production in most fungi. In the first optimization 

step, glucose (x1), mycological peptone (x 2), MnSO 4 (x 3) 

and KH 2PO 4 (x 4) were studied to investigate their effect on 

MnP production by fractional factorial design, 2 4_1 . The 

first order equation with the regression coefficients were 

computed to be y=0.7175+0.0625x 1+0.4318x 2-0.1201x 3- 

0.0364x 4 with a probability level of 91% for MnP 

production. Although glucose and KH2PO 4 positively 

affected the cell growth, they did not increase the enzyme 

levels. Among the response curves obtained for four media 

components, the highest response was obtained for 

MnSO4-peptone pair, which was chosen as main factors for 

central composite design. Effect of each media ingredient 

on fungal morphology was also followed. The optimum 

MnSO 4-peptone concentrations were found for maximum 

MnP production.


1189 - Expression studies of plant genes differentially 

expressed in leaf and root tissue of arbuscular 

mycorrhizal (AM) fungal colonised tomato plants 

J. Taylor & L.A. Harrier * 

Scottish Agricultural College, Kings Buildings, West Mains 

Road, Edinburgh, Lothian, Scotland, U.K. - E-mail: 

l.harrier@ed.sac.ac.uk 

Arbuscular mycorrhizal (AM) fungi of the unique phylum 

Glomeromycetes are a multifaceted group of mutualistic 

symbionts that are common to terrestrial ecosystems. The 

interaction between AM fungi and plant root systems is of 

environmental and agronomic importance. Understanding 

the molecular changes within the host plant upon AM 

fungal colonisation is a pre-requisite to a greater 

understanding of the mechanisms underlying the 

interaction. Differential mRNA display was conducted on 

leaf tissue of tomato plants colonised and non-colonised by 

the AM fungus Glomus mosseae and five putative 

differentially regulated cDNAs were identified. All cDNAs 

isolated shared high sequence similarity to known plant 

genes. Semi-quantitative RT-PCR was used to establish 

gene expression patterns for all five clones within leaf and 

root tissue of mycorrhizal and non-mycorrhizal colonised 

tomato plants. Differential regulation was observed for all 

five cDNAs. Down-regulation within the leaf tissue of 

mycorrhizal plants was observed for 4 out of the 5 cDNAs 

with an up-regulation observed only for one. Tissue 

specific regulation was observed for several cDNAs, with 

down-regulation observed in mycorrhizal leaf tissue and 

up-regulation observed within mycorrhizal root tissue as 

compared to non-mycorrhizal tissue. We thank the Scottish 

Executive Environmental Rural Affairs Division 

(SEERAD) for financial support. 

1190 - Application of AFLP markers to genetic study of 

an edible mushroom, Lentinula edodes: strain typing, 

genetic diversity, and genetic linkage map 

K. Terashima 1* , T. Matsumoto 2 , E. Hayashi 3 & Y. 

Fukumasa-Nakai 2 

1 Japan Science and Technology Corporation (JST), 

Honmachi 4-1-8, Kawaguchi, Saitama, 332-0012, Japan. - 

2 The Tottori Mycological Institute, Kokoge211, Tottori, 

689-1125, Japan. - 3 Forest Tree Breeding Center, Ishi, 

Juo, Taga, Ibaraki 319-1301, Japan. - E-mail: 

ktera@hal.ne.jp 

Lentinula edodes (shiitake) is one of the most popular 

edible mushrooms in east Asia (Japan, Korea and China). It 

is desirable to establish efficient breeding systems of this 

mushroom using molecular markers. In this study, we 

evaluated the usefulness of amplified fragment length 

polymorphism (AFLP) as genetic marker in L. edodes. 

Strain typing and genetic diversity: Six AFLP primer pairs 

reproducibly detected 179 polymorphic DNA fragments 

among 15 strains of L. edodes currently used for cultivation 

in Japan. These markers could differentiate all of the 

strains. Cluster analysis and principle coordinates analysis 

based on AFLP data revealed two groups which 

corresponded to those categorized on their fruiting season 

types. Genetic linkage map: A medium-dense genetic 

linkage map of L. edodes was constructed based on 203 

AFLP markers and two mating type factors. The 

segregation of these markers was generated from 95 

progeny of a single cross of two distantly related L. edodes 

strains. Segregation analysis showed that the map consisted 

of 11 linkage groups, and the total genetic distance of the 

map was 1956.7cM. The average rate of physical size to 

genetic distance could be roughly estimated to be less than 

18.4 kb/cM, which is low compared to the values obtained 

for other filamentous fungi. Seventeen of the AFLP 

markers showed highly distorted segregation ratios (χ 2 

values ≥ 6.63; P ≤ 0.01), and many of these were located in 

LG II (6 markers) and IV (6 markers). 

1191 - pH optimization of AM symbiosis with Glomus 

intraradices and carrot transformed roots in vitro 

P. Tiwari 1* , U.G. Reddy 1 , A. Prakash 2 & A. Adholeya 1 

1 

Centre for Mycorrhizal research, Tata Energy Research, 

Institute, Habitat Centre, Lodi Road, New Delhi-110003, 

2 

India. - School of Biotechnology, Department of 

Microbiology, Barkatullah University, Bhopal,Madhya 

Pradesh, India. - E-mail: pragati@teri.res.in 

AM fungi Glomus intraradices was introduced on M media 

with different pH regimes ranging from pH 4 to pH 13 in 

vitro. The symbiosis was found to express differently over 

the acid and alkaline range. The investigation was directed 

to study the difference in expression of AM fungi in terms 

of inter and intraradical spread, spore formation, mycelial 

and root biomass along with nutritional uptake in roots. pH 

8 was observed to be more supportive to maximum AMF 

symbiosis, also supporting maximum root biomass. pH 9 

supported more vegetative proliferation of the fungus. An 

increasing trend was observed in mycorrhizal colonization 

percentage (MCP) from pH 4 to 13. The study is significant 

in optimizing parameters responsible for optimum 

symbiosis and exploit its potential in mass inoculum 

production. 

1192 - Comparative symbiotic events of various coal ash 

amendments in vitro on AM fungi Glomus intraradices 

and Ri T-DNA transformed carrot roots 

P. Tiwari 1* , U.G. Reddy 1 , A. Prakash 2 & A. Adholeya 1 

1 

Centre for mycorrhizal research, Tata Energy Research 

Insitute, Habitat Centre, Lodi Road, New Delhi, 110003, 

2 

India. - School of Biotechnology, Department of 

Microbiology, Barkatullah University, Bhopal, Madhya 

Pradesh, India. - E-mail: pragati@teri.res.in 



In vitro mycorrhizal behaviour in the presence of three 

types of coal ash was studied. Minimal media was 

amended with 5% of three types of coal ashes with 

different physico-chemical properties i.e., ESP ash, pond 

ash and bottom ash obtained from the Korba Super 

ThermalPower Station located in Chattisgarh, India. A 

comprehensive elemental analysis was conducted to 

evaluate the absorption, translocation and tolerance of 

macro, microelements and heavy metals (Ca, Na, Mg, K, 

Zn, Fe, Cu, Mn, Cd, Co, Pb, Cr, Mo, As, Al, & B) in roots 

and extramatrical mycelial biomass. The effect of ashes on 

the mycorrhizal expression of Glomus intraradices was 

compared in terms of spore production, mycelial biomass, 

root biomass and extent of root colonization. Mycorrhizal 

expression measured by spread and internal and 

extramatrical proliferation was maximum in ESP ash and 

minimum in bottom ash. This is the first report of direct 

ash amendment with multiple heavy metal accumulation in 

AM fungal biomass and roots in vitro. 

1193 - Conidiation in Penicillium cyclopium is triggered 

by conidiogenone: an endogenous diterpene 

U. Ugalde 1* , T. Roncal 1 , S. Cordobes 1 & O. Sterner 2 

1 University of the Basque Country, P.O. Box 1072 San 

Sebastian 20080, Spain. - 2 Lund University, P.O. Box 124, 

Lund SE-221, Sweden. - E-mail: qppugmau@sc.ehu.es 

The filamentous fungus Penicillium cyclopium conidiates 

in submerged culture without nutrient limitation, in the 

presence of calcium ions according to a precisely timed 

programme. Conidiation could be prematurely induced in a 

supplemented medium which had previously supported 

growth, suggesting that a metabolite which influenced the 

process was produced. A diterpenoid compound with 

conidiation inducing activity, conidiogenone, was purified 

from the culture medium, along with conidiogenol, a 

derivative with delayed activity. Contrary to previous 

thought, the presence of calcium was demonstrated to only 

decrease the requirement of accumulated conidiogenone for 

the induction to proceed. In the light of the results a 

mechanism of conidiation induction is presented, according 

to which the mycelium constitutively produces a 

conidiation inducer (conidiogenone) that accumulates 

extracellularly. When a threshold concentration is reached 

induction takes place by the likely interaction with a 

specific cellular receptor. The results indicate that 

conidiogenone is both sufficient and necessary to induce 

conidiation. 

1194 - Isolation and characterization of the 

glyceraldehyde-3-phosphate dehydrogenase gene from 

Rhizomucor miehei 

C. Vágvölgyi * , M. Vastag, Zs. Kasza & K. Ács 

University of Szeged, Department of Microbiology, Szeged, 

P.O. Box 533, H-6701, Hungary. - E-mail: csaba@bio.uszeged.hu 

362 


Fungi belonging in the Rhizomucor genus has medical and 

biotechnological importance. Rhizomucor miehei isolates 

are well known as producers of rennin, which enzyme is 

used in cheese-industry. Until now little is known about the 

genetics of Rhizomucor strains. The aim of the present 

study was to clone the gpd gene from Rhizomucor miehei 

and characterize the structural gene and its adjacent 

regulatory elements (promoter and terminator regions) at 

sequence level. Glyceraldehyde-3-phosphate 

dehydrogenase is an important enzyme in both glycolysis 

and gluconeogenesis. It catalyses the oxidation and 

phosphorylation of the glyceraldehyde-3-phosphate to 1,3bis-phosphoglycerate. 

As a main housekeeping enzyme its 

gene shows a strong structural conservation by different 

prokaryotic and eukaryotic organisms. The Rhizomucor 

miehei gene encoding glyceraldehyde-3-phosphate 

dehydrogenase (gpd) was isolated from a genomic library 

by hybridization of the library clones with gpd-specific 

gene probe. The complete nucleotide sequence encodes a 

polypeptide chain of 337 amino acids interrupted by 5 

introns. The putative protein product of this gene shows a 

high degree of sequence identity with the corresponding 

gene products of various fungi. The promoter region, 

containing a consensus TATA box and a 246 nucleotid 

long termination region were also determined. This study 

was supported in by OTKA grant T 032738. 

1195 - Chrysosporium lucknowense, a new fungal host 

for protein production 

C.M.J. van Zeijl 1 , M. Heerikhuisen 1* , P.J. Punt 1 , M. 

Emalfarb 2 & R. Burlingame 2 

1 TNO Nutrition and Food Research Institute, 

Utrechtseweg48, Zeist 3704 HE, The Netherlands. - 

2 Dyadic International. Inc, Jupiter, Florida, U.S.A. - Email: 

Heerikhuisen@voeding.tno.nl 

Chrysosporium lucknowense, an ascomycetous fungus, not 

closely related to Aspergillus or Trichoderma, is being 

developed as a new fungal host for homologous and 

heterologous protein production. This thermophilic fungus 

is able to secrete large amounts of (hemi)cellulases. Mutant 

strains of C. lucknowense have been isolated with 

improved protein yields (per gram biomass) compared to 

available Aspergillus and Trichoderma strains. Large-scale 

fermentations were developed up to 150,000 litre cultures. 

Growth conditions of Chrysosporium strains are very 

versatile, with growth from acid to alkaline pH and at 

temperatures of 25-43 °C. In contrast to other industrial 

fungi, Chrysosporium has growth characteristics that make 

it amenable to microtiter plate based culture methods. The 

development of a versatile gene expression and High 

Throughput gene discovery system for this fungus will be 

discussed.


1196 - Quantitative transcript analysis and 

heterologous expression of copper-radical oxidases in 

the white rot basidiomycete Phanerochaete 

chrysosporium 

A.J. Vanden Wymelenberg 1* , P.J. Kersten 2 , R.A. 

Blanchette 3 & D. Cullen 2 

1 University of Wisconsin-Madison Department of 

Bacteriology and USDA Forest Products Laboratory, One 

Gifford Pinchot Drive, Madison, WI 53705, U.S.A. - 

2 USDA Forest Products Laboratory, One Gifford Pinchot 

Drive, Madison, WI 53705, U.S.A. - 3 University of 

Minnesota Department of Plant Pathology, 495 Borlaug 

Hall, 1991 Upper Buford Circle, St. Paul, MN 55108-6030, 

U.S.A. - E-mail: ajvanden@facstaff.wisc.edu 

The white rot basidiomycete Phanerochaete chrysosporium 

is able to degrade all major components of wood: cellulose, 

lignin and hemicellulose. The oxidative enzymes thought 

to be involved in lignin degradation by this model system 

include lignin peroxidases (LiP), manganese peroxidases 

(MnP), and the peroxide-generating enzyme glyoxal 

oxidase (GLOX). Recently, a draft P. chrysosporium 

genome sequence has been made publicly available 

(http://www.jgi.doe.gov/programs/whiterot.htm). Blast 

searches of this database unexpectedly revealed additional 

GLOX-like sequences. Comparisons of these sequences to 

GLOX and to related copper-radical oxidases showed close 

structural similarities around the active site. At least three 

of the GLOX-like sequences have a putative eukaryotic 

secretion signal as predicted by SignalP 

(http://www.cbs.dtu.dk/services/SignalP/). Transcripts 

were detected in various defined culture media and from P. 

chrysosporium-colonized wood. Quantitative transcript 

analyses and heterologous expression of these genes will 

be presented. 

1197 - Evidence for carbon source regulated PKA and 

PKC signaling in the duplication cycle, polarization and 

septum formation in Aspergillus nidulans 

A.P.F.C. Vanzela & S. Said * 

Faculdade de Ciências Farmacêuticas de Ribeirão Preto, 

Av. do Café s/n 14040-903- Ribeirão Preto/SP, Brazil. - Email: 

susaid@usp.br 

The effects of glucose and of a pectic substrate in the 

duplication cycle, spore polarization and septation of 

Aspergillus nidulans were tested in poor and rich media. 

Growth on poor conditions and on sodium polypectate 

slowed nuclear duplication and reduced the coupling of 

polarization to mitosis. Coupling of septation to the third 

mitosis was also reduced by changing growth conditions. 

When PKA and PKC activators were added to the media 

the results suggested a role for PKA in slowing the 

duplication cycle, while allowing polarization. Addition of 

a PKC activator to poor media uncoupled the first septum 

formation from the third mitosis in a carbon source- 

regulated manner, suggesting a role for PKC in 

coordinating cell cycle signals, growth and cytokinesis. 

1198 - Ultrastructural study of partial symbiont 

delichenisation process in lichen Peltigera aphthosa 

T.A. Vlassova 

Dept.of Plant Physiology, Fac.of Biology, Moscow State 

University, 119899 Vorobjovy Gory, Moscow, Russia. - Email: 

t_vlas@yahoo.com 

The thalli of the foliose lichen Peltigera aphthosa were 

kept in the different conditions. The influence of 

unfavorable conditions on lichen thalli resulted in their 

degradation associated with the ultrastructural changes, 

including destructive alterations of cell organells, 

thickening of cell walls of both phycobiont and mycobiont 

and loosening of contacts between the symbionts. These 

changes indicated some degree of delichenisation of lichen 

components. Within certain time limits the degradation was 

reversible: the symbiont cell structure and accordingly their 

functions and interaction could be reestablished by 

transferring to the favorable conditions. The phycobiont 

recovered more quickly than the mycobiont, whose 

nutrition depended sufficiently on the algal partner, 

demonstrating the dominant role of the photobiont in this 

situation. However, the prolonged action of unfaborable 

conditions led to the irreversible ultrastructural alterations: 

to the cell destruction and the disjunction of symbionts and 

accordingly to the pronounced delichenisation, to the 

degradation of the whole thalli and finally to their death. 

The cell ultrastructure and the viability of such thalli could 

not be restored. The reversible destruction of the lichen 

symbionts could be one of the mechanisms of the 

reversible delichenization. It may be considered as a 

constituent of normal lichen physiology ensuring survival 

of these organisms in the extreme environmental 

conditions. 

1199 - Comparative karyology of Agaricus bisporus 

(Lange) Imbach strains 

V.N. Volkova * , M.V. Kozlova & O.V. Kamzolkina 

Moscow State University, Moscow, Leninskiye Gory, 

Biological Department, Russia. - E-mail: 

agaricus@yandex.ru 

There was made the comparison of basidial 

morphogenesis, sporogenesis and nuclear behavior in 

vegetative mycelium in several strains of A. bisporus, 

which have different types of life cycle, by light 

microscopy (Feulgen method and DAPI) and electron 

microscopy. The next cultivated and wild strains were 

investigated: pseudohomothallic Bs26 (Agaricus bisporus 

var. bisporus), heterothallic Bs94 (A. bisporus var. 

burnettii Kerrigan et Callac), and primary homothallic 

Bs423 (A. bisporus var. eurotetrasporus Callac & 



Guinberteau). Nuclear number in vegetative mycelium 

varied from 1 to 8 per cell and most of the cells contained 4 

nuclei. We observed amitotic nuclear divisions and nuclear 

migration through dolipore septum. Morphometric 

measuring of basidia and nuclear apparatus were made. 

Some preliminary results were obtained for ploidy level 

and place of meiosis in examined strains. We have 

described in details basidium morphogenesis in all the 

strains by using light and electron microscopy. We've 

observed: asynchronous 2-nd meiotic division; 

asynchronous migration of postmeiotic nuclei into 

sterigmata; postmeiotic mitosis in basidium. Electron 

microscopy of basidial nuclei showed the presence of 

synaptonemal complexes (SC) in all the strains; in 

heterothallic and primary homothallic ones we revealed SC 

for the first time. This fact confirms the processing of 

meiotic division in basidia of all the strains. We've 

reconstructed life cycles of all investigated strains. 

1200 - Biochemical and molecular characterization of 

arginase from Agaricus bisporus: Expression study in 

sporophore formation and postharvest development 

M.J.M. Wagemaker 1* , C. van der Drift 1 , M.S.M. Jetten 1 , 

L.J.L.D. Van Griensven 2 & H.J.M. Op den Camp 1 

1 Department of Microbiology, University of Nijmegen, 

Toernooiveld 1, 6525 ED, The Netherlands. - 2 Wageningen 

UR, Plant Research International B.V., P.O. Box 16, 6700 

AA Wageningen, The Netherlands. - E-mail: 

matthe@sci.kun.nl 

An extensive survey of higher fungi reveals that members 

of the family Agaricaceae, including Agaricus bisporus, 

accumulate substantial amounts of urea in their fruit 

bodies. During postharvest storage the amount of urea even 

increases substantially. Such a large increase in urea 

content may affect the quality of the stored mushroom, e.g. 

by the formation of ammonia from urea through urease 

action. Production of urea is not limited to fruit bodies, but 

also occurs in the mycelium. Despite the abundance of urea 

in the edible mushroom A. bisporus little is known about its 

physiological role, although it was proposed that in fruit 

bodies urea is an end product of catabolic pathways. Two 

major mechanisms for urea formation can be envisaged: 

the ornithine cycle and nucleic acid degradation. Arginase 

is the ornithine cycle enzyme that catalyzes the hydrolysis 

of arginine to urea and ornithine. In the scope of the study 

on urea metabolism, the work on arginase was initiated 

with the isolation of the arginase cDNA, gDNA and 

promoter region. Sequence analysis revealed the cDNA 

encodes a 311 aa protein which is probably expressed in 

the cytosol. Expression of the cDNA in E. coli has been 

established as a His-tagged protein. The purified 

recombinant protein was used to determine enzyme 

kinetics. The characterized gene and enzyme are used as 

molecular markers to study expression and regulation 

during sporophore formation and postharvest development. 

364 


1201 - Effect of Glomus intraradices inoculation on 

phosphate uptake in lettuce with two different P 

sources 

Y.H. Wong * & W.K. Chan 

Department of Applied Biology and Chemical Technology, 

The Hong Kong Polytechnic University, Hung Hom, 

Kowloon, Hong Kong, China. - E-mail: 

00900991r@polyu.edu.hk 

The effect of Glomus intraradices inoculation on 

phosphate uptake in Lactuca sativa L. (lettuce) cultivated 

in sand was examined. Soluble KH2PO 4 (SP) and sparingly 

soluble Ca5(PO 4) 3OH (RP) were applied to the potting 

medium at the rates of 0, 25, 125 mgP/kg sand. The harvest 

at week 6 shown that among all the treatments, the nonmycorrhizal 

(NM) plants with 25 mg SP/kg sand had the 

highest yield in dry matter. However, within the RP 

treatments, the dry matter for the mycorrhizal (M) plants 

with 25 mg and 125 mg RP/kg sand were significantly 

higher (P


cooled and non-cooled mycelium P. ostreatus on isolated 

plant mitochondria (T. aestivum L.) was observed 

uncoupling of oxidative phosphorilation. Uncoupling of 

mitochondria oxidation and phosphorilation processes 

contribute to heat generation and reduce oxidative stress 

that may be important to survival of mycelium. The results 

acquired confirm a certain degree of similarity in 

mechanisms of plant and fungi cells protection at lowtemperature 

stress. However, unlike plants, mycelium of 

the fungi of boreal zone is able to survive at low negative 

temperatures without preliminary acclimation, and contains 

protection proteins in the conditions of 'comfortable' 

temperature. 

1203 - Alpha-galactosidase gene cloning from 

Aspergillus fumigatus 

B. Yalcindag 1* , Z.B. Ogel 1 , U. Bakir 2 & P. Biely 3 

1 Middle East Technical University, Department of Food 

Engineering, 06531, Ankara, Turkey. - 2 Middle East 

Technical University, Department of Chemical 

Engineering, 06531, Ankara, Turkey. - 3 Slovak Academy of 

Sciences, Institute of Chemistry, SK-84 238, Bratislava, 

Slovakia. - E-mail: banuy@metu.edu.tr 

In this study, molecular cloning studies were performed on 

the α-galactosidase gene of Aspergillus fumigatus IMI 

385708. Degenerate primers were designed based on amino 

acid sequence of the purified enzyme and conserved 

sequences among α-galactosidases of Aspergillus and 

Penicillium species. These primers were used in PCR to 

amplify α-galactosidase gene fragments. A 900 bp PCR 

product was selected as a probe for further studies. RNA 

dot blot analysis showed that the highest expression of αgalactosidase 

mRNA was on the fifth day of growth on 

locust bean gum. This information was employed in cDNA 

synthesis. The synthesized cDNA was ligated onto Lambda 

ZAP Express vector to construct the primary cDNA library. 

After amplification of the primary library, screening 

studies were performed. At the end of primary and 

secondary screening, a number of putative positive plaques 

were obtained for further characterization studies. 

1204 - Transformation of Neotyphodium lolii with 

plasmids containing a native promoter disturbs the 

symbiotic interaction with its host 

N. Zhang 1 , V. Scott 1 , T.H. Al-Samarrai 1 , Y.Y. Tan 1 , M.J. 

Spiering 1 , L. McMillan 1 , D.B. Scott 1 , M.J. Christensen 2 & 

J. Schmid 1* 

1 IMBS, Massey University, Palmerston North, New 

Zealand. - 2 2AgResearch Grasslands, Palmerston North, 

New Zealand. - E-mail: J.Schmid@massey.ac.nz 

Neotyphodium lolii is an endosymbiont of perennial 

ryegrass. It produces alkaloids neurotoxic to herbivores. 

We wanted to determine in planta expression patterns of 

the N.lolii 3-hydroxy-3-methylglutaryl-CoA reductase 

(HMG CoA reductase) gene, believed to be involved in 

neurotoxin synthesis. We transformed the endophyte with 

plasmids, in which DNA fragments upstream of the open 

reading frame of the HMG CoA reductase gene controlled 

expression of the GUS reporter gene. In laboratory 

cultures, >400 bp were required for expression and >=1100 

bp for maximum expression. Transformants often showed 

abnormal hyphal morphology when growing within 

ryegrass. This occurred significantly more often in 

transformants which carried additional functional HMG 

CoA reductase promoter sequences compared to 

transformants with HMG CoA reductase gene upstream 

sequences of

Genera Index 

Abortiporus: 932 

Abrothallus: 594 

Absidia: 445, 525, 1175 

Acanthodochium: 739 

Acarospora: 649, 734, 918 

Acaulospora: 520, 530, 580, 1141 

Achlya : 687, 691, 1177 

Acremonium: 525, 556, 996, 1129 

Acrocordiopsis: 452 

Acrodontium: 996 

Acytostelium: 613 

Aenigmatomyces : 655 

Agaricostilbum: 727 

Agaricus: 144, 165, 167, 170, 304, 

305, 546, 597, 862, 908, 915, 1110, 

1140, 1199, 1200 

Agrocybe : 95, 1171 

Ahtiana: 760 

Aigialus: 452 

Albatrellus: 541 

Aldona: 688 

Aldonata: 688 

Aleurodiscus: 41, 557 

Allocetraria: 760 

Alnicola: 979 

Alpova : 723 

Alternaria: 71, 109, 220, 499, 524, 

575, 600, 712, 793, 840, 848, 855, 

860, 896, 919, 963, 1016, 1044 

Amanita: 40, 50, 128, 139, 236, 473, 

485, 487, 500, 546, 602, 605, 614, 

1143 

Amarrendia: 40 

Amauroderma: 478, 502 

Ampelomyces: 434, 881, 903 

Amphinema : 465, 536, 984, 1005 

Amylascus: 295 

Amyloathelia: 41 

Amylosporus: 554 

Amylostereum : 22, 429, 430 

Anaptychia: 535 

Anavirga: 992 

Anguillospora: 451, 475 

Anthostomella: 153, 739 

Anthracoidea: 495, 572, 701, 1031 

Anthracophyllum: 473 

Antrodia: 503, 554 

Antrodiella: 277, 554 

Aphanomyces: 691 

Arachnomyces: 668 

Archaeospora: 231, 520 

Arctoparmelia: 561 

Armillaria: 355, 439, 602, 646, 938, 

1006, 1015 

Arrhenia: 48, 50 

Arthonia: 320, 918 

Arthrinium: 525, 595 

Arthrobotrys: 247, 250, 525, 1047 

Arthromyces: 1139 

Artomyces: 541 

Aschersonia: 718 

Ascobolus: 38, 39, 525, 589 

Ascochyta: 462, 749, 882, 888 

Ascodesmis : 414, 525, 542 

Aseroë: 679 

Ashbya: 335 

Aspergillus: 29, 30, 31, 69, 101, 120, 

134, 218, 219, 220, 227, 230, 266, 

268, 308, 312, 339, 340, 343, 406, 

424, 482, 483, 525, 529, 542, 618, 

662, 800, 801, 803, 810, 813, 840, 

842, 844, 847, 848, 853, 855, 856, 

862, 863, 872, 880, 885, 887, 894, 

898, 905, 906, 909, 914, 923, 927, 

937, 963, 968, 974, 1016, 1072, 

1094, 1095, 1115, 1117, 1119, 1121, 

1124, 1125, 1127, 1139, 1163, 1164, 

1178, 1186, 1187, 1195, 1197, 1203 

Aspicilia: 261 

Asterostroma: 696 

Astraeus: 752, 889 

Astrocystis: 739 

Athelia: 594 

Athelopsis: 41 

Atichia: 283 

Aulacostroma: 688 

Aureobasidium: 120, 575, 919, 1064, 

1100 

Auricularia: 167, 308, 955, 1171 

Aurificaria: 554 

Auriscalpium: 352 

Australoporus: 476 

Auxarthron: 671 

Badimia: 568 

Bankera: 214 

Bartalinia: 542 

Bathyascus: 56 

Battarrea: 679 

Battarreoides: 679 

Beauveria: 79, 183, 187, 556, 770, 

1049 

Beltrania: 524, 542 

366 Book of Abstracts 

Beltraniella: 542 

Beltraniopsis: 542 

Bensingtonia: 727 

Bifusella: 685 

Bionectria: 542 

Bipolaris: 524, 759, 823 

Biscogniauxia: 865 

Bjerkandera: 517 

Blastobotrys: 253 

Blastospora: 798 

Blumeria: 854 

Boletus: 139, 307, 500, 541, 571, 

605, 723, 889, 1084 

Botrydina: 51 

Botryosphaeria: 669, 765 

Botryotinia: 684 

Botrytis: 86, 87, 88, 173, 227, 792, 

897, 914, 991, 1040 

Bovista: 48, 495, 679 

Brachiosphaera: 475 

Brefeldiella: 283 

Brevicellicium: 536 

Bridgeoporus: 390 

Brooksia: 283 

Bryoria: 316 

Buellia: 734 

Byssochlamys : 121, 874 

Byssoloma: 625 

Byssostilbe: 3 

Cadophora: 633 

Caecomyces: 118 

Callistosporium: 785 

Calocera: 762 

Calocybe: 50 

Caloplaca: 561, 630, 667, 734, 766 

Calvatia: 48, 679 

Camarophyllopsis: 145, 146 

Camillea: 153, 154, 739 

Campanella: 785 

Campylospora: 475 

Cancellidium: 585 

Candelabrum: 585 

Candelariella: 535, 734 

Candida: 258, 333, 369, 375, 844, 

871, 875, 890, 904, 924, 996, 1144, 

1152 

Canoparmelia: 591 

Cantharellus: 467, 552, 737 

Capnodium: 282 

Caripia: 210, 501, 502, 503 

Catathelasma: 139

Cavernularia: 356 

Cenococcum: 500, 984, 988, 1023, 

1039, 1045 

Cephalosporium: 869, 932 

Ceraceomyces: 41 

Ceratobasidium: 15, 557, 815 

Ceratocystiopsis: 296, 426, 706 

Ceratocystis: 20, 195, 296, 300, 301, 

353, 464, 706, 722, 822, 933, 1003, 

1009, 1027, 1099 

Ceratorhiza: 460 

Cercophora: 1, 525 

Cercospora: 669, 753, 763, 780 

Cercosporella: 763 

Cerinomyces: 762 

Ceriporia: 554, 564, 721 

Ceriporiopsis: 1119, 1142 

Cerrena: 202, 1131 

Cetraria: 624 

Cetrariastrum: 776 

Chaetocladium: 229 

Chaetomium: 38, 525, 542, 862, 

1134 

Chaetoporellus: 554 

Chaetosphaeria: 992, 1015 

Chalara: 300, 595 

Chamaemyces: 657 

Chlamydomyzium: 251 

Chloridium: 992, 1015 

Chlorophyllum: 657 

Choanephora: 445 

Choiromyces: 292, 702 

Chroodiscus: 284, 320 

Chroogomphus: 723 

Chrysomyxa: 10, 798 

Chrysosporium: 483, 506, 507, 579, 

1195 

Chytridium: 1002 

Ciboria: 684 

Ciborinia: 684 

Cintractia: 12, 473 

Circinella: 643 

Cladia: 682, 918 

Cladonia: 318, 561, 591, 1062 

Cladosporium: 120, 176, 220, 477, 

482, 575, 588, 598, 618, 763, 840, 

848, 855, 887, 914, 919, 1016, 1031, 

1033, 1100, 1176 

Claustula: 44 

Clavaria: 523, 666 

Clavariadelphus: 666 

Clavatospora: 475 

Claviceps: 883 

Clavicorona: 352, 466 

Clavulina: 737, 988 

Clavulinopsis: 473, 523 

Clitocybe: 552 

Clitopilus: 474 

Coccodothis: 688 

Coccomyces,: 685 

Cocconia: 688 

Cochliobolus: 227, 374, 690, 823 

Coemansia: 643 

Coenogonium: 287 

Coleosporium: 10, 798 

Colletotrichum: 77, 477, 556, 581, 

598, 914 

Collybia: 50, 95, 197, 352, 466, 552, 

785, 1184 

Colpoma: 685 

Coltricia: 215 

Coltriciella: 215 

Concamerella: 776 

Conidiobolus: 575, 1102 

Conidiosporomyces: 641 

Coniophora: 1122 

Coniosporium: 678 

Coniothyrium: 825 

Conocybe: 557 

Coprinopsis: 39 

Coprinus: 75, 76, 304, 525, 552, 589, 

738, 862, 1012, 1067, 1111 

Corallomycetella: 3 

Cordyceps: 104, 108, 183, 248, 358, 

599, 713, 718, 770, 1032, 1061 

Coremiella: 595 

Coriolopsis: 501, 502, 554, 932 

Coriolus: 304, 602, 1115, 1131 

Cortinarius: 46, 50, 51, 139, 141, 

314, 396, 397, 398, 399, 401, 403, 

495, 547, 552, 597, 605, 627, 699, 

931, 984, 988, 993, 1039 

Cosmospora: 3 

Craterellus: 473 

Crepidotus: 557, 564 

Crinipellis: 43, 473 

Cronartium: 10, 798 

Cryphonectria: 518, 672, 673 

Cryptendoxyla: 412 

Cryptococcus: 257, 716, 729, 844, 

1064 

Cryptosporiopsis: 182, 1015, 1024 

Cumminsiella: 10 

Cunninghamella: 525 

Cuphophyllus: 142, 145, 146 

Curvularia: 477, 524, 759 

Cyclomyces : 215, 554 

Cycloschizon: 688 

Cyclostomella: 688 

Cylindrocarpon: 607 

Cylindrocladium: 477 

Cyllamyces : 118 

Cymatoderma: 501, 502, 503 

Cystodendron: 1015 

Cystoderma: 50, 738 

Cytonaema: 819 

Cytospora: 996 

Cyttaria: 37, 750 

Dacrymyces: 541, 557, 762 

Dacryomitra: 762 

Dacryopinax: 473 

Dactylellina: 247 

Dactylina: 760 

Dactylium: 862 

Daedalea: 501, 517, 554 

Daldinia: 24, 153, 158, 739, 769, 865 

Davisomycella,: 685 

Delitschia: 589 

Delphinella: 893 

Dematophora: 156 

Dendrothele: 41 

Dendryphiella: 56, 595 

Dermocybe: 699 

Diaporthe: 181, 751 

Diaporthopsis: 751 

Dibaeis: 918 

Dicephalospora: 479 

Dichomitus: 476 

Dictyocyclus: 688 

Dictyonema: 260 

Dictyopanus: 210 

Dictyophora: 679 

Dictyostelium : 613 

Didymella: 749, 882 

Diheterospora: 248 

Dimargaris: 643 

Dimerella: 287 

Dingleya: 292 

Diplodia: 996 

Dirina: 285 

Discosphaerina: 1064 

Dispira: 643 

Dothidasteroma: 688 

Dothidea: 669 

Drechmeria: 248 

Drechslera: 477, 524, 882 

Drechslerella: 247 

Duplicaria: 685 

Dyctiuchus: 691 

Earliella: 501 

Echinochaete: 501 

Echinodontium: 430, 469 


Echinopodospora: 525 

Echinosporangium: 786 

Elaphomyces: 379 

Ellisembia: 788 

Emericella: 1178 

Emericellopsis: 3 

Emmonsia: 507 

Endomyces: 863, 932 

Endophragmiella: 788 

Endophyllum: 10, 798 

Endothia: 672, 673 

Englerodothis: 688 

Entoleuca: 136 

Entoloma: 50, 495, 523, 544, 708 

Entomocorticium: 426 

Entomophthora: 186 

Entonaema: 153, 739 

Entrophospora: 580 

Entyloma: 12 

Epichloë: 203, 311, 325, 715, 770, 

883, 1182 

Epicoccum: 220, 914 

Epidermophyton: 309, 506, 507, 579 

Epithelopsis: 41 

Epulorhiza: 460 

Erichseniomyces: 624 

Erioderma: 25 

Erotium: 863 

Erratomyces: 12 

Erynia: 475 

Erythrobasidium: 689 

Esslingeriana: 760 

Esteya: 996 

Euepixylon: 739 

Eupenicillium: 65, 66, 67, 68, 482, 

525, 542 

Eurotium: 120, 482, 525, 542, 914 

Eutypa: 971 

Evernia: 1112 

Everniastrum: 776 

Exobasidium: 495 

Exserohilum: 524, 759 

Favolaschia: 210 

Favolus: 478 

Ferrarisia: 688 

Fibricium: 41 

Filobasidiella: 257, 729 

Fistulina: 473 

Flabellospora: 475 

Flagellospora: 451 

Flammulina: 164, 167, 169, 304, 

352, 559, 922, 1169, 1202 

Flavocetraria: 561 

Flavoparmelia: 561 

Fomes: 502, 503, 517, 971 

Fomitella: 502, 503 

Fomitiporia: 696 

Fomitopsis: 23, 517, 554, 602, 918 

Fulgensia: 1173 

Fulvidula: 675 

Fulvifomes: 696 

Funalia: 1073 

Fusarium: 16, 70, 74, 101, 112, 160, 

161, 162, 182, 227, 269, 310, 322, 

422, 424, 458, 477, 542, 556, 588, 

607, 629, 692, 707, 712, 731, 732, 

764, 767, 775, 790, 791, 797, 801, 

809, 815, 817, 832, 833, 837, 838, 

839, 840, 850, 853, 855, 858, 860, 

864, 869, 870, 878, 879, 880, 884, 

901, 910, 932, 944, 960, 968, 1037, 

1043, 1044, 1115, 1151 

Fuscidea: 570 

Fuscoporia: 696 

Fusicladium: 319, 636 

Gaeumannomyces: 113, 598, 930 

Galerina: 48, 50, 51, 52, 495 

Gamsylella: 247 

Ganoderma: 44, 165, 166, 167, 236, 

277, 304, 306, 357, 363, 476, 478, 

501, 502, 503, 517, 541, 602 

Gastrodia: 972 

Gastrosuillus: 723 

Gautieria: 141, 651 

Geastrum: 541 

Genea: 953 

Geniculosporium: 156, 739, 1053 

Genistellospora: 151 

Geoglossum: 6, 523 

Geomyces: 176, 507 

Geopora: 180, 495 

Geosmithia: 533, 996 

Gerhardtia: 663 

Gibberella: 85, 732, 764, 797, 858 

Gigaspora: 126, 129, 443, 445, 580, 

928, 958, 987, 1107, 1141, 1179 

Gigiaspora: 530 

Gilmaniella: 542 

Glaziella: 414 

Gleoporus: 541 

Gliocladium: 3, 111, 477, 920, 1101 

Gliophorus: 146 

Gloeocystidiellum: 213 

Gloeophyllum : 200, 553, 554, 557, 

1157 

Gloeoporus: 554 

Glomus: 96, 129, 135, 225, 231, 332, 

442, 445, 520, 530, 539, 580, 928, 


958, 987, 1022, 1025, 1069, 1082, 

1107, 1130, 1141, 1147, 1159, 1179, 

1189, 1191, 1192, 1201 

Glyphium: 761 

Gnomonia: 859 

Gomphidius: 307, 723 

Gomphus: 139, 651, 781 

Gondwanamyces: 300 

Gracilistilbella: 3 

Graphis: 591 

Graphium: 694, 706 

Grifola: 169, 637, 899 

Guepinia: 762 

Guignardia: 598, 1053 

Gyalecta: 287 

Gymnoascus: 618 

Gymnopilus: 398, 404, 675, 683, 744 

Gymnopus: 210, 726, 785 

Gymnosporangium: 10, 798 

Haematomma: 591, 918 

Halophytophthora: 55 

Halorosellinia: 452 

Halosphaeria: 54, 963 

Hamigera: 542 

Hapalopilus: 554 

Haptocillium: 248, 249 

Haptoglossa: 78 

Harpella: 150, 151 

Harposporium: 5 

Hebeloma: 27, 39, 50, 51, 402, 465, 

495, 953, 979, 1039, 1113 

Helicobasidium: 978, 1068 

Helicodendron: 606 

Helicoon: 606 

Heliscus: 1132 

Helvella: 479, 495 

Hemileia: 756 

Hemimycena: 626 

Hendersonia: 789 

Hericium: 608 

Heterobasidion: 16, 18, 22, 197, 205, 

432, 650, 822, 831, 843, 1001, 1024, 

1138, 1157 

Heteroconium: 176 

Heterodea: 682 

Heterodermia: 591, 642 

Hexagona: 501, 502, 503 

Hirsutella: 5, 183, 599, 1061 

Hortaea: 120 

Humaria: 314 

Humicola: 119, 122, 542, 932, 1052 

Humidicutis: 146 

Humiditis: 145 

Hyalopsora: 798

Hyalorbilia: 479 

Hyalorhinocladiella: 693 

Hyaloscypha: 479 

Hydnangium: 956 

Hydnellum: 214 

Hydnochaete: 215 

Hydnocristella: 651 

Hydnodon: 748 

Hydnotryopsis: 295 

Hydnum: 552, 737 

Hydropus: 541 

Hygroaster: 145 

Hygrocybe: 44, 142, 143, 145, 146, 

495, 523, 527 

Hygrophoropsis: 723 

Hygrophorus: 144, 145, 146, 314, 

541 

Hymenochaete: 41, 215, 501, 502, 

503, 517, 696 

Hymenogaster: 956 

Hymenopsis: 1033 

Hymenoscyphus: 61, 647, 677, 941, 

1033, 1050 

Hymenostilbe: 599 

Hyphoderma: 351, 736 

Hyphodontia: 41, 536, 557, 564 

Hypholoma: 304, 436, 541, 936, 

1055 

Hypocenomyce : 660 

Hypochniciellum: 41 

Hypocopra: 155 

Hypocrea: 72, 343 

Hypocrella: 718 

Hypocreopsis: 44 

Hypoderma: 685 

Hypogymnia: 537, 1086 

Hypotrachyna: 591 

Hypoxylon: 13, 153, 154, 157, 158, 

564, 592, 739, 1052 

Hypsizygus : 164, 167 

Hysterangium: 956 

Hysteropezizella: 595 

Hysterostomella: 688 

Idriella: 914 

Leproloma: 659 

Incrustocalyptella: 474 

Ingoldiomyces: 641 

Inocutis: 696 

Inocybe: 50, 51, 141, 314, 400, 495, 

541, 547, 1039 

Inocyclus: 688 

Inonotus: 553, 554, 696, 915 

Iodophanus: 295 

Irpex: 473 

Isoachlya: 687 

Jacobia: 626 

Junghuhnia: 501 

Kentingia: 688 

Kiehlia: 688 

Kirschsteiniothelia: 452 

Kluyveromyces: 343 

Knufia: 678 

Kodamaea: 1064 

Kompsoscypha: 479 

Kuehneola: 10 

Laboulbenia: 952 

Laccaria: 39, 50, 51, 487, 495, 549, 

956, 1039, 1160, 1161, 1162 

Lachnum: 611, 647 

Lactarius: 40, 50, 51, 139, 307, 495, 

498, 602, 614, 658, 740, 741, 779, 

988, 999, 1038 

Laetiporus: 631, 745 

Lamprospora: 48 

Langermannia: 541 

Lasiosphaeria: 1 

Laurera: 591 

Lecanicillium: 248, 249 

Lecanora: 535, 570, 591, 594, 628, 

659, 1112 

Leccinum: 656, 1075 

Lecidea: 734 

Lemonniera: 639 

Lentaria: 651 

Lentinellus: 352, 466, 557, 754 

Lentinula: 167, 169, 304, 877, 880, 

1171, 1190 

Lentinus: 166, 168, 202, 308, 364, 

501, 502, 503, 546, 670, 758, 1110, 

1171 

Lenzites: 478, 501, 502 

Lepiota: 50, 657 

Lepista: 304, 552, 557, 597 

Lepraria: 570, 659, 735 

Leprieuria: 153 

Leproloma: 570 

Leptodontidium: 176, 588, 943 

Leptogium: 1059 

Leptoglossum: 51 

Leptographium: 296, 693, 706, 725, 

778, 919, 1099 

Leptoporus: 631 

Leptosphaeria: 1052 

Leptosporomyces: 41 

Letrouitia: 697 

Leucoagaricus: 597, 657 

Leucocoprinus: 657 

Leucosporidium: 9 

Lewia: 712 

Libertella: 996 

Licheniconium: 594 

Lichina: 616 

Lignincola: 452, 747 

Linderia: 445 

Lindquistia: 157, 739 

Lineolata: 1052 

Linochorella: 789 

Linospora: 208 

Lirula: 685 

Lobaria: 25, 262, 521, 526, 705, 

1054 

Lojkania: 686 

Lopharia: 501, 502 

Lophiostoma: 452 

Lophiotrema: 595 

Lophodermium: 136, 685 

Loweporus: 478, 973 

Loxosporopsis: 288 

Lulworthia: 54, 452, 1052 

Lunulospora: 475 

Lycoperdon: 889 

Lyophyllum: 495 , 738 

Macentina: 796 

Macrolepiota: 557, 597, 657 

Macrophomina: 891 

Macrosporium: 499 

Magnaporthe: 269, 565 

Malassezia: 886 

Malbranchea: 668, 1134 

Mammaria: 542 

Marasmiellus: 466, 541, 557, 785 

Marasmius: 50, 304, 314, 455, 466, 

474, 557, 564, 602, 785, 913, 926 

Margaritispora: 639 

Marssonina: 902 

Massalongia: 1059 

Massarina: 452, 585, 598, 1052 

Mastigobasidium: 9 

Megasporoporia: 501, 502, 503 

Melampsora: 10, 208, 798 

Melampsoridium: 319, 798 

Melanaria: 288 

Melanconium: 319 

Melanoleuca: 50, 557 

Melanoporia: 517 

Meliola: 279 

Menispora: 1015 

Mensularia: 696 

Meripilus: 169, 541 

Metacapnodium: 281 

Metarhizium: 79, 183, 185, 187, 376, 

713, 834, 977, 1049, 1102 


Micarea: 625, 660 

Microascus: 525 

Microbotryum: 42 

Micromphale: 785 

Micromucor: 1154 

Micropeltis: 283 

Microporellus: 502 

Microporus: 478 

Microsporum: 309, 506, 507, 579 

Microsporus: 602 

Microtheliopsis: 284 

Microthyrium: 283 

Miyagia: 798 

Mollisia: 61, 595, 611, 633 

Monacrosporium: 337, 373, 1066 

Monilinia: 684 

Moniliophthora: 43 

Monosporascus: 835 

Morchella: 293, 724, 899 

Mortierella: 445, 588, 618, 786, 

1154 

Mucor: 445, 588, 618, 643, 1153 

Muellerella: 594 

Muhria: 659 

Multiclavula: 666 

Mutinus: 473, 679 

Myceliophthora: 579, 1134 

Mycena: 210, 314, 466, 474, 544, 

557, 602, 626 

Mycoaciella: 557 

Mycobilimbia: 660 

Mycogone: 862 

Mycosphaerela: 882 

Mycosphaerella: 595, 632, 669, 749, 

780, 882, 939 

Mycovellosiella: 763 

Myriangiella: 283 

Myriococcum: 190 

Myxarium: 557 

Myxotrichum: 412, 638, 671 

Myzocytiopsis: 251 

Naemacyclus: 685 

Naeviopsis: 595 

Nais: 1052 

Nectria: 3, 585, 809 

Nemania: 136, 739 

Nematoctonus: 1047 

Nematoloma: 95 

Neococcomyces,: 685 

Neocordyceps: 599 

Neocosmospora: 542 

Neohygrocybe: 146 

Neohygrophorus: 146 

Neolentiporus: 469 

Neopaxillus: 723 

Neosartorya: 525, 542 

Neotyphodium: 203, 311, 325, 883, 

1077, 1182, 1204 

Neovossia: 641 

Neozygites: 186 

Nephroma: 262 

Nephromopsis: 760 

Neurospora: 35, 196, 227, 228, 266, 

267, 268, 1109, 1166 

Nexomyces: 665 

Nigrofomes: 501, 502, 503 

Nigroporus: 502 

Nigrospora: 524, 542, 887 

Nodulisporium: 477, 524, 542, 581, 

739 

Nodulosphaeria: 595 

Nomuraea: 79, 183 

Nothocorticium: 41 

Nyctalis: 708 

Oberwinkleria: 641 

Ochrolechia: 288, 695 

Ochropsora: 10 

Oidiodendron: 81, 176, 330, 638, 

941, 949, 1015 

Oligoporus: 1157 

Oliveonia: 15 

Omphalina: 50, 51, 146, 326, 495 

Omphalotus: 723 

Onychocola: 668 

Ophiocordyceps: 599 

Ophiostoma: 206, 296, 297, 298, 

299, 426, 427, 693, 706, 709, 710, 

911, 919, 996, 1063, 1099 

Orbilia: 64 

Orphella: 148 

Ossicaulis: 564 

Oxyporus: 478, 564 

Pachyella: 479 

Pachypatella: 688 

Paecilomyces: 5, 79, 121, 183, 542, 

556, 996, 1049 

Palawaniella: 688 

Panaeolus: 557 

Pandora: 186 

Panellus: 210, 352 

Panus: 758 

Papulospora: 483, 862 

Paradoxa: 292 

Paraglomus: 231 

Parasitella: 229, 1174, 1175 

Parmotrema: 591, 1172 

Parmularia: 688 

Parmulariopsella: 688 


Parmulariopsis: 688 

Parmulina: 688 

Passalora: 753 

Paxillus: 131, 315, 329, 337, 500, 

723, 957, 1080, 1103, 1113, 1137 

Peltigera: 730, 1198 

Penicillium: 65, 66, 67, 68, 69, 73, 

101, 119, 120, 121, 218, 220, 340, 

406, 424, 425, 470, 482, 515, 525, 

533, 575, 588, 598, 618, 662, 746, 

799, 808, 810, 813, 816, 840, 846, 

848, 855, 862, 863, 874, 880, 885, 

887, 898, 917, 932, 963, 968, 996, 

1016, 1090, 1094, 1097, 1104, 1193, 

1203 

Peniophora: 557, 602, 787, 1181 

Penzigia: 157 

Perenniporia: 502, 503, 554, 557, 

973 

Periconia: 452 

Periconiella: 739 

Perischizon: 688 

Peronospora: 644 

Pertusaria: 288, 695 

Pesotum: 296, 298, 706 

Pestalotiopsis: 477, 542, 581, 598, 

895, 1053 

Petractis: 287 

Peziza: 39, 295, 479 

Phacopsis: 289 

Phaeoacremonium: 996 

Phaeocollybia: 547 

Phaeogalera: 51 

Phaeolus: 631, 696 

Phaeomarasmius: 400 

Phaeophyscia: 535, 561 

Phaeoramularia: 763 

Phaeoseptaria: 789 

Phaeosphaeria: 882, 939, 1052 

Phaeotheca: 120 

Phakopsora: 798 

Phallus: 541, 679 

Phanerochaete: 957, 1055, 1119, 

1181, 1196 

Phellinus: 23, 197, 277, 361, 478, 

501, 502, 503, 541, 554, 696, 915 

Phellodon: 214 

Phialocephala: 26, 61, 81, 176, 633, 

943, 1005 

Phialophora: 61, 176, 182, 638, 930, 

943, 1005, 1015, 1024, 1050 

Phlebia: 541, 564, 936, 1183 

Phlebiella: 536, 541 

Phlebiopsis: 22, 111

Phlebopus: 828 

Pholiota: 304, 515 

Phoma: 134, 176, 542, 598, 600, 

749, 751, 830, 1033, 1053 

Phomopsis: 181, 477, 581, 598, 600, 

751, 996 

Phragmidium: 10, 798 

Phragmocephala: 595 

Phycomyces: 1153 

Phylacia: 153 

Phyllachora: 845 

Phyllopsora: 591 

Phyllosticta: 477, 576 

Physalacria: 772 

Physisporinus: 501, 1142 

Physopella: 798 

Phytophthora: 19, 55, 173, 207, 338, 

463, 652, 711, 809, 815, 849, 850, 

851, 892, 900, 948 

Pichia: 1150 

Pileolaria: 798 

Pilobolus: 589 

Piloderma: 984, 988, 1045 

Piloporia: 469 

Piptocephalis: 522 

Piptoporus: 517 

Piriformaspora: 182 

Pisolithus: 440, 752, 953, 956, 1162 

Pisoolithus: 889 

Pityrosporum: 886 

Placopsis: 1059 

Platismatia: 1086 

Plectania: 479 

Pleospora: 48, 690 

Pleurotus: 165, 166, 167, 197, 199, 

202, 236, 302, 303, 304, 308, 357, 

360, 363, 541, 559, 562, 640, 670, 

814, 862, 877, 912, 915, 922, 932, 

951, 1021, 1035, 1070, 1091, 1093, 

1110, 1119, 1133, 1145, 1170, 1171, 

1202 

Plicaria: 295 

Ploioderma: 685 

Pluteus: 541, 557 

Pochonia: 248, 249, 250, 1118 

Podabrella: 663 

Podophacidium: 479 

Podoscypha: 502 

Podosordaria: 155, 157, 739 

Podosphaera: 881, 903 

Podospora: 1, 38, 194, 525, 589 

Pollaccia: 636 

Polycephalomyces: 3 

Polycyclina: 688 

Polycyclus: 688 

Polydesmia: 479 

Polyporus: 476, 478, 502, 503, 515, 

546, 758 

Polysphondylium: 613 

Porina: 320, 568 

Porodaedalea: 696 

Poronia: 155, 157, 589, 739 

Postia: 200 

Preussia: 525 

Protodaedalea: 517 

Protothyrium: 688 

Psathyrella: 557, 738, 1033 

Pseudeurotium: 588 

Pseudocercospora: 753, 992 

Pseudochaete: 215 

Pseudocolus: 679 

Pseudocyphellaria: 521 

Pseudogymnoascus: 588 

Pseudohygrocybe: 146 

Pseudolembosia: 688 

Pseudotulostoma: 379, 412 

Pseudozyma: 1064 

Psilocybe: 487 

Psoroma: 1059 

Puccinia: 10, 42, 44, 495, 620, 681, 

798, 820, 830, 934, 1014 

Pucciniastrum: 10, 798 

Pucciniostele: 798 

Pulvinula: 479 

Pycnoporellus: 631 

Pycnoporus: 478, 501, 502, 503, 932 

Pyrenochaeta: 1011 

Pyrenocollema: 616 

Pyrenographa: 452 

Pyrenopeziza: 902 

Pyrenophora: 676, 820, 882 

Pyricularia: 524, 565, 595 

Pythiopsis: 687 

Pythium: 90, 601, 654, 809, 815, 

822, 839, 840, 850, 917, 1087 

Quintaria: 452 

Raffaelea: 916 

Ramalea: 682 

Ramalina: 918, 1105 

Ramaria: 541, 552, 651 

Ramariopsis: 666 

Ramichloridium: 996 

Ramicola: 557 

Ramonia: 537 

Ramularia: 763, 780 

Ravenelia: 798 

Redellomyces: 292 

Restiosporium: 42 

Rhagadolobium: 688 

Rhexographium: 694 

Rhipidocarpon: 688 

Rhizina: 22 

Rhizocarpon: 734 

Rhizochaete: 41 

Rhizoctonia: 173, 174, 438, 460, 477, 

542, 815, 827, 840, 850, 914, 944, 

1034, 1079, 1123, 1158 

Rhizodiscina: 479 

Rhizomucor: 1194 

Rhizoplaca: 950 

Rhizopogon: 139, 141, 465, 723, 975, 

989, 1038 

Rhizopus: 445, 525, 717, 848, 914 

Rhizopycnis: 181 

Rhizostilbella: 3 

Rhodocollybia: 726, 785 

Rhodophyllus: 602 

Rhodosporidium: 9 

Rhodotorula: 9, 689, 716, 1064 

Rhodotus: 564 

Rhytisma: 495 

Rickenella: 50, 626 

Rigidoporus: 478, 501, 502, 517, 554 

Rimularia: 594 

Roccella: 285 

Roccellina: 285 

Rosellinia: 136, 153, 156, 739, 978, 

1010 

Rozites: 398 

Russula: 40, 50, 51, 139, 141, 381, 

495, 546, 547, 602, 605, 614, 621, 

623, 774, 931, 984, 1038, 1039, 1045 

Saccharomyces: 30, 193, 252, 253, 

327, 333, 335, 336, 341, 343, 366, 

368, 369, 371, 980, 1095, 1119, 

1144, 1178 

Saccobolus: 525, 589 

Saprolegnia: 687, 691 

Sapromyces: 55 

Sarcodon: 214 

Sarcosphaera: 295 

Scabropezia: 295 

Schizophyllum: 28, 193, 304, 357, 

359, 501, 502, 503, 637 

Schizopora: 478, 696 

Schizosaccharomyces: 30, 1095 

Schizothyrium: 283 

Sclerocystis: 530, 987 

Scleroderma: 487, 500, 679, 752, 

953, 999, 1162 

Sclerotinia: 89, 684 

Sclerotium: 1033 


Scoliciosporum: 625 

Scopulariopsis: 482, 525 

Scutellinia: 495, 564 

Scutellospora: 445, 530, 580, 987, 

1141, 1179 

Scutomollisia: 595 

Scytalidium: 122, 176, 190, 483, 542, 

871, 1116 

Scytinopogon: 666 

Selenosporella: 992 

Septobasidium: 104, 680 

Septoria: 454, 632, 669, 780, 789, 

820, 876, 882 

Septoriella: 789 

Serpula: 217, 609, 704 

Setchelliogaster: 956 

Sigmoidea: 56 

Simplicillium: 248, 575 

Sinotermitomyces: 663 

Sirobasidium: 13 

Sistotrema: 41, 737 

Skeletocutis: 554 

Smittium: 148 , 150, 151 

Soleella: 685 

Sordaria: 228, 525 

Sparassis: 541 

Spegazzinia: 482 

Sphaerophorus: 286 

Sphaeropsis: 16 

Sphaerosporella: 178 

Sphaerostilbella: 3 

Spilocaea: 636 

Spongipellis: 564 

Sporidesmium: 788, 1015 

Sporidiobolus: 9, 1064 

Sporobolomyces: 689, 716 

Sporormia: 589 

Sporormiella: 38, 525 

Sporotrichum: 483 

Stachybotrys: 218, 219, 220, 525, 

595 

Stachylina: 148, 150 

Stagonospora: 882 

Stecchericium: 502 

Steccherinum: 473, 557 

Stemphylium: 690 

Stenella: 753, 763 

Stephanoascus: 253 

Stereocaulon: 659, 918, 1112 

Stereum: 22, 41, 501, 502, 503, 517, 

602, 971 

Sticta: 521 

Stigmidium: 594 , 734 

Stilbella: 3, 122, 482, 1129 

Stilbocrea: 3 

Stilbum: 3, 727 

Strigula: 284 

Strobilomyces: 473 

Strongwellsea: 186 

Stropharia: 95, 165, 304, 541, 1093 

Suillus: 27, 304, 331, 465, 546, 723, 

827, 957, 975, 999, 1007, 1093, 

1113, 1126 

Swampomyces: 452 

Symphaeophyma: 688 

Syncephalastrum: 445 

Syncephalis: 522, 643 

Synnmukerjiomyces: 1134 

Taeniolella: 624 

Talaromyces: 65, 67, 121, 122, 190, 

482, 525, 542, 1033 

Tapesia: 61 

Taphrina: 208, 254, 757 

Teloschistes: 123 

Tephrocybe: 39 

Terfezia: 702 

Termitomyces: 542, 619, 663, 733 

Tetracladium: 451 

Tetrapyrgos: 785 

Thallomyces: 688 

Thamnomyces: 153, 739 

Thecotheus: 295 

Theissenia: 157 

Thekopsora: 10, 798 

Thelephora: 214 

Thermomyces: 190 , 1134 

Thielaviopsis: 300, 914 

Thraustochytrium: 1149 

Thraustotheca: 691 

Thysanophora: 983 

Thysanothecium: 682 

Tilletia: 12, 204, 641, 866 

Tolypocladium: 588, 770 

Tomentella: 214, 700, 953, 954, 984, 

1039, 1045 

Toninia: 660, 1173 

Torpedospora: 56 

Torrendia: 40 

Torrendiella: 37 

Torrubiella: 248 

Torula: 624, 1134 

Trachyspora: 10 

Trametes: 308, 478, 501, 502, 503, 

517, 553, 554, 602, 773, 918, 932, 

971, 1110, 1146, 1156, 1157 

Tranzschelia: 10 

Trechispora: 501, 502, 503 

Tremella: 13, 594, 915 


Tremolecia: 1114 

Trichaptum: 23, 501, 502, 503, 554, 

696 

Tricharia: 568 

Trichocladium: 963 

Trichoderma: 72, 101, 111, 112, 115, 

171, 172, 173, 174, 175, 217, 343, 

372, 525, 542, 585, 618, 634, 794, 

801, 812, 836, 862, 869, 896, 932, 

1015, 1016, 1101, 1158, 1168, 1195 

Trichoglossum: 523 

Tricholoma: 27, 165, 467, 546, 552, 

605, 648, 826, 899, 1171 

Trichomerium: 279 

Trichophaea: 122, 564 

Trichophaeopsis: 564 

Trichophyton: 309, 506, 507, 575, 

579, 1074, 1188 

Trichosporon: 904 

Trimmatostroma: 120, 624 

Triphragmium: 10 

Triscelophorus: 451, 475 

Trochodium: 798 

Trogia: 785 

Truncocolumella: 723 

Tryblidiopycnis: 1053 

Tubaria: 557 

Tuber: 132, 292, 702, 777, 953, 

1008, 1048, 1051, 1148 

Tubercularia: 3 

Tuberculina: 720 

Tubulicrinis: 41, 536 

Tuckermannopsis: 760 

Tuckneraria: 760 

Tulasnella: 15, 41 

Tulostoma: 541, 679 

Tylopilus: 139 

Tylospora: 984 

Tympanis: 992 

Typhula: 1120 

Tyromyces: 554 

Umbelopsis: 1015 

Uredinopsis: 798 

Uredo: 42, 728 

Urocystis: 674 

Urohendersoniella: 789 

Uromyces: 10, 645, 798, 1041 

Usnea: 290, 309, 555, 591, 1112, 

1172 

Ustilago: 267, 841 

Vanromburghia: 785 

Vararia: 41, 696 

Vascellum: 679 

Venturia: 208, 319, 636

Verrucaria: 616 

Verruculina: 963 

Verticillium: 183, 248, 249, 250, 777, 

783, 795, 809, 965, 966 

Volutella: 3 

Volvariella: 167, 304, 541 

Vouauxiella: 594 

Vulpicida: 760 

Wallemia: 120, 743, 914 

Wawelia: 155 

Wilcoxina: 178, 180, 1029 

Wolfiporia: 631 

Xanthomendoza: 703 

Xanthoparmelia: 261 

Xanthoria: 260, 661, 667, 703, 918, 

1165, 1180 

Xerocomus: 999 

Xeromphalina: 236, 357, 626 

Xeromyces: 117 

Xylaria: 152, 153, 154, 157, 477, 

542, 592, 1053 

Xylobolus: 517, 602 

Xyloclodium: 739 

Xylocoremium: 157 

Zamenhofia: 537 

Zygosaccharomyces: 117, 914 

Zygospermella: 589 

Zymonema: 507 


Family Index 

Acarosporaceae: 755 

Agaricaceae: 210, 622, 657, 1200 

Amanitaceae: 40, 379, 540 

Annulatascaceae: 382 

Arthrodermataceae: 4 

Ascobolaceae: 295, 414 

Ascodesmidaceae: 294, 414 

Asterinaceae: 688 

Bacidiaceae: 625 

Bionectriaceae: 3 

Bolbitiaceae: 738 

Boletaceae: 307, 379, 656 

Cantharellaceae: 379 

Carbomycetaceae: 414 

Catenariaceae: 245 

Cerinomycetaceae: 762 

Chaetosphaeriaceae: 348, 382 

Cladoniaceae: 682 

Clathraceae: 679 

Clavariaceae: 143, 527, 666 

Clavicipitaceae: 2, 3, 5, 108, 203, 

245, 248, 325, 715, 718, 770, 883 

Cochlonemataceae: 245, 655 

Coenogoniaceae: 287 

Coprinaceae: 51, 1012, 1185 

Corticiaceae: 41, 211, 212, 501, 502, 

503, 509, 787, 967 

Cortinariaceae: 307, 379, 396, 400, 

403, 540, 547, 675, 738 

Dacrymycetaceae: 762 

Delitschiaceae: 742 

Dermateaceae: 59, 61, 611 

Diatrypaceae: 157 

Discinaceae: 414 

Echinodontiaceae: 430 

Elaphomycetaceae: 379 

Ganodermataceae: 501, 502, 503, 

1106 

Geastraceae: 679 

Geoglossaceae: 143 

Gigasporaceae: 443 

Glaziellaceae: 294, 414 

Glomerellaceae: 348 

Gomphaceae: 212 

Gomphidiaceae: 622, 723 

Gyalectaceae: 287 

Gymnoascaceae: 4, 668 

Halosphaeriaceae: 53 

Hapalopilaceae: 721 

Harpellaceae: 151 

Helicocephalidaceae: 245 

Helotiaceae: 59, 61, 63, 647 

Helvellaceae: 414 

Hemiphacidiaceae: 59 

Hericiaceae: 212, 502 

Hyaloscyphaceae: 61, 62, 63, 64, 

611, 647 

Hydnaceae: 748 

Hydnodontaceae: 748 

Hygrophoraceae: 51, 145, 146 

Hymenochaetaceae: 501, 502, 503, 

696 

Hyphodermataceae: 245 

Hypocreaceae: 3, 245, 382 

Karstenellaceae: 414 

Lasiosphaeriaceae: 1, 382 

Lecanoraceae: 950 

Legeriomycetaceae: 148, 151 

Leotiaceae: 59, 64, 611 

Letrouitiaceae: 697 

Lobariaceae: 521 

Loramycetaceae: 59 

Lycoperdaceae: 679 

Melanommataceae: 382 

Meristacraceae: 245 

Metacapnodiaceae: 281, 678 

Micareaceae: 625 

Microascaceae: 348 

Morchellaceae: 414 

Mortierellaceae: 786 

Mucoraceae: 1175 

Mycenaceae: 210 

Myzocytiopsidaceae: 245 

Nectriaceae: 3 

Nidulariaceae: 679 

Onygenaceae: 4, 668 

Ophiostomataceae: 1063 

Orbiliaceae: 245, 246, 611 

Otideaceae: 294 

Parmeliaceae: 289, 555, 760, 776 

Parmulariaceae: 688 

Paxillaceae: 307, 379 

Peltigeraceae: 730 

Pertusariaceae: 288, 695 

Pezizaceae: 295, 414 

Phallaceae: 679 

Phragmidiaceae: 10 

Physciaceae: 642 

Pilocarpaceae: 625 

Piptocephalidaceae: 522 


Platystomaceae: 382 

Pleosporaceae: 109, 348 

Pleurotaceae: 210, 245 

Pluteaceae: 540, 733 

Podoscyphaceae: 501, 502, 503 

Polyporaceae: 501, 502, 503, 631, 

995, 1106, 1131 

Pyronemataceae: 294, 414 

Rhizinaceae: 414 

Rhizopogonaceae: 723 

Rhynchostomataceae: 761 

Rhytismataceae: 136 

Roccellaceae: 285 

Russulaceae: 307, 379, 738, 988 

Rutstroemiaceae: 59 

Saprolegniaceae: 687, 691, 698 

Sarcoscyphaceae: 294, 414 

Sarcosomataceae: 294, 414 

Schizophyllaceae: 501, 502, 503 

Sclerodermataceae: 679 

Sclerotiniaceae: 59, 64, 611, 647, 

684 

Sebacinaceae: 14, 1034, 1039 

Sordariaceae: 665 

Sporormiaceae: 742 

Stereaceae: 212, 430, 501, 502, 503 

Stereocaulaceae: 659 

Strophariaceae: 51, 1106 

Suillaceae: 723 

Tapinellaceae: 622 

Teloschistaceae: 661, 667 

Terfeziaceae: 702 

Thelephoraceae: 212, 700, 748 

Trapeliaceae: 287 

Trichocomaceae: 348, 555 

Tricholomataceae: 51, 146, 210, 455, 

540, 549 

Truncocolumellaceae: 622 

Tuberaceae: 292, 414, 702, 777 

Tubeufiaceae: 348, 382 

Tulostomataceae: 679 

Valsaceae: 555 

Verrucariaceae: 796 

Vibrisseaceae: 59 

Xylariaceae: 136, 153, 154, 155, 156, 

157, 158, 249, 382, 415, 558, 592, 

739, 769, 1028 

Zoopagaceae: 245

Order Index 

Agaricales: 40, 50, 51, 210, 211, 212, 

397, 399, 400, 403, 515, 547, 549, 

582, 675, 683, 726, 733, 772, 782, 

993, 1039, 1185 

Aphyllophorales: 23, 41, 49, 51, 211, 

469, 476, 478, 492, 501, 502, 503, 

666, 696, 773, 967, 981, 1185 

Arthoniales: 285 

Ascosphaerales: 107 

Asellariales: 149 

Auriculariales: 14, 492, 1039 

Boletales: 212, 547, 582, 622, 656, 

723, 993 

Botryobasidiales: 211 

Cantharellales: 212, 547, 582 

Capnodiales: 678, 761 

Ceratobasidiales: 15 

Chaetothyriales: 107, 555, 761 

Cyttariales: 750 

Dacrymycetales: 762 

Diaporthales: 2, 555 

Dothideales: 120, 281, 283, 454, 669, 

761, 780 

Eccrinales: 149 

Entomophthorales: 104, 186, 229 

Erysiphales: 273, 514 

Eurotiales: 107, 121, 533, 555 

Gautieriales: 622 

Geastrales: 622 

Glomales: 6, 225, 231, 441, 1107 

Gomphales: 547, 622, 651 

Gyalectales: 287 

Halosphaeriales: 57, 747 

Harpellales: 148, 149, 150, 151 

Helotiales: 61, 64, 413, 479, 595, 

633, 647, 684, 750, 943 

Hymenochaetales: 211, 215, 478, 

696 

Hypocreales: 2, 3, 108, 121, 348, 

382, 533, 770, 1129 

Hysteriales: 761 

Kickxellales: 148, 229 

Laboulbeniales: 104, 671, 952 

Lecanorales: 286, 356, 410, 521, 555, 

625, 659, 784 

Leotiales: 3, 59 

Lycoperdales: 210, 582, 622 

Microascales: 3, 300 

Microbotryales: 9 

Mortierellales: 229, 786, 1174 

Mucorales: 188, 229, 643, 717, 1175 

Mycosphaerellales: 595 

Myriangiales: 761 

Neocallimastigales: 118 

Onygenales: 4, 107 

Ophiostomatales: 671 

Ostropales: 287 

Patellariales: 761 

Peronosporales: 274, 514, 644 

Pezizales: 180, 246, 291, 294, 295, 

414, 479, 505, 777 

Phallales: 582 

Phyllachorales: 3, 248 

Pleosporales: 57, 595, 639, 742, 761 

Polyporales: 478, 515, 721 

Rhytismatales: 479, 685 

Russulales: 40, 547, 582, 658, 741, 

993, 1039 

Saccharomycetales: 253, 409 

Saprolegniales: 687 

Sclerosporales: 274 

Sordariales: 665 

Taphrinales: 254 

Teloschistales: 123 

Thelephorales: 214, 1039 

Tilletiales: 12, 641 

Tulasnellales: 15, 211 

Uredinales: 10, 42, 275, 514, 620, 

720, 934 

Ustilaginales: 12, 42, 701 

Verrucariales: 411 

Xylariales: 382, 505, 555, 592 

Zoopagales: 522, 655 


Author Index 

Aanen, D.K.: 402, 619, 663 

Abarenkov, K.: 214 

Abba', S.: 1135 

Abbasi, M.: 620 

Abdel-Gliel, M.: 801 

Abdel-Raheem, A.: 451 

Abdel-Wahhab, M.: 801 

Abdurahimova, Z.H.: 925 

Abesha, E.: 926 

Abu El-Naga, S.: 1013 

Ács, K.: 1194 

Adam, R.K.: 1152 

Adamcík, S.: 621 

Addy, H.D.: 81 

Adholeya, A.: 584, 1162, 1191, 1192 

Adler, P.H.: 150 

Adriaensen, K.: 331, 1007 

Aebi, M.: 75 

Aeny, T.N.: 823 

Agerer, R.: 622 

Aguirre, E.: 505 

Ahanjan, M.: 800 

Ahlén, C.: 221 

Ahmad, R.: 927 

Ahn, B.Y.: 486 

Ahrén, D.G.: 131, 337, 1066 

Aime, M.C.: 1067 

Aimi, T.: 1068 

Akiyama, M.: 928 

Al-Ansari, H.I.: 904 

Al-Masri, M.: 89 

Al-Samarrai, T.H.: 1204 

Alabouvette, C.: 112 

Alamouti, S.: 778 

Albers, A.R.: 1117 

Aldana-Gomez, R.: 623 

Alekhina, I.A.: 790, 791 

Alexander, I.J.: 1050 

Alizadeh, -: 794 

Allameh, A.: 1163, 1164 

Allaway, W.G.: 439 

Allen, M.F.: 97, 1000 

Allen, T.R.: 179 

Alleyne, A.T.: 1041 

Allmér, J.: 453 

Alonso, J.: 590 

Alstrup, V.: 624 

Amann, R.: 980 

Amerian, M.R.: 1069 

Amicucci, A.: 1148 

Ammirati, J. F.: 400 

Amra, H.: 801 

Andersen, B.: 71 

Andersen, H.L.: 625 

Andersen, R.A.: 1084 

Anderson, I.: 1050 

Anderson, J.B.: 369 

Anderson, R.C.: 929 

Andersons, B.: 1122 

Andrianova, T.V.: 137, 454 

Anthony, K.P.: 802, 840 

Antipov, A.: 1131 

Antonín, V.: 455, 626 

Antonini, D.: 456, 457, 627 

Antonini, M.: 456, 457, 627 

Antonsen, H.: 929 

Anusarnsunthorn, V.: 695 

Aoki, T.: 458 

Aragno, M.: 536, 1098 

Arakawa, M.: 978, 1010 

Aranyanart, P.: 767 

Araujo, W.L.: 556 

Archer, A.W.: 695 

Arenal, F.: 647 

Ariza, A.: 609 

Armaleo, D.: 1172 

Arnold, A.E.: 321 

Arnould, C.: 441, 1107 

Articus, K.: 290 

Arup, U.: 628, 766 

Ascaso, C.: 124, 459, 590, 610 

Ashford, A.E.: 439, 440 

Ashida, H.: 169, 170 

Ashkannejhad, S.: 975 

Askun, T.: 803 

Assefa, S.: 820 

Athipunyakom, P.: 460 

Atkinson, H.A.: 264 

Augustin, C.: 930 

Aust, H.J.: 1024 

Avery, S.V.: 327 

Avio, L.: 442 

Aviram, S.: 702 

Avis, P.G.: 931 

Awad, M.: 1013 

Ayres, M.P.: 426, 427 

Azevedo, J.L.: 556 

Baar, J.: 80, 1038 

Baars, J.J.P.: 165 

Babcock, C.: 461 

Baciarelli Falini, L.: 1051 

Badalyan, S.M.: 304, 364 

Badebo, A.: 820 

Bago, B.: 127, 270 

Bahyl, V.: 804 

Bainbridge, B.W.: 138 

Baker, C.J.: 353 

Bakir, U.: 1203 

Bakradze, M.: 202 

Balali, G.R.: 629 

Balasubramanian, S.: 1160 

Balci, Y.: 463 

Baldrian, P.: 932, 1070 

Baloch, E.: 320 

Balschmidt, L.: 630 

Banik, M.T.: 631 

Banke, S.: 632 

Banks, J.: 868 

Baptista, S.: 567 

Barakat, R.: 89 

Baral, H.-O.: 413, 647 

Barberis, G.: 617 

Bärlocher, F.: 994 

Barnes, I.: 464, 933, 1009, 1027 

Barreno, E.: 734 

Barrera-Saldaña, H.A.: 848 

Barreto de Oliveira, M.T.: 257 

Barroetaveña, C.: 465 

Barros, N.M.: 556 

Barry, K.M.: 21, 805, 806 

Barta, D.J.: 840 

Bartnicki-García, S.: 268 

Barve, M.: 749 

Bassiri, Sh.: 807 


Bauer, R.: 8, 9, 11, 720, 1034 

Baumgarten, A.: 192 

Baumgartner, M.: 769 

Beakes, G.W.: 78 

Beard, C.E.: 150 

Beauvais, A.: 31 

Bécard, G.: 129, 443 

Bech-Andersen, J.: 217 

Beck, A.: 323, 1173 

Beckett, R.P.: 1071 

Bedard, J.E.J.: 654 

Beecher, T.: 1140 

Begerow, D.: 11 

Beharav, A.: 968 

Belardinelli, M.: 308 

Bell, A.: 38 

Belova, N.V.: 466 

Benaraba, R.: 874 

Bencina, M.: 1072, 1124 

Bencivenga, M.: 1051 

Bendiksen, K.: 49 

Benhamou, N.: 90, 850 

Benítez, T.: 173 

Benson, E.E.: 609 

Benzie, I.F.F.: 167 

Berbee, M.L.: 109, 179, 350, 690 

Berch, S.M.: 179, 467 

Berdal, K.G.: 832 

Berg, G.: 966 

Bergero, R.: 226, 446 

Bermek, H.: 1073, 1074, 1188 

Bernard, M.: 29 

Berndt, R.: 275, 468, 756 

Bernhoft, A.: 808, 811 

Bernicchia, A.: 469 

Berreck, M.: 307, 1093 

Bertossa, R.C.: 75 

Bhadoria, P.B.S.: 560 

Bhattacharya, D.: 755 

Bhumiratana, A.: 1129 

Bianciotto, V.: 443 

Bielawski, L.: 1075 

Biely, P.: 1203 

Bills, G.F.: 633 

Binder, M.: 110, 209 

Birch, R.N.: 43 

Birzele, B.: 424 

Bissett, J.: 70, 72, 634 

Bisson, M.-C.: 919, 920 

Bjelland, T.: 1076 

Bjorvand Engh, I.: 770 

Bjureke, K.: 494 

Blackwell, B.: 70 

Blackwell, M.: 104, 184 

Blanchette, R.A.: 1196 

Blankenship, J.D.: 1077, 1182 

Blesdoe, C.: 1000 

Blom, J.: 341 

Bloomer, P.: 355 

Bock, A.: 128 

Boddy, L.: 322, 431, 608, 908, 936, 

957, 971, 1055, 1058 

Bødker, L.: 1037 

Bodles, W.J.A.: 809 

Boekhout, T.: 257, 377, 438, 470, 

471 

Boellmann, J.B.: 934

Bogomolova, E.V.: 1078 

Bohar, Gy.: 845 

Bojosi, B.W.: 600 

Bokhari, F.: 810 

Bol, R.: 143 

Bolchi, A.: 132 

Bolot, O.: 1014 

Bonati, C.: 1087 

Bondartseva, M.A.: 635 

Bonello, P.: 16 

Bonfante, P.: 126, 330, 443, 1135 

Boomsma, J.J.: 619 

Boonkerd, N.: 1171 

Boonpratuang, T.: 473, 474 

Boonyeun, N.: 475, 585 

Bordallo, J.J.: 250 

Borgen, T.: 47 

Borovskii, G.B.: 1202 

Boswell, G.P.: 1079, 1123 

Bothe, H.: 332 

Bottoli, A.P.F.: 75 

Bougoure, D.S.: 1089 

Boulay, H.: 676 

Bourett, T.M.: 269 

Bowden, R.L.: 959 

Bown, J.L: 1057 

Boyle, C.: 182, 830 

Braathen, O.-A.: 221 

Brachat, S.: 335 

Bran, M.C.: 139, 498 

Branco, S.M.: 567, 935 

Brand, J.H.: 1161 

Brandenburger, W.: 514 

Brandrud, T.E.: 399 

Branney, A.W.: 936 

Brasier, C.M.: 206, 207, 299 

Braun, S.: 662, 861, 907 

Braun, U.: 273, 636, 753 

Brceski, I.: 363 

Brechenmacher, L.: 129 

Brem, D.: 715 

Bremnes, N.B.: 811 

Breuil, C.: 778, 1099 

Breuninger, M.: 444 

Brezeanu, A.: 1147 

Brizzio, S.: 716 

Brodie, E.: 986 

Brofiga, G.: 870 

Brown, M.: 586, 826 

Brozová, J.: 812, 836, 837 

Bruggeman, J.: 937 

Bruhn, J.N.: 938, 1006 

Brun, A.: 329 

Brunner, K.: 175 

Bruns, T.D.: 223, 989 

Brusoni, M.: 821 

Brzostowski, A.: 1080 

Buchalo, A.S.: 637 

Buchan, A.: 939 

Buchanan, P.K.: 44, 476 

Buck, K.W.: 206 

Bücking, H.: 1081, 1082 

Buddie, A.: 609 

Büdel, B.: 287 

Bulat, S.A.: 790, 791 

Bullerman, L.B.: 898 

Bullock, S.: 797 

Bunyard, B.A.: 510, 940 

Buratti, C.: 575 

Burford, E.P.: 328, 1083 

Burgstaller, W.: 1104 

Burhenne, M.: 271, 513, 514 

Burlingame, R.: 1195 

Burmester, A.: 1175 

Burns, S.C.: 941, 949 

Burton, K.: 1140 

Busarakam, K.: 477 

Buscot, F.: 82, 91 

Bush, L.P.: 1077 

Bussey, H.: 32 

Bustan, A.: 1048 

Buswell, J.A.: 167 

Butler, G.: 333 

Butt, T.M.: 114, 187, 977 

Buyck, B.: 99, 381, 623 

Byrne, A.: 911 

Cabañes, F.J.: 909 

Caetano-Anollés, G.: 926 

Cafaro, M.J.: 104, 149 

Cairns, V.: 813 

Calderone, R.: 258 

Calduch, M.: 638 

Camaño Heredia, H.: 814 

Camarero, S.: 199 

Camargo-Ricalde, S.L.: 942 

Campbell, J.: 53, 639 

Campoamor, J.N.: 478 

Campos, R.A.: 556 

Cannon, P.F.: 348, 581, 688 

Cannon, R.D.: 924 

Cano, J.: 665 

Cantrell, S.A.: 145, 479 

Capelari, M.: 640 

Capello, S.: 480 

Capieau, K.: 897 

Carasan, E.M.: 1147 

Carballo, V.: 480 

Carbone, I.: 417 

Caretta, G.: 575, 1047 

Carlsen, T.A.: 943 

Carranza, J.: 380 

Carris, L.M.: 204, 641 

Carstensen, J.M.: 515, 516 

Carter, D.A.: 230 

Carter, M.D.: 1042 

Carter, V.: 898 

Carulli, I.: 821 

Caruso, A.: 481 

Casselton, L.A.: 1067 

Castaneda, A.: 729 

Castellano, M.A.: 232, 390 

Castlebury, L.A.: 2, 641, 681 

Castrejon, F.: 30 

Castro, B.: 722 

Cavalcante, U.M.T.: 539, 1179 

Cavalcanti, M.A.Q.: 482, 501, 502, 

503 

Cavaletto, J.R.: 669 

Cerminara, S.: 330 

Ceska, A.: 314 

Ceska, O.: 314 

Chabane, S.: 29 

Chabanenko, S.: 705 

Chague, V.: 88 

Chalot, M.: 329 

Chamswarng, C.: 477, 524 

Chan, W.K.: 496, 587, 1201 

Chandler, D.: 908 

Chang, D.C.N.: 944 

Chapman, B.: 467 

Charvat, I.: 931 

Chatagnier, O.: 441, 1107 

Chatamala, I.: 56 

Chaudhuri, S.: 945 

Chavarriaga, D.: 815 

Chen, A.W.: 306 

Chen, C.-J.: 13 

Chen, C.Y.: 686 

Chen, J.B.: 642 

Chen, K.Y: 483 

Chen, S.-F.: 484 

Chen, S.-R.: 717 

Chet, I.: 172, 174 

Chevalier, G.: 1008 

Chhetri, D.B.: 296, 646 

Chien, C.-Y.: 643 

Chirkova, J.: 1122 

Cho, D.H.: 485, 486 

Cho, W.K.: 486 

Choi, Y.J.: 644 

Choi, Y.K.: 731 

Choinowski, T.: 199 

Chong-Lopez, A.: 905 

Chou, L.C.: 944 

Christensen, M.: 67, 508, 519 

Christensen, M.J.: 1204 

Chrpova, J.: 901 

Chulze, S.N.: 878, 879 

Chung, J.Y.: 486 

Chung, W.H.: 645 

Ciardo, D.: 75 

Cifuentes, J.: 99, 480, 487, 491, 666, 

675, 748, 762, 781 

Ciliento, R.: 115, 171, 372 

Claassen, N.: 560 

Clapp, J.P.: 903, 1022 

Clark, A.G.: 418 

Clasen, P.-E.: 811, 909 

Clavería, V.: 946, 953, 954 

Cleef, A.M.: 470 

Clergeot, P.-H.: 75 

Clipson, N.: 986 

Codjia, J.T.C.: 490 

Coetzee, M.P.A.: 355, 646 

Cole, L.: 440 

Collado, J.: 647 

Collin-Hansen, C.: 1084 

Collins, M.D.: 961 

Colpaert, J.V.: 331, 1007 

Colquhoun, I.: 338 

Comandini, O.: 648, 741 

Conomikes, M.: 947 

Consiglio, G.: 627 

Cooke, D.E.L.: 207, 217, 948 

Cooper, J.A.: 243 

Corazza, L.: 880 

Cordobes, S.: 1193 

Cornejo, C.: 705 

Cortés, C.: 1158 

Costa, I.P.M.W.: 482 

Courbot, M.: 329 


Courtecuisse, R.: 98, 99, 105, 235 

Coutinho, T.A.: 430, 692, 765 

Cowen, L.E.: 369 

Crane, J.: 220 

Craven, K.D.: 203, 325 

Crawford, J.W: 1057 

Crewe, A.T.: 649 

Cripps, C.L.: 50, 52, 549 

Crittenden, P.D.: 123, 318 

Croneborg, H.: 488 

Crous, P.W.: 100, 276, 753, 765 

Cuero, R.: 1085 

Cullen, D.: 1196 

Curir, P.: 870 

Currah, R.S.: 26, 81, 176, 412, 668, 

671, 677, 1033 

Czederpiltz, D.L.L.: 631 

Czymmek, K.J.: 269 

D'Aquila, F.: 870 

Daboussi, M.J.: 159 

Dahlberg, A.: 24, 453, 488, 593 

Dahlkild, Å.: 285 

Dahlman, L.: 1086 

Dahms, T.: 1127 

Dai, Y.C.: 489, 650 

Daikoku, C.: 990 

Dalili, A.R.: 362 

Dammann-Tugend, U.: 182 

Daniëls, P.P.: 651 

Darrah, P.R.: 437 

Davidson, F.A.: 1079, 1123 

Davidson, J.M.: 652 

Davies, D.: 440 

Davies, D.R.: 118, 550, 1019 

Davies, N.W.: 805, 806, 818, 819 

Davies, P.W.: 949 

Davis, C.: 653 

Davis, D.J.: 1087 

Davydov, E.A.: 950 

de Cock, A.W.A.M.: 654 

de Gioia, T.: 951 

de Hoog, G.S.: 120 

De Kesel, A.: 490, 614, 779, 952 

de Koe, W.J.: 74 

de los Rios, A.: 124 

de Luna, E.: 666 

de Miguel, A.M.: 946, 953, 954 

de Román, M.: 946, 953, 954 

de Vera, J.-P.: 1088 

Deacon, L.J.: 138 

Dean, N.: 34 

Dearnaley, J.D.W.: 1089 

Debets, A.J.M.: 937 

Debuchy, R.: 194 

Decker, M.: 1090 

Degawa, Y.: 655 

Deising, H.B.: 77 

Del Frate, G.: 512, 1047 

Del Olmo, M.: 491 

del Río, J.C.: 1181 

Del Sorbo, G.: 1168 

Delaforge, M.: 874 

Delarea, J.: 1105 

Delaruelle, C.: 1008 

Delavy, D.: 1098 

Dell, B.: 615 

Deml, G.: 514 

den Bakker, H.C: 656 

Dennis, J.: 467 

DePriest, P.T.: 682, 824, 1062 

Descals, E.: 102 

Desjardin, D.E.: 99, 474, 679, 785 

Dethoup, T.: 542 

Deveci, T.: 1073 

Dhillion, S.S.: 237, 494, 929, 942 

Di Mario, F.: 562, 955, 1021 

di Menna, M.: 220 

Diaquin, M.: 31 

Díaz, G.: 498 

Diaz, M.: 257, 531 

Díaz-Godínez, G.: 1091, 1170 

Dickson, S.: 1092, 1130 

Didukh, M.: 915 

Didukh, M.Ya.: 657 

Dietrich, F.: 335 

Diez, J.: 956, 1008 

Dikshit, A.: 309, 871 

Dilip, A.: 891 

Dimou, D.M.: 557, 605 

Dobrowolski, M.P.: 338 

Dobson, A.: 868 

Dodd, J.C.: 1022 

Doi, S.: 918, 973 

Dombrink-Kurtzman, M.: 422 

Donatini, B.: 305 

Donnelly, D.P.: 431, 957 

Dornieden, Th.: 1108 

Dorph-Petersen, D.: 816 

Dott, W.: 662, 861, 907 

Douanla-Meli, C.: 492 

Douhan, G.W.: 958 

Dreisbach, T.A.: 232, 390, 586 

Dresler-Nurmi, A.: 1142 

Drint-Kuyvenhoven, A.: 1117 

Druzhinina, I.: 634, 1093 

Duckworth, J.C.: 234 

Dudzinski, M.J.: 464 

Due, M.J.: 1094 

Dueñas, M.: 691 

Dujon, B.: 334 

Duletic-Lausevic, S.: 363 

Duñabeitia, M.K.: 1023 

Duncan, J.M.: 207, 948 

Duncan, K.E.: 269 

Duplessis, S.: 125, 1135 

Dupré, C.: 1008 

Duran, A.: 30 

Dvorska, J.: 817 

Dyakov, Y.T.: 849 

Dyer, P.S.: 1095 

Easton, G.L.: 143 

Eberhardt, U.: 658 

Eberhart, J.L.: 141 

Egerton-Warburton, L.: 97, 1000 

Egger, K.N.: 964 

Eggleton, P.: 619 

Egli, S.: 133, 391, 493 

Ehlers, T.: 467 

Ehrig, F.: 1030 

Ehrman, J.: 994 

Eikenes, M.: 831 

Eilenberg, J.: 186, 1049 

Ekinci, M.S.: 550 

Ekman, S.: 356, 659 


El-Ghandour, I.: 1096 

Elad, Y.: 86, 991 

Elansky, S.N.: 892 

Elborne, S.A.: 47 

Elisashvili, V.I.: 202, 915 

Elix, J.A.: 695 

Ellis, C.J.: 318 

Eltem, R.: 803 

Elvebakk, A.: 1056 

Emalfarb, M.: 1195 

Erdogan, M.: 1073 

Eriksen, M.: 494 

Eriksson, O.E.: 405, 416 

Erland, S.: 315 

Ershad, -: 794 

Eskridge, K.M.: 1041 

Estrada-Torres, A.: 666 

Etayo, M.L.: 953 

Evans, S.E.: 234 

Eversmeyer, M.G.: 959 

Exeter, R.L.: 547 

Eyjolfsdottir, G.G.: 495 

Eyles, A.: 21, 818, 819 

Fabbri, A.A.: 308, 880 

Fackler, K.: 1136 

Falandysz, J.: 1075, 1080 

Falconer, R.E: 1057 

Falk, R.: 257 

Fanelli, C.: 308, 868, 880 

Farkas, E.: 660 

Farmer, M.J.: 496 

Farr, D.F.: 2, 242, 278 

Farron, G.: 1098 

Fatehi, J.: 859, 913, 960 

Fazli, A.: 1176 

Fedorenko, N.M.: 661 

Fehrmann, H.: 820 

Fekete, C.: 1066 

Fell, J.W.: 102, 256, 257, 471, 531 

Feng, G.: 496 

Fenton, J.G.: 1041 

Ferdman, Y.: 702 

Fernandes, M.J.S.: 482 

Fernandez, F.A.: 382 

Ferreira, P.: 199 

Ferrer, A.: 140, 497 

Feuerer, T.: 103 

Figliuolo, G.: 951 

Filipello Marchisio, V.: 872, 873 

Finlay, R.D.: 1005 

Fiorina, A.: 575 

Fischer, G.: 334 

Fischer, G.: 662, 861, 907 

Flåøyen, A.: 73, 808 

Fleet, C.: 1099 

Flegel, T.: 473, 474 

Fleming, L.V.: 234 

Flier, W.G.: 711 

Florack, D.E.A.: 166 

Flores, R.E.: 139, 498 

Florio, S.: 821 

Foley, A.J.: 961 

Fomina, M.: 328, 1100, 1101 

Føns, K.: 962 

Fonseca, A.: 254, 689, 757, 980 

Fontaine, T.: 29 

Foote, B.A.: 940

Forlani, G.: 565 

Forsby, A.: 842, 894 

Fossdal, C.G.: 822, 831 

Foures, A.: 874 

Fowler, T.J.: 193 

Franceschi, V.R.: 20 

Franco Molano, A.E.: 377, 470 

Franken, P.: 129 

Frankland, J.C.: 138 

Fraser, M.: 963 

Fredrickson, S.: 467 

Freimoser, F.M.: 185, 376, 1102 

Frey, B.: 263 

Frey, P.: 208 

Fricker, M.D.: 437 

Frieders, E.M.: 727 

Friman, E.: 1066 

Frisvad, J.C.: 67, 119, 719, 746, 868, 

1094 

Frödén, P.: 628 

Frøslev, T. G.: 663 

Fu, H.-M.: 717 

Fuchs, B.: 84 

Fujii, H.: 601, 1114 

Fujimura, K.E.: 180, 964 

Fujiyoshi, M.: 987 

Fukiharu, T.: 39 

Fukumasa-Nakai, Y.: 1145, 1190 

Fukushi, Y.: 839, 917 

Fungaro, M.H.P.: 640 

Gaal, Y.: 1096 

Gabriel, J.: 1070 

Gadanho, M.: 9, 664, 716 

Gadd, G.M.: 328, 1079, 1083, 1100, 

1101, 1115, 1123 

Gaffney, T.: 335 

Gafur, A.: 823, 1103 

Gaitan, A.: 722 

Galán, R.: 413, 647 

Galli, E.: 955 

Gallmetzer, M.: 1104 

Galtier, P.: 874 

Gams, W.: 101, 248, 349 

Gamundi, I.: 37 

Gannibal, Ph.B.: 499, 754 

Gao, X.D.: 34 

Garbelotto, M.M.: 205, 652 

García, D.: 665 

García-Pantaleón, F.: 480 

García-Sandoval, R.: 487, 666, 748 

Gargas, A.: 714, 735, 776, 824 

Garnica, S.: 403 

Gartland, J.S.: 217 

Garty, J.: 1105 

Gasparyan, A.V.: 304 

Gates, K.: 335 

Gauslaa, Y.: 262, 1180 

Gavric, O.: 1109 

Gavrilova, V.P.: 1106, 1131 

Gaya, E.: 667 

Gebhardt, S.: 500, 1060 

Geiser, D.M.: 160 

Gené, J.: 638 

Genre, A.: 443 

Gerhardson, B.: 859, 913, 960 

Gezahgne, A.: 825 

Ghignone, S.: 181 

Gianinazzi, S.: 129, 441, 496, 1107 

Gianinazzi-Pearson, V.: 129, 441, 

496, 1107 

Giasson, L.: 1144 

Gibas, C. F.: 668 

Gibertoni, T.B.: 501, 502, 503 

Gignac, M.: 919, 920 

Gijswit, G.: 471 

Gilbert, G.S.: 280 

Gill, W.: 826, 899 

Gioacchini, A.M.: 1148 

Giovannetti, M.: 442 

Girlanda, M.: 181, 226 

Girrbach, V.: 33 

Givnish, T.J.: 714 

Glass, N.L.: 196, 227 

Glew, K.A.: 504 

Gliech, M.: 271, 513 

Glina, I.: 1183 

Glockling, S.L.: 78, 251 

Goddard, M.R.: 367 

Goeker, M.: 274 

Gojkovic, Z.: 371 

Goktepe, G.: 302 

Goldberg, D.: 724 

Golldack, J.: 965 

Gollotte, A.: 441, 446, 1107 

Gómez, I.: 1158 

Gonçalves, P.: 567 

Gonthier, P.: 205 

González, D.: 748 

González, G.: 853 

Gonzalez, M.C.: 505 

González, V.: 647 

González-Gómez, R.: 942 

González-Jaén, M.T.: 732, 858 

Goodwin, S.B.: 620, 669 

Gorbushina, A.: 323, 1108 

Gorfer, M.: 1113 

Görke, C.: 134, 135, 468 

Gorovits, R.: 35 

Gorton, C.: 297 

Goto, H.: 1063 

Götz, M.: 182, 966 

Gough, K.: 1127 

Gouka, R.J.: 1139 

Gow, N.A.R.: 375, 386, 1152 

Goward, T.: 316 

Gräfenhan, T.: 271, 513 

Graham, S.W.: 26 

Grand, E.A.: 670 

Gravendeel, B.: 656 

Gravesen, S.: 218 

Gray, S.: 1140 

Grebenc, T.: 508 

Green, T.G.A.: 259 

Gregersen, R.: 521 

Greif, G.: 1118 

Greif, M.D.: 671 

Greslebin, A.G.: 41, 509, 967 

Grgurinovic, C.A.: 582 

Griffith, G.W.: 43, 118, 143, 145, 

322, 550, 1019, 1058, 1155 

Griffiths, A.: 1109 

Griffiths, H.: 1069 

Griffiths, R.: 322, 1058 

Griffiths, R.C.: 419 

Grishkan, I.: 968 

Grodzinskaya, A.A.: 307 

Grogan, P.: 223 

Grönberg, H.: 827 

Grossman, R.B.: 1077 

Grube, M.: 290, 320 

Gryta, H.: 27 

Gryzenhout, M.V.: 518, 672, 673 

Grzywnowicz, K.: 1110, 1111, 1183 

Guarro, J.: 638, 665 

Gucin, F.: 510 

Guevara, R.: 511 

Guglielminetti, M.: 512 

Guidot, A.: 24 

Guillen, F.: 199 

Guinko, S.: 828 

Guissou, K.M.L: 828 

Guldberg-Froslev, T.: 619 

Gulden, G.: 48 

Gulseren, I.: 1074 

Gulyamova, T.G.: 1128 

Gunawan, A.G.: 580 

Gunde-Cimerman, N.: 120, 303 

Gunn, L.V.: 1043 

Guo, L.-D.: 136 

Guo, L.: 674 

Guo, S.X.: 972 

Gure, A.: 829 

Guske, S.: 830 

Gutíerrez, A.: 1181 

Guyette, R.P.: 938 

Guzmán-Dávalos, L.: 675 

Ha, H.C.: 361 

Hagedorn, G.: 240, 247, 271, 513, 

514 

Haggblom, P.: 842, 894 

Hagn, A.: 969 

Haikara, A.: 868 

Hajslová, J.: 837 

Hall, B.D.: 400 

Hall, M.: 21 

Hallenberg, N.: 351, 736, 1045 

Halling, R.E.: 99, 354 

Halmschlager, E.: 463, 710 

Hamada, N.: 1112 

Hamari, Zs.: 529 

Hambleton, S.: 676, 677, 678 

Hammel, K.E.: 200 

Hampp, R.: 128, 1143 

Hanif, M.: 1113 

Hanlin, R.T.: 505 

Hansen, E.M.: 19, 652 

Hansen, K.: 295, 414 

Hansen, K.T.: 398 

Hansen, M.E.: 515 

Hansen, P.W.: 515, 516 

Hanson, Jr., R.W.: 727 

Hantula, J.: 970 

Hara, K.: 918, 1114 

Harding, S.: 1049 

Hardy, G.E.St.J.: 19, 338 

Hardy, G.P.M.A.: 341 

Harkness, J.: 690 

Harper, F.A.: 328, 1115 

Harrex, A.L.: 924 

Harrier, L.A.: 446, 1189 

Harrington, T.C.: 195, 300, 353 


Harrison, K.: 21 

Harrison, K.J.: 298 

Hasbay Ifrij, I.: 1116 

Haselwandter, K.: 84, 307, 777 

Hashidoko, Y.: 917 

Hashimoto, T.: 169, 170 

Hatakka, A.: 95, 1142, 1184 

Hattori, H.: 1114 

Hattori, T.: 517, 745 

Haug, I.: 648 

Haugen, P.: 755 

Hawksworth, D.L.: 350 

Hayashi, E.: 1190 

Hazard, C.: 975 

Heath, R.N.: 518 

Hedayati, M.T.: 362 

Hedger, J.N.: 43 

Hedjaroude, Gh.A.: 620 

Hedlund, K.: 532 

Heerikhuisen, M.: 1117, 1195 

Heilmann-Clausen, J.: 433, 519, 971 

Heiskanen, J.: 1020 

Hellwig, V.: 249, 1118 

Hemmes, D.E.: 679 

Hendrickx, P.M.: 165 

Henk, D.A.: 680 

Henkel, T.W.: 379 

Hennebert, G.L.: 346 

Hernandez, J.R.: 681 

Herre, E.A.: 224, 321 

Herrera-Estrella, A.: 174, 1158 

Herrero, M.L.: 601 

Heyser, W.: 1082, 1155 

Hibbett, D.S.: 110, 209 

Hickey, P.C.: 264 

Hietala, A.: 831 

Hijri, I.: 520 

Hilal, A.A.: 905 

Hildebrandt, U.: 332 

Hillier, S.: 1083 

Himelbrant, D.E.: 616 

Hinojo, M.J.: 855 

Hiranpradit, S.: 767 

Hiremath, S.T.: 1160 

Hjortstam, K.: 99 

Hladun, N.: 667 

Ho, I.C.: 602 

Hobbie, E.A.: 93 

Hodge, K.T.: 5, 718 

Hoekstra, R.F.: 937 

Hoffland, E.: 94 

Hoffmann, M.: 1012 

Hoffmann, N.: 682 

Hofrichter, M.: 95, 1184 

Hofstetter, R.W.: 426, 427 

Holdenrieder, O.: 1012 

Holec, J.: 683 

Holland, B.: 924 

Holm, A.C.: 521 

Holma, A.: 985 

Holmer, L.: 432 

Holmes, S.: 868 

Holst-Jensen, A.: 161, 398, 684, 704, 

707, 832, 884 

Homolka, L.: 773 

Honda, Y.: 1119 

Honegger, R.: 260 

Hong, I.P.: 972 

Hong, S.B: 644 

Honrubia, M.: 139, 498 

Hope, R.: 833 

Horak, E.: 46, 52 

Horikoshi, T.: 987 

Horisawa, S.: 973 

Horiuchi, H.: 1121, 1187 

Horn, B.W.: 974 

Hornok, L.: 1151 

Horton, T.R.: 180, 223, 975 

Horwitz, B.: 174 

Hoshino, T.: 601, 1120 

Hou, C.-L.: 685 

Houbraken, J.A.M.P.: 719 

Howard, R.J.: 269 

Howden-Chapman, P.: 220 

Hsiao-Man, H.: 522 

Hsieh, H.-M.: 157 

Hsieh, W.H.: 686 

Hsu, H.S.: 602 

Hu, G.: 185, 376, 834, 1102 

Huang, D.J: 483 

Huang, K.S.: 835 

Hughes, H.B.: 1152 

Hughes, K.W.: 146, 352, 670, 726 

Hughes, M.: 104 

Huhndorf, S.M.: 1, 382 

Humbel, B.M.: 438 

Hunt, A.F.: 123 

Husband, R.: 224 

Hussain, S.: 948 

Hüttl, R.F.: 500, 1030, 1036, 1060 

Huynh, T.T.: 976 

Hyde, G.: 440 

Hyde, K.D.: 100 

Hymes, M.J.: 1161 

H_sek, J.: 812, 836, 837 

Hysek, J.: 901 

Hywel-Jones, N.L.: 79, 121, 183, 

473, 474, 475, 585, 589, 591, 599, 

713, 770, 1061, 1129 

Höftberger, M.: 1165 

Högberg, N.: 197 

Högnabba, F.: 286 

Høiland, K.: 398, 926, 943, 1039 

Iakovlev, A.: 432 

Ibrahim, L.: 977 

Ichinomiya, M.: 1121 

Ikeda, K.: 978, 1010 

Ikekawa, T.: 164 

Imamura, A.: 979 

Imamura, C.: 1119 

Imamura, K.: 1153 

Imbabi, I.: 1014 

Inaba, S.: 687 

Inacio, C.A.: 688 

Inácio, J.: 689, 980 

Inderbitzin, P.: 690 

Ineichen, K.: 520 

Iniesta, E.P.: 691 

Ipulet, P.: 981, 982 

Iranpour, M.: 629 

Irbe, I.: 1122 

Irianto, R.S.B.: 806 

Ishimoto, H.: 839 

Isikhuemhen, O.S.: 302 


Ito, R.: 973 

Iturriaga, T.: 60, 99, 378, 479 

Iwamoto, S.: 596, 983 

Iwasaki, Y.: 1068 

Jackson, J.A.: 548 

Jackson, S.: 729 

Jacob, C.: 329 

Jacobs, A.: 692 

Jacobs, H.: 328, 1079, 1123 

Jacobs, K.: 298, 693, 694, 706 

Jacobson, D.J.: 196 

Jacobsson, S.: 144 

Jacques-Hernandez, C.: 848 

Jakobsen, I.: 92, 1130 

Jalink, L.M.: 523 

James, T.Y.: 28, 357 

Jansson, H.-B.: 250 

Jansson, J.K.: 115 

Jargeat, P.: 443 

Jariangprasert, S.: 695 

Jaroenthai, K.: 524 

Jeamjitt, O.: 525 

Jeffries, P.: 881, 903 

Jejelowo, O.A.: 802, 840 

Jenkinson, P.: 977 

Jensen, A.B.: 186 

Jensen, B.: 189 

Jeong, W.: 696 

Jernejc, K.: 1124, 1125 

Jervis, M.: 908 

Jestoi, M.: 880 

Jetten, M.S.M.: 1200 

Ji, L.: 841 

Jiang, Z.D.: 841 

Jimenez, C.: 30 

Jiménez, M.: 853, 855 

Jiménez-González, C.: 942 

Job, D.: 536, 1098 

Johannesson, H.: 24 

Johansson, J.F.: 984 

Johansson, S.: 568, 697 

Johansson, T.: 131, 337, 1066, 1137 

Johnson, J.E.: 236, 548 

Johnson, T.W.: 698 

Johnston, P.R.: 37 

Jokela, S.: 1126 

Jokinen, H.: 985 

Jones, A.W.: 143 

Jones, E.B.G.: 57, 585, 592, 713, 

747, 1053 

Jones, R.H.: 699 

Jonglaekha, N.: 388 

Jonsson, A.: 868 

Jonsson, E.: 842, 894 

Jonsson, N.: 868 

Joosten, V.: 1139 

Jordaan, A.: 600 

Jordal, J.B.: 142 

Jovel, E.: 1109 

Ju, Y.-M.: 157 

Juhász, Á.: 529 

Jukucs, E.: 700 

Juliano, S.A.: 929 

Jumpponen, A.: 177 

Jung, H.: 696, 1074, 1188 

Jüriado, I.: 526 

Jurkiewicz, A.: 1159

Juuti, J.T.: 701, 1126 

Kabrick, J.M.: 938 

Kachlishvili, E.: 202 

Kagan Zur, V.: 702, 1048 

Kainz, C.: 273 

Kaiser, W.J.: 749 

Kakishima, M.: 645, 983 

Kakuta, H.: 928 

Kaldenhoff, R.: 1143 

Källersjö, M.: 285 

Kalyanasundaram, I.: 906 

Kaminskyj, S.: 1127 

Kanda, H.: 601 

Kaneko, S.: 725, 1063 

Kanematsu, S.: 195 

Kano, S.: 1068 

Karadzic, B.: 1004 

Karaffa, L.: 343 

Karg, V.: 1093 

Karlshøj, K.: 1094 

Karlsson, M.: 18, 843 

Kärnefelt, I.: 703 

Kashiwagi, Y.: 847, 1186 

Kasza, Zs.: 1194 

Katan, T.: 991 

Kauff, F.: 287 

Kauhanen, M.: 970 

Kauserud, H.: 23, 704 

Kautman, J.: 528 

Kautmanova, I.: 527, 528 

Kawagishi, N.: 844 

Kawakami, S.: 169, 170 

Keblys, M.: 808 

Keller, C.: 705 

Keller, J.: 493 

Keller, N.P.: 312 

Kennedy, N.: 986 

Kerbalaeva, A.M.: 1128 

Kerényi, Z.: 1151 

Kersten, P.J.: 1196 

Kevei, F.: 529 

Kezman, M.: 363 

Khaksar, A.A.: 807 

Khardziani, T.: 202 

Kharwar, R.N.: 763 

Kidd, S.: 729 

Kiesling, T.: 531 

Kim, B.R: 793 

Kim, H.J.: 361 

Kim, J.-J.: 778 

Kim, S.Y.: 361 

Kinugasa, S.: 987 

Kiriaki, M.: 1120 

Kirisits, T.: 296, 298, 301, 463, 646, 

693, 694, 706, 709, 710, 1003 

Kirk, P.M.: 241, 345, 350 

Kirshner, B.: 86 

Kiss, L.: 434, 845, 903 

Kjøller, R.: 962, 988, 989 

Kladwang, W.: 1129 

Klamer, M.: 532 

Klassen, G.R.: 654 

Kleber, R.: 128 

Klemsdal, S.S.: 601, 790, 884 

Kleppe, K.: 1174 

Klepzig, K.D.: 426, 427 

Kleymann, G.: 1118 

Knecht, W.: 371 

Knight, K.B.: 768 

Knudsen, H.: 47 

Knudsen, L.M.: 1130 

Knutsen, A.K.: 161, 707, 832 

Kobayashi, H.: 708 

Koepke, L.: 504 

Kohler, A.: 125 

Kohlmeyer, J.: 102 

Kohn, L.M.: 369 

Kokaew, J.: 542 

Kolarík, M.: 533 

Kolesnik, I.: 86 

Kõljalg, U.: 96, 212, 214, 215, 737, 

1045 

Komine, M.: 918, 1114 

Kondo, R.: 721, 990 

Kondratyuk, S.Ya.: 661, 703, 705 

Kong, R.Y.C.: 713, 747 

Konrad, H.: 706, 709, 710 

Konstantinova, P.: 790 

Korf, R.P.: 58 

Korhonen, K.: 650 

Korolev, N.: 86, 991 

Koroleva, O.V.: 1106, 1131 

Korona, R.: 368 

Kosiak, B.: 161, 712 

Koster, B.: 438 

Kotter, S.: 1036 

Koukol, O.: 992 

Kovács, G.: 700 

Kovalenko, A.E.: 146, 239 

Kozlova-Zwinderman, O.: 266, 1072 

Kraigher, H.: 508 

Kramadibrata, K.: 580 

Kramer, C.L.: 959 

Kranabetter, M.: 467 

Krastina, I.: 993 

Krauss, G.-J.: 1132 

Krauss, G.: 994, 1132 

Krettek, R.: 995 

Krisai-Greilhuber, I.: 133, 777 

Kristensen, R.: 161 

Kroken, S.: 227 

Kroon, L.P.N.M.: 711 

Kruse, P.: 909 

Kubátová, A.: 996 

Kubicek, C.P.: 175, 343, 634, 790, 

884 

Kubota, R.: 1080 

Kudratov, I.: 535 

Kuehn, K.A.: 961 

Kües, U.: 75, 1012 

Küffer, N.: 536 

Kuijpers, A.F.A: 719 

Kullman, B.: 1133 

Kullnig-Gradinger, C.M.: 634, 790, 

884 

Kumar, J.V.: 302 

Kumar, R.: 1134 

Kunin, P.: 1105 

Kunito, T.: 1080 

Künzler, M.: 75 

Kure, C.F.: 846 

Kurtzman, C.P.: 253, 409, 471 

Kusumoto, K.: 847 

Kutil, B.L.: 311 

Kuwahara, M.: 1119 

Kuyper, T.W.: 94, 222, 365, 402, 

569, 648, 656 

Kuznetsov, S.A.: 892 

Kvaalen, H.: 831 

Kwan, Y.C.: 922 

Kwasna, H.: 712 

Kwon-Chung, K.J.: 257 

Kwong, T.F.N.: 713 

Kyllo, D.A.: 321 

LaCamera, S.: 129 

Lachance, M.A.: 255 

Lacourt, I.: 1135 

Lafontaine, I.: 334 

Laganà, A.: 456, 997 

Lamaipis, P.: 1136 

Lammers, P.: 127 

Landesman, E.O.: 1106 

Landeweert, R.: 94, 222, 569 

Landis, F.C.: 714 

Landvik, S.: 191, 515 

Lanfranco, L.: 126 

Lange, L.: 122, 406, 1129 

Lange, O.L.: 259 

Lange, T.: 182, 1024 

Langer, E.: 211, 492, 995 

Langkjaer, R.B.: 371 

Lanter, K.: 1087 

Lanzuise, S.: 171, 372, 1168 

Lapeyrie, F.: 1162 

Lapsker, Z.: 86 

Larralde-Corona, C.P.: 848 

Larsen, H.J.S.: 808 

Larsen, R.S.: 537 

Larsson, E.: 144, 212, 213, 215, 737 

Larsson, K.-H.: 99, 212, 213, 394, 

737 

Larsson, K.-J.: 569 

Læssøe, T.: 153, 244, 415, 663 

Latgé, J.P.: 29, 31 

Latouche, G.N.: 729 

Laubach, H.: 963, 1052 

Lavín, P.: 152 

Lavrova, O.I.: 849 

Lawrie, A.C.: 941, 947, 976 

Lazera, M.: 257 

Le Blaye, I.: 305 

Le Floch, G.: 90, 850 

Le Quéré, A.: 131, 1137 

Leacock, P.: 99 

Leake, J.R.: 957 

Leal Lara, H.: 814, 877, 912 

Learmonth, B.A.: 1089 

Lebedeva, E.: 1016, 1017 

Lebel, T.: 40, 774 

Lebrón, L.: 998 

Lee, C.-Z.: 923 

Lee, F.-L.: 717 

Lee, G.C.: 944 

Lee, H.S.: 922 

Lee, J.: 696 

Lee, J.S.: 361, 922 

Lee, M.W.: 972 

Lee, N.J.: 922 

Lee, S.: 100 

Lee, T.S.: 972 

Lees, A.K.: 948 


Legisa, M.: 1072, 1125 

Leifert, C.: 809 

Lentzsch, P.: 965, 1030, 1044 

Leonowicz, A.: 1146, 1183 

Lerch, A.: 335 

Leroy, P.: 329 

Lesage, G.: 32 

Leski, T.: 999, 1029 

Leslie, J.F.: 797 

Leuchtmann, A.: 715 

Lévesque, C.A.: 654 

Li, H.: 32 

Li, K.: 1074, 1188 

Li, X.: 496 

Li, Y.: 702 

Libkind, D.: 716 

Lichtwardt, R.: 147 

Lihra, T.: 919 

Likhitekaraj, S.: 851 

Lilleskov, E.A.: 223 

Lim, Y.: 696 

Lima, D.M.M.: 482 

Lin, Y.S.: 835 

Lind, M.: 18, 197, 1138 

Lindahl, A.: 1000 

Lindahl, B.D.: 317, 436, 569 

Lindberg, A.: 960 

Lindblad, I.: 538 

Linde, C.C.: 370 

Lingua, G.: 873 

Linovitska, V.M.: 637 

Lins, C.E.L.: 539 

Liou, G.-Y.: 717, 923 

Liu, C.C: 483 

Liu, M.: 718 

Liu, Y. J.: 400 

Liu, Yi: 75 

Lizon, P.: 564, 621 

Llorens, A.: 853 

Lobanov, A.L.: 239 

LoBuglio, K.F.: 246, 295 

Lodge, D.J.: 99, 145, 146, 998 

Logrieco, A.: 858 

Lokman, B.C.: 1139 

Lommel, M.: 33 

Loos, S.: 75 

Lopez Quintero, C.A.: 377, 470 

Lopez-Llorca, L.V.: 250 

Lorenzi, E.: 565 

Lorito, M.: 115, 171, 372, 1168 

Lotfi, A.: 1176 

Lottmann, J.: 966 

Lu, P.L.: 540 

Lu, Z.X.: 115 

Luangsa-ard, J.J.: 121 

Lücking, R.: 284 

Luecking, R.: 320 

Luis, P.: 91, 1008 

Lumbsch, H.T.: 288, 411, 1059 

Lumyong, P.: 615 

Lumyong, S.: 100, 615 

Lund, F.: 719 

Lunder, C.: 221 

Luoma, D.L.: 141 

Luppi, A.M.: 181 

Luszczyñski, J.: 541 

Lutz, M.: 720 

Lutzoni, F.: 107, 287, 326, 667, 730, 

755 

Lygis, V.: 1001 

Lökös, L.: 660 

Løvseth, A.: 832 

Mach, R.L.: 175, 790, 884 

Machado, C.: 883 

Maddani, M.: 887 

Mäder, P.: 520 

Maeda, T.: 928 

Maekawa, N.: 721, 736 

Magan, N.: 813, 833, 868, 878, 879, 

927, 1140, 1150 

Magyar, D.: 845 

Mahajna, J.: 921 

Mahmood, S.: 315 

Mahoney, D.P.: 38 

Maia, L.C.: 539, 1141, 1179 

Maier, W.: 10 

Maijala, P.: 1142 

Makselan, S.D.: 1087 

Man in't Veld, W.A.: 207 

Manjon, J.L.: 956 

Mannanov, R.N.: 852 

Mannini, M.: 457 

Manoch, L.: 121, 388, 460, 477, 524, 

525, 542 

Mariano, R.L.R.: 1179 

Marin, M.: 301, 722, 1003 

Marino, R.: 951 

Marjanovic, Z.: 1004, 1143 

Markov, G.: 1016 

Marquéz-Guzmán, J.: 762 

Marshall, C.: 220 

Martellos, S.: 512 

Martin, F.: 91, 125, 1008, 1135 

Martín, M.P.: 651, 723, 752 

Martineau, P.: 1144 

Martínez, A.T.: 199, 590, 1181 

Martínez, M.J.: 199, 590, 1181 

Martino, E.: 330 

Marvanová, L.: 102, 639 

Masaphy, S.: 724 

Massicotte, H.B.: 543 

Masuya, H.: 725, 1063 

Mata, J.L.: 726 

Mateo, R.: 853, 855 

Matheny, P. B.: 400 

Matsuda, Y.: 854, 864, 910 

Matsumoto, N.: 978, 1010 

Matsumoto, T.: 1145, 1190 

Mattey, M.: 342 

Mattsson, J.: 216, 221 

Matuszewska, A.: 1146, 1156 

Maximilian, R.C.: 1147 

May, G.: 192 

May, T.W.: 36, 44, 99, 233, 699, 774 

Mayer-Bartschmid, A.: 1118 

McCreadie, J.W.: 150 

McDonald, B.A.: 370, 632 

McDonald, J.: 70 

McDougall, K.D.: 900 

McKenzie, E.H.C.: 42, 100 

McLaughlin, D.J.: 727, 931 

McLean, C.B.: 179, 774, 941, 947, 

949, 976 

McMillan, L.: 1204 


McNew, D.L.: 195 

McPherson, M.A.: 26 

Medina, A.: 853, 855 

Mejía, L.C.: 321 

Melville, L.H.: 543 

Menard, P.: 32 

Mendoza-Mendoza, A.: 174, 1158 

Menkis, A.: 1005 

Mennicken, M.: 468, 728 

Menotta, M.: 1148 

Mensonides, F.I.C.: 341 

Mesjasz-Przybylowicz, J.: 1159 

Messias, A.S.: 539 

Messner, K.: 201, 1136 

Methvin, B.R.: 1042 

Meyer, W.: 257, 729 

Miadlikowska, J.: 730 

Micales, J.A.: 631 

Mihail, J.D.: 938, 1006 

Mihlan, M.: 85 

Mikhailova, I.: 705 

Miller Jr., O.K.: 51, 52 

Miller, A.N.: 1, 675 

Miller, C.L.: 504 

Miller, J.D.: 313 

Mills, D.: 1048 

Min, B.R.: 731 

Minato, K.: 169, 170 

Minibayeva, F.V.: 1071 

Minter, D.W.: 239, 378, 688 

Mirete, S.: 732 

Mishra, J.: 896 

Miskiewicz, A.: 544 

Misra, J.K.: 856 

Mitchell, D.: 467 

Mitchell, J.I.: 1065, 1149 

Mitchell, T.G.: 875 

Mitton, M.F.: 193 

Miyagawa, H.: 1112 

Miyazaki, A.: 1153 

Mizuno, M.: 169, 170 

Moeller, K.: 371 

Mogongwa, N.: 1085 

Mohammed, C.: 21, 612, 818, 819, 

826, 899 

Mohammed, C.L.: 805, 806 

Mokiou, S.: 1150 

Molina, R.: 232, 390, 586 

Momany, M.: 1178 

Moncalvo, J.-M.: 28, 145, 146, 210, 

236, 357, 397, 548 

Money, N.P.: 76, 265, 1087 

Montanini, B.: 132 

Montemartini, A.: 857 

Montijn, R.C.: 438 

Moon, C.D.: 203, 311, 325, 1182 

Moora, M.: 96 

Moore, D.: 387, 1170 

Moosavi, M.: 886 

Morales, O.: 139 

Morales-Torres, E.: 487 

Moran, M.A.: 939 

Morelle, W.: 31 

Moreno, G.: 956 

Moreta, J.I.L.: 939 

Moretti, A.: 858, 1151 

Morinaga, T.: 359, 1068

Morocko, I.: 859 

Moser, J.C.: 426, 427 

Moser, M.: 45, 46, 397, 401 

Moss, S.T.: 151 

Mossebo, D.C.: 733 

Mota, S.: 567 

Mouchacca, J.: 188 

Mouyna, I.: 29 

Mueller, G.M.: 98, 99, 105, 623, 675 

Mueller, M.: 860 

Mujim, S.: 823 

Mukagasana, R.: 72 

Mulè, G.: 858, 1151 

Muller, L.A.H.: 1007 

Müller, T.: 861, 907 

Müller, W.H.: 438 

Mullins, J.T.: 1177 

Munch, J.-C.: 82 

Munkasci, A.: 192 

Munro, C.A.: 1152 

Münzenberger, B.: 965, 1030, 1036 

Murat, C.: 1008 

Murayama, T.: 1153 

Mutch, P.: 446 

Muthumary, J.: 576 

Myburg, H.: 672, 673 

Myhre, E.: 61 

Nagy, Á.: 1154 

Nagy, N.E.: 822 

Nakabonge, G.: 1009, 1027 

Nakagiri, A.: 55 

Nakamura, H.: 978, 1010 

Nakasone, K.K.: 145 

Narayan, S.: 763 

Nash III, T.H.: 734 

Näsholm, T.: 1086 

Nasmetova, S.M.: 1128 

Nauta, M.M.: 523, 738 

Navarro-Gonzalez, M.: 1012 

Navarro-Rosinés, P.: 667 

Nazim, M.: 1013, 1014 

Neda, H.: 546 

Neely, R.K.: 961 

Nehls, U.: 128, 1143 

Neiva, R.: 567 

Nelsen, M.P.: 735 

Nenninger, A.: 43, 1155 

Nerud, F.: 773 

Neuner-Plattner, I.: 777 

Neves, S.P.B.: 566, 567 

Nevo, E.: 968 

Newcombe, G.: 208 

Newell, S.Y.: 939 

Ng, M.-L.: 168 

Nicholson, J.N.: 43 

Nicklin, J.L.: 282 

Nicolaisen, M.: 792 

Nicolotti, G.: 205 

Nielsen, C.: 186, 1049 

Nielsen, J.: 340 

Nielsen, K.: 792 

Nielsen, K.F.: 219, 515 

Nielsen, P.V.: 816, 863, 1090 

Nielsen, R.: 420 

Niemelä, P.: 970 

Niemelä, T.: 650 

Niessen, L.: 884, 885 

Nilsson, L.-O.: 315 

Nilsson, R.H.: 351, 736, 737 

Nirenberg, H.I.: 764, 775, 791, 884, 

966 

Nocentini, G.: 457 

Nolan, C.A.: 72 

Nonomura, T.: 854, 864, 910 

Noordeloos, M.E.: 626, 656, 738 

Nordén, B.: 395, 736 

Norvell, L.L.: 547 

Novotn, D.: 996, 1015 

Novozhilov, Yu.K.: 239 

Nowak, G.: 1146, 1156 

Nowak, M.: 1156 

Nugaeva, N.: 1016, 1017 

Nugent, L.K.: 739, 865 

Nuytinck, J.: 740, 741 

Nyberg, Å.: 742, 1018 

Nyborg, M.S.C.: 1157 

O'Brien, H.E.: 548 

O'Brien, O.M.: 603, 866 

O'Brien, P.A.: 338 

O'Dell, T.: 232, 390 

O'Donnell, K.: 162, 293, 764 

Oberwinkler, F.: 7, 13, 14, 135, 274, 

403, 468, 685, 728, 756 

Oehl, F.: 520 

Oertel, B.: 403 

Oftedal, T.: 684 

Ogawa, Y.: 983 

Ogel, Z.B.: 1116, 1203 

Ohenoja, E.: 49 

Ohki, S.T.: 601 

Ohkohchi, N.: 844 

Ohta, A.: 1121, 1187 

Ohtaka, N.: 1063 

Okada, G.: 743 

Okoth, S.A.: 867 

Old, K.M.: 464 

Oliver, A.-M.B.: 404, 744 

Oliver, S.G.: 252 

Olmedo-Monfil, V.: 174, 1158 

Olsen, M.: 868 

Olson, Å.: 18, 197, 843, 1138 

Olsson, C.: 1142 

Olsson, S.: 435 

Onguene, N.A.: 365 

Oosawa, K.: 928 

Op den Camp, H.J.M.: 1200 

Öpik, M.: 96 

Orlovich, D.A.: 404, 744 

Orlowska, E.: 1159 

Oros-Sichler, M.: 182, 1024 

Osmundson, T.W.: 549 

Osswald, W.: 19 

Ostle, N.: 143 

Ota, Y.: 745 

Otrosina, W.: 205 

Ott, S.: 1088, 1173 

Otto, P.: 283 

Ottonello, S.: 132, 292 

Ouellet, T.: 1085 

Outerbridge, R.: 467 

Ouvanich, W.: 851, 869 

Ouziad, F.: 332 

Overy, D.P.: 746 

Ozkose, E.: 118, 550, 1019 

Paavanen-Huhtala, S.: 791 

Padgett, D.E.: 102, 698, 875 

Padmanaban, A.: 551 

Page, O.M.: 759 

Pagel, M.: 106, 107 

Pajunen, E.: 868 

Paksiri, U.: 845 

Palfreyman, J.W.: 217 

Palm Hernandez, M.E.: 681 

Palm, M.E.: 345 

Palmqvist, K.: 1086 

Panaccione, D.G.: 883 

Pang, K.L.: 747 

Paoletti, M.: 206 

Paolocci, F.: 292 

Papouskova, L.: 901 

Pardo, A.G.: 1113 

Pareek, M.: 439 

Park, H.S.: 486 

Park, K.B.: 361 

Parmasto, E.: 215, 392 

Parrent, J.L.: 548 

Pascale, M.: 423 

Pasini, C.: 870 

Passavante, Z.: 482 

Patel, N.: 187 

Patiño, B.: 732 

Patiño-Conde, V.: 748 

Patra, M.: 309, 871 

Paul, B.: 1162 

Paulin, L.: 827, 1126 

Paulin-Mahady, A.E.: 300 

Paulus, B.: 100 

Pavlik, M.: 552, 804 

Pawlowska, T.E.: 225 

Peever, T.L.: 749 

Peintner, U.: 46, 397 

Pekcan, O.: 1073 

Peláez, F.: 647 

Pennanen, T.: 1020 

Penzina, T.A.: 553 

Percudani, R.: 292 

Perdomo, O.P.: 554 

Perez-Boada, M.: 199 

Pérez-Ramírez, L.: 487 

Perini, C.: 235, 456, 457, 571 

Perotto, S.: 226, 330 

Perry, B.: 294 

Persoh, D.: 289, 555, 734 

Persson, I.-L.: 1018 

Persson, J.: 1086 

Persson, S.-J.: 868 

Pesti, M.: 1154 

Peterbauer, C.K.: 175 

Petersen, J.H.: 244 

Petersen, R.H.: 146, 352, 670, 726 

Peterson, K.R.: 750 

Peterson, M.: 467 

Peterson, R.L.: 83, 178, 543 

Peterson, S.: 68 

Petrini, L.: 156 

Petzold, A.: 1174 

Pfeffer, P.E.: 127, 270 

Pfister, D.H.: 59, 246, 291, 479, 750 

Phan Van, T.: 804 

Philippsen, P.: 335 

Phillips, A.J.L.: 751 


Phosri, C.: 752 

Piacenti, D.: 115, 171, 372 

Picco, A.M.: 565 

Piepenbring, M.: 12, 272, 383, 685 

Piercey, M.M.: 26 

Pimentel, I.C.: 556 

Pintado, A.: 261 

Piontek, K.: 199 

Piskur, J.: 336, 371 

Pitt, J.I.: 65, 117, 230 

Plaingam, N.: 1053 

Platas, G.: 647 

Plemenitas, A.: 120, 303 

Podila, G.K.: 1160, 1161 

Poeggeler, S.: 228 

Pohleven, F.: 1136 

Polacheck, I.: 257 

Polemis, E.: 557, 605 

Polle, A.: 1103 

Pons, N.: 753 

Portales, J.M.: 389 

Porter, D.: 102 

Potacharoen, W.: 558 

Poteri, M.: 902 

Potila, H.M.: 985 

Prakash, A.: 1191, 1192 

Prásil, K.: 996 

Preisig, O.: 301, 722, 1003 

Prigione, V.: 872, 873 

Printzen, C.: 356 

Pritsch, K.: 82, 969 

Przybylowicz, W.: 1159 

Psurtseva, N.V.: 559, 754 

Puel, O.: 874 

Punt, P.J.: 1117, 1195 

Purvis, O.W.: 649 

Pystina, T.: 705 

Querejeta, J.I.: 97 

Quick, M.G.: 875 

Raja, H.: 102 

Rajchenberg, M.: 41, 99 

Rakhimova, E.: 876 

Rakshit, A.: 560 

Ramaswamy, A.: 312 

Rambold, G.: 410, 734 

Ramirez Carrillo, R.: 814, 877 

Ramirez, M.L: 878, 879 

Ramot, O.: 172 

Ramotshodi, G.: 692 

Rana, G.L.: 951 

Randlane, T.: 561, 760 

Rapanà, P.: 562, 955, 1021 

Raper, C.A.: 193 

Rasaee, M.: 886 

Raudaskoski, M.: 1113 

Rawal, R.D.: 895 

Ray, P.: 1162 

Razzaghi abyaneh, M.: 886, 1163, 

1164, 1176 

Read, N.D.: 196, 264, 266, 1072 

Reader, R.J.: 83 

Reblova, M.: 344 

Reddy, C.A.: 198 

Reddy, U.G.: 1162, 1191, 1192 

Redecker, D.: 231, 520 

Redhead, S.A.: 99, 210, 347, 352 

Reeb, V.: 107, 755 

Rees, B.J.: 404 

Rees, E.I.: 193 

Reeser, P.: 652 

Reich, P.: 931 

Reilla, M.: 1152 

Reiter, R.: 1165 

Renker, C.: 82 

Requena, N.: 444 

Rerkasem, B.: 615 

Resendez-Perez, D.: 848 

Reverberi, M.: 308 

Rexer, K.-H.: 14 

Rey, P.: 90, 850 

Reynolds, D.R.: 280, 283 

Rice, A.V.: 81 

Ricelli, A.: 308, 880 

Ridgway, H.J.: 563 

Riethmueller, A.: 274 

Rikkinen, J.: 281, 324 

Rinaldi, A.C.: 648, 741 

Rinaudo, V.: 442 

Rincón, A.M.: 173 

Ripková, S.: 564 

Ritschel, A.: 636, 756 

Ritz, K.: 1079, 1101, 1123 

Rizzo, A.: 868, 880 

Rizzo, A.M.: 151 

Rizzo, D.M.: 652, 958 

Roberson, R.W.: 268 

Robert, V.: 471 

Roberts, I.: 471 

Roberts, P.J.: 15, 574 

Roberts, R.G.: 71 

Roberts, S.K.: 1166 

Robinson, C.H.: 138 

Rodino, D.: 565 

Rodolfi, M.: 565 

Rodrigues, A.P.: 566, 567, 935 

Rodrigues, M.G.: 254, 757 

Rodríguez Hernández, M.: 238, 279 

Rodriguez, A.: 1022 

Rodriguez, N.: 1023 

Rodríguez, R.: 998 

Rodtong, S.: 388, 1171 

Rogers, H.: 936 

Rogers, J.D.: 152, 157 

Rogerson, A.: 963, 1052 

Rojas, E.: 321 

Rolen, T.: 758 

Romero, I.: 511 

Romero, J.: 1181 

Roncal, T.: 1193 

Roncero, C.: 30 

Rong, I.H.: 603, 759, 866, 1026 

Rong, R.P.: 759 

Rosendahl, S.: 619, 663, 764, 1025, 

1037, 1157, 1167 

Rosique, J.E.: 491 

Rosling, A.: 569 

Rossman, A.Y.: 2 

Rosu, A.: 1147 

Roth-Bejerano, N.: 702, 1048 

Rouland, C.: 619 

Roux, C.: 603, 1026 

Roux, J.: 464, 518, 825, 933, 1009, 

1027 

Rozkowicz, A.: 565 


Rubini, A.: 292, 1021 

Rubio, V.: 647 

Ruchikachorn, N.: 1028 

Rudawska, M.L.: 999, 1029 

Ruetters, H.: 1108 

Ruffini, A.: 716 

Ruiz-Boyer, A.: 380 

Ruiz-Dueñas, F.J.: 199 

Ruksawong, P.: 473, 474 

Rullkoetter, J.: 1108 

Rumberger, M.: 1030 

Rummo, S.: 215 

Rundberget, T.: 73 

Ruocco, M.: 115, 171, 372, 1168 

Rupricht-Robert, G.: 75 

Russell, J.C.: 881, 903 

Ruzieva, D.M.: 1128 

Ryan, M.J.: 609 

Ryvarden, L.: 99, 277, 394, 476, 554, 

758 

Røberg, K.: 398 

Römmert, A.-K.: 182, 1024 

Rønhede, S.: 568, 1167 

Saag, A.: 561, 760 

Saag, L.: 570 

Sáez, J.M.: 853, 855 

Said, S.: 1197 

Saikkonen, K.: 319 

Saito, M.: 445 

Saitta, A.: 605 

Sakai, K.: 721 

Sakamoto, Y.: 1169 

Sakaue, D.: 916 

Sakayaroj, J.: 56 

Saksirirat, W.: 1061 

Sakuma, Y.: 973 

Salcedo, I.: 1023 

Saleem, A.R.: 1174 

Salerni, E.: 469, 571, 575 

Salerno, M.I.: 90, 850 

Salinas, J.: 250 

Salo, V.: 572, 701, 1031, 1142 

Salvo, V.: 857 

Sampaio, E.V.S.B.: 539 

Sampaio, J.P.: 9, 664, 716 

Samson, P.: 1137 

Samson, R.A.: 69, 121, 719 

Samuels, G.J.: 344 

San Martín, F.: 152 

Sánches-Ballesteros, J.: 647 

Sánchez, C.: 1091, 1170 

Sanchis, V.: 927 

Sancho, L.G.: 261, 1059 

Sangvichien, E.: 388 

Sanjust, E.: 360 

Sankara, Ph.: 828 

Santerre, A.: 675 

Santiago-Mena, M.R.: 848 

Sanz, M.: 30 

Sarkar, S.: 196 

Sarova, J.: 882 

Sarrionandia, E.: 1023 

Sasa, M.: 515 

Sato, H.: 1032 

Satomi, S.: 844 

Satomura, T.: 987 

Sattarova, R.K.: 852

Sattayaphisut, W.: 1171 

Saunders, E.M.: 574 

Saunders, M.: 1172 

Savelkoul, H.F.J.: 166 

Savino, E.: 575, 821 

Sbrana, C.: 442 

Scala, F.: 115, 171, 372, 1168 

Scala, V.: 171, 372, 1168 

Schaper, T.: 1173 

Schardl, C.L.: 203, 311, 325, 883, 

1077, 1182 

Schatz, S.: 963, 1052 

Scheidegger, C.: 25, 263, 705, 1054 

Scherrer, S.: 260 

Schimek, C.: 1174 

Schloter, M.: 969 

Schmid, J.: 924, 1204 

Schmidbauer, N.: 221 

Schmidt, H.: 884, 885 

Schmit, J.P.: 98, 99, 102, 105 

Schmitt, I.: 288 

Scholler, M.S.: 247, 620, 934 

Schroeter, B.: 261, 263, 1059, 1165 

Schubert, K.: 636 

Schubert, P.: 965 

Schulte, A.: 323 

Schultze, K.: 1175 

Schulz, B.: 182, 830, 1024, 1065 

Schulz, M.J.: 1033 

Schumacher, T.: 23, 61, 684, 704, 

758, 770, 943, 1039 

Schurko, A.M.: 654 

Schützendübel, A.: 1103 

Schwärzel, H.: 965 

Schweiger, P.: 1092 

Scott, D.B.: 1204 

Scott, V.: 1204 

Screen, S.: 376 

Scrimgeour, C.M.: 123, 318 

Sdicu, A.-M.: 32 

Seiboth, B.: 343 

Seidl, M.T.: 396 

Seidl, V.: 175 

Seifert, K.A.: 3, 70, 298, 348, 676, 

678 

Seker, U.O.S.: 1188 

Selosse, M.-A.: 14, 1034 

Selvaggini, S.: 1152 

Sen, R.: 96, 130, 827, 1031, 1126 

Sengupta, A.: 945 

Senn-Irlet, B.: 391, 536 

Senthil Kumaran, R.: 576 

Senthilkumar, S.: 577 

Sentjurc, M.: 1136 

Sergeeva, L.E.: 578 

Serrano, E.: 444 

Seymour, R.L.: 698 

Shachar-Hill, Y.: 127, 270 

Shahi, S.K.: 309, 871 

Shams, M.: 886, 1163, 1164, 1176 

Shapiro, A.: 1177 

Sharif, S.: 1014 

Sharon, A.: 88, 89 

Sharples, G.P.: 525, 739, 865 

Shaw, B.D.: 1178 

Shearer, B.L.: 338 

Shearer, C.A.: 102, 639 

Sherrer, S.: 192 

Shi, Y.L: 167 

Shimazu, M.: 1032 

Shin, H.D.: 644 

Shinoda, I.: 1014 

Shnyreva, A.V.: 1035 

Shoemaker, R.A.: 676, 761 

Shokohi, T.: 800, 887 

Shoresh, M.: 172 

Shresta, G.: 889 

Shrestha, B.: 358 

Sierra-Galván, S.: 487, 762 

Siewers, V.: 87 

Sigler, L.: 668, 677 

Sihanonth, P.: 155, 388, 1028 

Silberstein, A.: 1036 

Silin, D.S.: 890 

Siller, I.: 597 

Silva, F.S.B.: 1141 

Silva, G.A.: 539, 1179 

Silvestri, M.: 470 

Sim, S.C.: 312 

Simanungkalit, R.D.M: 580 

Simmons, C.M.: 581 

Simpson, J.A.: 582 

Singh, K.G.: 871 

Singh, R.: 584 

Singh, R.K.: 763 

Singh, T.: 891 

Sinsin, B.: 614 

Sip, V.: 901 

Sirimungkararat, S.: 1061 

Sisakyan, S.H.: 364 

Sisti, D.: 1148 

Sisto, D.: 951 

Sivichai, S.: 56, 475, 558, 585, 591 

Six, D.L.: 428 

Skaar, I.: 73, 425 

Skovgaard, K.: 764, 1037 

Slatter, J.: 1149 

Slippers, B.: 430, 765 

Smalla, K.: 966 

Smirnov, A.N.: 892 

Smirnov, I.: 1016 

Smit, E.: 222, 1038 

Smith, F.A: 1092 

Smith, G.: 298 

Smith, J.E.: 180, 586 

Smith, M.: 1026 

Smith, S.E.: 1092, 1130 

Smits, M.: 94 

Smreciu, E.A.: 1033 

So, K.Y.: 587 

Soares, S.A.G.: 1179 

Sobczyk, K.: 1110 

Sogonov, M.V.: 588 

Solari, N.: 512 

Solhaug, K.A.: 1180 

Solheim, H.: 831, 893 

Soliman, S.: 1096 

Soliva, M.: 1054 

Solyakov, A.: 894 

Somrithipol, S.: 589 

Sonnenberg, A.S.M.: 165 

Sontirat, P.: 767 

Southworth, D.: 1000 

Spangler, R.: 192 

Spatafora, J.W.: 108, 180 

Spencer-Martins, I.: 689, 980 

Spender, S.W.: 83 

Speranza, M.: 590, 1181 

Sperisen, C.: 25, 1054 

Spiering, M.J.: 311, 1077, 1182, 

1204 

Spooner, B.M.: 64 

Srebotnik, E.: 201 

Sri-indrasutdhi, V.: 591 

Sridhara Gupta, K.: 895 

Srikitikulchai, P.: 592 

Srivastava, A.: 896 

St. Clair, L.L.: 768 

St. Leger, R.J.: 185, 376, 834, 1102 

Staats, M.: 1040 

Stadler, M.: 158, 249, 769, 1118 

Stajic, M.: 363, 915 

Stalpers, J.: 438, 471 

Starink-Willemse, M.: 780 

Starrett, M.C.: 941, 947 

Staszczak, M.: 1183 

Statzell-Tallman, A.: 531 

Stauffer, C.: 710 

Stchigel, A.M.: 638, 665 

Steadman, J.R.: 1041 

Steenkamp, E.T.: 692 

Steffen, K.T.: 95, 1184 

Steimel, J.: 195, 300 

Steinberg, G.: 267 

Steinnes, E.: 1084 

Stenlid, J.: 18, 22, 24, 197, 205, 432, 

453, 829, 843, 897, 1001, 1138 

Stensrud, Ø.: 770 

Stenström, E.: 897 

Stepanova, A.A.: 1185 

Stepanova, E.V.: 1106, 1131 

Stepinska, A.: 842 

Sterner, O.: 1193 

Stevens, K.J.: 83 

Stewart, A.: 563 

Stewart, W.S: 1069 

Stiles, J.: 898 

Stocchi, V.: 1148 

Stokland, J.N.: 394, 593 

Stott, A.: 143 

Stott, K.: 899 

Straatsma, G.: 133, 190 

Strahl, S.: 33 

Strani, P.: 442 

Strasser, H.: 114 

Strömmer, R.: 985 

Strunnikova, O.: 783 

Støen, M.A.: 237 

Suberkropp, K.: 1042 

Sugiyama, J.: 771 

Sugui, J.A.: 77 

Suh, S.-O.: 104 

Suhara, H.: 721, 990 

Suhr, K.I.: 816, 863 

Suija, A.: 594 

Suková, M.: 595 

Sullivan, L.: 948 

Summerbell, R.C.: 470, 668 

Summerell, B.A.: 797, 900, 1043 

Sung, G.-H.: 108 

Sung, J.M.: 358 


Susca, A.: 858 

Sutton, B.C.: 789 

Sutton, M.A.: 123 

Suwanarit, P.: 477, 524 

Suwanto, A.: 823 

Suyama, Y.: 983 

Suzuki, A.: 39 

Suzuki, K.: 596, 826, 916 

Suzuki, S.: 847, 1186 

Swann, E.C.: 727 

Sweigard, J.A.: 269 

Swift, S.R.: 264 

Sykorova, S.: 837, 901 

Syrjälä, L.: 902 

Szabo, L.J.: 727 

Szakacs, G.: 634 

Szedlay, Gy.: 597 

Szentivanyi, O.: 903 

Søchting, U.: 521, 537, 568, 630, 

697, 703, 766 

Söderström, B.: 131, 1092, 1137 

Sønstebø, J.H.: 1039 

Tabak, H.F.: 341 

Tabata, M.: 429 

Tagu, D.: 125 

Tahara, S.: 839, 917 

Taj-Aldeen, S.J.: 904, 905 

Takahashi, H.: 1119 

Takahashi, K.: 918 

Takamatsu, S.: 845 

Takashima, M.: 743 

Takeshita, N.: 1187 

Talla, E.: 334 

Tamai, Y.: 1169 

Tamasloukht, M.: 129 

Tamerler, C.: 1073, 1074, 1188 

Tan, T.K.: 54 

Tan, Y.Y.: 1204 

Tanabe, S.: 1080 

Tanabe, Y.: 771 

Tanahashi, T.: 1112 

Tanaka, C.: 39 

Tanaka, I.: 772 

Tang, A.M.C.: 598 

Tanticharoen, M.: 460, 558 

Tasanatai, K.: 599 

Tauschke, M.: 1044 

Taylor, A.F.S.: 984, 1005 

Taylor, D.L.: 223 

Taylor, J.: 1189 

Taylor, J.E.: 600 

Taylor, J.W.: 225, 227 

Teaumroong, N.: 1171 

Tedder, S.: 467 

Tedersoo, L.: 1045 

Tehler, A.: 6, 285 

Teixeira de Mattos, M.J.: 341 

Tellería, M.T.: 478, 651, 691, 723 

Tenreiro, R.: 664 

Terashima, K.: 1190 

Terekhova, V.A.: 1046 

Termorshuizen, A.J.: 393 

Theelen, B.: 257 

Thell, A.: 760 

Theodorou, M.K.: 118 

Theraisnathan, V.: 906 

Thienhirun, S.: 154 

Thißen, R.: 861, 907 

Thomsen, L.: 186 

Thormann, M.N.: 81, 1033 

Thorn, R.G.: 209, 245 

Thrane, U.: 70, 310, 884 

Tibbles, L.: 908 

Tibell, L.: 290 

Tichy, H.V.: 249, 1118 

Tillier, A.: 1034 

Timdal, E.: 408, 521 

Tirilly, Y.: 90, 850 

Tiwari, P.: 1191, 1192 

Tjahjono, B.: 823 

Tlalka, M.: 437 

Tojo, M.: 601 

Tokumasu, S.: 687, 983 

Tolpysheva, T.: 1071 

Tomati, U.: 308 

Tomimatsu, G.: 114 

Tommerup, I.C.: 338 

Tomsovsky, M.: 773 

Tonkin, J.E.: 774 

Torp, M.: 161, 425, 707, 775, 832, 

884, 909, 914 

Torres, N.V.: 339 

Tosi, S.: 1047 

Toyoda, H.: 854, 864, 910 

Tran-Dinh, N.: 230 

Trappe, J.M.: 99 

Tremblay, T.-L.: 1144 

Trembley, M.L.: 260 

Trest, M.T.: 776 

Triebel, D.: 273, 407, 720 

Trilles, L.: 257 

Trionfetti-Nisini, P.: 308, 880 

Troein, C.: 337 

Tronsmo, A.M.: 601 

Tschen, E.F.T.: 602 

Tschen, J.S.M.: 602 

Tsiklauri, N.: 202 

Tsuda, S.: 1120 

Tsukiboshi, T.: 645 

Tsuneda, A.: 678 

Tudzynski, B.: 85 

Tudzynski, P.: 87 

Tunlid, A.: 131, 337, 373, 1066, 

1137 

Turgeon, B.G.: 227 

Türk, R.: 1059, 1165 

Turnau, K.: 1159 

Tuthill, D.E.: 66 

Typas, M.: 783 

Typinski, T.: 1111 

Tømmervik, H.: 1056 

Tønsberg, T.: 356, 659 

Udovicic, F.: 40, 699 

Uehlein, N.: 1143 

Uetake, Y.: 445 

Ugalde, U.: 1193 

Uhlirova, J.: 528 

Uhlmann, E.: 135, 468 

Ulloa, M.: 505 

Ulrich, K.: 930 

Untereiner, W.A.: 4 

Upreti, D.K.: 309 

Urban, A.: 777 

Uzunovic, A.: 778, 911, 1099 


Vaccari, A.: 821 

Vach, M.: 812, 836 

Vágvölgyi, C.: 700, 1154, 1194 

Vainio, E.J.: 970 

Vajna, L.: 845 

Valcuvia, M.: 821 

Valencia del Toro, G.: 912 

Valenzuela, R.: 487 

van Aelst, A.C.: 438 

van Baarlen, P.: 1040 

van Breemen, N.: 94 

van Broock, M.: 716 

van den Hondel, C.A.M.: 1117, 1139 

van der Drift, C.: 1200 

van der Krift, T.P.: 438 

van der Linde, E.J.: 603, 866 

van der Merwe, P.C.: 1026 

Van Dijk, J.F.W.: 365 

van Driel, K.G.A.: 438 

Van Griensven, L.J.L.D.: 166, 190, 

1200 

Van Iperen, A.: 780 

van Kan, J.A.L.: 1040 

Van Rooij, P.: 779 

Van Ryckegem, G.: 604 

van Schöll, L.: 94 

van Tuinen, D.: 129, 441, 496, 1107 

van Zeijl, C.M.J.: 1195 

Vanden Wymelenberg, A.J.: 1196 

Vangronsveld, J.: 331, 1007 

Vani, A.: 895 

Vánová, M.: 837 

Vanzela, A.P.F.C.: 1197 

Vasiliauskas, R.: 22, 453, 1001, 1005 

Vastag, M.: 1194 

Vázquez, C.: 732 

Vedenyapina, E.: 1016, 1017 

Veenman, C.: 222, 1038 

Ventura, Y.: 1048 

Venturella, G.: 605, 769 

Verbeken, A.: 604, 658, 740, 741, 

779 

Verkleij, A.J.: 438 

Verkley, G.J.M.: 63, 780 

Verrips, C.T.: 1139 

Vestergaard, S.: 1049 

Vidal, G.: 487, 491 

Vilgalys, R.: 28, 46, 145, 146, 210, 

236, 357, 397, 548, 680, 1062 

Villareal, M.: 647 

Villarreal-Ruiz, L.: 1050 

Villegas, M.: 487, 666, 748, 781 

Vinay, M.: 1051 

Vinnere, O.: 913 

Virag, A.: 1109 

Visconti, A.: 423 

Viterbo, A.: 172 

Vlassova, T.A.: 1198 

Voegeli, S.: 335 

Vogel, C.: 963, 1052 

Vogel, R.F.: 884, 885 

Voglmayr, H.: 102, 606 

Voigt, K.: 229 

Volkmann, M.: 1108 

Volkmann-Kohlmeyer, B.: 102 

Vrijmoed, L.L.P.: 713, 747, 1053 

Vrålstad, T.: 61

Vukojevic, J.: 363 

Vurro, M.: 116 

Vyboh, J.: 804 

Vysotskaya, A.M.: 618 

Waalwijk, C.: 1151 

Wagemaker, M.J.M.: 1200 

Wahlström, K.: 829 

Waipara, N.W.: 220, 607, 914 

Wald, P.M.: 608 

Wallander, H.: 315 

Walser, J.-C.: 25, 705, 1054 

Walser, P.J.: 75 

Walsh, D.: 446 

Walsh, J.L.: 900 

Walther, G.: 782 

Walton, J.D.: 374 

Wang, C.S.: 187 

Wang, J.: 883 

Wang, P.-H.: 835, 1064 

Ward, T.: 162 

Wasser, S.P.: 163, 307, 637, 915, 

921, 968 

Watanabe, M.M.: 771 

Watanabe, T.: 1119, 1136 

Watkinson, S.C.: 437 

Watling, R.: 99, 385, 525, 752 

Webber, J.F.: 297, 299 

Weber, D.: 783 

Weber, N.S.: 180 

Wedin, M.: 286, 290, 649, 742, 784 

Weiner, S.: 1105 

Weir, A.: 104 

Weiß, M.: 9, 14, 403, 782, 1034 

Weisgram, M.: 201 

Weiss, M.: 271, 513 

Weller, D.M.: 113 

Wells, J.M.: 1055 

Wenehed, V.: 842, 894 

Werner, A.: 930 

Werner, K.: 1044 

Werner, S.: 77 

Wernitz, M.: 77 

Werth, S.: 25, 1056 

West, H.K.: 1055 

Weyers, H.S.: 1042 

Whalley, A.J.S.: 155, 384, 739, 752, 

865, 1028 

Whalley, M.A.: 154, 739 

Whipps, J.M.: 111 

White, M.M.: 104, 148 

White, N.A.: 217, 609, 1057 

Wiemken, A.: 520 

Wierzchos, J.: 124, 459, 590, 610 

Wijngaarden, P.J.: 937 

Wiklund, E.: 784 

Wilberforce, E.M.: 322, 1058 

Wilkinson, H.H.: 311, 312, 1182 

Williams, N.A.: 948 

Wilson, A.W.: 785 

Wilson, B.J.: 81 

Wilson, D.M.: 421 

Winder, R.S.: 314 

Wingfield, B.D.: 301, 355, 430, 464, 

646, 672, 673, 692, 722, 765, 933, 

1003, 1027 

Wingfield, M.J.: 296, 301, 355, 430, 

464, 518, 646, 672, 673, 692, 694, 

706, 722, 765, 825, 933, 1003, 1009, 

1027 

Wirtz, N.: 1059 

Wojciechowski, M.: 734 

Wolfe, K.H.: 333 

Wollweber, H.: 769 

Wong, S.: 333 

Wong, Y.H.: 1201 

Wongkaeo, A.: 1061 

Woo, S.: 115, 171, 372, 1168 

Woodward, S.: 17, 809, 815 

Wright, D.: 1137 

Wrzosek, M.: 786 

Wu, M.L.: 611 

Wu, Q.X.: 99 

Wu, S.H.: 787 

Wu, W.P.: 788, 789 

Wöllecke, J.: 500, 1036, 1060 

Wöstemeyer, J.: 1174, 1175 

Xiang, Q.: 196 

Xu, J.: 875 

Xu, J.L.: 841 

Xu, X.: 881 

Yahr, R.: 1062 

Yakovlev, A.Yu.: 1202 

Yakovleva, N.S.: 1106 

Yalcindag, B.: 1203 

Yamada, A.: 708 

Yamada, T.: 916 

Yamaji, K.: 917 

Yamamoto, Y.: 918, 1114 

Yamaoka, Y.: 725, 1063 

Yamato, M.: 916 

Yang, D.Q.: 919, 920 

Yang, S.-H.: 1064 

Yano-Melo, A.M.: 1141 

Yap, A.-T.: 168 

Yarden, O.: 35, 266 

Yassin, M.: 921 

Yavmetdinov, I.: 1131 

Yedidia, I.: 172 

Yee, M.: 612 

Yeh, Z.-Y.: 613 

Yetman, D.: 25 

Yli-Mattila, T.: 790, 791, 884 

Yoder, O.: 227 

Yohalem, D.S.: 792 

Yorou, S.N.: 490, 614 

Yoshino, T.: 928 

Yotsutani, Y.: 1068 

Young, J.P.Y.: 224 

Youpensuk, S.: 615 

Youssef, I.: 1013 

Yu, H.J.: 922 

Yu, J.: 1085 

Yu, S.H.: 793 

Yuan, G.-F.: 717, 923 

Yuan, Z.Q.: 612, 819 

Yumoto, I.: 1120 

Yumoto, Y.: 979 

Zabari, L.: 724 

Zafari, -: 794 

Zalar, P.: 120 

Zappatore, S.: 617 

Zare, -: 794 

Zare, R.: 248, 795 

Zavarzin, A.A.: 616 

Zeilinger, S.: 175 

Zelenko, S.D.: 796 

Zeller, K.A.: 797 

Zelter, A.: 266 

Zervakis, G.I.: 557, 605 

Zeyl, C.: 366 

Zhang, L.H.: 841 

Zhang, N.: 924, 1204 

Zhang, Y.: 788, 789 

Zhao, B.: 496 

Zhuang, J.Y.: 798 

Zhuang, W.-y.: 62 

Zipfel, W.: 270 

Zitzmann, W.: 1118 

Zjalic, S.: 308 

Zobel, M.: 96 

Zoina, A.: 115, 171, 372 

Zoller, S.: 326 

Zotti, M.: 617 

Zubek, Sz.: 1159 

Zuccaro, A.: 1065 

Zvereva, L.V.: 618 



Typ & Graf AS – Foto: Fjellanger Widerøe AS - Trykk: Printhouse AS, Oslo

Book of Abstracts (PDF) - International Mycological Association

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