The JapaneseSociety The Japanese Society for for Plant Plant Systematics Systematics ISSN 1346-7565 Acta Phytotax,GeobeL 58 (1): 19-32 (2007) Myxochinmys(Zingiberaceae), a new genus from Borneo ATSUKO TAKANoiand HIDETOSHI NAGAMASU2 iMtiseum Ri,ogo 669-154a Jinpan; Activities, ilyogo,6-chome,}Zryoigaoka, Skencla, andHuman ofNdture 2111te UhiversityMitseum, K),oto UhiversiijlSZik}'o, 606-8501,Jbpan K)?oto K),oto A new genusand species, filyxochtanzys muilerensis from CentralKalimantan, Borneo (Zingiberaceae) infiorescences with isdescribed, Thisnew species ischaracterized by indistinct congested pseudostems; from spirally arranged bracts; copious, highlyviscous, axjllary-pooled mucus secreted glandson imbricatebracts;and fiowerswith a concave labellum,versatile anthers, and a grooveforrned by the inner surface of thecorolla tube to envelope and holdthe style. Parallel orientation ofthe planeofdistichy of the leafyshoots, well-developcd staminodes, and labellumnot cennate with the filament suggest that Adyxochlanrysbelongsto the tribe Zingibereae,Among the generain Zingibereae,Adyxoehlanrys issimilarto Staphochlanrys inthat itlacksa distinct itisdistinct from pseudostem and spiral inflorescence; Sbaphochlaiays, however,inthatithas a groovewithin the corolla tube, concave labellum and versatile anthers, Phylogenetic analyses basedon DNA sequences of the nuclear internal transcribedspacer (ITS) K and maturase and Sbaphoehlamps. The ofthe'chloreplast (matlO similarities and DNA show similarity with Distichochlanv]s,Adyxochlanrys, differences betweenA()xeehldop;s and related generaare dis- cussed. Key words: Borneo,toxochlamys, mullerensis, toxochlanrys Sbaphoehlantys,Zingiberaceae,Zingibereae The Zingiberaceaecomprise ilyof53 and over genera 1200 a fampantropical species et al. (Kress molecular phylegcny,ncw genus, Alpinieae),and ZingiberoideaeHaask. the remaining Currentstudies genera), (containing on the mol- diversity is southern and ecular phylogenies of the Alpinioideae and some southeastern Asia(Larsen et al. 1998). TraditionallMlarge genera such as Aipinia,Amomum, and fburtribes (Alpinieae Meisn.,Globbeae Meisn., Etlingera 2004,Xia et aL 2004,Kresset (Pedersen Hedychieae Petersen, and Zingibereae Burtt & al, 2005)suggest thatthe Zingiberaceae are monoOlatubji) havebeenrecognized inthe familybased phyletic, butrequire redefined genericboundaries, et al. especially fbrAipinia and Amomum, which are on fioraland vegetative characters (Dahlgren 2002) with a center of 1985;Larsen et al. 199g).Kress et al. recent(2002) In Zingiberoideae, Williams polyphyletic, showed that Globbeae are mono(2004) apparently lyproposed a new classification ofthe Zingiberaceaeet al. basedon a molecular phylogeneticanalysis, They phyletic and sister to Zingibereae; however, the recognized fbursubfamilies: Siphonochiloideae W, generic1irnjts intheZingibereae remain unresolved. J. Kress (containing only Siphonochilus),[lamijiIn 2004, we explored Muller Range, Central oideae W, J. Kress (containing only 7Zimijia), Kalimantan, Indonesia and found an interesting of Alpinioideae Link (containing most of the fbrmer orientation ofthe planeof distichy ginger.Parallel NII-Electronic Library Service The JapaneseSociety The Japanese Society for for Plant Plant Systematics Systematics 20 APG the leafy shoots, well-developed staminodes, and labellum not connate to the filament suggest thatthe plantbelongsto the tribeZingibereae,but comparative morphological studies and molecular provideevidence against phylogenetic the placement of Xlo1. 58 54eC. Amplified productswere Microspin S-300 HR columns purifiedusing (Amersham Biosciences, Buckinghamshire,England,UK). Sequencingwas perfbrmedusjng ABI Prism(tm) Big Dye Terminator Cycle Sequencing Ready Reaction Kit (Applied Biosystems,FosterCity, Califbrnia, USA), Sequencingprimersincludedthe thisplantwithin any ofgenera inthis tribe.Thus, a new and new species, M genus, ILd),xochlantys, mullerensis, isdescribed here. In accordance with amplification primers as well as ITS2K (Rangsiniji Article42 of the InternationalCode of Botanical et al. 2000)and ITS3P (M611er & Cronk 1997) fbr Nomenclature(McNeill et aL 2006), the names of the ITS region and m5R, mSP2R, m5fa, m8R, the new genus and species are simultaneously datedusing the description provided below. vali- Materialsand Methods matK8R from Kress (all 2002), matK8 et aL (Steele & Vilgalys1994),and [EA05R (Takano & Okada 2002) fbrthe matK region. The productswere cleaned using the manufacturer's and were protocol, run on an ABI Prism'M 310 GeneticAnalyzer Biosystems Japan,Tokyo, Japan),Raw (Applied Descriptionofthe species The species description isbasedon studies of plants in the fieldand in the herbaria of BO, E, HYO, K, and KYO, The plant material studied includes pressed and driedspecimens, spirit-fixed floral material, and silica-gel dried tissueformolecular phylogenetic analysis. sequences were assembled and edited using the BioEditver. 5.0.9 software (Hall1999). DNA sequences were aligned by multiple alignments using the CLUSTAIIWI ,83 computer software package with defaultsettings et al. (Thompson 1994). Pdylageneticanalyses Pbylogenetic analysis For the DNA somal DNA of using DAC4 sequence clata analysis, we used nuclear ribothe internaltranscribed spacer ITS1, 5,8S,and (ITSs; of the chloroplast (matl() regions The ITS dataset was analyzed bothseparately and in combination with the matK dataset. To test dataincongruence, the incongruence lengthdifTerence (ILD) test(Fanis et aL 1995)was implemented as the partition homogeneity test option in PAUP" with 100 replicates foreach of two parti- ITS2)and maturase K DNA. The ITS and matK sequences inA(lyxochlanrys were generatedby the fo11owing method and deposited in theDNA tions. Data Bank of Japan (DDBJ). The sequences of Maximum parsimony (MP)was amalyzed using other taxawere obtained fromGenBank (AppendixPAUP" ver. 4,O. beta10 (Swoffbrd 2003). Heuristic rlbtal 1). with SIMPLE addition genomicDNAs were extracted fromsilica- searches were conducted driedtissuesby using the modified C[IIABmethod tree bisection-reconnection (TBR)branchswap& Doyle 1987, Takano & Okada 2002). (Doyle ping,and the MULPARS options, Only basesubITS sequences were amplified using the ITS5P and stitutionswere treatedas characters fbrphylogeny ITS8P prirners(M611er & Cronk 1997).The chlororeconstmction, Allthe gaps were treated as missing , plast mati(; region was amplified using trnKIF (Manos& Steele1997) and trnK2621 (Liston & Kadereit 1995).All amplifications were perfbrmed using TaKaRa Ex [ktqTM Bio Inc.,Otsu, ([lakara Shiga,Japan),at annealing temperatures of 50"to values. Support forbranches was bootstrap analysis estimated using 1000 replications 1985) by a heuristic search using SIM(Felsenstein PLE addition, NNT branch swapping, and the MULIIARS options, Three species ofSiphonochitus with NII-Electronic Library Service The JapaneseSociety The Japanese Society for for Plant Plant Systematics Systematics May 2007 were TAKANO selected et al. 2002). (Kress the outgroup as & NAGAMASU: 21 tauroehtam.v,y, gcn.nov, duobus;ovarium A Bayesian analysis was eonducted v, Central Kalimantan. Murung using Raya MrBayes basali. placentatione uniloculare, 7]tpus,INDONESIA, District, behind Tumbang Naan (O'09'03"S, 3,1.2(Huelsenbeck & Rohnquist 2001, from Sungai(River) Lapangan,a Rohnquist & Huelsenbeck 2003). The bestfitting branch of Sungai Joloi(OO11'OO"N 113031'59"E),ca, fbrBayesian analysis was selectsubstitution model 200 m from Muara Sopan (ee12'22"N 113e31'23"E), ed using a series of likelihood ratio testsas imple- December 21, 2004, H. Okada, H. Ndgamasu, H. mented in Modeltest v. 3.7 (Posada & Crandall 7lsukLu/a, A. lahano,&A. AJdiki, K7L671 (HoloBO; isoHYO, KYO), 1998).We ran over one million generations of fbur 113045'04"E) simultaneous cold, (one en route three heated) Matkov chains Rhizomatous herbs.Rhizome slender, elonby using MrBayes y, 3.1.2until the average standard gate,Leafy shoots close together. Leaves sessile, 3deviation of split firequencies became less 9,distichous, (ASDSF) to rhizome. Planeof leavesparallel than O.O05.The tree from every 100th generation Leafsheathopen to thcbase,17.5cm long,ca. 1 cm was retained (excluding a quarter of 1OO/total gen- wide; ligule, 5 mni long,Leafblades narrowly oboerations), and a 50% majority rule consensus tree vatc, to 50 cm long,ca. 7 cm wide, basecuneate, was constructed from these. For each ITS and the apex acute, upper surfhce sparsely hairy,uniformly combined dataset, the logstabilized before50 millower surface densety hairy,unifbrmly glandular; liongenerations in the Bayesian analysis (1,OOO,OOO Inflorescence terminal on a leafy shoot, 4glandular. forthe ITS dataset and 4,500,OOO fbrthe combined 15 cm long,fiowering from the baseto apex; scape dataset), The results of theBayesiananalyses are 2.5-4 cm long.Bracts spirally arranged, imbricate, reported as the posterior probabilities (PP; ca. 5 cm long, ca. 1.3cm wide, concave, uniforrnly Huelsenbeck & Rohnquist 2001),which are equal to puberuleus, with many glands,blood red, fi11ed the percentage is resolved. trees sampled of Only PP when scores a excess given clade of 509,6are with to bracts, ca. 3.5cm long,ca. 1.3 keeled,opposite inour shown trees, cm The result of exploratory analyses diMcult-to-align regions thatexcluded wide, resolved (not 117 (ITS, Calyx tubular, ca, 2.5 cm ment dataset excluding within the ITSs, regions 2 with results from the ambiguous align- Results ca. mullerensis gen.et 1,2) (Figs. 1.5cm wide, concave, gin. Lateralstaminodes sp. nov. ca. Herba fbliadisticha parallelead rhizoma. fbliatoterminalis, e basi ad dispositae, fioremsinfiorens; bracteae spiraliter rhizomatesa; Infiorescentiain surculo apicem mm 3 mm wide, subtendens, mucum gelatinum copiosum white. long,with and e:Tveloping long. Ovary gularem 3 unequal teeth, glabrous, Corollatube ca. 4.7 cm long,thick, with a groove fbrmed by innersurface enveloping style; lateral petalsovate, white with red apex, ca. 2 cm long,ca. O.8cm wide,slightly hooded;dorsal petalovate, ca. 2,1 cm long, ca, O.9 cm wide, slightly hooded, beaked,totally glal)rous. Labe]lum ca. 2.8cm long, A. [[lakano& Nagam. Myxochlamys or topologies;there- pairs)produced fbre,in thispaper,we present only ofthe base.Flowerssolitary. long,split along one side to open glabrous, shown) to base, apex more analyses Bracteoles2- transparent mucilage, colorless, a pinkishmarobovate, ca. 5 mm long, Antherwhite; filament ca. 5 groove style; ca, white 5 mm connected with to corolla tube thecae versatile; spur ca. 6 mm long, glabrous, unilocular, secer- basal,rudimentary septa as placentation present bracteolaebracteisoppositae, bicarinatae, ad longitudinal ridges inside ovary. Epigynousglands basinapertae. Florescorollae tubo et filamento intraproStylewhite, Stigmawhite, fundesulcato, stylo perlongitudinemin suLco involuto; 4-5 mm long,bilebed. basalibiis cup shaped, ciliate around mouth. Fruitunknown. labellum concavum; anthera versatilis calcaribus nentes; NII-Electronic Library Service The JapaneseSociety The Japanese Society for for Plant Plant Systematics Systematics 22 APG M)1.58 B A ([glgl) [!l) Ii I2mm cm D 5mm cm Ii Ii cm E F z1: ; '1tl N111 1l[t :11ll F 1 :i1t'ai E il'1 l .d /t t + icmI ] Fl1I, FiG.1.deochlanots mullerensis, A: Crosssection of ovary, B[ Epigynousgland. C: Anther D: Calyx. E:Bract, F: Bracteole. G: Flawer. H: Partof fiowershowing corolla tube with a furrowenclosing a style,lateral staminodes, dersalcorolla lobe,anther, and labellum (from spirit-fixedmaterial ofH/ Ohada et al. K767J). Distrihtttion: West and CentralKalimantan. JP8 CK); ibidem,CS Awmaek J99(K). wrst Kalimantan: Serawai,8 km northeast ofDesa Jelundung, BatuLmiang 112"36'7.1"E),AC Chureh & U]drMlabyar (O"30'57.9"S, Etymology:The highlyviscous mucus from glandson the bracts is pooledin the axils of the 17e2 eo). imbricatebracts,The name of the new genus is derived from these features slimy + ehlanurgMblecularpbylogenetic analyses (MJarocovering, using I71Sand mantle). combinedclatasets The ITS-1had a tota1 aligned lengthof 184 bp Additionalspecimens examined, INDONESIA. sequences ranged from 173 to 217 bp) CentralKalimantan:Sarnba, 1994-1995, Cuttingbleck (unaligned ofPT Handiyani(Oa43'22.5"S, 112'50'37,S"E),JKJti,'vie with a mean guanine-cytosine (GC)content of & A Ruskandi 5447 (BO);Near Muara Joloi (Oe02'S 53,59t6, the 5,8S region had an aligned lengthof 114"06'E), es Awmack 109 (K), ibidem, {:USAwmack 164 bp 158-164bp) and a GC content (range, of NII-Electronic Library Service The JapaneseSociety The Japanese Society for for Plant Plant Systematics Systematics May 2007 TZiXKANO & NAGAMASU: FIG.2.A(ytrochlam.vs murlerensis. "6,xochlanrys, gen.nov. 23 A/ Habitat. B: Close-upof fiower.C: Infiorescence (all photosby H.Okada.; H. Okada etal. KZ 67J). 51.79t6, and the ITS-2 had an aligned bp (range, 208-270bp)with GC a lengthef226 content In toxochtaop,s, the ITS-1 sequence vv'as of58.1%. The result ofBayesian inference ofphy(58,4%), logeny(posterior PP), howevegstrongprobahilities: 179 bp in ly supported lengthwith a GC content of52.5%, the 5.8S region was 164 bp inlength with a GC content of51,8%, and the ITS-2 region was 229 bp in Iength with a GC content of 59,89r6, Analysis of the ITS sequence data,excluding the ambiguously aligned regions by using 70 taxa, resulted in eight equally parsimonioustreesof 1508 steps (number of parsimony-informative characters,272; consistency index [CI] O.404;retention index [RI] O,622).In the strict consensus tree (Fig, 3), Myxochlamys is found in the Zingiberoideae clade, which is sister to the Scuphochlamysand Distichochlam.vs clades, but bootstrapsupport for this relationship is low = = and = 4). (Fig. Congruence of the datapartitions (chloroplast nuclear) could not be rejected by the ILD testlp (PP 1OO) this relationship = O,98),Analysis ofa 2227 characters combined fbr47 taxa matrix containing in 1617 equal- resulted ly parsimonioustrees of 1711 steps (number of characters, 401; consistency parsimeny-informative =: - index[CI] O.527;retention index[RI] O.603). The strictconsensus treeisshown in Fig.5,Both MP and Bayesiananalyses (Fig. 6)yieldedtreesof similar topology,but the Bayesian tree was more resolved inthe Zingiberoideaeclade. toxochtamys issister te Sbcrphochlam.vs and (BS!PP74.31100), Distichochlaiays issister to these genera (BS!PP 72.9flOO)(Figs, 5, 6). - - NII-Electronic Library Service The Japanese Society for TheJapaneseSocietyforPlantSystematics Plant Systematics 24 APG Vbl.S8 Gtobba talangensis Camptandraparvuta Caulokaempteriasaxicola Hanith'a aloifiora PJffgophyilumyunnanensis Curcumorpha longifiora 2 aoo.D7o.a Boesenbeigia iongifiora 100,O Curcumorphatongifiora7 59.899.9riOD.OBoesenbeigiapuicherrima Boesenbeigiagelatinosa Boesenbeigia rotunda Caulokaempieriathailandica Com"kaempferiatongipetiolata Comukaempteria a"rentifiora Zingibereliipticum Zingibergramineum Zingibersuiphureum Zingiberjunceum 98.850.758.0 95.4 Zlngiberoideae 9B,9 58,4 Clade 58.7 82.4 67.699,492.9 88.2 Zingiberwrayii ZingiberJongipeduncuiatum Zingibercoloratum MJorochlamys mullerensis Distichochlan?ys sp. Kress O1-6848 Distichochlamysciteaf Distichochiamyscitrea2 Scaphochlamyskunstierif Scaphochlamysbiloba7 Scaphochlamysbiioba2 Scaphochlamyskunstleri2 Scaphochlamysobcordata Kaerr7pteria elegans Kaernpteriaparvifiora Kaempferia rotunda 99.7 97.5 97.4 95.599.9 Kaempteria sp. kress98-6289 Cautieyagrac"is Cautieyaspicata Roscoea bhutanica Roscoeaputpurea Roscoea cautieoides Roscoeahumeana Hea)fchiumlongicomutum Hedychium viUosum Pommereschea iackneri Rhynchanthusbeesianus Smithatris scrpreneanae 8g.2 too,o too.o52.8 92.3 84.0 80,572.994.7 loe.o Curcuma bicolor Curcuma ecomata StahJianthusinvotucratus Curcuma harrnandii Curcumaparvifiora Curcuma thorelii Curcuma roscoeana Paracautleyabhath' Curcuma rubescens Hitcheniaglauca Hemiorchisrhodorrtrachis Gagnepainia hannandii loa.o93.a 55.793.78d,3 77.1 rioo.o 53.9loe.o Gagnepainia thoretiana AipiniablepharocaCyof Elettaria longituba Atnomum yunnanense Aipiniatonkinensis Aipiniaeiegans Renealrriiabattenbergiana Eiettariopsisstenosiphon Eiettariopsiskerbyi Tarnijia fiageliaris SiphonochiJusaethiopicus Siphonochilus kitVcfi Siphonochilusdecorus FIG,3,Strict consensus of eight maximum resulting from analy sis of the ITS parsimonioustrees of the subfamily Zingiberoideae sequence data(length1508,consistency index O.404,and retentjon index = e.622).Numbers above branchesindicateboot= strap x,alucs = (>50%). NII-Electronic Library Service The Japanese Society TheJapaneseSocietyfor for Plant PlantSystematics Systematics May 2007 llAKANO & NAGAMASU:A(yxochlanrys, 25 gen.nov. Gtobbatalangensis Oamptandiaparvula Cautokaempferiasaxicola PyrgophyitumHaniffiayunnanensis aJbifiera Boesenbergiageiatinosa Boesenbergiapulcherrima too73 10D Boesenbergiarongiflora Curcumorphatongifioraf ' Boesenberg aoo99CurcumorphaIongiftora2 Zlngiberoideae Claderioo iarot"nda ga riooCa"lokaempferiathaitandica Cornukaempferialongipetiolata Cornukaempferiaaurantifiora Zingibereliipticum 92 Zingibercoloratum leO Zingibertongipeduncuiatum Zingiberwray ii Zingibersuiphureum Zingibergrarnineum Zingiherl'unceum 5259 96 5360 nayxochiamysmullerensis Distichochlamyssp.KressOf-6848 aoo leo ioo DistichochtamyscitreaV Distichochtamyscitrea2 99 100 Scaphochlamysbitobaf Scaphochlamysbiloha2Scaphochtamyskunstferi7 rioo 100Scaphochtamyskunstleri2 63 Scaphochlamysobcordata Kaempferiaelegans Kaempferiaparvifiora 66 100 {ooaoo Kaempferiarotunda 100 Kaempferia kress98-6289 aooCautieyagracilis Cautleyaspicata 100 Roscoeabhutanica riootooRoscoeapurpurea looRoscoeacautleoides Hedychiumrongicotnututn 100Roscoeahurrieana 86 Hedyohiemvilros"m 100 beesianus 99RhynchanfhusPomrrierescheaJackneri Curcumabicolor CurcumaSmithatrissupraneanee ecomata leo C"rcumaharmandii 100100 60 StahJianthusinvolucratus Curcumapatvifiora ioe Curcumathoretii Curcumaroscoeana 9898100Paracautieyabhatii Curcumerubescens Hemiorchisthodorrhachis riDO GagnepainiaharmandiiHitcheniaglauca 92Gagnepainiathoretiana Alpiniablepharocatyx Reneatmia battenbergiana Elettariopsiskerbyi rioo Eiettariepsis stenosiphon 87 100 Eiettarialongituba 100 leo Amomumyunnanense 100 Arpiniatonkinensis Arpiniaelegans Tami'iafiageilaris100 99 Siphonochitusaethiopi Siphonochiiuskirkii Siphonochiiusdecorus O.fiSubstitutionsiSite Flci. 4. Bayesianconsensus phylogram based on branch lengthobtained than 5U%. (PP)higher posterior probabilities fromITS scqucnce data. Nurnbersabove branches indicate NII-Electronic Library Service The Japanese Society for TheJapaneseSocietyforPlantSystematics Plant Systematics 26 APG wa]1.58 Camptandtaparvuta Cauiokaempferiasaxicota RyngophWlumyunnanensis Boesenbeigiarotunda Boesenbeigia puichern'ma Curcumorpha tongifiora1 Curcumotphalongifiora 2 Cornukaempleriaautentifiora Zingtoerellpticum Zingtoergramineum Zingibersuiphureum Zingiber umayii Distichochlamyssp. Kress Ot-6848 IPlyxochlamysmutlerensis Scaphochlamys biloba f Scaphochlamyskunstieri1 Kaempforiaparvifiora Kaempferiarotunda Kaempforia sp. kTess98-6289 Cautleya gracilis Cautteyaspicata Roscoeacautleoides Roscoeapurpurea , Hedychium longicomutum Hedychiunivillosum Pommereschea lackneri Rhynchanthusbeesianus Curcuma bicoior Smithatrisstq)raneanae Curcuma roscoeana Hitcheniaglauca Curcuma thorelii Stahtianth"sinvoiucratus Gagnepainia thoreliana Hemiorehisrhodorri]achis Aipiniablepharocalyx Aipiniaelegans Amomum viitosum Amomum yunnanense Renea/mia battenbeigiana Elettariopsiskembw' Etettariopsisstenosiphon lamijiafiagelfaris Siphonochilusaethiopicus Siphonochilusdecorus Siphonochiluskirkii FIG,5,Strict consensus and es 1617 maximurn Zingiberoideae resultin.tr from analysis ofcembined parsimonious trees ofgubfamily ITS = L711,consistency indcx= O.527,and retention index= O.603).Numbers above branclidata(tength indicate bootstrapvalues (>50%). tnatK rcgion of sequence NII-Electronic Library Service The Japanese Society for TheJapaneseSocietyforPlantSystematics May 2007 Plant Systematics TAKANO 27 & NAGAMASU:ta･xochlam.vs, gen.nov. Arr]omum yunnanense Amomum villosum Aipiniaetegans Aipiniablepharocat)o( evettariopsisstenosiphon Elettariopsiskembyi Renealmiabattenbeigiana Gagnepainia thoreffana Stahlianthusinvolucratus Curcuma thoreM Hitcheniagiauca Curcuma roscoeana Smithatrissupraneanae Ryrgophytlum yunnanensis Camptandrapantula Rhynchanthusbeestanus 2ZlngiberaLdea Heclychiumvi"osum Hedychiumlongicornutum Roscoeaputpurea Cautleyaspicata Kaempleriasp. Kaempteria rotunda Keempteriaparvifiora ScaphochiamyskunstieriV Scaphochlamysbilobaf na)orochlamysmullerensis Distichochtarnys sp. Kress Of-6848 Zingiberwrayii Zingibersuiphureum Zjngibergremineum Zingiherettipticum Cornukaempferiaaurantifiora Curcumorpha tongifiora2 Curcumotpha iongifiora7 BoesenbeigiapulcheMma Boesenbergia totunda alhifioraCaulokaempteriasaxicola tamijiafiageliaris Siphonochiluskirfc" Siphonochilusdecorus Siphonochiiusaethiopicus O.1SubstitutionsiSite FJG,6. Bayesian consensus phylograrnbased on branch lcngthof combined branchesindicatc posterior prebubilities (PP)higherthan 50%, ITS and matK region sequence data.Numbers above NII-Electronic Library Service The JapaneseSociety The Japanese Society for for Plant Plant Systematics Systematics 28 APG Nlo1. 58 TABLE1 .Comparison of keycharacters betweentoxochlatays and related genera Arrangement Modeof ofbracts fiowering Flowerslst bracteoleCorollatube Thecae Anther Ovary SolitaryOpen to base, withagroove Basalspurs Oppositebract; formed by the keels2 innersurface Versatile IICucochtanozs Spiral Base-apex mutlerensis (indeteuninate) Unilocular with basal placentation Sbaphochlanrys Spiral Base-apex Usually Open to base, without such a With very Adnate in Oppositebract, groove short, free, (indeterminate) cincinni eften more or basalspurs lcsskeeled(2 (nospur in keelsonly inS. S.calctcola) Unilocular or trilocularwith basalplacentation catcicoia) Distichochtamp's Distichous Base-apex In Tubular; keels2 cincinni (indeterminate) withoutsucha Spursabsent Adnate Sbaphochlanrys are moderately basedon theirspecies alone Discussion Trilocular groove supported subclades sub(St]ophochlamrys MJtxochlanoLs mullerensis clade: BSIPP -= 67.61100; Distichochlatayssub- oideae clade: BS!PP isplacedintheZingiberin the phylogeneticanalyses basedon theITS - 95.4/100),and was tocrochlanrys placedoutside of these genera. In the combined was foundto be data(BSfPP 58.71100firomITS data,89.6flOO data set phylogeny,iLdyxochlantys from the combined data). sister to the Scaphochlamys clade In terms ofmorphology, (BSfPP 74,3/100)and Distichochlamys was the sister of A(lyxochlantys mullanensis could also be placed inthe tribeZingibereae, subfamily Zingiberoideae,inthe these two genera (BSfPP 72.9/100).The results classification system ofZingiberaceae by Kresset aL suggest thatthesethree generaare closely related, Morphologically,A(YxochlaTays issimilar to by yirtue of the parallelorientation of the (2002) of itsleafyshoots, its Scaphochtamys in that itlacksa distinct well-develplaneof distichy pseudostem,but the spiral arrangement of bracts oped latera1 staminodes, and 1ahellum not connate to difl At BO and K, most specimens of toxochlanrys the filament. The unilocular ovary with basal pla- fers. centation israther rare in this tribe, however; a without fiowerswere identified as Sbuphochianrys, transition in the ovary type within some genera, probablybased on inflorescenceand stem characteristics.Thereare also significant morphological e.g., Boesenbergia, Hbptochorema, and 5Zraphochlanrys, from trilocular differences, however,between 7VrlyxochlatTrys and with axile placentationto unilocular with freeor basalplacentation hasbeen Scaphocklamys,For example, flowers are borne reported singly inA{lyxochlanrys vs. bornein cincinni; corol& Nagamasu 2000), (Sakai latube with a grooveenveloping the style insidevs. Our molecular phylogeneticanalyses suggest the aMnity of Adyxochlaniys to Distichochlanrys Iackngsuch a groove;labellum concave vs. bilobed and Scophochlanqys (Figs. 3-6).Adyxochlanu,s is or entire, but never concave; anthers versatile vs. sister to the Distichochlannysand Scophochlanrys adnate, Distichochlanrys isunique and distinctfrom in that ithas tubular bracteoles clades as shown by theITS dataset analysis (Figs.?Lfyxochlanrys and 3-4),although the relationships among arranged bracts (Newman 1995). thesethree distichously Furthermore, DistichochlaTayshas adnate anthers subciades are unresolved in both the MP and lackingspurs, producesflowersin cineinni, and Bayesiantrees.Further,bothDistichochlanop?s and and the cembined ITS and matiZf region sequence = = = NII-Electronic Library Service The JapaneseSociety The Japanese Society for for Plant Plant Systematics Systematics 'rAKANO & NAGAMASU: May 2007 29 ndyxochlanv,s, gen.nov. 7Llyxochlainys Doyle, J,J,& J,L, Doyle.1987.A rapid DNA isolation has a flat, bilobedlabellum, while of freshleaftissue, procedure forsmall quantities hasversatile anthers with longspurs, producesa sinPhytochem. Bull. 19: 11-15. and hasa concave, entire labelgleflowerperbract, Fanis, J, S,,M. Kallerajo, A. G. Kluge & C. Bult.1995, lum, Comparison of key characters between Constructing a significance test fbrincongruence, Myxochtamys and the other genera is shown in Syst Biol.44:570-572. Table 1,Among these characters, the versatile an Felsenstein, J.1985.Confidence1imits on phylogenies: approach using the bootstrap,Evolution 39: 783anthers are rather rare in theZingiberaceaeand are 791, found only in five generabesidesMyxochtanv,s: Ha]1,T, A. 1999. BioEdit: a user-frielldly biological Camptandra, Cautleya,Curcuma, f]beracautlaya, sequence alignment editor and analysis program fbr and Roseoea(Larsen et al. 1998), The groovewithWindows 95f98fNT.Nucl. Acids Symp, Ser.41: inthecorolla tube isalso uncommon and so faris 95-98. known only in themonotypic Slddiochilusand a few Huelsenbeck,J. P. & F. Ronquist.2001. MRBAYES: 1980). Bayesian inferenceof phylogeny.Bioinformatics species of Heclychium and Zngiber (Smith 17:754-755, Myxochlamys might be placed in ScaphoKress, W. J,,L. M. Prince& K, J.Williams. 2002. The ehlantys as a peculiarspecies, but we hesitated to of the gingers phylegeny and a new classification expand the circumscription of the latter genus evidence from molecular data. (Zingiberaceae): becauseofpoor support from molecular phylogeAmer. J,Bot, 89: 1682-1696, netic analyses and such significant morphological A.-Z.Liu,M. F.Newman & , and differences, Furtherstudy of toxochlamys related genera isrequired beforefirm conclusions on itsplacementcan be reached, gratefu1 to Pro£ HiroshiOkada, Direetorof Betanical Gardens, Facultyof Science,Osaka City We are mo]ecular phylegeny ofAipinia Li, 2005. [lhe Q,-J. a (Zingiberaceae): Amer, J, po)yphyletic genusof gingers. Bot.92: 167-178, Larsen,K.,J.M. Loek,H. Maas & P.J.M. Maas. 1998. Kubitzki, K. (ed.), The families Zingiberaceae, 1}i: and generaofvascular plants.4 : 474-495.SpringerVerlag,Berlin. . & M, F, Newman. 2001. A new species of Distichochlanrysfrom Vietnam and some observacomplex and Universityforinviting us to panicipate in the Muller Range expedition. We also wish to express our cordial LIPI, Scientific Academy of thanks to Dr.Dedy Darnaedi, tions on genericlimits in Hedychieae (ZingiIndonesia, lndenesia, and Dr. Irawati, Directorof the Nat. Hist. Bull. Siam Soc, 49: 77-80. beraceae), Bogor Botanic Garden,LIPI, Indonesia, fbrtheir kind A, Liston, & J.W. Kadereit 1995.ChloroplastDNA assistance duringthe fieldtripto CcntralKalimantan, and long distance disevidence fbrintregression and Ms, Titien Praptosoewiryo who supported our fieldin the desert annual Senecio persal .flavus wotk. Thanks are also due to the directors and curators of Plant,Syst.Evol.197:33-41. (Asteraceae). BO, E, and K fbrailowing us to examine herbarium Manos, R S, & K. P. Steele. 1997,Phylogcnetic analyses specimens in their care. Dr, Resalind Searle provided of Hamamelididae bascdon plastid critical comments on an earli ¢ r vcrsion of the manusequence data. Amer. J.Bot, 84:1407-1419. script. Thisgtudy was partlysupported by research grants McNeill,J,,F. R, Barrie,H. M. Burdet, Vl Demoulin,D, from the Mitsubishi Foundation in the NaturalSciences L, Hawksworth, K. Marhold, D. H. Nicolson,J. and the Japan Societyfbr the Promotion of Science Prado, P. C. Silva,J.E. Skog,J.H, Wiersema & N. TropicalBio-resources ResearchFund. (JSPS) J, Turland (eds,), Inteniational code of botanical nomenclature Code), A, R, G. Gantner (Vienna References Verlag,Ruggell,Liechtenstein. M611er,M. & Q, C. B. Cronk. 1997. 0riginand relabased on & P,F.Yeo. 1985.The tionships of Sainipaulia Dahlgren,R. M, T.,H. T. Clifft,rd (Gesneriaceae) DNA internal transcribed spacer of the monocotyledons, Springer,New ribosomal families (ITS) Ybrk. sequences, Amer. J. Bet 84: 956-965. "higherT7 NII-Electronic Library Service The JapaneseSociety The Japanese Society for for Plant Plant Systematics Systematics 30 APG XioL58 Newman, M. E 1995, Distichochlam.vs, a new genus basedon ITS sequences of thenuclear rRNA cistron. from Vietnarn.Edinburgh J,Bot,52: 65-69, in:Wilson,K, L. & D. A. Monison (eds.), MonoNgamriabsaku1, C.,M. E Newman & Q,C, B. Cronk, cots-systematics and evolution, pp,71O-718.CSIRO 2001. The phylogeny of tribe Zingibereae Publishing,Collingwood,Australia. based on ITS (nrDNA) and trnL-F Smmb, R. M, 1980.A new genusofZingiberaceae fromN (Zingibcraceae) Burma. Notes Roy, Bot, Gard.Edinburgh38:13-17, (cpDNA)scquences. Edinburgh J, Bot. 60: 483507. Steele, K. R & R, Vilgalys,1994.Phylogenetic analysis of Pedersen,L, B. 2004. Phylogenetic analysis of the subPolemeniaceaeusing nucleotide sequences ef the familyAlpinioideae(Zingiberaceae), matK. Syst. Bot. 19: 126-142. particularly plastid gene Etlingera Gisckc,basedon nuclear and plastid DNA. Swofford,D. L. 2003, PAUP": phylogeneticanalysis Plant Syst.Evol.245:239-258, using parsimony ("and other methods), Version Posada, D. & K. A. CTandall, 1998,MODEL;I'EST: test4.0blO.SinauerAssociates, Sunderland. ing the modcl ofDNA substimtion. Bioinfbrmatics Takano,A, & H, Okada. 2002. Multipleoccurrences of 14:817-818, triploid formationin Globba (Zingiberaceae) from Rangsiniji, A.,M. F.Newman & Q.C. B. Cronk,2000,A molecular evidence. PlantSyst Evol.23Oi 143-159. study of the infrageneTie classification ofAlpinia Thompson, J.D., D. G. Higgins & T. J,Gibson.I994. based on the ITS region ofnuclear CLUSI[ALW: improvingthe sensitivity ofprogres(Zingiberaceae) rDNA and the trnL-F spacer ofchloroplast DNA, Ih: sive multiple sequence alignment through sequenee K. L. Wilson& D. A. Monison (eds.) Monocots-sysweighting, position-specific and weight gap penalties ternatics and evolution, pp. 695-709. CSIRO matrix choice. Nucl.Acids Res, 22: 4673-4680. Publishing,Collingwood, Australia. Williarns,K. J.,W. J.Kress & P. S. Mamos, 2004, Thc Ronquist,F. & J. R Huelsenbeck. 2003, MRBAYES 3: phylogenM evolution, and classification ofthe genus Bayesianphy]ogenetic inferenceunder mixed modGiobba and tribe Globbeae (Zingiberaceae): els. Bioinformatics 19: 1572-1574, appendages do matter, Amer. J.Bot.91: 100-114. Sakai,S. & H, Nagamasu. 2000. Systematicstudies of Xia,Yl M., W. J. Krcss & L. M. Prince.2004.A phyloBornean Zingiberaceae: III,7hmijia:a new genus. ofAmomum genetic analysis (Alpinioideae: EdinburghJ,Bot.57:245-255, Zingiberaceae) using ITS and mati< DNA sequence Searle, R. J. & T, A, J.Hedderson.2000.A preliminary data. Syst,Bot,29:334-344. phylogeny ofthe Hedychicae tribe(Zingiberaceae) ReceivedNbvember 2, 2006; accqpted.lanuar:y 10,2007 NII-Electronic Library Service The JapaneseSociety The Japanese Society for for Plant Plant Systematics Systematics May 2007 TAKANO Appendix 1.Genbank accesien numbers and & NAGAMASU: youcher specimens A45ixochlamys, gen.nov. used 31 in this study. Species ITS Aipiniablq;?harocalyx K. Sehum, AF47S709AF47S713AYT423S6AY769828AY352e12AY424744AY42474SAF47S725AF47S727AF47g730AY47873 AF478809AF478813 Kresset aJ, 2002 matK Aipiniaelegans K.Schum. AipiniatonkinensisGagncp. Referenees,iVouchcr Kresset aL 2e02 Kresset al. 200S AmomuntvillosumLour. AF47S824AY352042 Hanis et aL 200S(1TS),Kresset Amomum yunnanenseS, Q,Tong Xiaet aL 2004 BoesenbergiagelatinosaK.Larsen Ngainriabsakul et at. 2003 BoesenbergialongijZoraKuntze Ngamrtabsaku]et al. 2003 AF47Sg2SAF47S826AF47S83eAF47Sg31 Kresset al,2002 Boesenbenygi'apulchem'imaKuntze Boesenbergiarotunda(L,)Mansfi Kresset al. 2002 Kressetai, 2002 CanrptandraparvutaRicl1, CautohaenjleriasexicoJaK.Larscn Kresset CauiohaemnferiathaigandicaK.Larsen Ngamriabsakal et aL 2003 Cautieyagraeilis(Sm.)Dandy al, 2002 AF47S833AE478834AE47883S Kresset aL 2002 CautieyaspicataBaker Krcssetal.2002 Krcsset al. 2002 Coi'nuimeiqntleriaaut'antijioraJ,Mood&K,Larsen Cornukaemrv2irialongipetjolataJ.Mood&K.Larsen Curcuma bicolor J.Mood & K. Larsen Ngamriabsaku1et AF47S837 al. 2003 Ki'ess et al.2002 Curcumaecomata Craib CuffcumaharmandiiGagnep, Ngamriabsakulet al, 2003 CurcumaparvgeoraWall. tv'garuriabsaku1 et at. 2003 Kressetal. 2002 CITS) /Cuoet Ngamriabsaku]et CurcumaroscoeanaWal1. CurcumarubeseensRoxb. CurctfmathoreiiiGagnep, AB047741 Di,stichochlantys citrea M. F. Newman 2 Disiichochlanv,ssp.KressO1-6g4g al, unpublished (matK) Kressetal,2002 Kressetai.2002 Kresset al, 2002 NgamriabsakLiletaL2003 AF47S844 Eiettarialongituba(rudl.)Holtturn Eiettar'iopsis kerbyiR, M. Sm. (K.Schurn.)B. L. Burtt& R. M, Srri, Krcsset Sakaiet aJ. 2002 al. unpublished AF47SS45AF47gS47 Pedersen2004 (ITS) ,i Kresset Kresset al.2e02 Gagnepainia iJarmandii K. Schum. al. 2002 (matK) Williains et al, 2004 GagnepainiathorelianaK.Schum. AF47gS51 Globba taiangensis A, Takano et H, Okada filrnmpaatbijleraK.Larsen&J,Mood Hbd.vchiumlongicornuium Griff ex Baker Kressetal, 2e02 Williaills et aL 2004 AF47S855AF47SS60AF47SS61AY341090AF47Sg64 Kressetai.2e02 Kresset aL 2002 Hed),chiumvjllos"mWall, Kresset al, 2002 Williamsetal.2004 K, Schum, Kresset KlteiptriaelegansWatl, Kkeemnferiaparvt"of'aWal]. aL 2002 Ngamriabsaku1 etal. 2003 AB232052AF47gg68AF47SS69AB269791 Searle&HeddersonZOOO KlrtenrpteriarotundaL. Kaemnferiasp,kresb'9S-62SP Alyxochtano,s mullerensis A,Takano& 2003 AF47S841AF47SS42AF47S843 Krcsset al. 2002 Verma1 Curcumorpha longgeora(WalL) A, S, Rao & D. M. Vermu 2 Disiichochiantys citrea M, F.Ne-man 1 llbmiorchis fhoderrhachib' th't:henia glau:aWa[t. aL Ngamriabsakul et al, ?O03 Curcumotpha longijlora A. S. Rao & D. M, (Wa]l.) Eiettariopsis stenostPhon 2002 (matK) aL KressetaL 2e02 Kressetai.2002 Nagam, Thisstudy: H, Okada et al.KIL671 NgnmiTiabsakn1 et aL 2003 Paracazattq,a hAatii R, M. Sm. PommereschealackneriWittm, AF478877AF47SH78AF47S880AF47S88S Kresset al, 2002 P.Frgephytlune T. L. Wu & Z. Y. Chen )ninnanensis 1{eneainvia battenbejgiana Cummins ex Baker RkynckanthusbeesianusW.W.Srn, RoscoeabhutttnicaC.Ngamriabsaku1 RoseoeacautteoidesGagnep. Roh'coeahumeanaBalf.f, & W. W. Sm Ros'coeapurptfreaSm. thaphochtantvsbiloba(rudl,)HoltLam Kressetal. 2002 Kresset AF478887 2002 al. 20e3 Kresset at,2002 Wood et al. 2000 AF47gg88AY4788S9 Kresset al.2002 Krcsset al,2002 Scaphochlam.vsbiloba(Ridl.)Holttum2 Sbaphochlam.vslaunstleri(Bakei}Holttum aJ, Krcsset aL 2002 Ngamriabsaku]et Wood AY478890 et aL 2000 Kresset al.2002 NII-Electronic Library Service The JapaneseSociety The Japanese Society for for Plant Plant Systematics Systematics 32 APG Nio1.58 Appendix 1.continued, Species ITS SbcQvhochtafayskunstteri(Baker)Holttum AJ3SS2S7 AJ3gS2S6 Scaphochlan-'sobcordataP.Sirirugsu&K.Larsen Siphonochilusaethiopicus(Schweinf,)B,L,Burtt matK ReferenncesiVoucher Searle&1･Ledderson2000 Searle&Hedderson2000 M478893AF47g894AF478S95AF478896AF478897AF47gB9g Kresset aL 2002 Kress et aL 2002 Sij]honochilusdecoruh'([lruten)Lock AF478792 AF478793 Siphonochilusldrldi{Hook,)B,L.Burtt AF478794 Kresset Sinithatris supraneanae W. J.Kress&K, Larsen involttcratus ex Baker)R. M. Srn. Stahiiantkus (King AF478795 AF478796 Kresset al.2002 7lamijiafugellarisS.Sakai&Nagarn. AF478797 Z'ngibercoJorainmN.E,Br, AF414498 Kresset aL 2002 Pedemsen20pa CrTS) Znigber elij)tieum (S, Q.Tong & Y. M. Xia)S,Q,Tong & Y. M. Xia AF478799 ZingibergramineumNoronha Z'ngiberjunceumGagnep. AF478800 AY424774 Zingiberlvngipedunculatumrudl, AB0972S4 ft'ngibersuiphureumBurkillexl,Iheilade Z'ngiberwrayiiPrainexRidl, AIF478801 AF478802 Kresset AF4789elABOS8799 Kress et aL aL al. 2002 2D02 2002 Kresset aL 2002{ITS) iTamura et al. 2004(matK) Ngamriubsukuletal.2003 Sakaietal, unpublishcd AF4789e4AF478905 Kresset aL 2002 Kresset ai, 2e02 NII-Electronic Library Service