Research Article
Research Article
Hymenasplenium tholiformis (Aspleniaceae), a new fern species from southeastern Xizang, China based on morphological and molecular evidence
expand article infoYong-Lin Qiu, Ke-Wang Xu§, Wen-Bin Ju|, Wang-Lin Zhao#, Liang Zhang¤
‡ Yunnan University, Kunming, China
§ Nanjing Forestry University, Nanjing, China
| Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, China
¶ Tibet University, Lhasa, China
# Chinese Academy of Sciences, Linzhi, China
¤ Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China
Open Access


A new species of Aspleniaceae, Hymenasplenium tholiformis sp. nov., is described from Medog County in southeastern Xizang, China. The new species is morphologically similar to H. apogamum and H. szechuanense, but the former has ascending pinnae, pinna apex obtuse to rounded, pinna-marginal teeth entire, and veins terminating just below marginal teeth. Phylogenetic analysis based on five plastid markers confirmed that this new species represents a diverging lineage in the H. excisum subclade of Hymenasplenium.


Hymenasplenium, H. excisum subclade, Medog, pinna morphology


Hymenasplenium Hayata is one of two genera in the species-rich fern family Aspleniaceae, comprising more than 60 species worldwide (Murakami and Moran 1993; Murakami 1995; PPG I 2016; Schneider et al. 2017; Xu et al. 2018b). The genus is characterized by the stelar structure of the long-creeping rhizome, dorsiventrally symmetrical steles, swollen petiole bases, and unique rachis-costae structure (Murakami 1995; Xu et al. 2018b). Extensive cryptic speciation in Hymenasplenium has been demonstrated based on recent studies (Chang et al. 2018; Xu et al. 2018b), which resulted in the discovery of a large number of new species (Chang et al. 2018; Xu et al. 2018a, 2019a, 2019b).

Located in southeastern Xizang, Medog County is one of the biodiversity hotspots in China, which has rich plant diversity in the Eastern Himalaya (Sun and Zhou 1996). Recently, new taxa of seed plants have continuously added to the flora of this region (Li et al. 2018; Li et al. 2021; Tian et al. 2021; Ya et al. 2021; Zhou et al. 2021). Previous taxonomic studies on the ferns in Xizang have partially uncovered the fern diversity in Medog (Wu 1983; Ching and Ling 1984a, 1984b; Zhang et al. 2004); recent work has added more new species and new records to the fern flora of this region (Wei et al. 2018; Fan et al. 2021; Qiu et al. 2022). As a continuous effort to clarify the species diversity of pteridophytes in Medog, we conducted a 12-day fieldwork in Medog in October 2021. During that trip, we collected some specimens of a species of Hymenasplenium that were obviously different from all other species of the genus. Our later morphological study and phylogenetic analysis confirmed that these specimens represent a distinctive species.

Materials and methods

Morphological studies

Morphological characters of the new species were observed in the field. Herbarium specimens of Hymenasplenium at KUN and PYU were studied. Digital specimens of other related species of Hymenasplenium were examined from the online database CVH ( and JSTOR Global Plants ( Spore samples were taken from the type specimens and coated with gold particles using the BAL-TEC SCD 005 Cool Sputter Coater (BAL-TECAG., Liechtenstein) and imaged via a QUANTA 200 Scanning Electron Microscope (SEM; FEI Co., USA) at Yunnan University, Kunming, China.

Phylogenetic analyses

To clarify the phylogenetic position of the new species, we sampled representatives of all the three major clades of the genus and six subclades in the Old World clade (Xu et al. 2018b). DNA sequences from five plastid markers (atpB, psbA, rbcL, rps4 & rps4-trnS, and trnL & trnL-F) of 102 accessions representing 40 species of Hymenasplenium were sampled. Sixteen species of Asplenium were used as outgroups. Voucher information and GenBank accession numbers for each sampled taxon are provided in Appendix I.

Total genomic DNA was extracted from silica-gel-dried leaves using the TIANGEN plant genomic DNA extraction kit (TIANGEN Biotech., Beijing, China) following the manufacturers’ protocols. Five plastid markers (atpB, psbA, rbcL, rps4 & rps4-trnS, and trnL & trnL-F) were selected for amplification and sequencing. The PCR system uses the ready-to-use rapid PCR master mix gold Mix (green) 25 μl amplification system developed by Beijing Qingke Xinye Biotechnology Co., Ltd. The new sequences were viewed and edited using Sequencher v.4.14 (Gene Codes Corporation, Ann Arbor, Michigan). The total sequences were automatically aligned in MAFFT ver. 7 (Katoh and Standley 2013) and manually adjusted in BioEdit (Hall 1999). jModeltest2 (Darriba et al. 2012) was used to select the best-fitting likelihood model for maximum likelihood (ML; Felsenstein 1973) and Bayesian analyses. The Akaike information criterion (Akaike 1974) was used to select among models instead of the hierarchical likelihood ratio test, following Pol (2004) and Posada and Buckley (2004). Maximum likelihood (ML) bootstrapping was conducted with 1000 rapid bootstrap (BS) analyses followed by a search for the best-scoring tree in a single run in RAxML v. 8 (Stamatakis et al. 2008). Bayesian inference (BI) was conducted using MrBayes 3.1.2 (Huelsenbeck and Ronquist 2001) with two runs of four Markov chain Monte Carlo chains, each beginning with a random tree and sampling one tree every 1000 generations for 10,000,000 generations. Both ML and BI analyses were conducted on Cipres (Miller et al. 2010).

Results and discussion

Morphological comparison

Like most species in Hymenasplenium, the new species have long-creeping rhizome, once-pinnate laminae, asymmetrical pinnae, reddish-brown rachis, and elliptic to reniform spores, but can be distinguished from other species in the genus by the combined characters of pinna apex obtuse to rounded, ascending pinnae, relatively fewer pairs of pinnae, and pinna-marginal teeth entire, veins terminating just below marginal teeth. The pinna shape of H. tholiformis is most distinct in the genus, with acroscopic margins curved and irregularly toothed, basiscopic margins truncate or slightly curved and entire, and pinna apex obtuse to rounded. A few species in the genus sometimes also have pinna apex obtuse, for example, H. apogamum (N.Murak. & Hatan.) Nakaike, H. szechuanense (Ching) Viane & S.Y. Dong (Table 1), but none of them have obviously curved margins on the acroscopic side of pinnae.

Table 1.

Comparison of morphological characters to differentiate Hymenasplenium tholiformis, H. szechuanense, H. apogamum, and H. pseudobscurum.

Characters H. tholiformis H. szechuanense H. apogamum H. pseudobscurum
Size of lamina 13–16 × 3–5 cm 15–25 × 3–5 cm 10–20 × 3–5 cm 20–25 × 5–10 cm
Number of lateral pinnae 15–21 pairs 20–25 pairs 15–25 pairs 15–30 pairs
Pinna shape trapeziform to trapeziform-lunate trapeziform quadrangular-trapeziform trapeziform-falcate
Size of middle pinnae 2.5–3 × 0.6–1 cm 1.5–2.3 × 0.7–1 cm 2–3.5 × 0.6–1 cm 2.5–4 × 0.8–1.8 cm
Shape of pinna apex obtuse to rounded truncate to obtuse obtuse to subacute obtuse to subacute
Stipe color shiny, black purple shiny, dark purple shiny, purple not shiny or purple
Rachis color purple purple purple grayish green
Teeth entire retuse entire entire
Sori position medial inframedial medial supramedial
Indusium single single single double
Number of basalveins lacking 3–4 3–4 1–2 3–5

Molecular phylogenetic analyses

The alignment of five plastid markers was 5,309 bp, of which 3,848 sites were identical, 938 characters were parsimony informative, and 523 variable characters were parsimony-uninformative. A total of 12 sequences are newly generated for this study (Appendix I). The monophyly of Hymenasplenium was confirmed by our reconstructed phylogeny. Hymenasplenium tholiformis was strongly supported as a member of the H. excisum subclade in the Old World clade (Xu et al. 2018b). Its affinity with H. pseudobscurum Viane was weakly supported (Fig. 2). To date, only three species have been recognized in the H. excisum subclade (Xu et al. 2018b); our discovery adds one more species to this subclade. Within the subclade, H. tholiformis is different from H. obscurum (Blume) Tagawa and H. pseudobscurum by having black-purple to dark purple petioles and pinna apex obtuse to rounded, and different from H. excisum (C. Presl) S. Lindsay by having smaller habit and truncate or slightly curved margins of the basiscopic side of pinnae.

Taxonomic treatment

Hymenasplenium tholiformis Liang Zhang, W.B. Ju & K.W. Xu, sp. nov.


China. Xizang: Medog County, Beibeng Xiang, Xirang, ca. 600 m from the Yarlung Zangbo River, 29°11'17.63"N, 95°03'42.27"E, 720 m elev., 28 Oct 2021, Liang Zhang & Wen-Bin Ju 4781 (holotype: KUN1543824!; isotypes: CDBI!, KUN!).


Hymenasplenium tholiformis is morphologically most similar to H. szechuanense, but different by having larger pinnae (middle pinnae 2.5–3 cm vs. 1.5–2.3 cm), extremely ascending upper pinnae (vs. spreading or slightly ascending), curved margins of acroscopic side of pinnae (vs. truncate), and pinna-marginal teeth entire and veins terminating just below marginal teeth (vs. pinna-marginal teeth retuse to emarginate and veins terminating just below these notches).


Plants perennial, 20–36 cm. Rhizome long-creeping, ca. 2 mm in diam., apex scaly; scales dark brown, lanceolate or narrowly triangular, 0.5–1 × 0.2–0.3 mm, margins entire; roots yellowish brown when dried. Fronds remote, 10–12 mm apart, subglabrous; stipe shiny, black purple, 8–13 cm long, base ca. 2–3 mm in diam., with scales similar to those on rhizome; lamina herbaceous, once pinnate, narrowly oblong to lanceolate, 13–16 × 3–5 cm, base truncate and slightly reduced, apex acuminate to caudate; rachis 0.5–1 mm in diam., wingless, narrowly grooved adaxially, shiny, glabrous, black purple to dark purple; pinnae 15–21 pairs, trapeziform to trapeziform-lunate, basal pinnae nearly opposite, spreading or slightly ascending, upper pinnae alternate, extremely ascending, middle pinnae alternate, ascending, 2.5–3 × 0.6–1 cm, dimidiate, pinna asymmetrical, base largest, upper part of pinna enlarged, similar width as, or slightly wider than, the middle part of pinna, apex obtuse to rounded, acroscopic margins curved and irregularly toothed, teeth entire, basiscopic margins truncate or slightly curved and entire (Fig. 1). Veins visible on both sides of pinnae, free, forking and terminating in marginal teeth, 3–4 basal basiscopic veins lacking. Sori medial, linear or semi-elliptic, 5–8 pairs per pinna (Fig. 1), 2–3 mm long; indusia persistent, brown, linear or semi-elliptic, membranous, entire, opening toward costa. Spores elliptic to reniform, perispore fimbriate-alate, 43–47 μm in diam. (Fig. 1); 32 spores per sporangium.

Figure 1. 

Hymenasplenium tholiformis sp. nov. A habit B crozier C adaxial lamina D abaxial lamina E adaxial pinnae F abaxial pinnae, showing sori G rhizome scales H spore surface I spores. Photos credit: A–F by L. Zhang; G–I by W.-Z. Ma & Y.-L. Qiu.

Distribution and conservation assessment

Hymenasplenium tholiformis is endemic to Medog County. Currently, only one large population with ca. 35 individuals was found. According to IUCN Red List criteria B2a or D (IUCN 2022), this species should be listed as critically endangered (CR). More extensive fieldwork at low elevations in nearby mountains will be needed to accurately assess its conservation status.

Figure 2. 

Maximum likelihood phylogeny of Hymenasplenium based on five plastid markers (atpB, psbA, rbcL, rps4 & rps4-trnS, and trnL & trnL-F). The numbers associated with branches are maximum likelihood bootstrap support (MLBS) and Bayesian posterior probability (BIPP). The asterisk indicates MLBS = 100, BIPP = 1.00. The subclades are indicated following Xu et al. (2018b).


Hymenasplenium tholiformis was observed in a shady place at the bottom of a large rock in the disturbed forest, at an elevation of 720 m, ca. 600 m from the Yarlung Zangbo River. High humidity and cool conditions are important for the growth of the new species.


The specific epithet alludes to dome shape of pinna apex.

Vernacular name

yuan ding mo ye tie jiao jue (圆顶膜叶铁角蕨; Chinese name).


In Medog County, ferns are highly diverse along the Yarlung Zangbo River and its tributaries. In this region, at elevations between 650 m and 4500 m, we have discovered four species in three subclades of Hymenasplenium, including H. cheilosorum (Kunze ex Mett.) Tagawa in the H. cheilosorum subclade, H. obliquissimum (Hayata) Sugim. in the H. obliquissimum subclade, and H. excisum and H. tholiformis in the H. excisum subclade. Of the four species, H. tholiformis is distributed at the lowest elevation, while H. obliquissimum is at the highest elevation between 2100 m to 2250 m.


The research was supported by the Second Tibetan Plateau Scientific Expedition and Research (STEP) program (2019QZKK0502) and the National Natural Science Foundation of China (#32100167). We thank Dr. Bo Xu and Dr. Meng Li for their assistance in the field.


  • Chang Y-F, Hori K, Murakami N, Cao L-M, Lu SG, Schneider H (2018) Validation of Hymenasplenium laterepens (Aspleniaceae): Evidence from morphology and molecular analyses. Phytotaxa 374(4): 277–290.
  • Ching R-C, Ling Y-X (1984a) A supplement to the Flora of Xizang (Tibet) (1). Acta Pytotaxonomica Snica 22: 193–204.
  • Ching R-C, Ling Y-X (1984b) A supplement to the Flora of Xizang (Tibet) (2). Acta Pytotaxonomica Snica 22: 397–408.
  • Darriba D, Taboada GL, Doallo R, Posada D (2012) jModelTest 2: More models, new heuristics and parallel computing. Nature Methods 9(8): e772.
  • Fan X-P, Zhang L, Zhang L-B, Zhang L (2021) Selliguea wusugongii (Polypodiaceae), a new fern species from southeastern Xizang, China based on morphological and molecular evidence. Phytotaxa 480(1): 57–68.
  • Felsenstein J (1973) Maximum likelihood and minimum-steps methods for estimating evolutionary trees from data on discrete characters. Systematic Biology 22(3): 240–249.
  • Hall TA (1999) BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95–98.
  • IUCN (2022) Guidelines for Using the IUCN Red List Categories and Criteria. Version 15.
  • 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.
  • Li J-W, Ya J-D, Ye D-P, Liu C, Liu Q, Pan R, He Z-X, Pan B, Cai J, Lin D-L, Jin X-H (2021) Taxonomy notes on Vandeae (Orchidaceae) from China: Five new species and two new records. Plant Diversity 43(5): 379–389.
  • Miller MA, Pfeiffer W, Schwartz T (2010) Creating the CIPRES Science Gateway for inference of large phylogenetic trees. Proceedings of the Gateway Computing Environments Workshop (GCE), 14 November 2010, New Orleans, LA, 1–8.
  • Murakami N (1995) Systematics and evolutionary biology of the fern genus Hymenasplenium (Aspleniaceae). Journal of Plant Research 108(2): 257–268.
  • Murakami N, Moran RC (1993) Monograph of the neotropical species of Asplenium sect. Hymenasplenium (Aspleniaceae). Annals of the Missouri Botanical Garden 80(1): 1–38.
  • Posada D, Buckley TR (2004) Model selection and model averaging in phylogenetics: Advantages of Akaike information criterion and Bayesian approaches over likelihood ratio tests. Systematic Biology 53(5): 793–808.
  • PPG I (2016) A community‐derived classification for extant lycophytes and ferns. Journal of Systematics and Evolution 54(6): 563–603.
  • Qiu Y-L, Zhang L, Zhang L-B, Zhou X-M, Zhang L (2022) Athyrium aberrans (Athyriaceae), a new species of the lady ferns from southeastern Xizang, China, based on morphological and molecular evidence. Phytotaxa 533(3): 165–172.
  • Schneider H, Liu H-M, Chang YF, Ohlsen D, Perrie LR, Shepherd L, Kessler M, Karger DN, Hennequin S, Marquardt J, Russell S, Ansell S, Lu NT, Kamau P, Lóriga J, Regalado L, Heinrichs J, Ebihara A, Smith AR, Gibby M (2017) Neo‐and Paleopolyploidy contribute to the species diversity of Asplenium – The most species‐rich genus of ferns. Journal of Systematics and Evolution 55(4): 353–364.
  • Sun H, Zhou Z (1996) The characters and origin of the flora from the big bend gorge of Yalutsangpu (Brahmabutra) river, eastern Himalyas. Yunnan Zhi Wu Yan Jiu 18: 185–204.
  • Tian D-K, Wang W-G, Dong L-N, Xiao Y, Zheng M-M, Ge B-J (2021) A new species (Begonia giganticaulis) of Begoniaceae from southern Xizang (Tibet) of China. PhytoKeys 187: 189–205.
  • Wei H-J, Zhou X-L, Jin D-M, Yan Y-H (2018) New records of pteridophytes in Xizang. Guihaia 38: 397–407.
  • Wu Z-Y (1983) Flora Xizangica Vol.1. Science Press, Beijing.
  • Xu K-W, Zhang L, Lu NT, Zhou X-M, He H, Luong TT, Knapp R, Liao W-B, Zhang L-B (2018a) Nine new species of Hymenasplenium (Aspleniaceae) from Asia. Phytotaxa 358(1): 1–25.
  • Xu K-W, Zhou X-M, Yin Q-Y, Zhang L, Lu NT, Knapp R, Luong TT, He H, Fan Q, Zhao W-Y, Gao X-F, Liao W-B, Zhang L-B (2018b) A global plastid phylogeny uncovers extensive cryptic speciation in the fern genus Hymenasplenium (Aspleniaceae). Molecular Phylogenetics and Evolution 127: 203–216.
  • Xu K-W, Chen C-W, Kamau P, Liao W-B, Zhang L-B (2019a) Four new species of the fern genus Hymenasplenium (Aspleniaceae) from Africa and Asia. Phytotaxa 416(1): 34–42.
  • Xu K-W, Lorence D, Wood KR, Liao W-B, Zhang L-B (2019b) A revision of the Hymenasplenium unilaterale subclade (Aspleniaceae; Pteridophyta) with the description of nine new species. Phytotaxa 419(1): 1–27.
  • Ya J-D, Zhang T, Pandey TR, Liu C, Han Z-D, Ye D-P, He D-M, Liu Q, Yang L, Huang L, Zhang R-Z, Jiang H, Cai J (2021) New contributions to Goodyerinae and Dendrobiinae (Orchidaceae) in the flora of China. Plant Diversity 43(5): 362–378.
  • Zhang X-C, Shen Z-H, Dong S-Y (2004) Additions to the pteridophyte flora of Xizang (II). Bulletin of Botanical Research 24: 274–277.

Appendix I

Voucher specimens and GenBank accession numbers for DNA sequences used in this study. Information is presented in the following order: species, voucher, locality, GenBank numbers for rbcL, trnL intron & trnL-F spacer, rps4 & rps4-trnS, atpB, psbA. “*” represents the newly published sequences in this study; “–” no data. Herbaria acronyms follow Index Herbariorum (Thiers 2016).

Asplenium aegaeum Lovis, Reichst. & Greuter in Reichst.; Jermy 9181 (BM); Crete; AY300103 (Schneider et al. 2005); AY300050 (Schneider et al. 2005); AY549774 (Schneider et al. 2005); –; –. A. aureum Cav.; Hughes 64 (BM); Belize; AF240651 (Pinter et al. 2002); AF240667 (Pinter et al. 2002); AY549759 (Pinter et al. 2002); –; –. A. cuspidatum Lam.; Grantham & Parsons 0233090 (UC); Costa Rica; AY300111 (Schneider et al. 2004); AY300058 (Schneider et al. 2004); AY549760 (Schneider et al. 2004); –; –. A. dielfalcatum Viane; Wood 7826 (PTBG); Hawaii; AY549738 (Schneider et al. 2005); AY549841 (Schneider et al. 2005); AY549787 (Schneider et al. 2005); –; –. A. erosum Maxon; Unknow; –; KX397706 (Germain-Aubrey,C. C et al., Unpublished); –; –; –; –. A. flabellifolium Cav.; Holmes 4/4/99 (BM); Australia; AY300115 (Schneider et al. 2005); AY300062 (Schneider et al. 2005); AY549779 (Schneider et al. 2005); –; –. A. hastatum Klotzsch ex Kunze; Viane 10182C; Merida, Venezuela; GU929869 (Leroux et al. 2011); –; –; –; –. A. juglandifolium Lam.; Viane 10667; Puerto Rico; GU929870 (Leroux et al. 2011); –; –; –; –. A. normale D. Don; Ohlsen 296 (MELU); Queensland, Australia; KP774926 (Ohlsen et al. 2015); KP851904 (Ohlsen et al. 2015); KP835419 (Ohlsen et al. 2015); –; –. A. polyodon G. Forst; Perrie NC 124 (WELT); New Caledonia; KP774900 (Ohlsen et al. 2015); KP835397 (Ohlsen et al. 2015); KP835433 (Ohlsen et al. 2015); –; –. A. subglandulosum subsp. papaverifolium (Kunze) Salvo, Prada & Consuelo Díaz; Prada & Consuelo Díaz Ohlsen 442 (MELU); Victoria, Australia; KP774929 (Ohlsen et al. 2015); KP851909 (Ohlsen et al. 2015); KP835456 (Ohlsen et al. 2015); –; –. A. tenerum G. Forst; Ohlsen 265 (MELU); Queensland, Australia; KP774858 (Ohlsen et al. 2015); KP835346 (Ohlsen et al. 2015); KP835437 (Ohlsen et al. 2015); –; –. A. trichomanes subsp. inexpectans Lovis; Vogel I-46-B04; France; AY549743 (Schneider et al. 2005); AY549846 (Schneider et al. 2005); AY549792 (Schneider et al. 2005); –; –. A. varians Wall. ex Hook. & Grev.; Fraser-Jenkins 10046–10047 (BM); China; AY300147 (Schneider et al. 2005); AY300094 (Schneider et al. 2005); –; –; –. A. wrightii D. C. Eaton ex Hook.; Cranfill TW040 (UC); Taiwan, China; AY549730 (Schneider et al. 2005); AY549833 (Schneider et al. 2005); AY549766 (Schneider et al. 2005); –; –. A. yunnanense Franch.; Fraser-Jenkins 10044–10045 (BM); China; AY300149 (Schneider et al. 2005); AY300096 (Schneider et al. 2005); AY549803 (Schneider et al. 2005); –; –. Hymenasplenium adiantifrons (Hayata) Viane & S. Y. Dong; Knapp 3609 (P); Taiwan, China; –; MH065559 (Xu et al. 2018); MH065322 (Xu et al. 2018); –; –. H. adiantifrons; Knapp 3904 (P); Taiwan, China; –; MH065560 (Xu et al. 2018); MH065323 (Xu et al. 2018); –; –. H. apogamum N. Murak. & Hatan.; Zhang et al. 7750 (CDBI, MO, VNMN); Thua Thien-Hue, Vietnam; MH065437 (Xu et al. 2018); MH065604 (Xu et al. 2018); MH065376 (Xu et al. 2018); MH065518 (Xu et al. 2018); –. H. apogamum; Chang 1040 (HITBC); Yunnan, China; MH884808 (Chang et al. 2018); –; MH884830 (Chang et al. 2018); –; MH884838 (Chang et al. 2018). H. cardiophyllum (Hance) Nakaike; Sysu 2; Cult. in Sun-yat Sen University; MH065387 (Xu et al. 2018); MH065534 (Xu et al. 2018); MH065306 (Xu et al. 2018); MH065454 (Xu et al. 2018); –. H. cardiophyllum; Zhang et al. 6588 (CDBI, MO, VNMN); Lang Son, Vietnam; –; MH065584 (Xu et al. 2018); –; MH065501 (Xu et al. 2018); –. H. cheilosorum (Kunze ex Mett.) Tagawa; Xu GX022 (SYS); Guangxi, China; MH065381 (Xu et al. 2018); –; MH065343 (Xu et al. 2018); MH065448 (Xu et al. 2018); –. H. cheilosorum; Xu 102 (SYS); Hainan, China; MH065385 (Xu et al. 2018); –; MH065346 (Xu et al. 2018); MH065452 (Xu et al. 2018); –. H. cheilosorum; Xu 299 (SYS); Guangxi, China; MH065402 (Xu et al. 2018); –; MH065350 (Xu et al. 2018); MH065473 (Xu et al. 2018); –. H. cheilosorum; Xu 322 (SYS); Yunnan, China; MH065405 (Xu et al. 2018); –; MH065352 (Xu et al. 2018); MH065476 (Xu et al. 2018); –. H. cheilosorum; Zhang et al. 8629 (CDBI, MO, PHH); Lam Dong, Vietnam; MH065415 (Xu et al. 2018); –; MH065357 (Xu et al. 2018); MH065487 (Xu et al. 2018); –. H. cheilosorum; Jin & Zhang 11348 (CDBI); Yunnan, China; –; MH065571 (Xu et al. 2018); MH065359 (Xu et al. 2018); MH065489 (Xu et al. 2018); –. H. chingii K. W. Xu, Li Bing Zhang & W. B. Liao; Jin & Zhang 11347 (CDBI); Yunnan, China; MH065428 (Xu et al. 2018); MH065586 (Xu et al. 2018); MH065364 (Xu et al. 2018); MH065502 (Xu et al. 2018); –. H. chingii; Zhang et al. 9455 (CDBI); Guizhou, China; MH065430 (Xu et al. 2018); MH065588 (Xu et al. 2018); MH065333 (Xu et al. 2018); MH065504 (Xu et al. 2018); –. H. delitescens (Maxon) L. Regalado & Prada; Jones 1078 (MO); Panama; MH065443 (Xu et al. 2018); MH065609 (Xu et al. 2018); MH065338 (Xu et al. 2018); MH065522 (Xu et al. 2018); –. H. distans Li Bing Zhang, K. W. Xu & Liang Zhang; Zhang et al. 6296 (CDBI, MO, VNMN); Hoa Binh, Vietnam; –; MH065591 (Xu et al. 2018); MH065335 (Xu et al. 2018); MH065507 (Xu et al. 2018); –. H. excisum (C. Presl) S. Linds.; Brownsey & Perrie FIJI 190 (WELT); Fiji; KP774884 (Ohlsen et al. 2015); KP851914 (Ohlsen et al. 2015); KP851882 (Ohlsen et al. 2015); –; –. H. excisum; Zhang et al. 8039 (CDBI, MO, VNMN); Quang Nam, Vietnam; MH065419 (Xu et al. 2018); MH065573 (Xu et al. 2018); MH065361 (Xu et al. 2018); –; –. H. excisum; Xu 100 (SYS); Hainan, China; MH065383 (Xu et al. 2018); MH065531 (Xu et al. 2018); MH065344 (Xu et al. 2018); MH065450 (Xu et al. 2018); –. H. excisum; Xu 110 (SYS); Hainan, China; MH065384 (Xu et al. 2018); MH065532 (Xu et al. 2018); MH065345 (Xu et al. 2018); MH065451 (Xu et al. 2018); –. H. excisum; Xu 198 (SYS); Hainan, China; MH065394 (Xu et al. 2018); MH065541 (Xu et al. 2018); MH065341 (Xu et al. 2018); MH065462 (Xu et al. 2018); –. H. excisum; Zhang et al. 6714 (CDBI, MO, VNMN); Bac Kan, Vietnam; MH065436 (Xu et al. 2018); MH065603 (Xu et al. 2018); MH065375 (Xu et al. 2018); MH065517 (Xu et al. 2018); –. H. filipes (Copel.) Sugim., Murakami 596902 (TI); Kagoshima, Japan; U30605 (Hasebe et al. 1995); –; –; –; –. H. filipes; Murakami 92-J012; Kagoshima, Japan; AB016176 (Murakami et al. 1998); –; –; –; –. H. furfuraceum (Ching) Viane & S. Y. Dong; Xu 304 (SYS); Yunnan, China; MH065406 (Xu et al. 2018); MH065553 (Xu et al. 2018); MH065353 (Xu et al. 2018); MH065477 (Xu et al. 2018); –. H. furfuraceum; Xu 307 (SYS); Yunnan, China; MH065409 (Xu et al. 2018); MH065557 (Xu et al. 2018); MH065320 (Xu et al. 2018); MH065481 (Xu et al. 2018); –. H. furfuraceum; Chang1072 (HITBC); Yunnan, China; MW194198 (Zhang et al. 2021); –; MW194221 (Zhang et al. 2021); –; MW194182 (Zhang et al. 2021). H. hastifolium Ke Wang Xu, Li Bing Zhang &W. B. Liao; Xu 282-2 (SYS); Guangxi, China; MH065397 (Xu et al. 2018); MH065546 (Xu et al. 2018); MH065312 (Xu et al. 2018); MH065468 (Xu et al. 2018); –. H. hastifolium; Xu 282-1 (SYS); Guangxi, China; MH065398 (Xu et al. 2018); MH065547 (Xu et al. 2018); MH065313 (Xu et al. 2018); MH065469 (Xu et al. 2018); –. H. hoffmannii (Hieron.) L. Regalado & Prada; Croat & Zhu 76985 (MO); Canal, Panama; MH065441 (Xu et al. 2018); MH065607 (Xu et al. 2018); MH065378 (Xu et al. 2018); –; –. H. hoffmannii; Monro & Douglas 3463 (MO); Santa Ana, El Salvador; MH065442 (Xu et al. 2018); MH065608 (Xu et al. 2018); MH065337 (Xu et al. 2018); MH065521 (Xu et al. 2018); –. H. hondoense (N.Murak. & Hatan.) Nakaike; Murakami 596920 (KYO); Kouchi, Japan; AB014705 (Murakami et al. 1999); –; –; –; –. H. hondoense; Chang1062 (HITBC); Yunnan, China; MH884814 (Chang et al. 2018); –; MH884833 (Chang et al. 2018); –; MH884840 (Chang et al. 2018). H. laetum (Sw.) L. Regalado & Prada; Jones 1155 (TUR); –; –; –; –; KM114105 (Lehtonen,S et al., Unpublished); –. H. laterepens N. Murak. & X. Cheng ex Y. Fen Chang & K. Hori; Chang 2655 (HITBC); Yunnan, China; MH884806 (Ohlsen et al. 2015); –; –; –; –. H. laterepens; Chang 1039 (HITBC); Yunnan, China; MH884807 (Chang et al. 2018); –; MH884829 (Chang et al. 2018); –; MH884837 (Chang et al. 2018). H. latidens (Ching) Viane & S. Y. Dong; Knapp 3672 (P); Taiwan, China; –; MH065566 (Xu et al. 2018); –; –; –. H. latidens; Xu 309 (SYS); Yunnan, China; MH065407 (Xu et al. 2018); MH065554 (Xu et al. 2018); MH065318 (Xu et al. 2018); MH065478 (Xu et al. 2018); –. H. latidens; Y.-F. Chang1029 (HITBC); Yunnan, China; MW194204 (Zhang et al. 2021); –; MW194219 (Zhang et al. 2021); –; MW194180 (Zhang et al. 2021). H. murakami-hatanakae Nakaike; Ranker 2072 (COLO); Taiwan, China; EF452140 (Schuettpelz et al. 2007); – (Schuettpelz et al. 2007); – (Schuettpelz et al. 2007); EF452020 (Schuettpelz et al. 2007); –. H. murakami-hatanakae Nakaike; C.-C. Chen 1089 (HITBC); Taiwan, China; MH884823 (Chang et al. 2018); –; –; –; –. H. neocaledonicum Li Bing Zhang & K. W. Xu; PerrieNC177 (WELT); New Caledonia; KP774896 (Ohlsen et al. 2015); KP851915, (Ohlsen et al. 2015); KP851878 (Ohlsen et al. 2015); –; –. H. ngheanense Li Bing Zhang, K. W. Xu & N. T. Lu; Zhang et al. 6532 (CDBI, MO, VNMN); Phu Tho, Vietnam; MH065426 (Xu et al. 2018); MH065583 (Xu et al. 2018); MH065331 (Xu et al. 2018); MH065500 (Xu et al. 2018); –. H. ngheanense; Zhang et al. 7163 (CDBI, MO, VNMN); Nghe An, Vietnam; MH065439 (Xu et al. 2018); MH065605 (Xu et al. 2018); MH065336 (Xu et al. 2018); MH065519 (Xu et al. 2018); –. H. obliquissimum (Hayata) Sugim.; Murakami & J. Yokoyarna 94-T628; Chiang Mai, Thailand; AB016178 (Murakami et al. 1998); –; –; –; –. H. obliquissimum; lwatsuki et al. 94-V302; Hoang Lien Son, Vietnam; AB016187 (Murakami et al. 1998); –; –; –; –. H. obliquissimum; Ju & Deng 12689 (CDBI); Sichuan, China; –; MH065597 (Xu et al. 2018); MH065370 (Xu et al. 2018); MH065511 (Xu et al. 2018); –. H. obliquissimum; Chang1292 (HITBC); Tibet, China; MW194212 (Zhang et al. 2021); –; MW194232 (Zhang et al. 2021); –; MW194192 (Zhang et al. 2021). H. obscurum (Blume) Tagawa; Xu 324 (SYS); Yunnan, China; MH065404 (Xu et al. 2018); MH065552 (Xu et al. 2018); MH065351 (Xu et al. 2018); MH065475 (Xu et al. 2018); –. H. obscurum; Zhang et al. 7715 (CDBI, MO, VNMN); Thanh Hoa, Vietnam; MH065411 (Xu et al. 2018); MH065567 (Xu et al. 2018); MH065354 (Xu et al. 2018); MH065483 (Xu et al. 2018); –. H. obscurum; Wu et al. WS2176 (MO); Xiangkhoang, Laos; ON859869 *; ON859878 *; ON859875;ON859872 *; –. H. obscurum; Wu et al. WS2552 (MO); Louangphrabang, Laos; ON859870 *; ON859879 *; ON859876*;ON859873 *; –. H. perriei Li Bing Zhang & K. W. Xu; Brownsey & Perrie FIJI 13 (WELT); Fiji; KP774885 (Ohlsen et al. 2015); KP851918 (Ohlsen et al. 2015); KP851880 (Ohlsen et al. 2015); –; –. H. phamhoanghoi Li Bing Zhang, K. W. Xu & T. T. Luong; Zhang et al. 8819 (CDBI, MO, PHH); Khanh Hoa, Vietnam; MH065432 (Xu et al. 2018); MH065590 (Xu et al. 2018); MH065334 (Xu et al. 2018); MH065506 (Xu et al. 2018); –. H. pseudobscurum Viane; Knapp 4133 (P); Taiwan, China; –; MH065565 (Xu et al. 2018); –; –; –. H. pseudobscurum; Xu 004 (SYS); Hong Kong, China; MH065380 (Xu et al. 2018); MH065530 (Xu et al. 2018); MH065342 (Xu et al. 2018); MH065447 (Xu et al. 2018); –. H. pseudobscurum; Fraser-Jenkins30715; Bhutan; MH884826 (Chang et al. 2018); –; MH884834 (Chang et al. 2018); –; –. H. pseudobscurum; Chang 1010 (HITBC); Yunnan, China; MH884827 (Chang et al. 2018); –; MH884835 (Chang et al. 2018); –; MH884841 (Chang et al. 2018). H. pseudobscurum; Chang 1047 (HITBC); Yunnan, China; MH884828 (Chang et al. 2018); –; MH884836 (Chang et al. 2018); –; MH884842 (Chang et al. 2018). H. quangnamense Li Bing Zhang, K. W. Xu & Liang Zhang; Zhang et al. 7884 (CDBI, MO, VNMN); Quang Nam, Vietnam; MH065412 (Xu et al. 2018); MH065568 (Xu et al. 2018); MH065355 (Xu et al. 2018); MH065484 (Xu et al. 2018); –. H. quangnamense; Schuettpelz 874; Malay Peninsula; MH884824 (Chang et al. 2018); –; –; –; –. H. queenslandicum Li Bing Zhang & K. W. Xu; Ohlsen 252 (MELU); Queensland, Australia; KP774849 (Ohlsenet al. 2015); KP851916 (Ohlsenet al. 2015); KP851879 (Ohlsen et al. 2015); –; –. H. quercicola (Ching) Viane & S. Y. Dong; Chang1064 (HITBC); Yunnan, China; MW194205 (Zhang et al. 2021); –; MW194220 (Zhang et al. 2021); –; MW194181 (Zhang et al. 2021). H. retusulum (Ching) Viane & S. Y. Dong; Jiang 00371 (SYS); Yunnan, China; MH065379 (Xu et al. 2018); MH065529 (Xu et al. 2018); MH065304 (Xu et al. 2018); MH065446 (Xu et al. 2018); –. H. retusulum; Xu 311 (SYS); Yunnan, China; MH065408 (Xu et al. 2018); MH065556 (Xu et al. 2018); MH065319 (Xu et al. 2018); MH065480 (Xu et al. 2018); –. H. riparium (Liebm.) L. Regalado & Prada; Murakami & Grayum 281 (KYO); Virgen del Socorro, Costa Rica; AB014708 (Murakami et al. 1999); –; –; –; –. H. sinense K. W. Xu, Li Bing Zhang & W. B. Liao; Xu PB001 (SYS); Yunnan, China; MH065386 (Xu et al. 2018); MH065533 (Xu et al. 2018); rps4 & MH065347 (Xu et al. 2018); MH065453. (Xu et al. 2018); –. H. sinense; Xu 134 (SYS); Jiangxi, China; MH065388 (Xu et al. 2018); MH065535 (Xu et al. 2018); MH065348 (Xu et al. 2018); MH065455 (Xu et al. 2018); –. H. sinense; Xu GX018 (SYS); Guangxi, China; –; –; MH065349 (Xu et al. 2018); MH065465 (Xu et al. 2018); –. H. speluncicola Li Bing Zhang, K. W. Xu & H. He; Zhang et al. 5494 (CDBI, MO); Guangxi, China; –; MH065592 (Xu et al. 2018); MH065366 (Xu et al. 2018); MH065508 (Xu et al. 2018); –. H. speluncicola; Zhang et al. 6035 (CDBI); Guizhou, China; –; MH065594 (Xu et al. 2018); MH065368 (Xu et al. 2018); MH065509 (Xu et al. 2018); –. H. subnormale (Copel.) Nakaike; Knapp 4525 (P); Taiwan, China; –; MH065612 (Xu et al. 2018); –; MH065526 (Xu et al. 2018); –. H. subnormale; Knapp 4509 (P); Taiwan, China; –; MH065613 (Xu et al. 2018); –; MH065527 (Xu et al. 2018); –. H. szechuanense (Ching) Viane & S. Y. Dong; Chang1099 (HITBC); Sichuan, China; MW194207 (Zhang et al. 2021); –; MW194223 (Zhang et al. 2021); –; MW194183 (Zhang et al. 2021). H. szechuanense; Chang1098 (HITBC); Sichuan, China; MW194206 (Zhang et al. 2021); –; MW194222 (Zhang et al. 2021); –; MW194193 (Zhang et al. 2021). H. tholiformis Liang Zhang, W.B. Ju & K.W. Xu; Zhang Liang 4781 (KUN); Tibet, China; ON859868 *; ON859877 *; –; ON859871 *; ON859874 *. H. triquetrum (N. Murak. & R. C. Moran) L. Regalado & Prada; Sylvestre 2208 (RB); Brazil; KT329398 (Mynssen et al. 2016); –; –; –; –. H. triquetrum; Jimenez 5153 (MO); La Paz, Bolivia; MH065444 (Xu et al. 2018); MH065610 (Xu et al. 2018); MH065339 (Xu et al. 2018); MH065523 (Xu et al. 2018); –. H. unilaterale (Lam.) Hayata; Viane 8344; Reunion; GU929873 (Leroux et al. 2011); –; –; –; –. H. unilaterale; Hemp A. 18 (BM); Kenya; AF240652 (Pinter et al. 2002); AF525232 (Pinter et al. 2002); –; –; –. H. vanuatuense Li Bing Zhang & K. W. Xu; Ohlsen 392 (MELU); Tanna, Vanuatu; KP774898 (Ohlsen et al. 2015); KP851917 (Ohlsen et al. 2015); KP851881 (Ohlsen et al. 2015); –; –. H. wangpeishanii Li Bing Zhang & K. W. Xu; Zhang et al. 9665 (CDBI); Guizhou, China; MH065423 (Xu et al. 2018); MH065577 (Xu et al. 2018); –; MH065494 (Xu et al. 2018); –. H. wangpeishanii; Zhang et al. 9455 (CDBI); Guizhou, China; MH065430 (Xu et al. 2018); MH065588 (Xu et al. 2018); MH065333 (Xu et al. 2018); MH065504 (Xu et al. 2018); –. H. wildii (Bail.) D.J.Ohlsen; Ohlsen 246 (MELU); Queensland, Australia; KP774927 (Ohlsen et al. 2015); KP851919 (Ohlsen et al. 2015); KP851877 (Ohlsen et al. 2015); –; –. H. wuliangshanense (Ching) Viane & S. Y. Dong; Y.-F. Chang1102 (HITBC); Yunnan, China; MW194208 (Zhang et al. 2021); –; MW194224 (Zhang et al. 2021); –; MW194184 (Zhang et al. 2021). H. wuliangshanense; Chang1104 (HITBC); Yunnan, China; MW194209 (Zhang et al. 2021); –; MW194225 (Zhang et al. 2021); –; MW194185 (Zhang et al. 2021).