A peer-reviewed open-access journal PhytoKeys 201: 23-34 (2022) & doi: 10.3897/phytokeys.201.849 1 | 46P h y toKe y S https:/ / Pp hyto keys -pen soft.net Launched to accelerate biodiversity research Lepisorus medioximus (Polypodiales, Polypodiaceae), a new species from Shan State of Myanmar Tao Fujiwara'”, Phyo Kay Khine’, Kiyotaka Hori, Thant Shin*, Noriaki Murakami!, Harald Schneider’ | Makino Herbarium, Tokyo Metropolitan University, 1-1 Minami-osawa, Hachioji, Tokyo 192-0397, Japan 2 Center for Integrative Conservation, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sci- ences, Menglun 666303, China 3 ‘The Kochi Prefectural Makino Botanical Garden 4200-6 Godaisan, Kochi 781-8125, Japan 4 Forest Research Institute, Yezin, Nay Pyi Taw, Myanmar Corresponding author: Tao Fujiwara (tao. fujiwara@gmail.com) Academic editor: Blanca Ledn | Received 6 April 2022 | Accepted 22 May 2022 | Published 20 June 2022 Citation: Fujiwara T,; Khine PK, Hori K, Shin T; Murakami N, Schneider H (2022) Lepisorus medioximus (Polypodiales, Polypodiaceae), a new species from Shan State of Myanmar. PhytoKeys 201: 23-34. https://doi. org/10.3897/phytokeys.201.84911 Abstract A new species of the species-rich fern genus Lepisorus (Polypodiales, Polypodiaceae) has been found to occur in Shan state, Myanmar. Lepisorus medioximus is described based on morphological characters and phylogenetic evidence. Phylogenetic analyses showed that the specimens of L. medioximus formed a dis- tinct clade nested in the Pseudovittaria clade. The morphological comparison demonstrated that the spe- cies is distinct from phylogenetically related species, namely L. elegans, L. contortus, and L. tosaensis, in the morphology of the rhizome scales, size, and shape of the lamina, position of sori, and paraphyses. Keywords Epiphyte, integrative taxonomy, micromorphology, overlooked species diversity, phylogeny Introduction The genus Lepisorus (J. Sm.) Ching (Polypodiaceae) occurs throughout Eastern and South- ern Asia with range extensions towards the Pacific islands including Hawai'i and towards tropical Afromadagascar (Ching 1933; Zink 1993; Wang et al. 2012). Taking into ac- count the various taxonomic studies and the recent proposal to expand the generic circum- scription by including all genera of the tribe Lepisoreae such as Lemmaphyllum C. Presl, Copyright Tao Fujiwara 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. 24 Tao Fujiwara et al. / PhytoKeys 201: 23-34 (2022) Lepidomicrosorium Ching & K.H. Shing, Neocheiropteris Christ, Neolepisorus Ching, Para- gramma (Blume) T: Moore, and Tricholepidium Ching (Zhao et al. 2020) Lepisorus can be currently recognized as one of the most species-rich genera among genera in Polypodi- aceae, comprising ca. 90 species in 18 sections (PPGI 2016; Fujiwara et al. 2020; Zhao et al. 2020). Whereas the core of the genus Lepisorus (Lepisorus s.s.) is easily recognized by its unique suite of morphological characters including creeping rhizomes covered by clathrate scales, simple leaves, and sori covered with scale-like paraphyses (Ching 1933; Qi et al. 2013; Zhao et al. 2020), some controversy still exists concerning the broader circumscrip- tion to avoid the need to recognize the genus El/ipinema Li Bing Zhang & Liang Zhang (Zhang et al. 2020). Despite significant progress (Wang et al. 2010a; Wei et al. 2017; Zhang et al. 2020; Zhao et al. 2020), taxonomic uncertainty is arguably not restricted to the generic classification but affects the estimation of the total species diversity that is expected to be still underestimated due to the difficulty in the taxonomic classification of this genus. Several characters utilized as key information on species delimitation show high variation within some species (Wang et al. 2010b). In particular, the identification of species relies on a few diagnostic characters such as the shape of the lamina, position of sori, and structure of the rhizome scales and paraphyses. Unfortunately, these characters are hardly diagnosable in the field. As a consequence, some species have been frequently misi- dentified or overlooked, as exemplified by the recent reclassification of Japanese L. thunber- gianus (Kaulf.) Ching and relatives (Fujiwara et al. 2018), and the rediscovery of Lepisorus cespitosus Y.X. Lin previously known only as type specimens (Fujiwara et al. 2020). Yunnan has been increasingly recognized as the diversity hotspot of Lepisorus s.s (Wang et al. 2012; Fujiwara et al. 2020). While sufficient explorations of the Lepisorus diversity have been made available for China and India, this cannot be claimed for the regions south and southwest of Yunnan, namely Laos and Myanmar. ‘These two coun- tries as well as Thailand show extremely low species diversity of the genus despite being expected to harbor a notable diversity of this genus (Suppl. material 1: Table S1). This can be attributed to not only the difficulty in the taxonomic classification of this genus but also the underestimation of whole fern species diversity due to fewer flora surveys previously conducted. ‘Thus, overlooked species are expected to occur in these regions. The Shan state of Myanmar is the focus of this study. The Shan state covers 155,800 km? which is almost a quarter of the whole area of Myanmar and is mainly comprised of a hilly plateau bordering Yunnan, China in the north, Laos in the east, and Thailand in the south. Shan state has been in historical times much less surveyed than Yunnan Province of China although the latter is known for its rich diversity of ferns including Lepisorus. Thus, we expect to retrieve not only new records but also some new fern species that are putative endemics to Shan State. To make this expecta- tion tested, floristic inventories were carried out across the Shan state in September 2019. Two unusual specimens of Lepisorus were collected in Pin Laung Township, Ka Thaung (upper) located in the southern part of the state, which were recognized as a putative new species. This proposal was studied by consulting checklists of Myanmar and adjacent areas (Dickason 1946; Lindsay and Middleton 2012; Qi et al. 2013; Khine et al. 2017; Khine and Schneider 2020; Hori 2021; Vongthavone et al. 2021), and careful comparison of morphological characters with previously described species A new fern species of Lepisorus from Myanmar 25 by consulting specimens and species protologues (e.g., Ching 1933; Bir and Trikha 1969; Yu and Lin 1996, 1997; Lin 2000; Qi et al. 2013). Besides morphological diag- nostics, we employed DNA sequences to identify genotypic evidence supporting these two specimens as distinct species from any other species that are previously described. Materials and methods Morphology The morphology of the two specimens of Lepisorus sp. (Hori et al. 108225 and 108229) was compared to descriptions and specimens of species sharing similarities in the main diagnostic features, namely rhizome scales, the size and shape of the lamina, the posi- tion of sori, and paraphyses. The morphological observation was conducted using a stereomicroscope. Voucher specimens were deposited in MBK, HITBC, and RAF. DNA extraction and chloroplast DNA region sequencing Total DNA for each of the two specimens was extracted from silica dried leaves using the CTAB method according to Doyle and Doyle (1987). Four plastid regions, rbcL gene, rbcL-atpB intergenic spacer, rps4-trnS intergenic spacer, and trnL-trnF region including the ¢rnL intron and the #nL-trnF intergenic spacer were amplified according to the pro- tocol provided (Wang et al. 2010b) using ExTaq (TaKaRa Bio Inc., Shiga, Japan). The PCR products were purified using Illustra ExoStar 1-Step (GE Healthcare, Wisconsin, USA) and used as templates for Sanger sequencing. Reaction mixtures for sequencing were prepared using the SuperDye v3.1 Cycle Sequencing Kit (ADS). The reaction mix- tures were analyzed using an ABI 3130 Genetic Analyzer (Applied Biosystems). Phylogenetic analyses To unveil the phylogenetic position of the new Lepisorus, a genus-level phylogeny was re- constructed incorporating a total of 88 species of Lepisorus including species representing clades previously treated as distinct genera, namely Lemmaphyllum, Lepidomicrosorium, Neolepisorus, Neocheiropteris, Paragramma, and Tricholepidium, retrieved from the se- quence matrices assembled in previous studies (Wang et al. 2010b; Fujiwara et al. 2018, 2020; Zhao et al. 2020) (Suppl. material 2: Table S2). Leptochilus ellipticus (Thunb.) Noot., Microsorum punctatum (L.) Copel., and Bosmania membranacea (D.Don) Tes- to were included as outgroup taxa. Sequences of each plastid region were separately aligned using MAFFT (Katoh and Standley 2013) followed by manually editing in Aliview (Larsson 2014), and subsequently merged into a concatenated matrix using SequenceMatrix (Vaidya et al. 2011). GIR +I +G was adopted as the best substitu- tion model as selected based on AIC using jModelTest 2.1.10 (Darriba et al. 2012). Phylogenetic hypothesis reconstruction was performed with three different methods: maximum likelihood (ML), Bayesian inference (BI), and maximum parsimony (MP). 26 Tao Fujiwara et al. / PhytoKeys 201: 23-34 (2022) ML analyses were performed using IQ-TREE v.1.6 (Nguyen et al. 2015) with default settings. 1000 ultrafast bootstrap replicates were conducted to infer node robustness (Hoang et al. 2018). For the BI method, MrBayes 3.2.6 (Ronquist et al. 2012) was em- ployed by applying two runs of four MCMC chains for 100,000,000 generations with samples taken every 1000 generations. Tracer 1.6 (Rambaut and Drummond 2013) was used to evaluate the samples trees with a focus on convergence. ‘The first 25% were discarded as burn-in. The MP analysis was performed using a heuristic approach with TBR branch-swapping, as implemented in MEGAX (Kumar et al. 2018). Ten initial trees were generated by the addition of randomly selected sequences. ‘The robustness of each branch was assessed by bootstrap analysis calculating 1000 replicates. Results and discussion The combined dataset of rbcL, rbcL-atpB, rps4-trnS, and trnL-F contained 4,617 bp of which 744 sides were parsimoniously informative. The optimal log-likelihood for the reconstructed phylogeny inferred by the ML method was In = -21430.138. The to- pologies were congruent among the phylogenetic hypothesis obtained using the three distinct phylogenetic inference approaches. The result showed that two specimens of Lepisorus from the Shan state of Myanmar formed a clade with a bootstrap value of 100% (ML ultrafast bootstrap value = 100%, MP bootstrap value = 100%) and BI posterior probability of p = 1.0. This clade was nested in the subclade of the sect. Pseudovittaria clade (Fig. 1) (Zhao et al. 2020) that included L. contortus (H. Christ) Ching, L. elegans Ching & W.M.Chu, L. lineariformis Ching & S. K. Wu, and L. nyala- mensis Ching & S. K. Wu. While the latter two species were highly distinct from the new species in their linear to linear-lanceolate lamina, the accumulated substitution event causing a rather long branch separated the two specimens from L. elegans—the morphologically most similar species of this clade (Fig. 1). The morphological comparison revealed that the new species was similar to L. elegans, L. contortus, and L. tosaensis (Makino) H. It6, species from the sect. Pseudovittaria, consistent with the result of phylogenetic analyses. However, the species was discernible from the similar species in lanceolate, pale brown rhizome scales, very short stipe, lanceolate lamina widest at the proximal 1/3, sori located closely to the cos- ta and restricted to the 3/4 distal part of the lamina, and ovate-lanceolate paraphyses (Table 1 and Fig. 2). Lepisorus medioximus T.Fujiw., K.Hori & Khine, sp. nov. urn:lsid:ipni.org:names:77300053-1 Figs. 273 Diagnosis. The new species differs from similar species, Lepisorus elegans and L. contor- tus, by the combination of the following morphological characteristics: the lanceolate laminae with the widest at proximal 1/3 of the lamina, sori closer to costa, sori on distal “196/" A new fern species of Lepisorus from Myanmar 27 ht. a 84/68/.93ll..... eye 99/93/" appt Pp B7/G6/.98) fb ‘hone eee 86/50/.71 91/36/.59 = */99/* ve er “pepe if “pry 83/-/* 95/74/.97 94/80/.97 1.0.2... 93/71/" rr ne ai ae “py “ 9155) po 90/59/0.9 pepe 98/93/") "94s" “pepe S7I611.53 ~ Pe Ee ag ee ne oem See OR "199/" 94/61/" /98/" 0.01 Lepisorus luchunensis Qi 097 Lepisorus suboligolepidus Zhang 4537 Lepisorus lewisii ¥. Liu 05620 Lepisorus oligolepidus Zhang 5082 Lepisorus tosaensis $.Fujimoto 2005042904 Lepisorus obscurevenolusus Zhang 4151 Lepisorus subconfluens Zhang 4518 Lepisorus angustus Z. H. Shen S25 Lepisorus heterolepis Zhang 5064 Lepisorus tibeticus Zhang 4694 Lepisorus mikawanus T. Fujiwara 16120817 Lepisorus nigripes T. Fujiwara 16040919 Lepisorus kuratae T. Fujiwara 16040927 Lepisorus thunbergianus K. Ohora 2005042404 Lepisorus medioximus Hori et al. 108225 Lepisorus medioximus Hori et al. 108229 Lepisorus elegans Zhang 4444 Lepisorus lineariformis Zhang 4437 Lepisorus nyalamensis Zhang 5276 Lepisorus contortus Zhang 5204 Lepisorus confluens C. D. Xu s.n. Lepisorus sordidus Zhang 0612 Lepisorus sinensis Shui 81069 Lepisorus medogensis Z. D. Fang XZ266 Lepisorus loriformis Zhang 4440 Lepisorus sublinearis Shui 81060 Lepisorus xiphiopteris C. D. Xu A0303 Lepisorus pseudonudus Zhang 4249 Lepisorus stenistus Z. D. Fang XZ 412 Lepisorus hachijoensis Zhang 4358 Lepisorus onoei Zhang 4352 Lepisorus monilisorus H. M. Zhang 20050117 Lepisorus boninensis TBG cult.54022 Lepisorus cespitosus FRG-201901-170 Lepisorus miyoshianus C. C. Liu DBO6104 Lepisorus annamensis D. Li 873 Lepisorus henryi Shui 80679 Lepisorus spicatus Ranker 1915 Lepisorus validinervis Daniele 2205 Lepisorus mucronatus Jaman 5891 Lepisorus platyrhynchos Kreier s.n. Lepisorus ussuriensis var. distans S. Fujimoto SF05051602 Lepisorus ussuriensis var. ussuriensis B. D. Liu s.n. Lepisorus pseudoussuriensis Crantill TWo9s Lepisorus annuifrons K. Kokubo s.n. Lepisorus uchiyamae S. Fujimoto 2005042902 Lepisorus yamaokae TNS766444 Lepisorus affinis Znang 4219 Lepisorus subsessilis Zhang 1075 Lepisorus kuchenensis J. M. Xi 08188 Lepisorus nudus F. J. 1749 Lepisorus macrosphaerus Zhang 4794 Lepisorus kawakamii Ranker 2051 Lepisorus megasorus Crantill TWO69 Lepisorus asterolepis Zhang 5171 Lepisorus marginatus Zhang 3360 Lepisorus accedens FJ11-129 Schneider H. Lepisorus albertii Znang 4325 Lepisorus pumilus M. Z. Wang 60667 Lepisorus clathratus Zhang 4515 Lepisorus patungensis Zhang 3413 Lepisorus thaipaiensis G. Y. Rao 2005_045A Lepisorus hsiaowutaiensis Q. R. Liu s.n. Lepisorus likiangensis Zhang 5117 Lepisorus soulieanus Zhang 5168 Lepisorus variabilis US3540435 Lepisorus waltonii Zhang 4639 Lepisorus rotundus RV7675 Lepisorus schraderi RV8253 Lepisorus perrierianus EB245 Lepisorus bampsii R.Viane 11233 Lepisorus vesiculari-paleaceus RV11233 Lepisorus excavatus Hemp 3561 Lepisorus bicolor Zhang 5157 Lepisorus morrisonensis Zhang 5113 Lepisorus scolopendrius Zhang 2295 Lepisorus scolopendrius Zhang 4659 Lepisorus buergerianus Zhang 4111 Lepisorus subhemionitideus 1290 Zhang X. C. Lepisorus palmatopedatus Zhang 4482 Lepisorus maculosus Shui 80596 Lepisorus normalis Wade 2649 Lepisorus fortunei GC. C. Chen 1012 Lepisorus zippelii Wade 1794 Lepisorus ensatus Zhang 3611 Lepisorus carnosus Zhang 4364 Lepisorus pyriformis JRH2242 Jiang R.H. Lepisorus rostratus wxp108 Wei X.P. et al Lepisorus squamatus 3925 PE Lepisorus longifolius Schneider s.n. Lepisorus jakonensis C.F. Zhao LZ8 Leptochilus ellipticus Zhang 1923 Microsorum punctatum Zhang 4194 Bosmania membranacea Li 95 Pseudovittaria Sclerophylion Obtusi Caespes Drymotaenium Belvisia Ussurienses Lepisorus Weatherbya Hymenophyton Pachyphylion _ EE ne | 1| Lepidomicrosorium ‘] Neocheiropteris ] Tricholepidium Neolepisorus Lemmaphyllum *] Paragramma J Pseudoclathrati Figure |. Maximum likelihood tree of Lepisorus based on the combined dataset of rbcL, rbcL-atpB, rps4- trnS and trnL-F. The number on each branch indicates support values as follows: ML bootstrap support/ MP bootstrap support/BI posterior probability. The classifications of genus and section for Lepisorus fol- lows Zhao et al. (2020) and Fujiwara et al. (2020). 3/4 of the lamina, and ovate-lanceolate, ovate to orbicular clathrate paraphyses with entire margins. The species is discernible from L. tosaensis by pale-brown lanceolate rhizome scale with a narrow opaque band, remote fronds, and lanceolate leaf scales. Type. Myanmar. Shan state: Pin Laung Township, Ka Thaung upper, 19°57'58.5"N, 96°31'09.1"E, alt. ca. 904 m, 26 Sep. 2019, K. Hori, PK. Khine [“Kine”], T. Fujiwara, M. Nagashima, PP. Shwe & A.K. Moe 108225 (holotype: MBK0328223!, isotype: HITBC! and RAF!). 28 Tao Fujiwara et al. / PhytoKeys 201: 23-34 (2022) Table |. Comparison of morphological characters between Lepisorus medioximus and three related species. Lepisorus medioximus Lepisorus tosaensis Rhizome | Lanceolate, pale brown, Lanceolate, yellow- Broadly lanceolate, pale- | Lanceolate or broadly scale iridescent, clathrate brown, iridescent, almost | brown, clathrate with — | lanceolate, iridescent, with short and narrow, | clathrate, sometimes with | narrow, brown opaque | opaque dark brown dark brown opaque narrow, brown opaque _ | band, margin denticulate, with clathrate band, margin entire to band, margin entire to lumina small margin, lumina small subentire, lumina large | subentire, lumina large Fronds Remote, 0.5-1.5 cm | Remote, 0.5—2 cm apart | Remote, 0.5—2 cm apart | Fronds clustered apart Stipe Stipe short, straw- Stipe straw-colored to Stipe normally straw- | Stipe straw-colored, colored to deep brown, | deep brown, 1-5 cm long | colored, less often brown, 1-3 cm long 0.40.8 cm long 2-5 cm long Laminae Lanceolate, widest at the Lanceolate, widest at Linear-lanceolate to Lanceolate to broadly proximal 1/3 of lamina, middle, base cuneate, lanceolate, widest at lanceolate, widest at base cuneate, slightly slightly decurrent, apex middle, base cuneate, | middle, base cuneate, decurrent, apex long long caudate decurrent, apex shortly decurrent, apex caudate acuminate acuminate Leaf scale Lanceolate, brown, Lanceolate, brown, Ovate, pale brown, Ovate, brown, clathrate clathrate clathrate clathrate Sori On distal 3/4 of lamina, | Restricted to distal 1/3— | Restricted to distal half, Restricted to distal close to costa, orbicular | 1/2 of lamina, midway slightly closer to costa, | half of lamina, close to ovate between costa and orbicular, or slightly ovate] to costa, orbicular margins, orbicular Paraphyses| Ovate-lanceolate, ovate | Orbicular, brown, lumina Orbicular, brown, Orbicular, brownish, to orbicular, brown, small, usually opaque, | clathrate with center dark | clathrate, central clathrate, lumina large, sometimes clathrate, brown, thick and opaque, | lumina small, margin margin entire margins with awn-spines margin denticulate denticulate Description. Plant epiphytic. Rhizomes long creeping, 0.10—0.15 cm in diam., densely scaly, sometimes naked when old; Rhizome scales lanceolate, pale-brown, iridescent, clathrate with short and narrow, dark brown, opaque center band, 2.1— 2.8 mm long x 0.4—0.6 mm wide, margin entire to subentire, apex acuminate, lumina large. Fronds remote, up to 1.5 cm apart; stipe short, 0.4-0.8 cm long, 0.6-1.0 mm diam., straw to dark brown colored; Lamina lanceolate, abaxially grayish-green, adaxi- ally light green when fresh, 8—16 cm long x 0.9-1.5 cm wide, widest at proximal 1/3 of lamina, thinly leathery, adaxially glabrous, abaxially sparsely scaly, lamina base at- tenuate, decurrent, apex long caudate; costa raised on both sides, veinlets obscure; Leaf scales lanceolate, brown, clathrate, 0.8—-1.4 mm long x 0.1—0.3 mm wide, margin denticulate, apex acuminate; Sori on distal 3/4 of lamina, very close to costa, orbicular or elliptic, 0.17—0.35 mm long x 0.12—0.23 mm wide, occasionally sunken on abaxial side of lamina; Paraphyses ovate-lanceolate, ovate to orbicular, brown, clathrate, lu- mina large, margin entire, 0.19-0.28 mm in diam. Etymology. The epithet ‘medioximus’ refers to the sori attached to the middle location on lamina. Distribution. This species is only known from the type locality in Myanmar, Shan state. A new fern species of Lepisorus from Myanmar 29 a a* Figure 2. Living plant of Lepisorus medioximus T.Fujiw., K.Hori & Khine A habit B adaxial side of lamina C abaxial side of lamina with sori D rhizome scale E leaf scale F paraphyses. Habitat. Epiphyte on tree trunks and branches in evergreen to sub-evergreen forest. Additional specimens examined (paratypes). Myanmar. Shan state: Pin Laung Township, Ka Thaung upper, 19°57'58.5"N, 96°31'09.1"E, alt. ca. 904 m, 26 Sep. 2019, K. Hori, PK. Khine [“Kine”], T. Fujiwara, M. Nagashima, P.P. Shwe & A.K. Moe 108229 (MBK 0328227!, HITBC! and RAF!). Note. Until now, we have not discovered additional specimens from other locali- ties despite our exhaustive search focusing on herbarium specimens collected in all 30 Tao Fujiwara et al. / PhytoKeys 201: 23-34 (2022) 0.5mm 0.25 mm Figure 3. Lepisorus medioximus T.Fujiw., K.Hori & Khine (holotype, Hori et al. 108225 = MBK0328223) A habit B rhizome scale C leaf scale and D paraphyses. parts of Myanmar and the Yunnan province of China. We specifically checked not only the Myanmar Lepisorus specimens deposited to the Makino Botanical Garden (MBK), the Institute of Botany, Chinese Academy of Sciences at Beijing (PE) and the Royal Botanic Gardens (K) but also the Lepisorus specimens of Dickason collec- tion deposited in the United States National Herbarium (US), the Natural History Museum (BM), and Naturalis Biodiversity Center (L). Given the observation of more than 50 individuals of the species at the type locality, we expected this spe- cies to be abundant in this poorly collected area. Further inventories in Shan state and the adjacent areas should be necessary to find new localities of the species and evaluate the conservation status of the species. Reflecting our limited knowledge, the IUCN red list status of this species is given as “Data Deficient” instead of “Critical Endangered”. ‘The latter status would assume a restriction of this species range to the two localities recorded. A new fern species of Lepisorus from Myanmar 31 Acknowledgements This study was conducted under a Memorandum of Understanding between the For- est Department, Ministry of Natural Resources and Environmental Conservation, Myanmar, and the Kochi Prefectural Makino Botanical Garden, Japan. The authors greatly appreciate the help in coordinating the expeditions from Dr. Nyi Nyi Kyaw, Director General, Forest Department, and Dr. Thaung Naing Oo, Director of the Forest Research Institute, Forest Department, Ministry of Natural Resources and Environmental Conservation, Myanmar. The authors also thank the Myanmar-Jap- anese Cooperative Program for allowing us to study the specimens collected in Shan State. Fieldwork was performed by the joint research between the Kochi Prefectural Makino Botanical Garden, Japan, and Xishuangbanna Tropical Botanical Garden, China, and was financially supported through the JICA grassroots program and post- doctoral orientation training in Yunnan Province in 2018 (Y7YN021B14). This re- search received partial financial support through the Kochi Prefectural Government. References Bir SS, Trikha CK (1969) Taxonomic revision of the Polypodiaceous genera of India IV: Polypodium lineare complex and allied species. Bulletin of the Botanical Survey of India 11: 260-276. Ching RC (1933) The studies of Chinese ferns IX. Bulletin of the Fan Memorial Institute of Biology. Botany 4: 47-106. Darriba D, Taboada GL, Doallo R, Posada D (2012) jModelTest 2: More models, new heu- ristics and high-performance computing. Nature Methods 9(8): 772-772. https://doi. org/10.1038/nmeth.2109 Dickason FG (1946) The ferns of Burma. The Ohio Journal of Science 46: 109-141. Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin 19: 11-15. Fujiwara T, Serizawa S, Watano Y (2018) Phylogenetic analysis reveals the origins of tetraploid and hexaploid species in the Japanese Lepisorus thunbergianus (Polypodiaceae) complex. Journal of Plant Research 131(6): 945-959. https://doi.org/10.1007/s10265-018-1061-6 Fujiwara T, Liu H, Khine PK, Zuo A, Zhang S, Xing D, Schneider H (2020) Rediscovery of Lepisorus cespitosus supported the floristic affinities between western Yunnan and southeast Tibet. Plant Systematics and Evolution 306(5): e77. https://doi.org/10.1007/s00606-020- 01701-8 Hoang DT, Chernomor O, Von Haeseler A, Minh BQ, Vinh LS (2018) UFBoot2: Improving the ultrafast bootstrap approximation. Molecular Biology and Evolution 35(2): 518-522. https://doi.org/10.1093/molbev/msx28 1 Hori K (2021) Lycophytes & Pteridophytes. In: Fujikawa K, Baba Y, Shin T, Moe AZ, Mizu- kami H (Eds) Taxonomic Enumeration of Natma Taung National Park Vol. 1., 49-102. D2 Tao Fujiwara et al. / PhytoKeys 201: 23-34 (2022) Katoh K, Standley DM (2013) MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Molecular Biology and Evolution 30(4): 772-780. https://doi.org/10.1093/molbev/mst010 Khine PK, Schneider H (2020) Data on pteridophyte species diversity and status of the International Union for Conservation of Nature in each political unit of Myanmar. Data in Brief 30: 105503. https://doi.org/10.1016/j.dib.2020.105503 Khine PK, Fraser-Jenkins C, Lindsay $, Middleton D, Miehe G, Thomas P, Kluge J (2017) A contribution toward the knowledge of ferns and lycophytes from northern and northwest- ern Myanmar. American Fern Journal 107(4): 219-256. https://doi.org/10.1640/0002- 8444-107.4.219 Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution 35(6): 1547-1549. https://doi.org/10.1093/molbev/msy096 Larsson A (2014) AliView: A fast and lightweight alignment viewer and editor for large data- sets. Bioinformatics (Oxford, England) 30(22): 3276-3278. https://doi.org/10.1093/bio- informatics/btu531 Lin YX (2000) Lepisorioideae. Flora of China. Science Press, Beijing, 32-115. Lindsay S, Middleton DJ (2012 onwards) Ferns of Thailand, Laos and Cambodia. http://rbg- web2.rbge.org.uk/thaiferns/ Nguyen LT, Schmidt HA, Von Haeseler A, Minh BQ (2015) IQ-TREE: A fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Molecular Biology and Evolution 32(1): 268-274. https://doi.org/10.1093/molbev/msu300 PPGI (2016) A community-derived classification for extant lycophytes and ferns. Journal of Systematics and Evolution 54(6): 563-603. https://doi.org/10.1111/jse.12229 Qi X-P, Zhang X-C, Lin Y-X, Gilbert MG, Hovenkamp PH (2013) Lepisorus (J. Smith) Ching. In: Wu ZY, Raven PH, Hong DY (Eds) Flora of China Vol. 2—3 (Pteridophytes). Science Press, Beijing & Missouri Botanical Garden Press, St. Louis, 808-824. Rambaut A, Drummond AJ (2013) Tracer v1.6. http://beast.bio.ed.ac.uk/software/tracer/ Ronquist E, Teslenko M, van der Mark P, Ayres DL, Darling A, Hohna S, Larget B, Liu L, Suchard M, Huelsenbeck JP (2012) MrBayes 3.2: Efficient Bayesian phylogenetic infer- ence and model choice across a large model space. Systematic Biology 61(3): 539-542. https://doi.org/10.1093/sysbio/sys029 Vaidya G, Lohman DJ, Meier R (2011) SequenceMatrix: Concatenation software for the fast assembly of multi-gene datasets with character set and codon information. Cladistics 27 (2): 171-180. https://doi.org/10.1111/j.1096-0031.2010.00329.x Vongthavone T, Tagane S, Phonepaseuth P, Souvannakhoummane K, Souladeth P (2021) Twelve new records of Pteridophytes from Bolaven Plateau, southern Laos. Thai Forest Bulletin (Botany) 49: 182-190. https://doi.org/10.20531/tfb.2021.49.2.05 Wang L, Wu Z, Xiang Q, Heinrichs J, Schneider H, Zhang X (2010a) A molecular phylogeny and a revised classification of tribe Lepisoreae (Polypodiaceae) based on an analysis of four plastid DNA regions. Botanical Journal of the Linnean Society 162(1): 28-38. https://doi. org/10.1111/;.1095-8339.2009.01018.x A new fern species of Lepisorus from Myanmar 33 Wang L, Qi X, Xiang Q, Heinrichs J, Schneider H, Zhang X (2010b) Phylogeny of the paleotropical fern genus Lepisorus (Polypodiaceae, Polypodiopsida) inferred from four chloroplast DNA regions. Molecular Phylogenetics and Evolution 54(1): 211-225. https:// doi.org/10.1016/j.ympev.2009.08.032 Wang L, Schneider H, Zhang X, Xiang Q (2012) The rise of the Himalaya enforced the diver- sification of SE Asian ferns by altering the monsoon regimes. BMC Plant Biology 12(1): e210. https://doi.org/10.1186/1471-2229-12-210 Wei XP, Wei R, Zhao CF, Zhang HR, Zhang XC (2017) Phylogenetic position of the enig- matic fern genus Weatherbya (Polypodiaceae) revisited: Evidence from chloroplast and nu- clear gene regions and morphological data. International Journal of Plant Sciences 178(6): 450-464. https://doi.org/10.1086/692088 Yu SL, Lin YX (1996) Research on taxonomy of genus Lepisorus (Smith) Ching in China. Bulletin of Botanical Research 16: 3-24. Yu SL, Lin YX (1997) A study on systematics of the genus Lepisorus (Polypodiaceae). Zhiwu Fenlei Xuebao 35: 341-347. Zhang L, Zhou XM, Liang ZL, Fan XP, Thi Lu N, Song MS, Knapp R, Gao XE Sun H, Zhang LB (2020) Phylogeny and classification of the tribe Lepisoreae (Polypodiaceae; pteridophyta) with the description of a new genus, Ellipinema gen. nov., segregated from Lepisorus. Molecular Phylogenetics and Evolution 148: 106803. https://doi.org/10.1016/j. ympev.2020.106803 Zhao CF, Wei R, Zhang XC, Xiang QP (2020) Backbone phylogeny of Lepisorus (Polypodi- aceae) and a novel infrageneric classification based on the total evidence from plastid and morphological data. Cladistics 36(3): 235-258. https://doi.org/10.1111/cla.12403 Zink MJ (1993) Systematics of the fern genus Lepisorus (J. Smith) Ching (Polypodiaceae- Lepisoreae). Inaugural-Dissertation, Universitat Ziirich, ADAG Administration und Druck AG, Ziirich. Supplementary material | Table S1 Authors: Tao Fujiwara, Phyo Kay Khine, Kiyotaka Hori, Thant Shin, Noriaki Murakami, Harald Schneider Data type: Excel datasheet Explanation note: A checklist for Lepisorus s.s. (sensu stricto) species occurrence in Yunnan and the adjacent regions of Indochina. Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODDbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/phytokeys.201.84911.suppl1 34 Tao Fujiwara et al. / PhytoKeys 201: 23-34 (2022) Supplementary material 2 Table S2 Authors: Tao Fujiwara, Phyo Kay Khine, Kiyotaka Hori, Thant Shin, Noriaki Murakami, Harald Schneider Data type: Excel datasheet Explanation note: Summary table for the information of species used in the phyloge- netic analyses in this study. Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODDbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/phytokeys.201.84911.suppl2