Oreocharis jasminina (Gesneriaceae), a new species from mountain tops of Hainan Island, South China

Shao-Jun Ling; Shu-Ping Guan; Fang Wen; Yu-Min Shui; Ming-Xun Ren

doi:10.3897/phytokeys.157.50246

Abstract

A new species of Gesneriaceae, Oreocharis jasminina S.J.Ling, F.Wen & M.X. Ren from Hainan Island, south China, is highlighted and described. The new species is distinguished by its actinomorphic corolla, narrow floral tube and ovate anthers hidden in the floral tube. The new species also showed clear geographic and altitudinal isolation from the three currently-recognised Oreocharis species on the Island. Molecular phylogenetic analysis, based on nuclear ITS1/2 and plastid trnL-trnF sequences, supported the delimitation of the new species, which forms a single lineage with all the other Oreocharis species from Hainan Island. The roles of geographic and floral isolation in the evolution of the new species and its affinities are discussed.

Keywords

Hainan Island, new taxon, Oreocharis

Introduction

The Oreocharis Bentham was recently re-circumscribed to a large genus by including ten more genera and over 135 species, based mainly on molecular phylogenetic studies (Möller et al. 2011, 2016; Xu et al. 2017; Möller 2019; Wen et al. 2019). The enlarged genus was predominantly distributed in China with some species in India, Myanmar, Thailand and Vietnam (e.g. Li and Wang 2005; Möller and Clark 2013; Möller et al. 2018). Regardless of the limited differences in habit and fruit structure, Oreocharis shows a strikingly-high diversity in floral syndromes (Li and Wang 2005; Wei 2010; Möller and Clark 2013).

As one of the globally-important biodiversity hotspots, Hainan Island harbours about 4000 seed-plant species, of which ca. 500 are endemics (Francisco-Ortega et al. 2010) and which are concentrated in the south-central mountains. Gesneriaceae, in Hainan Island especially, includes a high ratio of species endemism, eight out of the total of 24 species being endemic (Ling et al. 2017a). Currently, three taxa of Oreocharis are recorded on Hainan Island and all of them are Hainan-endemic and monophyletic, i.e. O. dasyantha Chun, O. dasyantha Chun var. ferruginosa Pan and O. flavida Merrill (Li and Wang 2005; Ling et al. 2020), while each of these species shows considerable variations in morphological traits (Wei 2010; Ling et al. 2020a).

During several fieldwork trips in the past three years, we found that some populations of Oreocharis on mountain tops in Hainan Island showed obvious differences in various morphological characters. After careful literature studies (Pan 1987; Li and Wang 2005; Wei 2010) and morphological and molecular examinations, we are convinced that populations from the mountain tops of Mt. Yingge and Mt. Limu represent a new species, which we report and describe here.

Materials and methods

Morphological observations

The field study and conservation on Gesneriaceae were undertaken by two of the authors (SJL and MXR) over a long period of time, especially focusing on the Hainan-endemic species (Ling et al. 2017a, b; Xing et al. 2018; Li et al. 2019). Morphological observations and measurements were carried out, based on living plants during fieldwork. All available specimens of Oreocharis species, stored in the herbaria in China (PE, KUN, IBK and IBSC), were examined. We also downloaded all Oreocharis specimens from JSTOR Global Plants (http://plants.jstor.org), and Chinese Virtual Herbarium (http://www.cvh.ac.cn) to compare detailed morphological traits between the proposed new species with the currently-accepted species of Oreocharis. Specifically, we compared morphological traits of the possible new species with all the three currently-recognised Oreocharis species from Hainan Island, i.e. O. dasyantha, O. dasyantha var. ferruginosa and O. flavida. The specimens of new species were collected over the past two years and deposited in the herbarium of Hainan University (HUTB) and Kunming Institute of Botany, Chinese Academy of Sciences (KUN).

Taxonomic sampling, DNA extraction and molecular markers

The leaf samples of O. dasyantha, O. dasyantha var. ferruginosa, O. flavida and the putative new species were collected in the field and dried in a vascular bag with silica gel. Total genomic DNA extraction was conducted using CTAB methods (Doyle and Doyle 1987). One nuclear ribosomal DNA (nrDNA) sequence, the ITS region comprising spacer 1, the 5.8S gene and spacer 2 (White et al. 1990) and one chloroplast DNA (cpDNA) intron-spacer region trnL-trnF (Taberlet et al. 1991) were used in this study. Laboratory procedures followed Ling et al. (2020) and newly-acquired sequences were deposited in GenBank (Table 2).

Alignments and phylogenetic analyses

According to Möller et al. (2011), Chen et al. (2014) and Ling et al. (2020), Oreocharis sinohenryi (Chun) Mich.Möller & A.Weber which had the closest phylogenetic relationships with the Hainan Oreocharis taxa was used as outgroup with sequences (Genbank with accession numbers HQ632913 and HQ633009). The original chromatograms from both directions of the ITS1/2 and trnL-trnF sequences were evaluated using Bioedit (Hall 1999) for base confirmation and contiguous sequences editing, then we manually aligned sequences, where necessary, using MEGA v.6.5 (Kumar et al. 2008) and ambiguous positions were excluded from the alignments. The ITS1/2 and trnL-trnF were concatenated to a single matrix after a congruency test by PAUP* 4.0a164 (Swofford 2003). Bayesian Inference (BI) analysis was conducted using MrBayes version 3.1.2 (Huelsenbeck and Ronquist 2001) and Maximum Likelihood (ML) analysis was performed using MEGA v.6.5 (Kumar et al. 2008). Both procedures followed the Ling et al. (2020), based on the combined ITS1/2 and trnL-trnF sequences.

Results

Phylogenetic reconstruction

The combined ITS1/2 and trnL-trnF datasets were 640 and 818 bp long, amongst which 64 and 17 were polymorphic sites and 27 and 6 were parsimony-informative sites, respectively. The aligned dataset was 1458 bp long and a total number of 81 polymorphic sites were measured, of which 33 were parsimony-informative sites. There was no significant incongruence, based on the incongruence length difference (ILD) test between the ITS1/2 and trnL-trnF (p > 0.05).

Molecular phylogeny recognised the individuals from different mountains and these were grouped as separate lineages. The putative new species from Mt. Limu and Mt. Yingge is accepted as a new species with PP (posterior probability) = 1 and BS (bootstrap value) = 100% (Fig. 1). All the Oreocharis species from Hainan Island form a single lineage with relatively-high support (Fig. 1).

Figure 1.

Molecular phylogeny of Hainan Oreocharis taxa with outgroup O. sinohenryi, based on the combined nuclear ribosomal DNA (nrDNA) sequence ITS1/2 and chroloplast gene trnL-trnF data matrices. Posterior probability (PP) and Bootstrap value (BS) are showed above branches.

Taxonomic treatment

Oreocharis jasminina S.J.Ling, F.Wen & M.X. Ren, sp.nov.

urn:lsid:ipni.org:names:77211189-1

Figs 2, 3

Diagnosis

Oreocharis jasminina has the closest phylogenetic relationship with O. dasyantha, O. dasyantha var. ferruginosa and O. flavida with very high support values, all being Hainan-endemic and monophyletic. O. jasminina can be easily distinguished from them by having: (1) a long and narrow floral tube (both O. dasyantha and O. dasyantha var. ferruginosa have conical floral tubes, O. flavida has campanulate-tubular floral tube); (2) yellow and actinomorphic corolla (both O. dasyantha and O. dasyantha var. ferruginosa are zygomorphic with orange-red to yellow corolla, O. flavida is actinomorphic with orange corolla); (3) didynamous stamens with ovate anthers hidden in the floral tube (both O. dasyantha and O. dasyantha var. ferruginosa have exposed didynamous stamens with ovate anthers, O. flavida has four equivalent stamens with horseshoe-shaped anthers included in the floral tube) (Table 1, Fig. 4).

Figure 2.

Oreocharis jasminina sp. nov. A Habitat B habit C face view of corolla D lateral view of corolla E opening flower showing stamens and staminode F stamens and staminode G pistil and sepals H fruit pods I adaxial leaf surface J abaxial leaf surface.

Figure 3.

Oreocharis jasminina sp. nov. (all drawings based on the holotype S.J.Ling 20181126–01 in HUTB, drawn by S.P. Guan). A Habit B face view of corolla C lateral view of corolla D opening corolla showing pistil and stamens E pistil F stamens and staminode.

Figure 4.

Three formerly-recognised Oreocharis taxa in Hainan Island. Oreocharis dasyantha (A, B), Oreocharis dasyantha var. ferruginosa (C, D) and Oreocharis flavida (E, F).

Type

China. Hainan: Qiongzhong County (琼中县), Limu Mountain, 1350 m a.s.l., on moist rocks, 26 Nov 2018, S.J.Ling 2018112601 (holotype: HUTB!; Isotypes: HUTB!, KUN!).

Description

Perennial herb, rhizomatous, leaves basal; 4.0–10.0 cm long, 2–3 mm in diameter, densely pale brown villous or woolly; leaf blade ovate to broadly ovate, rarely elliptic or obovate, 6–11 × 4–8 cm, adaxially densely grey to brown pubescent, abaxially sparsely to densely grey or grey-brown pubescent, sparsely brown villous along veins which are adaxially sunken and abaxially ridged, lateral veins 6–7 on each side of midrib, base often cordate to rounded, margin nearly entire to shallowly crenate, apex rounded. Cymes axillary, 2–3, inflorescence 3–10-flowered; Peduncle 9–16 cm long, sparsely pale grey villous; bracts 2, linear to narrowly triangular, outside densely villous, apex acuminate, cuneate to triangular, margin entire; pedicel 1.5–2.2 cm long, densely pale brown villous to woolly. Calyx 5-lobed, divided to base, lobes green, narrowly lanceolate, 9–11 × ca. 2 mm, apex acuminate, margin entire, outside villous, inside glabrous. Corolla yellow, 1.7–2.2 cm long, outside pubescent; tube thin tubular, 1.8–2.1 cm × 3–4.5 mm, limb barely 2-lipped, adaxial lip shallowly 2-lobed from near base, abaxial 3-lobed slightly equal. Stamens 4, 8–9 mm long, included, adnate to corolla 4–5 mm from base; filaments slender, pubescent; anthers ovate, 2-loculed, dehiscing transversely; staminode 1, adnate to corolla 2–4 mm from base, ca. 2 mm. Disc ca. 1 mm high, entire. Pistil ca. 7 mm long; ovary cylindrical, ca. 5 mm long, glabrous. Stigma 2, equal, suborbicular. Capsula linear, 3–4 cm long, glabrous to sparsely puberulent.

Table 1.

Download as

CSV

XLSX

Diagnostic morphological characters of Oreocharis jasminina sp. nov. and all the three currently-recognised species in Hainan Island.

Characters	Oreocharis jasminina sp. nov.	O. dasyantha	O. dasyantha var. ferruginosa	O. flavida
Corolla colour	yellow	orange-red to yellow	orange-red to yellow	orange
Corolla tube	narrowly tubular,1.7–2.2 cm long 1.8–2.2 cm × 3–4.5 mm	conical,1.6–2.4 cm long 0.9–2 cm × 6–7 mm	conical, ca.1.6 cm, tube 9–1.1 mm	campanulate-tubular,1.7–1.9 cm long 1.6–1.8 cm × 6–8 mm
Corolla symmetry	actinomorphic	zygomorphic	zygomorphic	actinomorphic
Leaf blade shape	ovate to broadly ovate, rarely elliptic or obovate	ovate-elliptic to broadly ovate	ovate-elliptic to broadly ovate	ovate-elliptic to broadly ovate, rarely broadly elliptic
Leaf base shape	cordate to rounded	oblique, cuneate to subrounded or cordate	sometimes oblique, cuneate to subcordate	oblique, subrounded
Leaf base margin	nearly entire to shallowly crenate, apex rounded	serrulate or crenate-serrate, apex acute to rounded	crenate-serrate	shallow crenate
Stamens	included, didynamous, staminode 1	exposed, didynamous, staminode absent	exposed, equivalent, staminode absent	included, equivalent, staminode 1
Anthers	ovate, 2-loculed, dehiscing transversely	broadly oblong, 2-loculed, dehiscing longitudinally	broadly oblong, 2-loculed, dehiscing longitudinally	horseshoe-shaped,1-loculed, dehiscing transversely
Filaments	pubescent	pubescent	pubescent	glabrous
Pistil	ca. 9 mm long	ca. 22mm long	ca. 22mm long	ca.9 mm long

Phenology

Oreocharis jasminina flowers from September to December and fruits from November to January.

Distribution and habitat

Oreocharis jasminina is currently only found in cloud forests on the mountain tops of Mt. Limu and Mt. Yingge, in the middle of Hainan Island. The habitat of O. jasminina is on the moss layer on wet rocks under cloud forests.

Etymology

The specific epithet refers to the yellow and narrowly tubular corolla of this new species.

Vernacular name

迎春花马铃苣苔 (Yíng Chūn Huā Mǎ Líng Jù Tái) is the Chinese name for Oreocharis jasminina, the first three characters meaning ‘winter jasmine’, indicating its similar floral syndromes to Jasminum nudiflorum Lindl. The last four characters are the Chinese name for Oreocharis.

Conservation status

Oreocharis jasminina is, so far, known only from the two locations with about 800–1000 individuals. The populations are under threat due to the restricted and fragmented habitat. Therefore, we propose that O. jasminina should be considered as ‘Vulnerable’ (VU), according to the IUCN Red List Categories and Criteria (IUCN 2012).

Table 2.

Download as

CSV

XLSX

List of Hainan Oreocharis taxa and outgroup O. sinohenryi used in the phylogenetic analysis, including respective Genbank accession and voucher numbers.

Species	trnL-trnF	ITS1/2	Voucher Number
O. dasyantha Chun (Mt. Bawang)-1	MK587993	MK587954	S.J.Ling & M.X. Ren 2015011803 (HUTB)
O. dasyantha Chun (Mt. Bawang)-2	MK587994	MK587954	S.J.Ling & M.X. Ren 2015011804 (HUTB)
O. dasyantha Chun (Mt. Jianfeng)-1	MK587995	MK587955	S.J.Ling 2015102201 (HUTB)
O. dasyantha Chun (Mt. Jianfeng)-2	MK587996	MK587955	S.J.Ling 2015102202 (HUTB)
O. dasyantha Chun var. ferruginosa Pan (Mt. Jianfeng)-1	MK587954	MK587956	S.J.Ling 2015102203 (HUTB)
O. dasyantha Chun var. ferruginosa Pan (Mt. Jianfeng)-2	MK587954	MK587957	S.J.Ling 2015102204 (HUTB)
O. flavida Merrill (Mt. Qixian)	MK587947	MK587990	S.J.Ling 2018112901 (HUTB)
O. flavida Merrill (Mt. Wuzhi)	MK587989	MK587943	S.J.Ling 2018112902 (HUTB)
O. jasminina (Mt. Yingge)-1	MK587987	MK587948	S.J.Ling 2018112601 (HUTB)
O. jasminina (Mt. Yingge)-2	MK587988	MK587950	S.J.Ling 2018112602 (HUTB)
O. jasminina (Mt. Limu)-1	MK587981	MK587949	S.J.Ling 2018112603 (HUTB)
O. jasminina (Mt. Limu)-2	MK587982	MK587953	S.J.Ling 2018112604 (HUTB)
O. sinohenryi (Chun) Mich.Möller & A.Weber	HQ632913	HQ633009	M.Möller MMO 07-1150 (E)

Key to Oreocharis jasminina and its closely-related and sympatric species in Hainan Island

1	Anthers horseshoe-shaped, 1-loculed, dehiscing transversely	*O. flavida*
–	Anthers broadly oblong, 2-loculed, dehiscing longitudinally	2
2	Stamens included, floral tube thin tubular, corolla yellow	*O. jasminina*
–	Stamens exposed, floral tube conical, corolla orange-red	3
3	Leaf blade adaxially grey pubescent, base oblique, subrounded to cordate, margin serrulate; petiole to 14.5 cm, densely pale brown villous; cymes 1–3(or 4)-flowered; corolla 1.7–2.4 cm, tube 1.1–2 cm	*O. dasyantha*
–	Leaf blade adaxially grey to brown pubescent and villous, base sometimes oblique, cuneate to subcordate, margin crenate-serrate; petiole to 6 cm, densely pale brown woolly; cymes 3–8-flowered; corolla ca. 1.6 cm, tube 9–11 mm	O. dasyantha var. ferruginosa

Discussion

Our former study showed the new species O. jasminina and the three other Hainan-endemic taxa are homologous, indicating these species in Hainan Island had a common origin (Ling et al. 2020). The new species is only found on mountain tops higher than 1200 m in two mountains, Mt. Limu and Mt. Yingge, located at the middle of Hainan Island. These mountain tops likely formed island-like habitats because the deep and wide valleys interrupted gene flows, resulting in population differentiation and speciation (Shen et al. 2017; Ling et al. 2017a, b; Xing et al. 2018). Such ‘sky islands’ may be the main reason for the origin and maintenance of this Hainan-endemic alpine species (Robin et al. 2015; Ling et al. 2017a).

The new species also shows a clear geographic isolation from the three currently-recognised Oreocharis taxa on Hainan Island. The new species O. jasminina was only found in Mt. Limu and Mt. Yingge in the middle of the island, while O. dasyantha and O. dasyantha var. ferruginosa are restricted to the west side of the Island and O. flavida was only found in the east side (Fig. 5). They are isolated by a large river, the Changhua River (the second largest river on Hainan Island). Li et al. (2019) found that the geographic isolation by the Changhua River is a driving force for the great population differentiation in the two Hainan-endemic Gesneriaceae species, Primulina heterotricha (Merr.) Yan Liu and Metapetrocosmea peltata (Merr. et Chun) W. T. Wang. Thus, the geographic isolation by rivers or valleys may also play a key role in the evolution of O. jasminina and other Hainan-endemic Oreocharis taxa. However, the relative contributions of such geographic isolation and altitudinal differentiation are still in need of further experimental examination.

Figure 5.

Geographic distribution of Oreocharis jasminina sp. nov. and the three congeners on Hainan Island. ★ O. jasminina sp. nov. ∆ O. dasyantha ◊ O. dasyantha var. ferruginosa □ O. flavida.

Floral symmetry is widely recognised as a key trait in pollination and taxonomy. Normally, the zygomorphic corolla possesses higher pollen-transfer efficiency than the actinomorphic corolla (Sargent 2004). Oreocharis jasminina has yellow actinomorphic corolla with a long and narrow floral tube, differing from O. dasyantha and O. dasyantha var. ferruginosa (both have zygomorphic corolla). Although O. flavida has an actinomorphic flower, its campanulate corolla with four equivalent stamens and horseshoe-shaped anthers make it distinct from the new species O. jasminina (Table 1).

Floral shape was expected to be a vital factor in generating floral isolation and evolutionary shifts (Castellanos et al. 2004; Muchhala 2007). Generally, the floral shape has a strong connection with the expected pollinators in Gesneriaceae, for example, bees or hummingbirds for tubular flowers, bats for campanulate flowers and subcampanulate flowers having generalised pollination systems (Martén-Rodríguez et al. 2009). O. jasminina has thin-tubular corolla (Fig. 1), differing from O. dasyantha, O. dasyantha var. ferruginosa (both are conical corolla) and O. flavida (campanulate-tubular corolla), indicating a possible pollination mechanism associated with the long-tongued butterflies and moths. Such distinctive morphological differences indicate different pollination adaptation and clear reproductive isolation amongst these taxa, suggesting O. jasminina should be treated as a new species.

Acknowledgements

This work was funded by the Innovative Team Program of Hainan Natural Science Foundation (2018CXTD331, 2018CXTD334), National Natural Science Foundation of China (41871041) and the Postgraduate Innovation Project of Biological Science of Hainan University.

References

Castellanos MC, Wilson P, Thomson JD (2004) ‘Anti-bee’ and ‘pro-bird’ changes during the evolution of hummingbird pollination in Penstemon flowers. Journal of Evolutionary Biology 17(4): 876–885. https://doi.org/10.1111/j.1420-9101.2004.00729.x

Chen WH, Shui YM, Yang JB, Wang H, Nishii K, Wen F, Zhang ZR, Möller M (2014) Taxonomic status, phylogenetic affinities and genetic diversity of a presumed extinct genus, Paraisometrum W.T. Wang (Gesneriaceae) from the Karst Regions of Southwest China. PLoS ONE 9(9): e107967. https://doi.org/10.1371/journal.pone.0107967

Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin 19(1): 11–15.

Francisco-Ortega J, Wang ZS, Wang FG, Xing FW, Liu H, Xu H, Xu WX, Luo YB, Song XQ, Gale S, Boufford DE, Maunder M, An SQ (2010) Seed plant endemism on Hainan Island: A framework for conservation actions. Botanical Review 76(3): 346–376. https://doi.org/10.1007/s12229-010-9055-7

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.

Huelsenbeck JP, Ronquist F (2001) MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics (Oxford, England) 17(8): 754–755. https://doi.org/10.1093/bioinformatics/17.8.754

IUCN (2012) IUCN Red List Categories and Criteria, Version 3.1 (2^ndedition). IUCN Species Survival Commission, Gland & Cambridge. https://www.iucnredlist.org/technical-documents/categories-and-criteria

Kumar S, Ne iM, Dudley J, Tamura K (2008) MEGA: a biologist-centric software for evolutionary analysis of DNA and protein sequences. Briefings in Bioinformatics 9(4): 299–306. https://doi.org/10.1093/bib/bbn017

Li ZY, Wang YZ (2005) Plants of Gesneriaceae in China. Henan Science & Technology Publishing House, Zhengzhou, 14–67.

Li G, Ling SJ, Chen WF, Ren MX, Tang L (2019) Effects of geographic isolation caused by Changhua River on genetic diversity of Hainan-endemic Metapetrocosmea peltata (Gesneriaceae). Guihaia.

Ling SJ, Meng QW, Tang L, Ren MX (2017a) Gesneriaceae on Hainan Island: distribution patterns and phylogenetic relationships. Biodiversity Sciences 25: 807–815. https://doi.org/10.17520/biods.2016360

Ling SJ, Meng QW, Tang L, Ren MX (2017b) Pollination syndromes of Chinese Gesneriaceae: a comparative study between Hainan Island and neighboring regions. Botanical Review 83: 59–73. https://doi.org/10.1007/s12229-017-9181-6

Ling S-J, Qin X-T, Song X-Q, Zhang L-N, Ren M-X (2020) Genetic delimitation of Oreocharis species from Hainan Island. In: Shui Y-M, Chen W-H, Ren M-X, Wen F, Hong X, Qiu Z-J, Wei Y-G, Kang M (Eds) Taxonomy of Gesneriaceae in China and Vietnam. PhytoKeys 157: 59–81. https://doi.org/10.3897/phytokeys.157.32427

Martén-Rodríguez S, Almarales-Castro A, Fenster CB (2009) Evaluation of pollination syndromes in Antillean Gesneriaceae: Evidence for bat, hummingbird and generalized flowers. Journal of Ecology 97(2): 348–359. https://doi.org/10.1111/j.1365-2745.2008.01465.x

Möller M (2019) Species discovery in time: An example from Gesneriaceae in China. Guangxi Sciences 26(1): 1–16.

Möller M, Clark L (2013) The state of molecular studies in the family Gesneriaceae: A review. Selbyana 31: 95–125.

Möller M, Middleton D, Nishii K, Wei YG, Sontag S, Weber A (2011) A new delineation for Oreocharis incorporating an additional ten genera of Chinese Gesneriaceae. Phytotaxa 23(1): 1–36. https://doi.org/10.11646/phytotaxa.23.1.1

Möller M, Wei YG, Wen F, Clark JL, Weber A (2016) You win some, you lose some: Updated generic delineations and classification of Gesneriaceae-implications for the family in China. Guihaia 36(1): 44–60.

Möller M, Atkins HJ, Bramley GLC, Middleton DJ, Baines R, Nguyen VD, Bui HQ, Barber S (2018) Two new species of Oreocharis (Gesneriaceae) from Northern Vietnam. Edinburgh Journal of Botany 75(3): 309–319. https://doi.org/10.1017/S0960428618000148

Muchhala N (2007) Adaptive trade-off in floral morphology mediates specialization for flowers pollinated by bats and hummingbirds. American Naturalist 169: 494–504. https://doi.org/10.1086/512047

Pan KY (1987) Taxonomy of the genus Oreocharis (Gesneriaceae). Journal of Systematics and Evolution 25(4): 264–293. http://www.jse.ac.cn/CN/Y1987/V25/I4/264

Robin VV, Vishnudas CK, Gupta P, Ramakrishnan U (2015) Deep and wide valleys drive nested phylogeographic patterns across a montane bird community. Proceedings of the Royal Society B: Biological Sciences 282: 20150861. https://doi.org/10.1098/rspb.2015.0861

Sargent RD (2004) Floral symmetry affects speciation rates in angiosperms. Proceedings of the Royal Society B: Biological Sciences 271(1539): 603–608. https://doi.org/10.1098/rspb.2003.2644

Shen ZH, Yang MZ, Feng JM, Li XH, Peng PH, Zheng Z (2017) Geographic patterns of alpine flora in China in relation to environmental and spatial factors. Biodiversity Sciences 25(2): 182–194. https://doi.org/10.17520/biods.2017014

Swofford DL (2003) PAUP*: Phylogenetic analysis using parsimony (* and other methods) Version 4.0b10. Sinauer Associates, Sunderland, Massachusetts.

Taberlet P, Gielly L, Pautou G, Bouvet J (1991) Universal primers for amplification of three non-coding regions of chloroplast DNA. Plant Molecular Biology 17(5): 1105–1109. https://doi.org/10.1007/BF00037152

Wei YG (2010) Gesneriaceae of South China. Guangxi Science and Technology Publishing House, Nanning, 20–52.

Wen F, Li S, Xin ZB, Fu LF, Hong X, Cai L, Qin JQ, Pan B, Pan FZ, Wei YG (2019) The updated plant list of Gesneriaceae in China under the new Chinese naming rules. Guangxi Sciences 26(1): 37–63.

White TJ, Bruns TD, Lee SB, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR Protocols 1990: 315–322. https://doi.org/10.1016/B978-0-12-372180-8.50042-1

Xing EN, Xu ST, Ren MX (2018) Age structure and gene flows of fine-scale populations of Oreocharis dasyantha (Gesneriaceae), an alpine herb endemic to Hainan Island. Journal of Tropical Biology 9(1): 37–46.

Xu WB, Guo J, Pan B, Zhang Q, Liu Y (2017) Diversity and distribution of Gesneriaceae in China. Guihaia 37(10): 1219–1226.

Appendix 1

Download as

CSV

XLSX

The Oreocharis specimens we checked in this study.

Voucher Number	Species	Voucher Number	Species
PE00030859	Oreocharis rhytidophylla	bm000041491	Oreocharis aurantiaca
IBSC0004917	Oreocharis henryana	gh00353683	Oreocharis dentata
A00353713	Oreocharis magnidens	k000858130	Oreocharis maximowiczii
bm000041734	Oreocharis benthamii	e00087519	Oreocharis tubicella
e00067455	Oreocharis cavaleriei	e00087520	Oreocharis nemoralis
a00025113	Oreocharis flavida	IBSC0004912	Oreocharis aurea
bm000041721	Oreocharis georgei	p04060117	Oreocharis forrestii
KUN1219176	Oreocharis cordato-ovata	e00135096	Oreocharis amabilis
e00087535	Oreocharis dasyantha	e00135074	Oreocharis bodinieri
bm000041708	Oreocharis cinnamomea	gh00353695	Oreocharis benthamii var. reticulata
PE00030854	Oreocharis tubiflora	p04060171	Oreocharis delavayi
IBSC0004920	Oreocharis xiangguiensis	CSFI028502	Oreocharis brachypodus
PE19401111	Oreocharis amabilis	PE02052999	Oreocharis heterandra
IBSC0550960	Oreocharis sericea	IBSC0550860	Oreocharis cordatula
IBSC0550891	Oreocharis georgei	IBSC0550875	Oreocharis elliptica
GZTM0075588	Oreocharis primuloides	JIU63907	Oreocharis speciosa
PE02052990	Oreocharis argyreia var. angustifolia	PE02053568	Oreocharis eximia
IBK00054784	Oreocahris auricula	PE01909883	Oreocharis mileensis
KUN1219104	Oreocharis hekouensis	WUK0494363	Oreocharis saxatilis
PE02053062	Oreocharis concava	PE01486523	Oreocharis rosthornii
PE02106072	Oreocharis begoniifolia	KUN1385365	Oreocharis nanchuanica
KUN1385575	Oreocharis urceolata	HITBC106680	Oreocharis longifolia
PE00030861	Oreocharis rotundifolia	IBSC0548683	Oreocharis chienii
PE02053433	Oreocharis acaulis	KUN1385156	Oreocharis bullata
PE02241281	Oreocharis burttii	PE02053072	Oreocharis cinerea
KUN1220227	Oreocharis convexa	IBK00054466	Oreocharis cotinifolia
PE00155697	Oreocharis craibii	WUK0350789	Oreocharis crenata
IBSC0550709	Oreocharis dalzielii	PE02106079	Oreocharis dinghushanensis
IBSC0551649	Oreocharis esquirolii	PE02052984	Oreocharis fargesii
IBSC0649611	Oreocharis flabellata	PE02053533	Oreocharis gamosepala
PE02052812	Oreocharis giraldii	PE02106025	Oreocharis glandulosa
PE02052995	Oreocharis humilis	PE01548041	Oreocharis jiangxiensis
PE02021009	Oreocharis lancifolia	FJSI004239	Oreocharis leiophylla
PE02053066	Oreocharis leucantha	IBSC0550069	Oreocharis lungshengensis
IBSC0551655	Oreocharis mairei	PE02053564	Oreocharis minor
WUK0160594	Oreocharis muscicola	IBSC0548476	Oreocharis notochlaena
PE02106041	Oreocharis obliqua	PE02052801	Oreocharis obliquifolia
PE02088092	Oreocharis obtusidentata	PE02053064	Oreocharis pankaiyuae
PE01270485	Oreocharis primuloides	WUK0213194	Oreocharis pumila
PE02053576	Oreocharis pinnatilobata	KUN1241303	Oreocharis primuliflora
PE02053532	Oreocharis rhombifolia	PE00030693	Oreocharis ronganensis
PE00030747	Oreocharis sichuanensis	IBSC0550081	Oreocharis sichuanica
IBK00054319	Oreocharis sinensis	IBK00207093	Oreocharis sinohenryi
PE02053579	Oreocharis stenosiphon	IBSC0548730	Oreocharis stewardii
PE02053570	Oreocharis trichantha	HEAC0016525	Oreocharis villosa
PE02053561	Oreocharis wangwentsaii	PE02053077	Oreocharis wanshanensis
Y. M. Shui et al. B2014-299 (KUN)	Oreocharis synergia	Y.M.Shui et al. N699 (KUN)	Oreocharis ninglangensis
PE-02114626	Oreocharis duyunensis	IBSC0825078	Oreocahris ovata
KUN1219115	Oreocharis acutiloba	PE00030682	Oreocharis agnesiae
PE00140281	Oreocharis billburttii	PE02025205	Oreocharis elegantissima
IBSC0649550	Oreocharis latisepala	PE00030685	Oreocharis parva
Z.K. Wu et al.C2016055 (KUN)	Oreocharis parvifolia	PE02025202	Oreocharis pinfaensis
IBSC0548691	Oreocharis shweliensis	PE01909893	Oreocharis tongtchouanensis
Y.M.Shui, Y.K.Sima & W.H.Chen B2013-258 (KUN)	Oreocharis crispata	Y.M.Shui et al. 91309 (KUN)	Oreocharis jinpingensis
Bo Pan & M. Q. Han HMQ859 (IBK)	Oreocharis purpurata	Yun-Hong Tan 3308 (HITBC)	Oreocharis tsaii
Averyanov, L., Hiep, N.T., Khang, N.S., Thang, N.D. & Qui, L.D. CPC 7019 (KUN)	Oreocharis blepharophylla	Jia-Mei Li and Yao-Guang Zhang 1606151 (HEAC)	Oreocharis zhenpingensis
Bo Pan & Jia-Jia Wei et al. GY002 (IBK)	Oreocharis curvituba	C.Z. Yang et al. 35042620140913001 (FNU)	Oreocharis striata
Y.M. Shui et al. B2013-551 (KUN)	Oreocharis longituba	Averyanov, L., Hiep, N.T., Khang, N.S., Thang, N.D. & Qui, L.D. CPC 7175 (KUN)	Oreocharis argyrophylla
Y.M. Shui et al. B2013-550	Oreocharis grandiflora	T.V. Do 57 (VNMN)	Oreocharis caobangensis
L.H. Yang et al. YLH197 (IBSC)	Oreocharis pilosopetiolata	Li-Hua Yang et al. YLH285 (IBSC)	Oreocharis uniflora
Ying Guo C2015005 (KUN)	Oreocharis panzhouensis	L. E. Yang 60 (KUN)	Oreocharis rubrostriata
Yan Liu and Wei-Bin Xu 08018 (IBK)	Oreocharis dayaoshanioides	Yun-Hong Tan 6925 (HITBC)	Oreocharis glandulosa
PE 02053063	Oreocharis farreri	Lin Qin-Wen et al. 0016 (FAFU)	Oreocharis baolianis
IBSC0548624	Oreocharis guileana	IBK00054993	Oreocharis dasyantha var. ferruginosa