Research Article |
Corresponding author: Li-Hua Yang ( l.h.yang@163.com ) Academic editor: Yu Shui
© 2020 Li-Hua Yang, Fang Wen, Hang-Hui Kong, Zhi-Xia Sun, Lan-Ying Su, Ming Kang.
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.
Citation:
Yang L-H, Wen F, Kong H-H, Sun Z-X, Su L-Y, Kang M (2020) Two new combinations in Oreocharis (Gesneriaceae) based on morphological, molecular and cytological evidence. 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: 43-58. https://doi.org/10.3897/phytokeys.157.32609
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The newly-circumscribed genus Oreocharis is recently enlarged by incorporating ten other genera with high floral diversity. In this study, our morphological, molecular and cytological evidence supports our adding two species from other two different genera (Boeica and Beccarinda) to Oreocharis. The special corolla shape (campanulate or flat-faced) and related short filament of these two new combinations, Oreocharis guileana and O. baolianis, further enrich the diversity of floral characters of the enlarged Oreocharis. Meanwhile, some supplementary and amended descriptions of these two species are made here. Our morphological, molecular and geographical data indicate that O. guileana is related to O. pilosopetiolata to a certain extent. For O. baolianis, however, our current dataset does not allow conclusions on the species relationship within Oreocharis.
Beccarinda baolianis, Boeica guileana, ITS, Oreocharis, taxonomy, trnL-F
Recent phylogenetic studies on the Old World Gesneriaceae have greatly advanced our understanding of species relationships and generic delimitations in this plant group (
Boeica guileana was first described in 1977 by Burtt based on cultivated plants without description of mature fruits. The species has been regarded as endemic to Hong Kong for a long time (
Beccarinda baolianis was newly described in 2016 from southern China, based on two specimens (
Summarising, the generic placements of Boe. guileana and Be. baolianis need reconsideration. Therefore, this study aims to clarify the generic status of these two species and discuss their species relationships in the enlarged Oreocharis using cytological, molecular and morphological analyses.
The previous descriptions of Boe. guileana and Be. baolianis were based on only one or two specimens from a single location; thus, the described characters do not reflect the whole range of variation and other characters were not described at all. For example, the dehiscent way of fruit of both of these two species had not been mentioned in the original protologue. To revise and amend the morphological descriptions and clarify the morphological affinities between these two species and its congeners, detailed morphological studies were made on living plants in the field (Shenzhen and Fujian, China) and on plants cultivated in greenhouses (South China Botanical Garden, SCBG and Gesneriad Conservation Center of China, GCCC). Furthermore, dried specimens were investigated. Checking of specimens was undertaken at IBSC, IBK, SZG and FAFU. Additionally, high-resolution images of specimens were critically checked using the web service of E (http://data.rbge.org.uk/search/herbarium/) and HK (http://www.herbarium.gov.hk/index.aspx).
In the recent classification of Gesneriaceae, Boeica and Beccarinda were deposited in subfamily Didymocarpoideae, tribe Trichosporeae, subtribe Leptoboeinae and the enlarged Oreocharis was deposited in subtribe Didymocarpinae (
The plastid trnL-F and nuclear ribosome internal transcribed spacer (ITS) sequences were used for the phylogenetic analysis. Most of these DNA sequences were acquired from GenBank and the sequences of six species, Boe. guileana, Boe. ornithocephalantha F. Wen, T.V. Do & Y.G. Wei (
Sequence matrices of trnL-F and ITS were separately aligned using the programme MUSCLE implemented in the software MEGA7 (
Four species, Boe. guileana, Boe. stolonifera K.Y. Pan, Be. baolianis and Be. tonkinensis (Pellegr.) B.L. Burtt, were investigated cytologically. The plants for chromosome studies were collected from the field and cultivated in Gesneriad Conservation Center of China (GCCC). Actively growing root tips were collected and pretreated with 2 mM 8-hydroxyquinoline at 20 °C for about 2 hrs, then fixed with Farmer’s solution (absolute alcohol: glacial acetic acid 3:1) at 4 °C for about 2 hrs. After hydrolysis for 30 min in 1 M HCl at room temperature, followed by washing through several changes of distilled water, the roots were transferred to carbol fuchsin for about 2 hrs.
The results of morphological observation indicated that both of Boe. guileana (Fig.
The aligned trnL-F and ITS datasets were 966 and 824 characters long, thereof, 195 and 138 were parsimony-uninformative and 139 and 388 parsimony-informative characters, respectively. The combined dataset was 1790 characters long with 527 (29.4%) parsimony-informative characters. The incongruence length different (ILD) test showed no significant incongruence between the ITS and trnL-F (p = 0.53).
In the combined DNA sequence analysis, the BI tree was less resolved but congruent (virtually identical) where the ML branches received bootstrap support > 50% (Fig.
Bayesian (> 50%) tree resulting of the combined nuclear (ITS) and plastid (trnL-F) data matrices. Posterior probability (PP) from the BI analysis are indicated above branches and Bootstrap value (BS) from the ML analysis are indicated below. The asterisk indicates a BS < 50. The dash indicates the topological discordance between ML and Bayesian tree. The two species, O. baolianis and O. guileana, are highlighted in bold.
In the four investigated species, the somatic chromosomes were determined to be diploid with 2n = 34 in Boe. guileana (Fig.
Traditionally, the classification of the Old World Gesneriaceae is heavily based on floral characters (e.g.
In the present study, we show a similar finding for Boe. guileana and Be. baolianis. We combine detailed morphological studies, cytological examinations and phylogenetic analyses to show that Boe. guileana and Be. baolianis are two species belonging to the enlarged Oreocharis. The morphological characters of these two species, i.e. growth habit, anther and fruit dehiscence are clearly distinct from its original genera. Boeica guileana and Be. baolianis are both rosette plants with inconspicuous rhizome (Figs
After incorporation of ten additional genera, the genus Oreocharis becomes a large group with high flower diversity. Flowers of this group are extremely variable in corolla shape, colour, number of stamens and anthers being free or fused (
≡Boeica guileana B.L.
China. Hong Kong: New territories, Ma On Shan, 690 m alt., ravine in montane forest, on rocks in humid shade, July 1974, Guile; culture in R.B.G. Edinb. 1976, C.8467 (holotype: E!).
Perennial herbs with inconspicuous rhizome. Leaves in basal rosette, 8–22; petiole 1–3 cm long, with densely white villous; leaf blade elliptic to ovate or obovate, 2–6 × 1.1–3.2 cm, white villous and pubescent on both sides, abaxially more densely villous along veins, base cuneate, margin serrate, apex acute to rounded; lateral veins 4–6 on each side of midrib. Cymes 1–5, axillary, 1–4-flowered. Peduncles 2.5–5.5 cm long, with brown villous; bracts 2, lanceolate, 2.5–3 × ca. 1 mm, margins entire, outside brown villous; pedicel 1–2 cm long, with brown villous. Calyx 5-lobed near base, lobes equal, lanceolate to linear, 3–5 × ca. 1 mm, outside covered by brown villous, margin entire. Corolla blue-purple, outside with sparely-brown villous; tube short and not swollen, 1.5–2.5 mm long; adaxial lip 2-lobed near base, lobes obovate-oblong, margin entire or slightly erose, apex obtuse, 5–7 × 4–5 mm; abaxial lip 3-lobed near base, lobes obovate-oblong, margin entire or slightly erose, apex obtuse, 9–12 × 4.5–6 mm. Stamens 4, adnate to corolla base, filaments linear, with sparsely pubescent, adaxial stamens ca. 3 mm long, abaxial stamens ca. 4 mm long, anthers dorsifixed, free, ovate-oblong, dehiscing longitudinally, glabrescent; staminode absent; disc not obvious; pistil 8–12 mm long; ovary conical, 2–3.5 mm, with densely-white villous; style 6–8 mm long, with pubescent. Stigma 1, disc-shaped. Capsule ca. 1.5 cm long, villous, dehiscing loculicidally to base, initially on one side, valves 2, capsule straight in relation to pedicel, not twisted.
Oreocharis guileana was once recognised as an endemic species and only recorded at Ma On Shan, Hong Kong. However, recent field works reveal that it can be found from several sites in Shenzhen, such as Wu Tong Shan, Pai Ya Shan, Tian Xin Shan, Dakeng reservoir and Xigong village (Fig.
China. Shenzhen Special Economic Zone, Pingshan District, Tian Xin Shan, alt. 300 m, 22 May 2017, 114°25'E, 22°41'N, L.H. Yang & F. Wen YLH383 (IBSC!); in the same place, alt. 300 m, 20 October 2005, S.Z. Zhang et al. 4658 (SZG!); 20 April 2005, S.Z. Zhang et al. 0384 (SZG!); 5 November 2004, S.Z. Zhang et al. SCAUF490 (SZG!), SCAUF491 (SZG!), SCAUF583 (SZG!); Longgang District, Pai Ya Shan, alt. 600 m, 25 October 2006, G.D. Wang et al. 6896 (SZG!); 13 October 2005, S.Z. Zhang et al. 4533 (SZG!); 17 December 2005, S.Z. Zhang et al. 2144 (SZG!); 8 June 2005, S.Z. Zhang et al. 2340 (SZG!); Xi Chong, Xigong village, 8 November 2004, S.Z. Zhang et al. SCAUF469 (SZG!), SCAUF470 (SZG!); Luohu District, Wu Tong Shan, alt. 900 m, 7 October 2005, S.Z. Zhang et al. 4288 (SZG!); 23 March 2005, Team of Flora of Shenzhen 013556 (SZG!); Longgang District, Dakeng reservoir, 16 July 2005, S.Z. Zhang et al. 2932 (SZG!).
This work was supported by the National Natural Science Foundation of China (31900178, 31970231) and Biological Resources Programme, Chinese Academy of Sciences (KFJ-BRP-017-03). We thank Dr. Michael MÖller for the provision of some ITS sequences and Dr. Annemarie Heiduk for the modification of English.
Taxon, voucher information and GenBank accession number of the samples used in the phylogenetic analyses.
Data type: phyogenetic