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Research Article
Liparis macrosepala (Orchidaceae), a new species from southwest China with its phylogenetic position
expand article infoZhengwei Wang‡, Yi Zhang§, Ze Zhang§, Xiaochen Li‡, Zhijin Wu‡, Lan Yan§, Aixian Lu‡|, Chengzhi Xie‡, Chao Hu‡, Weichang Huang‡|
‡ Eastern China Conservation Centre for Wild Endangered Plant Resources, Shanghai Chenshan Botanical Garden, Shanghai, China
§ Yunnan Yelantang Biological Technology Co., Ltd., Kunming, China
| Fujian Agriculture and Forestry University, Fuzhou, China
¶ Hainan University, Haikou, China

Corresponding author: Chao Hu ( hc320320@163.com )

Corresponding author: Weichang Huang ( hwc_zx@126.com )

Academic editor: Timothée Le Péchon
© 2022 Zhengwei Wang, Yi Zhang, Ze Zhang, Xiaochen Li, Zhijin Wu, Lan Yan, Aixian Lu, Chengzhi Xie, Chao Hu, Weichang Huang.
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: Wang Z, Zhang Y, Zhang Z, Li X, Wu Z, Yan L, Lu A, Xie C, Hu C, Huang W (2022) Liparis macrosepala (Orchidaceae), a new species from southwest China with its phylogenetic position. PhytoKeys 210: 67-77. https://doi.org/10.3897/phytokeys.210.87033
Open Access

Abstract

A new orchid species, Liparis macrosepala, is illustrated and described from Yunnan Province, China, based on morphological and molecular analyses. This plant is characterised by the ovoid-fusiform, slightly compressed pseudobulbs with 4 or 5 leaves with slightly crisped margins on their apical half, dorsal sepal heart-shaped, lip with a bituberculate basal callus and a thickened folded lateral lobe on each side, centrally with one cavity with slightly raised margins, the column with a single pair of broadly triangular, obtuse wings. Maximum Likelihood and Bayesian Inference analyses of combined nrITS and plastid matK DNA sequences place this species in section Cestichis.

Keywords

Liparis section Cestichis, molecular phylogeny, morphology, matK, nrITS

Introduction

The genus Liparis Rich. (Epidendroideae, Malaxideae, Malaxidinae) comprises about 320 species distributed worldwide with more than 70 species in China (Pridgeon et al. 1999; Chen et al. 2009; Tian et al. 2015; Huang et al. 2018; Ya et al. 2021). Species from this genus are terrestrial, lithophytic, epiphytic and rarely mycoheterotrophic, with inflorescences laxly or densely many-flowered, lip often reflexed and usually with a basal callus, lacking a spur, column winged at apex and sometimes at base and four pollinia in two pairs (Chen et al. 2009).

During our field surveys in Xishuangbanna, Yunnan, China, an unknown species was found. In this paper, we analysed the morphological differences of the newly-found species and its allied species and the phylogenetic position of the new entity is also discussed, based on molecular evidence from nrITS and plastid matK. After careful morphological comparison and phylogenetic analyses, we concluded that this species is new to science.

Material and method

Morphological observations

Materials of the new species were collected from Xishuangbanna, Yunnan, China during a field expedition. Morphological characters were observed, measured and photographed based on five living individuals under a stereomicroscope (SZX16-6151, Olympus, Japan) and photographed with a digital camera (D750, Nikon, Japan). A voucher specimen, designated as the holotype, was deposited at Shanghai Chenshan Herbarium (CSH). Conservation assessment has been conducted following IUCN guidelines (IUCN 2019).

Taxonomic sampling

DNA sequences of nrDNA ITS and plastid matK of the new species were sequenced and sequences of the same markers for 82 related species were downloaded from GenBank, including five outgroup species from other subtribes (Table 1).

Table 1.
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Taxon sampling in this study.

Species Name nrITS matK
1 Acanthophippium mantinianum L.Linden & Cogn. AF521081 AF263618
2 Collabium simplex Rchb.f. EF670387 AY557200
3 Crepidium acuminatum (D.Don) Szlach. KJ459274 KJ459304
4 Crepidium bahanense (Hand.-Mazz.) S.C.Chen & J.J.Wood MH116611 MH117500
5 Crepidium bancanoides (Ames) Szlach. AB290885 AB290893
6 Crepidium brevidentatum (Schweinf.) M.A.Clem. & D.L.Jones AB290886 AB290894
7 Crepidium resupinatum (G.Forst.) Szlach. JN114483 JN004403
8 Dendrobium dixanthum Rchb.f. KY966535 KY966825
9 Dienia cylindrostachya Lindl. JN114491 JN004422
10 Eria ferruginea Lindl. AF521071 AF263660
11 Eulophia graminea Lindl. MH768269 MH767976
12 Liparis macrosepala Z.W. Wang, Y. Zhang & W.C. Huang ON642332 ON642331
13 Liparis anopheles J.J.Wood AY907075 AY907139
14 Liparis assamica King & Pantl. KJ459276 KJ459306
15 Liparis aureolabella J.D. Ya & Z.D. Han MN065679 MN065733
16 Liparis auriculata Blume ex Miq. AB289458 KF262076
17 Liparis balansae Gagnep.-1 KF589874 KF589880
18 Liparis balansae Gagnep.-2 KJ459277 KJ459307
19 Liparis bingzhongluoensis X.H. Jin MW169041 MW169042
20 Liparis bistriata E.C.Parish & Rchb.f. KJ459279 KJ459309
21 Liparis bootanensis Griff KJ459280 KJ459310
22 Liparis bracteata T.E.Hunt AY907076 AY907140
23 Liparis brunnescens Schltr. AY907098 AY907165
24 Liparis condylobulbon Rchb.f. AY907080 AY907144
25 Liparis cordifolia Hook.f. KJ459282 KJ459312
26 Liparis delicatula Hook.f. KJ459283 KJ459313
27 Liparis distans C.B.Clarke KJ459284 KJ459314
28 Liparis disticha (Thouars) Lindl. AY907081 AY907145
29 Liparis elliptica Wight KJ459285 KJ459315
30 Liparis fissilabris Tang & F.T.Wang KJ459286 KJ459316
31 Liparis fissipetala Finet KJ459287 KJ459317
32 Liparis formosana Rchb.f. AY907082 AY907147
33 Liparis fujisanensis F.Maek. ex Konta & S.Matsumoto EU024936 EU024937
34 Liparis gibbosa Finet-1 AY907083 AY907148
35 Liparis gibbosa Finet-2 AY907084 AY907149
36 Liparis glossula Rchb.f. KJ459289 KJ459319
37 Liparis guangxiensis C.L.Feng & X.H.Jin KF589875 KF589881
38 Liparis japonica (Miq.) Maxim. AY907086 AY907151
39 Liparis koreana (Nakai) Nakai EU017422 EU017444
40 Liparis kumokiri F.Maek. AY907087 AY907152
41 Liparis latifolia Lindl. AY907088 AY907153
42 Liparis latilabris Rolfe KJ459291 KJ459321
43 Liparis liliifolia (L.) Rich. ex Lindl. AY907090 AY907156
44 Liparis loeselii (L.) Rich. AY907091 AY907157
45 Liparis makinoana Schltr. EU017405 EU017428
46 Liparis mannii Rchb.f. KJ459293 KJ459323
47 Liparis meihuashanensis S.M.Fan MF959772 MF959773
48 Liparis mengziensis J.D. Ya & Lei Cai MN065734 MN065678
49 Liparis nanlingensis H.Z.Tian & F.W.Xing AB701346 /
50 Liparis napoensis L.Li, H.F.Yan & S.J. Li-1 MT012899 MT019986
51 Liparis napoensisL.Li, H.F.Yan & S.J. Li -2 MT012900 MT019987
52 Liparis nervosa (Thunb.) Lindl. AY907092 AY907158
53 Liparis nugentiae F.M.Bailey AY907093 AY907159
54 Liparis odorata (Willd.) Lindl. KJ021033 KJ021029
55 Liparis pandurata Ames AY907094 AY907160
56 Liparis pauliana Hand.-Mazz. AY907096 AY907163
57 Liparis petiolata (D.Don) P.F.Hunt & Summerh. MW186826 MW187482
58 Liparis resupinata Ridl. KJ459297 KJ459327
59 Liparis somae Hayata-1 MT012898 MT019985
60 Liparis somae Hayata-2 MT012897 MT019984
61 Liparis sootenzanensis Fukuy. KJ021034 KJ021030
62 Liparis stricklandiana Rchb.f.-1 MT012903 MT019990
63 Liparis stricklandiana Rchb.f. -2 KJ459298 KJ459328
64 Liparis sula N.Hallé AY907104 AY907171
65 Liparis terrestris J.B.Comber AY907105 AY907172
66 Liparis truncicola Schltr. AY907106 AY907173
67 Liparis viridiflora (Blume) Lindl.-1 MT012902 MT019989
68 Liparis viridiflora (Blume) Lindl.-2 MT012901 MT019988
69 Malaxis brachypoda (A.Gray) Fernald AY907108 AY907175
70 Malaxis monophyllos (L.) Sw. MW181626 MW187483
71 Malaxis soulei L.O.Williams AY907119 AY907186
72 Malaxis abieticola Salazar & Soto Arenas AY907129 AY907196
73 Oberonia acaulis Griff. KY242066 KY241943
74 Oberonia brunoniana Wight JN114623 JN004516
75 Oberonia equitans (G.Forst.) Mutel AY907130 AY907198
76 Oberonia heliophile Rchb.f. AY907131 AY907199
77 Oberonia iridifolia Roxb. ex Lindl. AY907132 AY907200
78 Oberonia mucronata (D.Don) Ormerod & Seidenf JN114640 JN004534
79 Oberonia neocaledonica Schltr.-1 AY907133 AY907201
80 Oberonia neocaledonica Schltr.-2 AY907134 AY907202
81 Oberonia padangensis Schltr. AY907135 AY907203
82 Oberonia wappeana J.J.Sm. AY907138 AY907206
83 Oberonioides pusillus (Rolfe) Marg. & Szlach. KJ527610 KJ459302

Phylogenetic analyses

DNA sequences were aligned using the MAFFT programme in Geneious v. 2020.2.4 (https://www.geneious.com, accessed on 10 March 2021). Phylogenetic analyses were conducted using Maximum Likelihood (ML) and Bayesian Inference (BI) in RAxML v.7.0.4 (Stamatakis 2006) and MrBayes v.3.2.6 (Huelsenbeck and Ronquist 2001; Ronquist et al. 2012), respectively. The appropriate DNA substitution model under AIC criteria was estimated using jModelTest 2.1.10 (Posada 2008). ML analyses were conducted with bootstrap values calculated by running 1,000 replicates. For BI analysis, four chains were run with random initial trees, each for 1,000,000 generations, until the average standard deviation of the split frequency values was less than 0.01 to ensure convergence, sampling trees every 1,000 generations. After the first 20% of samples were discarded as burn-in, the remaining replicates were used to estimate the posterior probabilities.

Results

Phylogenetic analyses

The length of nrITS matrix was 792 bp including 262 parsimony-informative sites and for matK, the length and parsimony-informative sites were 1443 bp and 120, respectively. Both analyses (MP and BI) recovered similar relationships. The ML tree with bootstrap percentages, on which the posterior probabilities from the BI analysis were also indicated, is shown in Fig. 1.

Figure 1. 

Maximum Likelihood tree of Liparis and its allied genera in subtribe Malaxidinae inferred from the combined analysis of nrITS and matK. ML bootstrap values (MLBP)/Bayesian posterior probabilities (PP) are indicated above the branches, respectively. The sectional taxonomy of Liparis follows Garay and Romero-Gonzalez (1999) and Li et al. (2020).

The phylogenetic analyses indicate that Liparis is not monophyletic, being mingled with species of other genera of Malaxideae. This result agrees with what was found in previous studies (Cameron 2005; Margońska et al. 2012; Tang et al. 2015; Li et al. 2020; Kumar et al. 2022). The new species, henceforth referred to as Liparis macrosepala Z.W. Wang, Y. Zhang & W.C. Huang, is grouped with species in Liparis sect. Cestichis Thouars ex Pfitzer as the sister of a clade consisting of L. delicatula Hook.f., L. fissipetala Finet, L. assamica King & Pantl. and L. resupinata Ridl.

Morphological comparisons

Liparis is defined as species with racemose inflorescences, resupinate lip lacking a spur, column without a conspicuous foot and four pollinia in two pairs with small viscidium, but no caudicle. The morphology of Liparis macrosepala is in accordance with the characteristics of sect. Cestichis like the slightly flattened, narrowly winged rachis with alternating bracts. The morphological characters can distinguish Liparis macrosepala from its close relatives L. delicatula, L. fissipetala, L. assamica and L. resupinata.

Taxonomic treatment

Liparis macrosepala Z.W. Wang, Y. Zhang & W.C. Huang, sp. nov.

urn:lsid:ipni.org:names:77306143-1
Figs 2, 3 Chinese name: 大萼羊耳蒜

Type

China. Yunnan Province (云南), Xishuangbanna (西双版纳), Mengla County (勐腊县) epiphyte on the tree trunk, 1620 m elev., 23Nov 2021, Zhengwei Wang, Xiaochen Li, Yu Zhang& Zhijin Wu, WZW04247 (holotype: CSH!)

Figure 2. 

Morphology of Liparis macrosepala. A plants in situ B flowering plant C pseudobulbs and leaves D inflorescence E flowers, front view F flowers, side view G perianth dissection H column from side I lip in oblique view J anther cap and pollinia. Photographs by Weichang Huang.

Diagnosis

Liparis macrosepala is characterised by the ovoid-fusiform, slightly compressed pseudobulbs with 4 or 5 alternate leaves on their apical half, these with slightly crispate margins, dorsal sepal ovate with cordate base, broadly elliptic, ca. 4 mm long, 2 callus-shaped and thickened folds, base with 2 oblong lobes on both sides, centrally with 1 thickened, concave callus, column with a single pair of arcuate wings.

Figure 3. 

Liparis macrosepala A flower, front view B flower, side view C lip, side view D inflorescence E column, side view F lip and column, side view G lip, back view H lip, front view I flowering plant J pollinia and anther cap K perianth dissection L column and ovary, oblique view M ovary, transection. Drawn by Lan Yan.

Epiphytic herbs. Roots slender, flexuose. Pseudobulbs clustered, ovoid-fusiform, slightly compressed laterally, 1–2 × 0.5–1 cm, upper half with 4–5 widely spaced leaves. Leaf blade ovate-oblong, 1.8–2.3 × 0.8–1.2 cm, apex acuminate, base contracted into a short petiole, articulate, margins of their apical half slightly crispate. Peduncle 7–10 cm long, with several sterile bracts 2–5 mm long; raceme with 7–10 flowers arranged in zigzag manner. Floral bracts broadly ovate with cordate base, 2–3 × 1–1.5 mm, acute. Flowers greenish-orange; pedicel and ovary ca. 7 mm long. Dorsal sepal broadly ovate with cordate base, 3.2–5 × 3–3.6 mm, 1-veined, abaxially carinate, apex acute; lateral sepal oblong-ovate or ovate-lanceolate, 5–6 × ca. 0.6 mm long, abaxially slightly carinate. Petals narrowly linear, 3–4 × ca. 0.2 mm; lip elliptic, 2–3 × ca. 1 mm, apex apiculate, base bearing a bituberculate callus, then expanded on each side into a thickened, folded, rounded lobe, with 1 excavation with raised margins between the lobes. Column straight, ca. 2 mm long, with a pair of subtriangular, obtuse wings on each side near the middle and a ridge on the back of the column. Anther cap hemispherical, pale yellow; pollinia 4 in 2 pairs with one pollinium of each pair smaller than the other, waxy, brownish, with minute apical viscidium.

Phenology: Flowering in November–December.

Distribution and habitat

It is found on tree trunks on a limestone ridge-top evergreen broad-leaved forest at an elevation of 1500–1700 m in Mengna County, Xishuangbanna Autonomous Prefecture, Yunnan Province, People’s Republic of China. The habitat presents a tropical monsoon climate.

Etymology

The species epithet refers to the large and conspicuous dorsal sepal of the flower.

Taxonomic notes

Liparis macrosepala differs from L. delicatula in its 4 to 5 leaves with slightly crispate margins on their apical half and single pair of wings on the column. Its entire, not Y-shaped petals and sessile lip (i.e. without a claw) easily distingush L. macrosepala from L. fissipetala. The dosal sepal of L. assamica is narrowly ovate-oblong, in contrast with the heart-shaped dorsal sepal of Liparis macrosepala. Liparis resupinata is distinguished from L. macrosepala by its 10–50-flowered raceme and the column with a single pair of broad wings, each with a retrorse thread. The main differences between these closely-related species, according to our phylogenetic analyses, are summarised in Table 2.

Table 2.
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Comparison of L. macrosepala and related species.

Characters L. delicatula L. fissipetala L. assamica L. resupinata L. macrosepala
Pseudobulbs oblong or cylindrical-fusiform 5–9 3–5 mm ovoid, 8–10 mm long ovoid-fusiform, slightly compressed 1.5–2.5 cm × 6–10 mm subcylindrical or ± spindle-shaped, 1.8–5 cm × 3–6 mm ovoid-fusiform, slightly compressed, 1–2 cm × 0.5–1 cm
Leaf 2 or 3, margin flat 3 or 4, strongly crisped-margined 3 or 4, apical half slightly crisped-margined 3 or 4, margin slightly serrate 4 or 5, apical half slightly crisped-margined
Scape 2–5 cm, several to 10-flowered, flowers white 5–10 cm long, with 10–15 flowers, flowers yellow, 10–13 cm, more than 10-flowered, flowers orange 7–18 cm, 10–50-flowered, flowers pale green or greenish-yellow 7–10 cm, more than 10-flowered, flowers greenish-orange
Bracts ovate-lanceolate, 2–3 mm ovate-lanceolate, 1.5–3.5 mm lanceolate, 2–3 mm lanceolate, 3–5 mm broadly ovate, 2–3 mm
Dorsal sepal ovate-oblong, 2.5–3 × 1.5–1.8 mm oblong-lanceolate, 3–4 × 0.8–1 mm narrowly ovate-oblong, 4.8–5.8 × ca. 1.6 mm oblong or elliptic-oblong, ca. 4 × 1.8 mm broadly ovate, ca. 3.2–5 ×3–3.6 mm
Petals narrowly linear-lanceolate, 2.5–3 × ca. 0.5 mm, entire narrow linear, 4–5 mm long, Y-shaped narrowly linear, 5–5.5 × ca. 0.7 mm, entire narrowly linear, ca. 3.5 × 0.3 mm, entire narrowly linear, 3–4 × ca. 0.2 mm, entire
Lip broadly elliptic or orbicular, ca. 2.5 mm, base with an orbicular, auriculate, callus-shaped fold on either side, with a concave callus near base epichile broadly oblong or subsquare, 1.5–2 × 1–1.5 mm, base with two auricles on both sides; claw short, with a fleshy callus centrally near base broadly obovate-oblong, ca. 4 × 2.7 mm, with two callus-shaped thickened folds, two suborbicular lobes on both sides, centrally with one concave callus near base broadly elliptic-oblong or broadly ovate-oblong, 2.5–3 mm, with two lateral splits below middle; two suborbicular lobes, centrally with one bilobed callus near base broadly elliptic, ca. 2–3 mm long, two callus-shaped and thickened folds, base with two oblong lobes on both sides, centrally with one bituberculate callus near base
Column ca. 2.2 mm, two pairs of wings ca. 1.5 mm, broadly winged with two horn-like appendages ca. 2 mm, two pairs of wings ca. 2.8 mm, a pair of wings, each with a retrorse thread ca. 2 mm, a single pair of subtriangular wings

Conservation assessment

The new species was found in a ridge-top evergreen broad-leaved forest on a limestone mountain. Despite numerous surveys in the areas, only six mature individuals were found without fruits or evidence of cross-pollination.

This extremely small effective population occurs in a touristic zone which is a serious threat to the survival of the species. Consequently, the species can be assessed as Critically Endangered (CR, D), based on current information and following IUCN guidelines (IUCN 2019).

Acknowledgements

This study was supported by grants from the National Wild Plant Germplasm Resource Centre for Shanghai Chenshan Botanical Garden (ZWGX2102), the Science and Technology Commission, Shanghai Municipality (19390743600) and the project of Shanghai Landscaping and City Appearance Administrative Bureau (G192424, G202401). We are grateful to Mingzhong Huang for his help with the literature and Hongjin Wei for his fieldwork. We are grateful to Dr. Pankaj Kumar for his help in improving the language and describing the conservation status. We are also grateful to the reviewers for their valuable views in improving this article.

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