PhytoKeys 75: 31–55 (2016) Borneocola (Zingiberaceae), a new genus from Borneo doi: 10.3897/phytokeys.75.9837 RESEARCH ARTICLE http://phytokeys.pensoft.net A peer-reviewed open-access journal 31 Launched to accelerate biodiversity research Borneocola (Zingiberaceae), a new genus from Borneo Yen Yen Sam1, Atsuko Takano2, Halijah Ibrahim3, Eliška Záveská4, Fazimah Aziz5 1 Forest Research Institute Malaysia, 52109 Kepong, Selangor, Malaysia 2 Museum of Nature and Human Activities, Hyogo 6 chome, Yayoigaoka, Sanda, Hyogo 669-1546, Japan 3 Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia 4 Institute of Botany, University of Innsbruck, Austria 5 Department of Aquatic Science, Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia Corresponding author: Yen Yen Sam (samyen@frim.gov.my) Academic editor: Pavel Stoev | Received 12 July 2016 | Accepted 9 November 2016 | Published 29 November 2016 Citation: Sam YY, Takano A, Ibrahim H, Záveská E, Aziz F (2016) Borneocola (Zingiberaceae), a new genus from Borneo. PhytoKeys 75: 31–55. doi: 10.3897/phytokeys.75.9837 Abstract A new genus from Borneo, Borneocola Y.Y.Sam, is described here. The genus currently contains eight species previously classified as members of the Scaphochlamys Baker. The finding is based on the results of the morphological and molecular studies of Scaphochlamys throughout its geographical range and its closely allied sister groups, Distichochlamys M.F.Newman and Myxochlamys A.Takano & Nagam. Borneocola is nested within the tribe Zingibereae and its monophyly is strongly supported by both ITS and matK sequence data. The genus is characterised by several thin, translucent and marcescent floral bracts, absence of coloured streaks on the labellum and capitate stigma with two dorsal knobs. The genus is distributed in northwest Borneo and all species are very rare and highly endemic. Keywords Distichochlamys, Myxochlamys, Scaphochlamys, morphology, phylogeny, taxonomy Copyright Yen Yen Sam et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 32 Yen Yen Sam et al. / PhytoKeys 75: 31–55 (2016) Introduction Southeast Asia is the centre of diversity for the family Zingiberaceae. Here, new taxa are continuously being discovered and named, both at the generic and specific levels. Several of the recent discoveries were further supported by the phylogenetic analyses which give a better understanding of the evolutionary relationships within the family (Kress and Larsen 2001; Kress et al. 2010; Leong-Škorničková et al. 2011). During the revision of the genus Scaphochlamys throughout its entire geographical range by the first author, some distinctive morphological traits were observed in several Bornean species, suggesting they might represent a separate group from the Peninsular Malaysian taxa. This hypothesis was confirmed by the phylogenetic analyses which are presented here and the eight species previously included in the genus Scaphochlamys are recircumscribed in this paper as a new genus, Borneocola Y.Y.Sam. The genus Scaphochlamys was described by Baker (1892) in the Flora of British India with Scaphochlamys malaccana Baker from Mt. Ophir (now known as Gunung Ledang), Peninsular Malaysia, chosen as the type species. Holttum (1950) carried out the first comprehensive revision of the genus in which he recognised 19 species, all of which were recorded in the peninsula. When Smith (1987) reviewed the tribe Hedychieae in Borneo, she applied the generic delimitation defined by Holttum and recognised five Scaphochlamys species in Borneo. Out of the five, S. polyphylla and S. petiolata were formerly placed in the genus Haplochorema K.Schum. Sakai and Nagamasu (2006) discovered that H. gracilipes K.Schum. also have the characteristics of Scaphochlamys and effected the transfer. Recent years have seen a surge in the new species discovered from Borneo bringing the total number of Bornean Scaphochlamys to 14 (Poulsen and Searle 2005, Meekiong et al. 2011, Ooi and Wong 2014; Meekiong 2015). Distichochlamys M.F.Newman and Myxochlamys A.Takano & Nagam. are sister genera to Scaphochlamys with several unique characteristics clearly separating them from Scaphochlamys (Newman 1995, Searle and Hedderson 2000, Kress et al. 2002, Ngamriabsakul et al. 2004, Takano and Nagamasu 2007). However, the distinction, based on morphological characters, became ambiguous as several taxa described recently exhibit exceptions to the usual generic characters. For example, S. calcicola A.D.Poulsen & R.J.Searle, a species named in 2005 from Sarawak, has a distichous inflorescence, a distinguishing character for the genus Distichochlamys M.F.Newman. Larsen and Newman (2001) also reported another Scaphochlamys species with a distichous inflorescence from north Peninsular Malaysia. A current study on the morphology of Scaphochlamys also revealed that some species display the characteristics of Distichochlamys and Myxochlamys. To test the validity of the current generic concept of Scaphochlamys and closely related genera Distichochlamys and Myxochlamys, we have examined their relationship by utilising ITS and matK markers together with the analysis of the morphology across these genera. Borneocola (Zingiberaceae), a new genus from Borneo 33 Materials and methods Morphological study The morphological study was based on living plants in the forest, cultivated plants in the nursery of the Forest Research Institute Malaysia and specimens in the herbaria of AAU, BKF, C, FI, E, K, KEP, KLU, PSU, SAN, SAR and SING. A total of 372 herbarium specimens were examined in this study which includes 29 Scaphochlamys species and four Borneocola species (the types of another four Borneocola species were not yet deposited in the herbaria).The morphological characters examined in the study were habit; position of the rhizome, thickness and colour; height of leafy stem, its base (whether swollen to form a bulbous base); distance between leafy stems; characters of bladeless sheath such as colour, indumentum, number and length; ligule length, indumentum and shape; petiole length, indumentum, whether channelled or rounded in cross section; number of leaves per leafy stem; lamina colour on both surfaces, size, shape, venation, texture, indumentum, apex and base; length of the inflorescence and infructescence, arrangement of the floral bracts on the rachis, characters of floral bracts and bracteoles (colour, indumentum, texture, shape); size, colour and shape of calyx, floral tube, corolla lobes, labellum, staminodes, stamen, ovary. DNA extraction, amplification and sequencing Fresh leaves from the cultivated plants or silica-dried materials from plants collected in the field were used for genomic DNA extraction. For the ITS, the genomic DNA was extracted using the DNeasy Plant Mini Kit (Qiagen, Valencia, California, USA) following the manufacturer’s protocol. Two primers, ITS 5P (5’-GGAAGGAGAAGTCGTAACAAGG-3’) and ITS 8P (5’-CACGCTTCTCCAGACTACA3’) (Moller and Cronk 1997) were used to amplify the ITS region during the polymerase chain reaction (PCR). The thermal cycle of PCR for the amplification of the ITS sequences is initial denaturation at 94°C for 2 minutes, 40 cycles of denaturation at 94°C for 30 seconds, primers annealing at 48°C for 2 minutes, an extension at 72°C for 45 seconds and final extension at 72°C for 7 minutes. The PCR products were then purified using MinElute Gel Extraction Kit (Qiagen, Valencia, California, USA). For the matK, the protocols for DNA extraction, condition, purification and DNA sequencing were described previously by Takano and Nagamasu (2007). The PCR and sequencing primers for matK (cpDNA) were TA-240f (5’-GGGAAA GGATGGGGTCTCCCG-3’), TA-150r (5’-CTCAAGGAGTTTTGTGGTTC-3’), TA-470F (5’-CCCTCTCCCGTCCATATGGA-3’) (all three were designed in the present study), matK8 (Steele and Vilgalys 1994), m5r (Kress et al. 2002), matK8r (Ooi et al. 1995), trnK2621 (Liston and Kadereit 1995), TA-10F, TA-05R, TA-02F and TA-02R (all from Takano and Okada 2002). 34 Yen Yen Sam et al. / PhytoKeys 75: 31–55 (2016) Sequence alignment and phylogenetic analysis Raw sequence data were assembled and edited manually using BioEdit software ver. 7.2.5 (Hall 1999). DNA sequences were aligned with the CLUSTALW 1.83 software package, with default settings and multiple alignments (Thompson et al. 1994). Alignments of the matK sequences of cpDNA and the ITS sequences of nrDNA were combined. Gaps were deleted. A total of 100 individuals including 54 taxa of Scaphochlamys and allied species were used. The three Siphonochilus species were used as an outgroup (Kress et al. 2002). Materials, accession numbers for the sequences, vouchers and references to the literature are presented in Table 1 at the end of this paper. Three datasets which comprise ITS, matK and ITS+matK combined, each containing 82, 78, and 61 taxa, were constructed. These three datasets were analysed using three methods: maximum parsimony, maximum likelihood and Bayesian analysis. A maximum parsimony (MP) analysis was performed with MEGA 6 (Tamura et al. 2013). Heuristic searches were conducted with RANDOM addition, SPR branch swapping and MULPARS options. Support for each branch was estimated with a bootstrap analysis, with 1000 replications (Felsenstein 1985), in a heuristic search with RANDOM addition and TBR branch swapping. The maximum likelihood (ML), based on the Tamura-Nei model (Tamura and Nei 1993), was also determined with MEGA 6 (Tamura et al. 2013). Neighbor-Join and BioNJ algorithms were applied to a matrix of pairwise distances estimated with the maximum composite likelihood approach; then, the topology that had the best log likelihood value was selected. Bootstrap analysis under the MP criterion was conducted with “fast” stepwise, addition searches, with 1000 replicates. In addition, a Bayesian analysis was carried out with MrBayes software ver. 3.1.2 (Huelsenbeck and Rohnquist 2001; Rohnquist and Huelsenbeck 2003). The best fitting substitution model (the GTR+G model for nrDNA datasets, the GTR+G model for cpDNA datasets and the GTR+I+G model for cpDNA+nrDNA datasets) was selected for Bayesian analysis based on a series of hierarchical likelihood ratio tests, implemented in MrModeltest software ver. 2.3 (Nylander 2004). The analysis was performed with the selected model and two simultaneous runs of two million generations with four chains, sampling every 100 generations. Each analysis reached stationarity (i.e. when the average standard deviation of split frequencies between runs was ≤ 0.01) well before the end of the run. Burn-in trees were discarded and the remaining trees and their parameters were saved. A 50 % majority rule consensus tree was constructed. The results of the Bayesian analysis were reported as the posterior probability (PP; Huelsenbeck and Rohnquist 2001), which is equal to the percentage of phylogenetic trees sampled when a given clade was resolved. Only PP scores above 50 % are shown. Borneocola (Zingiberaceae), a new genus from Borneo 35 Table 1. List of accession details, vouchers and references used in the phylogenetic analyses. No Subfamily Tribe 1 2 Alpinia blepharocalyx K.Schum. Alpinia elegans K.Schum. 3 Alpinioideae 4 Link 5 Species Amomum villosum Lour. Alpinieae A.Rich. matK AF478709 AF478809 AF478713 AF478813 – AF478824 Amomum yunnanense S.Q.Tong AY352012 – Elettariopsis kerbyi R.M.Sm. AF414496 AF478845 Renealmia battenbergiana Cummins ex Baker Siphonochilus aethiopicus 7 (Schweinf.) B.L.Burtt Siphonochileae Siphonochileae Siphonochilus decorus 8 W.J.Kress W.J.Kress (Druten) Lock Siphonochilus kirkii 9 (Hook.) B.L.Burtt Tamijioideae Tamijieae 10 Tamijia flagellaris S.Sakai & Nagam. W.J.Kress W.J.Kress 11 Gagnepainia thoreliana K.Schum. Zingiberoideae Globbeae 12 Hemiorchis rhodorrhachis K.Schum. Haask. Meisn. 13 Mantisia wengeri C.E.C.Fischer 14 Boesenbergia pulcherrima Kuntze 15 Boesenbergia rotunda (L.) Mansf. 16 Borneocola biru (Meekiong) Y.Y.Sam Borneocola calcicola 17 (A.D.Poulsen & R.J.Searle) Y.Y. Sam 18 Borneocola sp. FRI 50295 19 Borneocola sp. S 99106 Borneocola stenophyllus 20 (Ooi & S.Y.Wong) Y.Y.Sam Borneocola petiolatus 21 (K.Schum.) Y.Y.Sam 22 Borneocola reticosus (Ridl.) Y.Y.Sam 23 Camptandra parvula Ridl. 24 Zingiberoideae Zingibereae Caulokaempferia saxicola K.Larsen Haask. Meisn. 25 Cautleya gracilis (Sm.) Dandy 26 Cautleya spicata Baker Cornukaempferia aurantiflora 27 J.Mood & K.Larsen Curcuma bicolor 28 J.Mood & K.Larsen 6 29 Curcuma roscoeana Wall. 30 Distichochlamys citrea M. F. Newman Distichochlamys citrea M. F. Newman 2 31 ITS References/ Voucher Kress et al. 2002 Kress et al. 2002 Harris et al. 2000 (ITS), Kress et al. 2002 (matK) Xia et al. 2004 Pedersen 2004 (ITS)/Kress et al. 2002 (matK) AF478779 AF478880 Kress et al. 2002 AF478792 AF478893 Kress et al. 2002 AF478793 AF478894 Kress et al. 2002 AF478794 AF478895 Kress et al. 2002 AF478797 AF478898 Kress et al. 2002 AF478752 AF478851 Kress et al. 2002 Williams et al. AY339706 AY341090 2004 – AF478871 Kress et al. 2002 AF478725 AF478825 Kress et al. 2002 AF478727 AF478826 Kress et al. 2002 – LC148403 FRI 50290 (KEP) LC148062 LC148380 FRI 50290 (KEP) LC148085 LC148404 FRI 50295 (KEP) LC148086 LC148405 S 99106 (SAR) LC148084 LC148400 FRI 50288 (KEP) LC148075 LC148395 FRI 50291 (KEP) LC148078 AF478730 AY478732 AF478734 AF478735 LC148398 AF478830 AF478831 AF478833 AF478834 FRI 50294 (KEP) Kress et al. 2002 Kress et al. 2002 Kress et al. 2002 Kress et al. 2002 AF478736 AF478835 Kress et al. 2002 AF478737 AF478837 Kress et al. 2002 Kress et al. 2002 (ITS)/Cao et al. unpublished (matK) Ngamriabsakul et AY424757 – al. 2004 Ngamriabsakul AB552946 AB552951 24 (E) AF478739 AB047741 36 No Yen Yen Sam et al. / PhytoKeys 75: 31–55 (2016) Subfamily Tribe 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 Zingiberoideae Zingibereae 50 Haask. Meisn. 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 References/ Voucher Adele Smith 18 Distichochlamys sp. AS18 AB552947 AB553309 (E) Distichochlamys sp. Kress01-6848 AF478745 AF478844 Kress et al. 2002 Haniffia albiflora K.Larsen & J.Mood AF478756 AF478855 Kress et al. 2002 Hedychium longicornutum AF478761 AF478860 Kress et al. 2002 Griff. ex Baker Hedychium villosum Wall. AF478762 AF478861 Kress et al. 2002 Hitchenia glauca Wall. AF478765 AF478864 Kress et al. 2002 Searle and Kaempferia parviflora Wall. – AB232052 Hedderson 2000 Kaempferia rotunda L. AF478767 AF478868 Kress et al. 2002 Kaempferia sp. Kress98-6289 AF478768 AF478869 Kress et al. 2002 Myxochlamys mullerensis Takano and AB245522 AB269791 Nagamasu 2007 A.Takano & Nagam. Nagamasu 8274 Myxochlamys nobilis Nagam. ined. AB552948 AB553310 (BO, KYO) Pommereschea lackneri Wittm. – AF478877 Kress et al. 2002 Pyrgophyllum yunnanense AF478777 AF478878 Kress et al. 2002 (Gagnep.) T.L.Wu & Z.Y.Chen Rhynchanthus beesianus W.W.Sm. AF478784 AF478885 Kress et al. 2002 Roscoea cautleoides Gagnep. AF478736 AF478887 Kress et al. 2002 Roscoea purpurea Sm. AF478787 AF478888 Kress et al. 2002 Scaphochlamys abdullahii LC148054 – FRI 44375 (KEP) Y.Y.Sam & Saw Scaphochlamys abdullahii LC148055 LC148374 FRI 50198 (KEP) Y.Y.Sam & Saw Scaphochlamys atroviridis Holttum LC148056 – FRI 68924 (KEP) Scaphochlamys baukensis Y.Y.Sam LC148057 – FRI 68955 (KEP) Scaphochlamys biloba (Ridl.) Holttum LC148059 – FRI 46606 (KEP) Scaphochlamys biloba (Ridl.) Holttum LC148081 – FRI 50224 (KEP) Scaphochlamys biloba (Ridl.) Holttum LC148083 – FRI 66331 (KEP) Scaphochlamys biloba AF478788 AY478889 Kress et al. 2002 (Ridl.) Holttum 1 Scaphochlamys biloba AF202416 – Wood et al. 2000 (Ridl.) Holttum 2 Scaphochlamys breviscapa Holttum – LC148377 FRI 50269 (KEP) Scaphochlamys breviscapa Holttum LC148060 LC148376 FRI 44984 (KEP) Scaphochlamys burkillii Holttum – LC148379 FRI 68928 (KEP) Scaphochlamys burkillii Holttum LC148061 – FRI 46504 (KEP) Scaphochlamys concinna Searle and AJ388283 – Hedderson 2000 (Baker) Holttum Scaphochlamys concinna LC148063 LC148381 FRI 50351 (KEP) (Baker) Holttum Scaphochlamys cordata LC148064 – FRI 44306 (KEP) Y.Y.Sam & Saw Scaphochlamys endauensis – LC148383 FRI 50243 (KEP) Y.Y.Sam & Ibrahim Scaphochlamys endauensis LC148080 – FRI 50218 (KEP) Y.Y.Sam & Ibrahim Scaphochlamys erecta Holttum LC148065 – FRI 44987 (KEP) Scaphochlamys grandis Holttum – LC148384 FRI47184 (KEP) Scaphochlamys grandis Holttum LC148066 LC148385 FRI 50171 (KEP) Species ITS matK Borneocola (Zingiberaceae), a new genus from Borneo No Subfamily Tribe 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 Zingiberoideae Zingibereae Haask. Meisn. 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 37 References/ Voucher Scaphochlamys johorensis Y.Y.Sam LC148082 – FRI 66566 (KEP) Scaphochlamys klossii (Ridl.) Holttum LC148067 LC148387 FRI 50238 (KEP) Scaphochlamys kunstleri AF478789 AY478890 Kress et al. 2002 (Baker) Holttum Anon C 8003 & Scaphochlamys kunstleri (Baker) AB552950 AB553312 C. Ngamriabsakul Holttum var. rubra C.K.Lim 25 (E) Scaphochlamys kunstleri (Baker) – LC148388 FRI 68926 (KEP) Holttum var.kunstleri Scaphochlamys kunstleri (Baker) LC148068 – FRI 68936 (KEP) Holttum var. kunstleri Scaphochlamys kunstleri var. speciosa – LC148389 FRI 68936 (KEP) C.K.Lim Scaphochlamys lanceolata LC148069 LC148390 FRI 50130 (KEP) (Ridl.) Holttum Scaphochlamys laxa Y.Y.Sam & Saw – LC148391 FRI 68961 (KEP) Scaphochlamys longifolia LC148070 LC148392 FRI 47065 (KEP) (Ridl.) Holttum Scaphochlamys malaccana Baker – LC148393 FRI 50203 (KEP) Scaphochlamys malaccana Baker LC148071 – FRI 50208 (KEP) Scaphochlamys minutiflora – LC148394 3175 Jenjitt.& K.Larsen Scaphochlamys obcordata Searle and AJ388286 – Hedderson 2000 P.Sirirugsa & K.Larsen Scaphochlamys oculata LC148072 LC148396 FRI 50262 (KEP) (Ridl.) Holttum Scaphochlamys pennipicta Holttum LC148073 – FRI 50261 (KEP) Scaphochlamys perakensis Holttum LC148074 – FRI 50214 (KEP) Scaphochlamys polyphylla LC148076 LC148397 FRI 50289 (KEP) (K.Schum.) B.L.Burtt & R.M.Sm. Scaphochlamys pusilla Y.Y.Sam LC148077 – FRI 50260 (KEP) Scaphochlamys rubromaculata – LC148399 FRI 50178 (KEP) Holttum Scaphochlamys rubromaculata LC148079 LC148378 FRI 50172 (KEP) Holttum Scaphochlamys samunsamensis MK 2344 – LC148401 Meekiong & Hidir (HUMS) Scaphochlamys sp.nov. – LC148402 FRI 68983 (KEP) Scaphochlamys sub-biloba – LC148375 FRI 75334 (KEP) (Burkill ex Ridl.) Holttum Scaphochlamys sylvestris LC148087 – FRI 50197 (KEP) (Ridl.)Holttum Scaphochlamys tenuis Holttum LC148088 – FRI 47233 (KEP) Schaphochlamys cf. gracilipes K.Meekiong – LC148386 (K.Schum.) S.Sakai & Nagam. (HUMS) Smithatris supraneanae AF478795 AF478896 Kress et al. 2002 W.J.Kress & K.Larsen Stahlianthus involucratus AF478796 AF478897 Kress et al. 2002 (King ex Baker) R.M.Sm. Zingiber gramineum Noronha AF478800 AF478902 Kress et al. 2002 Zingiber sulphureum AF478801 AF478904 Kress et al. 2002 Burkill ex I.Theilade Zingiber wrayii Prain ex Ridl. AF478802 AF478905 Kress et al. 2002 Species ITS matK 38 Yen Yen Sam et al. / PhytoKeys 75: 31–55 (2016) Results Phylogenetic analyses The ITS datasets for 82 individuals with 29 taxa of Scaphochlamys and 6 taxa of Borneocola contained 786 characters after alignment, which decreased to 769 after gaps were deleted; 319 of these were parsimony-informative. Likelihood analysis resulted in a ML tree with –lnL = 10438.212. Parsimony analysis produced three parsimonious trees with 1865 steps, a consistency index (CI) of 0.391 and retention index (RI) of 0.609. The ML, MP and Bayesian trees had similar topology; the ML tree is shown with bootstrap (BS) and MP-BS, and Bayesian Posterior Probability (PP) support in Figure 1 below. Scaphochlamys formed a well supported clade (ML-BS/MP-BS/BA-PP support, 84/87/1.00). Each Myxochlamys and Borneocola consisted of a well supported subclade and became sisters to each other and they also became sistersto the Scaphochlamys clade. Distichochlamys species formed a well supported subclade and became sister to the Myxochlamys + Scaphochlamys + Borneocola clade (ML-BS/MP-BS/BA-PP 99/100/1.00). The matK datasets for 78 individuals including 25 taxa of Scaphochlamys and 7 taxa of Borneocola contained 1,599 characters after alignment; 182 of these were parsimony-informative. Likelihood analysis resulted in a ML tree with -lnL = 5952.438. Parsimony analysis produced ten parsimonious trees with 557 steps, a consistency index (CI) of 0.613 and retention index (RI) of 0.080. The ML, MP and Bayesian trees had similar topology; the ML tree is shown with BS and MP-BS, PP support in Figure 2 below. Each of the two Myxochlamys species and seven Borneocola species formed a strongly supported subclade and became sisters to each other. Scaphochlamys became sister to them, but bootstrap or probability support was weak. The Distichochlamys species formed a well supported subclade, but all the genera that belong to subfamily Zingiberoideae became sisters to Scaphochlamys + Myxochlamys + Borneocola clade and not only to Distichochlamys. The combined ITS and matK datasets for 61 individuals including 13 taxa of Scaphochlamys and 6 taxa of Borneocola, resulted in 2,336 characters, 488 of these were parsimony-informative (Figure 3 below). Likelihood analysis resulted in a ML tree with -lnL = 16671.531. Parsimony analysis produced the most parsimonious trees with 2247 steps, a CI of 0.440 and a RI of 0.635. The ML, MP strict consensus and Bayesian trees had almost the same topology; the ML tree is shown with MP-BS, MLBS and BA/PP support in Figure 3. Two Myxochlamys species and six Borneocola species formed a strongly supported subclade each and became sisters to each other. Scaphochlamys became sister to them and the bootstrap or posterior probability support was moderate. Distichochlamys species formed a well supported subclade and became sister to Scaphochlamys + Myxochlamys + Borneocola clade. Borneocola (Zingiberaceae), a new genus from Borneo 39 Morphology The Borneocola and Scaphochlamys species look similar in their vegetative morphologies. They are mostly small-sized gingers without the conspicuous pseudostem, with one to several leaves arranged spirally and tightly on a very short stem at the base. So far, all the Borneocola species examined are unifoliate. Similarly, most of the Scaphochlamys species also bear one leaf except for several species which have leafy shoots composed of multiple leaves, for example, S. grandis, S. lanceolata, S. kunstleri, S. malaccana and S. minutiflora. The basal part of the leaves is covered with a few bladeless sheaths which are rather different for both groups in terms of their texture and colour. For Scaphochlamys, the sheaths are coriaceous, green, green with a red tinge or red and mostly persistent until the end of flowering (Figure 4A, B). On the other hand, the sheaths of Borneocola are thinner in texture with a lighter shade of green or brown. The thin sheaths normally dry up early (Figure 4C) and sometimes they are completely shredded during the time of flowering. The inflorescences of Borneocola and Scaphochlamys are terminal, stalked and consisted of few to many floral bracts. The differences lie in the characteristics of the floral bracts and flowers. Borneocola species have thin, translucent, early decaying and marcescent floral bracts. The colours of the bracts can be pink, pale brown, pale or light green (Figure 5A). On the contrary, the bracts of Scaphochlamys are coriaceous and sometimes hard in texture. They are usually green, green tinged red, red or reddish brown and remain fresh throughout the flowering (Figure 5B, C). Besides the characteristics of the floral bracts, the variegation on the labellum can give a quick guide to the two genera. Most Scaphochlamys have white flowers with a yellow median band and lilac, purple, red streaks or patches flanking the band on the labellum (Figure 5D, E). However, there is no such variegation on the labellum of Borneocola (Figure 5F). The whole labellum of Borneocola is pale pink, lilac, violet or white with a light yellow or greenish yellow median band. Both Borneocola and Scaphochlamys have a long slender floral tube which is mostly puberulent externally in Borneocola (except for B. calcicola) but glabrous for Scaphochlamys. Another marked difference observed is in the stigma shape. Scaphochlamys has a funnel-shaped or beak-like stigma (Figure 6A, B) while it is almost oblate with two dorsal knobs in Borneocola (Figure 6C). Taxonomic treatment Borneocola Y.Y.Sam, gen. nov. urn:lsid:ipni.org:names:77158811-1 Diagnosis. Similar to Scaphochlamys and Myxochlamys. Borneocola has thin, translucent and marcescent floral bracts, absence of coloured streaks on labellum and two dorsal knobs on the stigma versus the coriaceous and persistent floral bracts, coloured streaks 40 Yen Yen Sam et al. / PhytoKeys 75: 31–55 (2016) Scaphochlamys atroviridis FRI68924 Scaphochlamys grandis FRI50171 Scaphochlamys erecta FRI44987 99/100/1.00 Scaphochlamys abdullahii FRI50198 19 Scaphochlamys breviscapa FR44984 Scaphochlamys johorensis FRI66566 Scaphochlamys rubmaculata FRI50172 70 74 Scaphochlamys cordata FRI44306 Scaphochlamys malaccana FRI50208 1 Scaphochlamys endauensis FRI50218 Scaphochlamys baukensis FRI68955 Scaphochlamys sylvestris FRI50197 Scaphochlamys abdullahii FRI44375 29 Scaphochlamys tenuis FRI47233 60/78/0.98 Scaphochlamys perakensis FRI50214 Scaphochlamys biloba2 Scaphochlamys biloba1 Scaphochlamys biloba FRI50324 --/--/1.00 Scaphochlamys biloba FRI46606 59/82/0.96 84/87/1.00 Scaphochlamys biloba FRI66331 71/95 Scaphochlamys kunstleri var. rubra Scaphochlamys kunstleri var. speciosa FRI68936 88/97/1.00 Scaphochlamys kunstleri var. kunstleri 54 Scaphochlamys obcordata 51/72/87 Scaphochlamys malaccana FRI50224 75/74/0.54 93/84/0.5 Scaphochlamys lanceolata FRI50130 Sarawak 70/41/-Scaphochlamys polyphylla FRI50289 Scaphochlamys klossii var. klossii FRI50238 20/--/-92/61/-Scaphochlamys pennipicta FRI50261 Scaphochlamys concina FRI50351 Scaphochlamys oculata FRI50262 Scaphochlamys pusilla FRI50260 71/59/-Scaphochlamys concina Scaphochlamys sp. FRI46504 Scaphochlamys longifolia FRI47065 85/96/1.00 58/82/1.00 20 Zingiber gram ineum Vietnam 99/99/1.00 Borneo K aem pferia sp. Kress98-6289 Zingiber wray ii Peninsular Malaysia + Borneo K aem pferia rotunda 100/100/1.00 31/15/-- Borneocola Distichochlamys 57/68/0.99 65/83/1.00 Myxochlamys M yxochlamys mullerensis Myxochlamys nobilis Borneocola sp. FRI50295 Borneocola calcicola FRI50290 59/66/0.71 Borneocola petiolatus FRI50291 70/77/1.00 Borneocola reticosus FRI50294 97/96/1.00 Borneocola stenophyllus FRI50288 Borneocola sp. S99106 99/100/1.00 Distichochlamys sp. Kress01-6848 Distichochlamys citrea 90/99/0.99 Distichochlamys citrea2 55/89/-Distichochlamys sp. AS18 Scaphochlamys 100/100/1.00 78/80/1.00 Zingiber s ulphureum B oesenbergia rotunda Cornuk aem pferia aurantiflora 16/35/-- B oesenbergia Haniffia albiflora pulc herrim a Rhy nc hanthus bees ianus 96/99/1.00 Cautleya grac ilis Cautleya s pic ata Ros c oea c autleoides 98/98/1.00 Ros c oea purpurea 93/92/1.00 Hedychium longic ornutum Hedychium villos um 81/76/1.00 100/100/1.00 G agnepainia thoreliana Hem iorc his rhodorrhac his 99/100/1.00 Cam ptandra parvula P y rgophy llum y unnanens e Caulok aem pferia s ax ic ola Curc uma bicolor 47/59/-- S m ithatris s upraneanae S tahlianthus involucratus 100/99/1.00 Curc uma ros c oeana 83/87/1.00 84/92/0.95 Hitc henia glauc a A lpinia elegans 97/99/1.00 A m omum yunnanensis A lpinia blepharoc aly x E lettario ps is k erby i 83/89/1.00 Renealm ia battenbergiana 65/59/0.97 Tam ijia flagellaris S iphonochilus aethiopic us 81/93/1.00 100/100/1.00 96/98/0.97 S iphonochilus k irk ii S iphonochilus dec orus 0.05 Figure 1. Molecular phylogenetic analysis of the ITS sequence data by the Maximum Likelihood method. Numbers above branches indicate bootstrap values of ML and MP and posterior probability of Bayesian Analysis. Borneocola (Zingiberaceae), a new genus from Borneo 41 Scaphochlamys breviscapa FRI50172 Scaphochlamys grandis FRI50171 Scaphochlamys rubromaculata FRI50178 Scaphochlamys endauensis FRI50218 Scaphochlamys grandis FRI48184 Scaphochlamys malaccana FRI50203 77/82/0.97 80/85/1.00 65/- /0.98 77/83/1.00 Scaphochlamys endauensis FRI50243 Scaphochlamys polyphylla FRI50289 Scaphochlamys rubromaculata FRI50172 54/64/0.97 22/31/0.92 99/98/1.00 55/67/1.00 Scaphochlamys 49/49/0.65 29/--/0.89 Scaphochlamys breviscapa FRI50269 Scaphochlamys concina FRI50351 Scaphochlamys longifolia FRI47065 Scaphochlamys burkilli FRI68928 Scaphochlamys biloba FRI75334 Scaphochlamys sp. FRI68983 88/85/0.98 Scaphochlamys kunstleri var. speciosa FRI68936 Scaphochlamys kunstleri var. kunstleri Scaphochlamys kunstleri var. rubra Scaphochlamys kunstleri var. kunstleri FRI68926 Scaphochlamys abdullahii FRI50198 Scaphochlamys biloba Scaphochlamys breviscapa FRI44984 --/--/0.85 Scaphochlamys minutiflora 3175 Scaphochlamys sub-biloba FRI75334 Scaphochlamys laxa FRI68961 Sarawak Scaphochlamys cf. gracilipes 52/--/0.99 --/--/0.79 Myxochlamys mullerensis Myxochlamys nobilis 100/99/1.00 85/94/0.99 Borneocola stenophyllus FRI50288 Borneocola biru 96/88/1.00 88/93/1.00 70/78/0.99 Borneocola petiolatus FRI50291 Borneocola reticosus FRI50294 Borneocola sp. S99106 B oesenbergia pulc herrim a B oesenbergia / /0.67 Borneo 93/96/1.00 Borneocola Borneocola calcicola FRI50290 Borneocola sp. FRI50295 Sarawak Myxochlamys Scaphochlamys samunsamensis MK 2344 Scaphochlamys klossii var. klossii FRI50238 80/88/1.00 Scaphochlamys lanceolata FRI50130 Scaphochlamys oculata FRI50262 64/68/0.99 Peninsular Malaysia + Borneo 72/77/1.00 Sarawak rotunda K aem pferia rotunda 54/54/0.51 Cornuk aem pferia aurantiflora Haniffia albiflora Vietnam Distichochlamys sp. Kress01-6848 Distichochlamys sp. AS18 Distichochlamys citrea Distichochlamys K aem pferia sp. Kress98-6289 99/99/1.00 85/88/1.00 S m ithatris s upraneanae 54/62/0.81 S tahlianthus involucratus Curc uma bic olor Zingiber gram ineum Zingiber s ulphureum 86/88/1.00 Zingiber wray ii 94/97/1.00 72/83/0.99 Ros c oea c autleoides Ros c oea purpurea 67/77/1.00 Cautleya grac ilis Cautleya s pic ata Hedychium longic ornutum 99/99 Hedichium villos um Cam ptandra parvula P y rgophy llum y unnanens e Curc uma ros c oeana 97/98/1.00 Hitc henia glauc a Rhy nc hanthus bees ianus 51/53/1.00 Caulok aem pferia s ax ic ola G agnepainia thoreliana 99/99/1.00 Hem iorc his rhodorrhac his Mantisia wengeri 65/63/0.76 Elettariopsis kerbyi Renealm ia battenbergiana 99/99/1.00 A lpinia blepharoc aly x 82/78/-86/83/1.00 A lpinia elegans A m omum villos um Tam ijia flagellaris S iphonochilus aethiopic us 100/100/100 98/98/0.99 S iphonochilus k irk ii S iphonochilus dec orus 0.005 Figure 2. Molecular phylogenetic analysis of the matK sequence data by the Maximum Likelihood method. Numbers above branches indicate bootstrap values of ML and MP and posterior probability of Bayesian Analysis. 42 Yen Yen Sam et al. / PhytoKeys 75: 31–55 (2016) Scaphochlamys grandis FRI50171 93/83/0.70 Scaphochlamys breviscapa FRI44984 --/92/-- Scaphochlamys klossii var. klossii FRI50238 94/86/0.93 92/94/0.68 Scaphochlamys polyphylla FRI50289 Scaphochlamys oculata FRI50262 Scaphochlamys concina FRI50351 96/99/1.00 Scaphochlamys longifolia FRI47065 96/97/1.00 93/99/1.00 Scaphochlamys kunstleri var. speciosa FRI68936 Sarawak Scaphochlamys Scaphochlamys lanceolata FRI50130 --/67/-- Scaphochlamys kunstleri var. kunstleri 80/82/0.99 Scaphochlamys kunstleri var. rubra Scaphochlamys biloba Myxochlamys mullerensis 99/100/1.00 Myxochlamys nobilis Borneocola stenophyllus FRI50288 Borneocola sp. S99106 95/96/1.00 Borneocola petiolatus FRI50291 51/51/- Borneocola reticosus FRI50294 Distichochlamys sp. AS18 B oesenbergia pulc herrim a B oesenbergia rotunda K aem pferia rotunda 99/100/1.00 97/99/1.00 Vietnam Distichochlamys sp. Kress01-6848 99/99/1.00 Distichochlamys Distichochlamys citrea 99/100/1.00 --/--/0.93 Borneocola Borneocola sp. FRI50295 99/99/1.00 Borneo Borneocola calcicola FRI50290 28 88/85/1.00 89/91/1.00 Myxochlamys 95/97/1.00 66/85/1.00 Peninsular Malaysia +Borneo Scaphochlamys rubromaculata FRI50172 74/--/0.61 K aem pferia sp. Kress98-6289 K aem pferia parviflor a --/--/0.97 Cornuk aem pferia aurantiflora Zingiber gram ineum -/-/0.89 Zingiber s ulphureum 99/100/1.00 91/97/1.00 30 Zingiber wray ii Haniffia albiflora 99/100/1.00 Cautleya grac ilis 99/99/1.00 Cautleya s pic ata Ros c oea c autleoides Ros c oea purpurea 5 91/98/1.00 16 Hedychium longic ornutum 99/100/1.00 Hedichium villos um 14 P om m eres c hea lac k neri 95/94/1.00 Rhy nc hanthus bees ianus 83/89/1.00 Caulok aem pferia s ax ic ola G agnepainia thoreliana 99/100/1.00 Hem iorc his rhodorrhac his P y rgophy llum y unnanens e 11 Cam ptandra parvula -/-/0.97 --/59/0.68 -/-/0.53 Curc uma bic olor S m ithatris s upraneanae 99/100/1.00 81/84/0.99 99/99/1.00 76/67/0.97 S tahlianthus involucratus Curc uma ros c oeana Hitc henia glauc a E lettariopsis kerbyi Renealm ia battenbergiana 99/100/0.98 77/81/-99/99/1.00 A lpinia blepharoc aly x A lpinia elegans A m omum villos um Tam ijia flagellaris S iphonochilus aethiopic us 97/98/0.97 99/100/1.00 96/98/1.00 S iphonochilus dec orus S iphonochilus k irk ii 0.01 Figure 3. Molecular phylogenetic analysis of the ITS+matK sequence data by the Maximum Likelihood method. Numbers above branches indicate bootstrap values of ML and MP and posterior probability of Bayesian Analysis. Borneocola (Zingiberaceae), a new genus from Borneo A B 43 C Figure 4. Bladeless sheaths A Green and coriaceous in Scaphochlamys klossii (Peninsular Malaysia) B Red and coriaceous in Scaphochlamys abdullahii (Peninsular Malaysia) C Papery and marcescent in Borneocola calcicola (Sarawak). (Photographs by Y.Y. Sam). on labellum and absence of dorsal knobs on the stigma in Scaphochlamys. The mucilage on the floral bracts and the versatile anther of Myxochlamys are absent in Borneocola. Type species. Borneocola reticosus (Ridl.) Y.Y.Sam, comb. nov. Gastrochilus reticosa Ridl., J. Straits Branch Roy. Asiat. Soc. 44: 195 (1905). Description. Terrestrial rhizomatous herb, evergreen, rarely exceeding 50 cm in height. Rhizome creeping on the ground, terminal decumbent, rhizome elements short or long; roots fine, extensive, not tuberous. Leafy stem unifoliate, enclosed by a few bladeless sheaths at base, bladeless sheaths linear, papery, glabrous to hairy, light green or light brown, decaying early, leaf sheath glabrous or hairy, base swollen, margin thin and narrow; ligule membranous, inconspicuous, decaying early; petiole channelled in cross section, glabrous, lamina narrowly ovate to elliptic, rarely oblong, asymmetric, margin entire, smooth. Inflorescence flowering from base to apex; peduncle short, usually hidden within leaf sheath; spike composed of compact rachis and 2–5 (–13) fertile bracts, bracts spirally and closely overlapping (rarely distichous), boat-shaped, 2-keeled, pink, pale brown, pale or light green, thin, translucent, glabrous or hairy, decaying early, marcescent, amplexicaul at the base of the bract, cincinni compact, 2–3 flowers in each cincinnus. First bracteole directly opposite floral bract and enclosing all the flowers and subsequent bracteoles, linear-shaped, 2-keeled, shorter than bracts, rarely same length. Flowers thin, delicate, ephemeral. Calyx tubular, splitting unilaterally on one side, floral tube long slender, usually puberulent externally, inner surface with a groove enveloping the style, corolla lobes 3, triangular ovate, translucent, glabrous, dorsal lobe apex hooded, lateral lobes 2, narrower than dorsal lobe. Staminodes elliptic to 44 Yen Yen Sam et al. / PhytoKeys 75: 31–55 (2016) A B D E C F Figure 5. A–C Floral bracts A Green and coriaceous in Scaphochlamys klossii B Red and coriaceous in Scaphochlamys pusilla C Scarious and marcescent in Borneocola petiolatus D–F Variegation on labellum D White labellum with purple lines beside the median band in Scaphochlamys malaccana E White labellum with red streaks beside the band in Scaphochlamys concinna F Lilac labellum without coloured streaks beside the band in Borneocola petiolatus. (Photographs by Y.Y. Sam) A B C Figure 6. Stigma A Funnel-shaped in Scaphochlamys endauensis B Beak-like in Scaphochlamys biloba C Capitate in Borneocola petiolatus. (Photographs by A & C N.M. Aidil, B Y.Y. Sam) narrowly obovate, white, light yellow or green, spreading laterally, lined with translucent veins from base to apex, covered with glandular hairs on adaxial surface. Labellum obovate, flat, bilobed distally, rarely entire, translucent veins spread from base to Borneocola (Zingiberaceae), a new genus from Borneo 45 apical part, pale white, pink, lilac or violet, median band light yellow or greenish yellow, without coloured streaks or patches beside the band, adaxial surface covered with glandular hairs. Stamen bends forward over labellum, usually white and covered with glandular hairs on abaxial surface, filament grooved, short, anther theacae 2, linear, dehiscing along entire length, basal ending with or without short spur, anther crest short, not recurved. Ovary ellipsoid, glabrous or sparsely hairy, unilocular, with basal placentation, less than 10 locules. Stigma small, held at apex of thecae, near oblate, 2 dorsal knobs, ostiole forward facing, ciliate. Epigynous glands 2, filiform, yellow. Fruit a capsule, ellipsoid or ovoid. Seeds not seen. Etymology. This new genus is named after the island of Borneo and -cola (Latin) means dweller or inhabitant. This is to recognise the extremely rich and unique biodiversity that is found in Borneo. Distribution. Borneo. The genus is currently known to occur only in the northwest and possibly central Borneo. Eight species are recorded from Sarawak, Malaysia and many more are undescribed. Key to Borneocola species (modified from Ooi and Wong 2014) 1 – 2 – 3 – 4 – 5 – 6 – 7 – Adaxial lamina with distinctly raised tessellate venation ................................ .................................................................................6 Borneocola reticosus Adaxial lamina without distinctly raised tessellate venation .........................2 Lamina broadly oblanceolate and elliptic to oblong, lateral veins conspicuously raised on adaxial surface ................................. 1 Borneocola argenteus Lamina linear, lanceolate to broadly ovate and elliptic, lateral veins not conspicuously raised on adaxial surface .............................................................3 Lamina linear to very narrowly lanceolate, < 3 cm wide ................................ ........................................................................... 8 Borneocola stenophyllus Lamina lanceolate, ovate to elliptic, > 3 cm wide ........................................4 Leaf sheath plus petiole < 10 cm long; lamina apex broadly acute to rounded, base cuneate ...................................................................4 Borneocola iporii Leaf sheath plus petiole > 10 cm long; lamina apex acute to acuminate, base attenuate, cordate, rounded or truncated ....................................................5 Lamina lanceolate to ovate, < 7 cm wide .....................................................6 Lamina elliptic to broadly elliptic and ovate, > 7 cm wide...........................7 Basal lamina rounded to truncated; inflorescence stalk 3–9 cm long; labellum pale lilac..................................................................5 Borneocola petiolatus Basal lamina attenuate; inflorescence stalk 1–2 cm long; labellum purplish blue .................................................................................2 Borneocola biru Inflorescence about 3 cm long; labellum pale pink ........................................ ....................................................................7 Borneocola salahuddinianus Inflorescence > 7 cm long; labellum white ................3 Borneocola calcicola 46 Yen Yen Sam et al. / PhytoKeys 75: 31–55 (2016) 1. Borneocola argenteus (R.M.Sm.) Y.Y.Sam, comb. nov. urn:lsid:ipni.org:names:77158824-1 Scaphochlamys argentea R.M.Sm., Notes Roy. Bot. Gard. Edinburgh 44: 209 (1987). Scaphochlamys depressa Mas Izzaty, A.Ampeng & K.Meekiong, Folia Malaysiana 14(2): 19 (2013). Type. MALAYSIA. Sarawak, First Division, Lundu, near foot of Gunung Perigi, 6 Aug 1962, Burtt B2700 (holotype: E!). Notes. It is one of the most distinctive species, either in the field or herbarium sheet. This dainty plant has a long creeping rhizome and well spaced leafy shoots, prominently stiff lateral veins raised on its adaxial lamina, both on living plants and dried specimens. Scaphochlamys depressa Mas Izzaty, Ampeng & Meekiong is unmistakably the same as S. argentea with its prominent raised lateral veins, broadly elliptic lamina and well spaced leafy shoots. Meekiong (2015) explained that the inflorescence of S. depressa which exerted from the petiole is different from S. argentea where the inflorescence emerges from the base of the petiole. This observation is incorrect as all gingers have terminal inflorescences. 2. Borneocola biru (Meekiong) Y.Y.Sam, comb. nov. urn:lsid:ipni.org:names:77158812-1 Scaphochlamys biru Meekiong, Folia Malaysiana 16(1): 37 (2015). Type. MALAYSIA. Sarawak, Kuching Division, Matang Wildlife Centre, 21 May 2014 Meekiong et al. s.n. (holotype: SAR; isotype: Herbarium, Universiti Malaysia Sarawak. Types not yet deposited as of 5 May 2016). Notes. Borneocola biru is the most recent species described from Sarawak. It has a deep purplish blue labellum, different from all other Borneocola species which are white or in lighter shades. 3. Borneocola calcicola (A.D.Poulsen & R.J.Searle) Y.Y.Sam, comb. nov. urn:lsid:ipni.org:names:77158813-1 Scaphochlamys calcicola A.D.Poulsen & R.J.Searle, Gard. Bull. Singapore 57: 29 (2005). Type. MALAYSIA. Sarawak, Kuching Division, Bau area, Gunung Tai Ton, 1°24'N, 110°8'E, 20 June 2003, Poulsen, Jugah & Clausager 2022 (holotype: SAR!; isotypes: AAU, E!, K!, L). Notes. Borneocola calcicola is the largest amongst the Borneocola species. Poulsen and Searle (2005) observed that the distichous inflorescence is one of the character- Borneocola (Zingiberaceae), a new genus from Borneo 47 istics of the plant. However, a recent collection of B. calcicola, Sam FRI 50290, from Seromah, Bau, showed spirally arranged floral bracts. There was a mixture of spirally and distichously arranged floral bracts in its population in Bau, Sarawak. 4. Borneocola iporii (Meekiong & A.Ampeng) Y.Y.Sam, comb. nov. urn:lsid:ipni.org:names:77158814-1 Scaphochlamys iporii Meekiong & A.Ampeng, Folia Malaysiana 12(1): 19 (2011). Type. MALAYSIA. Sarawak, Kapit, Lanjak Entimau Wildlife Sanctuary, Bukit Menyarin, 3 April 2008, Meekiong MK1839 (holotype: SAR; isotype Herbarium, Universiti Malaysia Sarawak. Types not yet deposited as of 5 May 2016). Notes. Borneocola iporii is a small ginger creeping on the humus rich forest floor. It is most similar to B. argenteus with both having a unifoliate shoot, leafy shoots far apart, broad lamina, short inflorescence and compact rachis. However, the conspicuously raised lateral veins of B. argenteus can readily distinguish it from B. iporii. 5. Borneocola petiolatus (K.Schum.) Y.Y.Sam, comb. nov. urn:lsid:ipni.org:names:77158815-1 Haplochorema petiolatum K.Schum. in Engler, Pflanzenr. IV, 46 (Heft 20): 90 (1904). Scaphochlamys petiolata (K.Schum.) R.M.Sm., Notes Roy. Bot. Gard. Edinburgh 44: 210 (1987). Type. MALAYSIA. Sarawak, First Division, Mt. Singhi (= Gunung Singai), Dec 1892, Haviland 2026 (lectotype: K! designated by Searle 2010; isolectotype: E!, SAR!). Notes. Borneocola petiolatus is distinguished by its long petiole and narrow leaves from the other species. Its lamina length is almost 3 times the width (12–21.5 × 3.1–7.1 cm). Smith (1987) found that B. petiolatus has small inflorescences as in B. argenteus. However, both can be easily separated by their leaf characters. Borneocola petiolatus has much longer petioles compared to B. argentea (12.7–31.5 cm versus 3–6 cm). Borneocola argenteus also has prominently raised lateral veins on the adaxial surface of lamina, more conspicuous on dried specimens than fresh materials. This character is lacking in B. petiolatus. 6. Borneocola reticosus (Ridl.) Y.Y.Sam, comb. nov. urn:lsid:ipni.org:names:77158816-1 Gastrochilus reticosa Ridl., J. Straits Branch Roy. Asiat. Soc. 44: 195 (1905). Boesenbergia reticosa (Ridl.) Merr., Bibl. Enum. Born. Pl. 122 (1921). Scaphochlamys reticosa (Ridl.) R.M.Sm., Notes Roy. Bot. Gard. Edinburgh 44: 209 (1987). 48 Yen Yen Sam et al. / PhytoKeys 75: 31–55 (2016) Type. Cultivated in Singapore Botanic Gardens, originally from Borneo, Sarawak, First Division, Bidi, 22 Nov 1904, Ridley s.n. (holotype: SING!). Notes. Borneocola reticosus is chosen as the type species as it is the easiest to recognise in the genus. Its reticulate lamina readily distinguishes it from other Borneocola species. 7. Borneocola salahuddinianus (Meekiong, A.Ampeng & Ipor) Y.Y.Sam, comb. nov. urn:lsid:ipni.org:names:77158817-1 Scaphochlamys salahuddiniana Meekiong, A.Ampeng & Ipor, Folia Malaysiana 12(1): 22 (2011). Type. MALAYSIA. Sarawak, Kapit, Ulu Katibas, Lanjak Entimau Wildlife Sanctuary, Bukit Sepali, 30 April 2008, Meekiong MK1856 (holotype SAR; isotype Herbarium, Universiti Malaysia Sarawak.. Types not yet deposited as of 5 May 2016). Note. Borneocola salahuddinianus is unique amongst the Bornean species with its broadly elliptic or ovate lamina held by a long slender petiole. It is doubtful that B. salahuddinianus is a lithophyte as observed by Meekiong et al. (2011). The plants are more of an opportunist growing on humus-rich substrate accumulated on the rocks. 8. Borneocola stenophyllus (I.H.Ooi & S.Y.Wong) Y.Y.Sam., comb. nov. urn:lsid:ipni.org:names:77158818-1 Scaphochlamys stenophylla I.H.Ooi & S.Y.Wong, Willdenowia 44(2): 241-245 (2014). Type. MALAYSIA. Sarawak, Kuching Division, Bau, Gunung Buan, 1°33'28.9"N, 10°08'35.2"E, 92 m, 21 Nov 2013, Ooi Im Hin & Jepom ak Tisai OIH74 (holotype: SAR. Type not yet deposited as of 5 May 2016). Note. Borneocola stenophyllus is another new species recently discovered from Sarawak. Its grass-like leaves instantly separate it from other species in the genus. Incompletely known species Scaphochlamys anomala (Hallier f.) R.J.Searle, Edinburgh J. Bot. 67: 85 (2010). Kaempferia anomala Hallier f., Bull. Herb. Boissier 6: 357 (1898). Gastrochilus anomalum (Hallier f.) K.Schum. in Engler, Pflanzenr. IV, 46 (Heft 20): 92 (1904). Boesenbergia anomala (Hallier f.) Schltr., Repert. Spec. Nov. Regni Veg. 12: 315 (1913). Gastrochilus hallieri (Hallier f.) Ridl., J. Straits Branch Roy. Asiat. Soc. 32: 109 (1899), nom. illegit. Borneocola (Zingiberaceae), a new genus from Borneo 49 Type. INDONESIA. Cultivated in Bogor, originally from Liang Gagang, Kalimantan Borneo, Hallier s.n. (original material: BO, specimen lost; lectotype (designated by Searle, 2010) Figure drawn from original Hallier’s material and published as t. IX, fig. 3, Bull. Herb. Boissier 6: 357 (1898). Notes. The type, the only specimen ever collected, was lost. However, Hallier (1898) gave a very detailed description and drawing of the plant and this has convinced Searle (2010) to place it in the genus Scaphochlamys. The drawing, which is based on the type specimen and designated by Searle as the lectotype, is the only material that gives a glimpse of the appearance of the species. In the drawing, the flower and spirally arranged floral bracts are typical of both Scaphochlamys and Borneocola. Until another specimen is collected and is available for close examination, we prefer to retain this imperfectly known species in Scaphochlamys. Discussion The phylogenetic analyses confirm the distinctive character of Borneocola and Scaphochlamys and their placement in the tribe Zingibereae (Figures 1, 2, 3). The Borneocola species form a monophyletic group which is sister to Myxochlamys. It is surprising to find Borneocola having a closer affinity to Myxochlamys than to Scaphochlamys, considering it shares more morphological similarities with Scaphochlamys than with Myxochlamys. Morphologically, Myxochlamys is very different from Borneocola. There are two Myxochlamys species named so far: M. amphiloxa and M. mullerensis (Takano and Nagamasu 2007; Searle and Newman 2010) and a third undescribed species, also from Borneo. All three Myxochlamys species are very robust plants that can attain a height of 70 cm. Most Borneocola species examined so far are small-sized (not more than 50 cm tall), except for B. calcicola which can grow to 60 cm tall. Myxochlamys has 3–10 large leaves (50–60 cm long) in each shoot whereas Borneocola are unifoliate and the leaves are small (less than 20 cm long except for B. calcicola). The leaves of Myxochlamys are sessile compared to the conspicuously stalked leaves in Borneocola. The most marked difference is in the inflorescence structure. Borneocola has small inflorescences consisting of less than 15 fertile bracts but Myxochlamys has large torch-like inflorescences with easily more than 40 bracts. The bracts of Borneocola are membranous and marcescent, often measuring less than 2 cm long (except for B. calcicola measuring 2.5–3.2 cm long). By contrast, the floral bracts of Myxochlamys are coriaceous, persistent, measuring 2.5–5 cm long and most notably are covered with transparent slimy mucilage. In addition, the unique versatile anthers of Myxochlamys, a rare feature in the Zingiberaceae, are distinct from the adnate anthers in Borneocola and also from all its sister genera. Based on morphological features, Myxochlamys is more similar to Scaphochlamys, the closest being S. grandis. Both have large sessile leaves and decurrent lamina base, large, coriaceous 50 Yen Yen Sam et al. / PhytoKeys 75: 31–55 (2016) and persistent floral bracts,their bracts being concave with reflexed and spreading apices. Based on morphology, Borneocola is also similar to Distichochlamys. However, Distichochlamys is distinguished from Borneocola, Myxochlamys and Scaphochlamys by its unique tubular bracteoles, floral tube without a groove on the inner surface and trilocular ovary (Newman 1995). Other characteristics such as distichous floral bracts, 2-keeled bracteoles, thecae without basal spurs have been observed in the three closely allied sister genera in this study (Table 2). Haplochorema K.Schum. is another small-sized genus endemic to Borneo, which can be mistaken for Borneocola. It has short and few-flowered inflorescences as in Borneocola but its flowers appear somewhat quadrate with the labellum and lateral staminodes held flat, more resembling Kaempferia L. Haplochorema has distichous floral bracts, single-flowered cincinni and the flowering proceeds from apex to base, to name some of the characters which distinguish it from Borneocola. In fact, the genus is more allied to Boesenbergia Kuntze than Borneocola. Borneocola is morphologically most similar to Scaphochlamys but both can be distinguished by the texture of the bladeless sheath and floral bracts, variegation on the labellum, indumentum on the floral tube and the stigma shape. The current study recognises eight Borneocola species while S. gracilipes, S. polyphylla B.L.Burtt & R.Sm., S. limiana Meekiong & K.Yazid and S. samunsamensis Meekiong & Hidir from Borneo remain in the genus Scaphochlamys. There are no recent collections of S. gracilipes but the lax inflorescence and persistent floral bracts in the type specimens clearly distinguish it from the Borneocola species. Scaphochlamys polyphylla, S. limiana and S. samunsamensis can be readily distinguished from the Borneocola species by their papery bladeless sheath and large, green or green tinged red, coriaceous floral bracts. This shows that the distinct morphologies that separate Borneocola and Scaphochlamys are significant and are also supported by the phylogenetic analyses (Figures 1, 2 and 3). An anatomical study on the leaves also discovered some characteristics that separate Borneocola from Scaphochlamys (Norhati, pers. comm). The morphology of Borneocola is very similar to Scaphochlamys but, combining both, necessitates synonymising Myxochlamys and possibly Distichochlamys and this will result in a very heterogenous genus. A similar situation is observed in the naming of Newmania N.S. Lý & Škorničk, a genus very similar in morphology to Haniffia Holttum but appears as its sister group in the molecular phylogenetic analyses. The authors decided against placing Newmania under Haniffia which would create a heterogenous group. The current description of Borneocola is further supported by the chromosome number with 2n=10 (Šída et al., unpublished data), different from Distichochlamys (2n=26) and Scaphochlamys (2n=28). Such significant differences in molecular data and chromosome number have conclusively supported the circumscription of the new genus Borneocola. Table 2. Comparison between the morphological characters of Borneocola, Distichochlamys, Myxochlamys and Scaphochlamys. Floral bracts Flowers First bracteole Floral tube Labellum Thecae Anther Ovary Chromosome number Geographical distribution Borneocola to 50(–60) cm Distichochlamys to 60 cm Myxochlamys 70 cm Scaphochlamys to 100 cm 1 1–3 3–10 1–7 Papery, drying fast 6–37 × 1–18; petiolate 3–11.5 3–13 Papery, decaying fast 15–28 × 8.3–14.5; petiolate to 15.5 7–13 Not mentioned 50–65 × 7–17; sessile 6.5–18 c. 40 Coriaceous, persistent 9–50 × 3–24; petiolate or sessile 4–28 4–44 Spiral, rarely distichous Distichous Spiral Spiral, rarely distichous Coriaceous, with mucilage; persistent Solitary Open to base, 2-keeled With a groove in inner surface, glabrous externally Not bilobed, entire, concave; without coloured streaks beside median band Coriaceous, without mucilage; persistent In cincinni Open to base, 2-keeled With a groove in inner surface, glabrous externally Bilobed, rarely entire, not concave; with coloured streaks beside median band Spurs absent or with short free basal spurs Adnate Thin, translucent, without mucilage; Without mucilage; persistent drying fast In cincinni In cincinni Open to base, 2-keeled Tubular, 2-keeled With a groove in inner surface, Without a groove in inner glabrous to puberulent externally surface, glabrous externally Bilobed, rarely entire, not concave; Bilobed, not concave; without coloured streaks beside median without coloured streaks beside band median band Spurs absent or with short free basal Spurs absent spurs Adnate Adnate Trilocular with axile Unilocular with basal placentation placentation 2n=10 (Šída et al., unpublished data) 2n=26 Borneo Vietnam Spurs present and long Versatile Unilocular with basal placentation — Borneo Unilocular with basal placentation Borneocola (Zingiberaceae), a new genus from Borneo Morphology Plant height Number of leaf in each leafy shoot Bladeless sheath Leaf (cm) Inflorescence height (cm) Number of floral bracts Arrangement of floral bracts 2n=28 Southern Thailand, Peninsular Malaysia, Sumatra, Borneo 51 52 Yen Yen Sam et al. / PhytoKeys 75: 31–55 (2016) Key to the genera of the Zingibereae tribe in Borneo 1 – 2 – 3 – 4 – 5 – 6 – 7 – 8 – Inflorescence arising directly from the rhizome on a leafless shoot...............2 Inflorescence emerging at the terminal of the leafy shoot ............................3 Distinct swelling at the base of the petiole; anther with long extended crest wrapped around the style ...............................................................Zingiber No swelling at the base of the petiole; anther crest short, not long extended and not wrapped around the style ................................................... Haniffia Flowers with versatile anther .......................................................................4 Flowers with adnate anther .........................................................................5 Inflorescence with few to many floral bracts, bracts mucilage ....Myxochlamys Inflorescence with one single large floral bract, bracts not mucilage .............. ................................................................................................ Camptandra Flowers opening from top to bottom of inflorescence .................................6 Flowers opening from bottom to top of inflorescence .................................7 Flowers appearing quadrate with the two petaloid staminodes .....Haplochorema Flowers no quadrate appearance, staminodes not petaloid........Boesenbergia Flowers with long narrow corolla lobes and long exserted stamens ................ .................................................................................................. Hedychium Flowers without such features .....................................................................8 Floral bracts coriaceous and persistent, labellum with coloured streaks on both sides of the median band ..............................................Scaphochlamys Floral bracts thin, translucent and marcescent, labellum without coloured streaks on both sides of the median band ....................................Borneocola Acknowledgements The authors wish to thank the Forestry Department of Peninsular Malaysia and its State Forestry Departments, Sarawak Forestry Department, Department of Wildlife and National Parks and Johor Parks Corporation for permission to conduct research in their forests; herbaria of AAU, BKF, C, FI, E, K, KEP, KLU, PSU, SAN, SAR and SING for the loan of specimens; Lucy Chong and Julia Sang from the Sarawak Forestry Corporation, Wong Sin Yeng and Peter C. Boyce from Universiti Malaysia Sarawak, Sarawak for their hospitality and logistics help in Sarawak. We are also grateful to Kalu Meekiong from the Universiti Malaysia Sarawak, Sarawak and John Mood for the leaf samples for the molecular study; J.F. Veldkamp for advice on the genus and epithet names; Jana Leong-Škorničková, Richard C.K. Chung, Leng Guan Saw, Ruth Kiew and the reviewers for their critical comments and suggestions to improve the manuscript. 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