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Research Article
Five new and noteworthy species of Epidendroideae (Orchidaceae) from southwestern China based on morphological and phylogenetic evidence
expand article infoJi-Dong Ya‡§, Wan-Ting Wang|‡, Yun-Long Liu‡, Hong Jiang, Zhou-Dong Han‡, Ting Zhang‡, Hua Huang#, Jie Cai‡, De-Zhu Li‡‡#
‡ Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China
§ Southwest Forestry University, Kunming, China
| University of Chinese Academy of Sciences, Beijing, China
¶ Yunnan Academy of Forestry and Grassland, Kunming, China
# Kunming Institute of Botany, Chinese Academy of Sciences, Lijiang, China

Corresponding author: De-Zhu Li ( dzl@mail.kib.ac.cn )

Academic editor: Vincent Droissart
© 2023 Ji-Dong Ya, Wan-Ting Wang, Yun-Long Liu, Hong Jiang, Zhou-Dong Han, Ting Zhang, Hua Huang, Jie Cai, De-Zhu Li.
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: Ya J-D, Wang W-T, Liu Y-L, Jiang H, Han Z-D, Zhang T, Huang H, Cai J, Li D-Z (2023) Five new and noteworthy species of Epidendroideae (Orchidaceae) from southwestern China based on morphological and phylogenetic evidence. PhytoKeys 235: 211-236. https://doi.org/10.3897/phytokeys.235.111230
Open Access

Abstract

Five new orchid species from southwestern China’s Yunnan Province and the Tibetan Autonomous Region, Neottia lihengiae, Neottia chawalongensis, Papilionanthe motuoensis, Gastrochilus lihengiae, and Gastrochilus bernhardtianus, are described and illustrated. To confirm their identities, and to resolve phylogenetic relationships, we sequenced the complete plastomes of these taxa with their congeneric species, adding new plastomes of three Neottia species, two Papilionanthe species and nine Gastrochilus species. Combined with published plastid sequences, our well-resolved phylogeny supported the alliance of N. lihengiae with the the N. grandiflora + N. pinetorum clade. Neottia chawalongensis is now sister to N. alternifolia, while P. motuoensis is closely related to P. subulata + P. teres. Conversely, phylogenetic analyses based on complete plastomes and plastid sequences showed inconsistent relationships among taxa in the genus Gastrochilus, but the two new species, G. lihengiae and G. bernhardtianus were supported by all datasets.

Key words

Neottia, Gastrochilus, Papilionanthe, Plastid phylogenomics, Taxonomy, Tibet, Yunnan

Introduction

The Himalaya and Hengduan Mountains of southwestern China are iconic biodiversity hotspots of global significance (Liu et al. 2022). However, the species diversity of this region remains inadequately understood. In recent years, a considerable number of new plant species were described from these regions (Cai et al. 2019). Taking the Orchidaceae as an example, early floristic accounts of Tibet (Xizang) in the “Flora Xizangica” included only 64 genera totaling 191 species (Wu 1987). In contrast, a number of new species and new geographic records were reported over the last three decades (Wang et al. 2018; Liu et al. 2020a; Li et al. 2022, 2023), increasing the number of orchids in Tibet to 110 genera with 491 species in the latest checklist (Wang et al. 2023). Long-term and in-depth field investigations are still required to meet the urgent challenge of conserving rare species in these mountainous regions as their habitats undergo rapid changes.

The Orchidaceae is one of the largest families of angiosperms in the world, with approximately 190 genera and 1600 species in China (Jin et al. 2019). The Hengduan Mountains and the Himalaya, particularly counties of Gongshan, Shangri-La, and Lijiang (Yunnan) and Motuo (Medog) in Tibet (Xizang) are particularly rich in orchids (Zhang et al. 2015).

The genus Neottia Guett. a member of the Neottieae, was first established in 1754 consisting of a few, small, mycoheterotrophic orchids (Chen 1999). Recent molecular studies indicate that the autotrophic genus Listera Brown should be submerged within Neottia s.l. The monophyletic tribe Neottieae becomes the sister to the majority of the remaining members of the largest subfamily, Epidendroideae in the Orchidaceae (Pridgeon et al. 2005; Chen et al. 2009; Chase et al. 2015; Zhou and Jin 2018). There are 73 accepted species in Neottia s.l. distributed widely across the temperate and subarctic regions of the Northern Hemisphere, but the genus extends to northwest Africa, with a few species native to alpine regions in subtropical eastern Asia (Pridgeon et al. 2005; Chen et al. 2009; POWO 2023). Morphologically, Neottia is readily distinguished from other terrestrial epidendendroids by its two opposite to nearly opposite leaves (when present) or it is leafless. Each resupinate flower contains a curved column containing two sectile but naked pollinia (indicative of basal epidendroids).

The small genus Papilionanthe Schltr., a member of Vandeae (subtribe Aeridinae), was first described by Schlechter in 1915 based on Vanda teres (Roxb.) Lindl. published previously in genus Vanda R.Br. The genus Papilionanthe s.s. is distinguished from other genera in subtribe Aeridinae by multiple characters. It has fleshy and terete leaves, and a short inflorescence arising from a node opposite the leaf. The trilobate labellum is spurred. Its mid-lobe is often dilated and 2- or 3-lobed at its apex. The subterete and short column has a short foot. Pollinia are attached to a broadly triangular or subquadrate stipe which, in turn, is attached to a large and cellular viscidium (Chen et al. 2009; Pridgeon et al. 2014). Ten Papilionanthe species are recognized currently distributed from India eastward to south-central China and Malesia (POWO 2023).

The genus Gastrochilus D. Don is also a member of the subtribe Aeridinae and was established in 1825. It is characterized by monopodial growth, erect or pendulous stems and short axillary inflorescences. The labellum has a saccate hypochile. Two porate and globose pollinia are borne on a slender stipe (Chen et al. 2009; Pridgeon et al. 2014; Liu et al. 2019). Recent molecular studies of Gastrochilus show that traditional classification based on morphological characters is well supported (Liu et al. 2019). The genus consists of ca. 70 species, distributed through subtropical Asia, from Sri Lanka and India into the Himalaya, eastwards to southern China, southern Japan and southwards to the Philippines and Indonesian archipelago. Appoximately 40 species are found in China (Chen et al. 2009; Liu et al. 2019; POWO 2023).

We collected specimens of five previously unidentified species during our field surveys in Yunnan and Tibet from 2016–2023. Following a review of the literature (see Pearce and Cribb 2002; Chen et al. 2009; Raskoti 2009; Jin and Pang 2016; Averyanov et al. 2018; Wu et al. 2019; Mu et al. 2020; Liao et al. 2022), morphological studies of herbarium specimens and plastid phylogenomic analyses, we concluded that these specimens are new to the genera Neottia (Orchidaceae: Epidendroideae, Neottieae), Papilionanthe and Gastrochilus (Orchidaceae: Epidendroideae, Vandeae, Aeridinae) respectively. These species are analyzed and described below.

Materials and methods

Morphological studies

Living plants and herbarium specimens were collected in the field in the Hengduan Mountains of northwestern Yunnan and the Himalaya of southeastern Tibet. Morphological characters and measurements of the specimens described here were based on at least 5 living specimens first observed in the field then cultivated plants in the greenhouse. Voucher specimens are deposited in the Herbarium of the Kunming Institute of Botany, Chinese Academy of Sciences (KUN) and the Herbarium of the Yunnan Academy of Forestry and Grassland (YAF).

Taxon sampling, DNA extraction, sequencing, assembling and annotation

To clarify the phylogenetic relationships of five potentially new species with closely related species, we sampled and sequenced plastomes of 17 accessions representing three Neottia species, two Papilionanthe species and nine Gastrochilus species. Including those retrieved from the National Centre for Biotechnology Information (NCBI) database, our dataset comprises 412 plastid genes of a total of 83 accessions.

Total genomic DNA was extracted from silica-dried tissue using the Plant Genomic DNA Kit (Tiangen Biotech, Beijing, China). Libraries for pair-end 150 bp sequencing with 200–400 bp insert size were conducted on a BGISEQ-T7 platform at BGI Shenzhen (China) for genome skimming, producing approximately 2Gbp high-quality reads per sample. The plastomes of Neottia ovata (L.) Hartm. (NC_030712) and Gastrochilus formosanus (Hayata) Hayata (MN124435) were used as references for the assembling of the clean reads (Feng et al. 2016; Liu et al. 2020b). Complete plastomes and the nuclear internal transcribed spacer (ITS) assembly were conducted using the Getorganelle toolkit (Jin et al. 2020). The parameters used were R = 15, k = 21,45,65,85,105,127, F = embplant_pt; R = 7, k = 21,45,65,85,105,127, F = embplant_nr, respectively. Assembled plastid genomes were annotated by PGA GENEIOUS R9.0.2 (Biomatters Ltd. Auckland, New Zealand) using the plastome of Neottia ovata, Holcoglossum amesianum (Rchb. F.) Christenson (NC_041511.1) and Gastrochilus formosanus (Li et al. 2019).

Phylogenetic analysis

For Neottia, a total of 22 taxa were included in the analysis of the data set comprising two plastid DNA (matK, rbcL) and nuclear ribosomal (nr) ITS sequences, Cephalanthera longifolia (L.) Fritsch was used as the outgroup based on Zhou and Jin (2018). For Papilionanthe, phylogenomic analysis was implemented based on nrITS and six plastid markers (matK, trnL-trnF, psbA-trnH, atpI-atpH, trnS-trnfM and rbcL) from 6 Papilionanthe species, including two newly sequenced species, and a total of 12 accessions representing four genera. They were all analyzed with Ascocentrum ampullaceum (Roxb.) Schltr. as the outgroup (Zhang et al. 2013). For Gastrochilus, eight publicly available plastome sequences of Gastrochilus species were obtained from GenBank (Suppl. material 1) along with nine newly sequenced Gastrochilus species (n = 12 plants). Therefore, a total of 17 Gastrochilus plastomes were included in this study with Pomatocalpa spicatum Breda as the outgroup (Liu et al. 2019). DNA sequences obtained from nrITS and chloroplast matK, trnL-trnF, psbA-trnH, psbM-trnD were combined as a data matrix. Voucher information and GenBank accession numbers are provided in Suppl. materials 1–4. The plastid genes were aligned individually using MAFFT v7.308 (Katoh and Standley 2013), and the alignment of ITS and plastid genes are available in ScienceDB, after which alignment columns used Gblocks 0.91(Castresana 2000). Before phylogenetic analysis, the best-fit Akaike Information Criterion (AIC) model was selected in JModelTest v2.1.10 (Darriba et al. 2012). Phylogenetic trees were constructed by maximum likelihood (ML) in RaxML v8.2.11 (Stamatakis 2014) with 1,000 bootstrap replicates and Bayesian inference (BI) methods in Mrbayes v3.2.6 (Ronquist et al. 2012). For Bayesian inference, two separate Markov chain Monte Carlo (MCMC) chains were run for 200,000 generations with mixed nucleotide substitution models and 25% (50,000) of the trees were deleted as burn-in, and the results of two independently run computations were merged to produce a Bayesian consistent tree and a posterior probability value (PP) for each branch.

Results and discussion

Characteristics of the plastomes

All newly sequenced plastomes were assembled completely and can be accessed from GenBank (Table 1). Their genome features were found to be nearly identical, and gene content is conserved with an identical set of 68 annotated unique protein-coding genes (except for Neottia) and 29–30 tRNA genes and 4 rRNA genes. While, all three newly sequenced Neottia plastome sizes ranged from 155,447–156,082 bp, their genomes were composed of an LSC region (84,270–84,930 bp), SSC region (17,875–18,113 bp) and two IR copies (26,367–26,682 bp), with 74–80 unique genes. Their overall G/C content was almost identical (37.5–37.6%). The total plastome lengths of Papilionanthe teres (Roxb.) Schltr. and the putative new species, P. motuoensis ranged from 147,829–148,619 bp. Among all Gastrochilus plastomes, plastome sizes ranged from 146,615 to 148,552 bp. The genomes were composed of a large single repeat region (LSC) (84,710–85,682 bp), a small single repeat region (SSC), region (10,357–11,173 bp) and two inverted repeat (IR) copies (25,767–26,007 bp). Their overall G/C content was nearly identical (36.6–36.8%).

Table 1.
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Summary of plastomic data and nrITS sequences for Neottia, Papilionanthe and Gastrochilus species.

Species GenBank accession number Raw data Genome size (bp) LSC SSC IR Number of unique protein coding genes Number of tRNAs Number of rRNA ITS GenBank accession number Sequence length [bp]
Neottia chawalongensis OR786306 1.40/1.38G 155447 84581 18113 26367 119 30 4 OR073413 625
N. lihengiae OR002177 4.71/4.70G 155600 84270 17969 26682 114 30 4 OR073414 623
N. sp. OR002178 4.05/3.81G 156082 84930 17875 26639 109 30 4 623
Papilionanthe motuoensis OR772949 1.64/1.58G 148,619 84,574 12,055 25,945 107 30 4 OR073415 668
P. teres OR772950 1.00/1.02G 147,829 85,680 11,445 25,352 101 29 4 OQ991258 662
Gastrochilus bernhardtianus OR772951 1.17/1.10G 147,078 84,845 10,357 25,938 101 29 4 OR073405 398
G. bernhardtianus OR002167 1.70/1,72G 146,615 84,710 10,371 25,767 101 29 4 OR073404 654
G. fargesii OR002175 1.55/1.54G 148,552 85,682 11,132 25,951 110 30 4 656
G. distichus OR002170 873/893MB 147,834 85,063 11,113 25,829 101 29 4 OR073407 409
G. distichus OR002171 975/914MB 147,826 85,010 11,112 25,852 101 29 4 OR073406 654
G. gongshanensis OR002173 1.46/1.48G 147,728 84,936 11,032 25,880 101 29 4 OR073411 410
G. gongshanensis OR786306 1.83/1.88G 147,794 85,026 11,032 25,867 110 30 4 OR073412 655
G. lihengiae OR002168 1.60/1.59G 147,940 84,863 11,165 25,956 101 29 4 OR073408 656
G. lihengiae OR002169 2.01/2.00G 147,934 84,829 11,173 25,966 101 29 4 655
G. nanchuanensis OR002176 1.56/1.46G 148,001 84,942 11,045 26,007 110 30 4 OR073410 89
G. sp. OR002172 1.69/1.73G 147,706 84,938 11,032 25,867 110 30 4 621
G. sp. OR002174 1.70/1.80G 147,708 84,938 11,032 25,869 101 29 4 OR073409 654

Phylogenetic relationships within Neottia

Phylogenetic relationships based on combined nrITS and plastid DNA (matK, rbcL) data indicated that Neottia s.l. is monophyletic with moderate support (BP = 84, PP = 0.9993). Within the sampled species, the widespread and temperate N. ovata diverged initially, which is consistent with the previous study by Zhou and Jin (2018), followed by the clade of N. cordata (L.) Rich. and N. smallii (Wiegand) Szlach. The newly discovered species N. chawalongensis (Fig. 1) is sister to N. alternifolia (King & Pantl.) Szlach. (PP = 1, BP = 97). Together, they constitute the sister clade of N. meifongensis (H.J.Su & C.Y.Hu) T. C. Hsu & S. W. Chung. The second new species, N. lihengiae (Fig. 1) is clustered with N. pinetorum (Lindl.) Szlach. the unidentified Neottia sp. and N. wardii (Rolfe) Szlach. (PP = 0.92, BP = 91).

Figure 1. 

Phylogenetic relationships of Neottia species based on the nrITS, matK and rbcL. The ML and BI trees have the same topology. Numbers at nodes are Bayesian posterior probabilities and bootstrap percentages, respectively. “*” represents 100% support with newly sequenced species are shown in bold italics.

Phylogenetic relationships of Papilionanthe

In the overall matrix of Papilionanthe, 75 sequences were obtained (13 nrITS sequences and 14 matK, 14 trnL-trnF, 11 psbA-trnH, 11 atpI-atpH, 9 trnS-trnfM, and 3 rbcL sequences, respectively), and the combined dataset of 7 markers comprised 8558 aligned nucleotides, 790 bp from nrITS and 7768 bp from plastid regions, respectively.

The concatenated tree of nrITS and its plastid data show that Papilionanthe is monophyletic. The main clade of Papilionanthe is divided into two subclades (Fig. 2). In the first, P. biswasiana (Ghose & Mukerjee) Garay and P. hookeriana (Rchb.f.) Schltr. are sister species. In the second subclade, P. uniflora (Lindl.) Garay diverged first while the newly sequenced P. teres is well supported as sister to P. subulata (Willd.) Garay (BP = 99, PP = 1). Collectively it is sister to the new species P. motuoensis (see below) (BP = 99, PP = 1). The BI and maximum likelihood (ML) trees yield the same topology. The posterior probabilities and bootstrap probabilities values are high, indicating a high degree of confidence for the result. However, this topology is inconsistent with the previous study by Zhang et al. (2013). The difference between these topologies was the position of P. hookeriana and P. biswasiana, now consisting of a sister group in this study.

Figure 2. 

Phylogram of the genus Papilionanthe based on ML and BI analyses of the combined nrITS and plastid matK, trnL-trnF, psbA-trnH, atpI-atpH, trnS-trnfM, rbcL sequences. The ML and BI trees are identical and the BP and PP values are given beside the nodes. “*” indicates 100% bootstrap support.

Phylogenetic relationships of Gastrochilus

A total of 41 Gastrochilus species were included in this study to represent all six sections, 12 accessions representing nine species were newly generated in this study. Their relationships were confirmed using a combined dataset of nrITS and plastid matK, trnL-trnF, psbA-trnH, psbM-trnD sequences (Fig. 3a). The detailed sequence information is listed in Suppl. material 4, and the best-fit model selected by jModeltest is given in Table 2. Both RAxML and Bayesian inference (BI) analyses of the concatenated sequence supermatrix produced similar topologies for the Gastrochilus species. Gastrochilus s.l. is strongly supported as monophyletic with high posterior probabilities (PP) and bootstrap probabilities (BP) (Fig. 3a). The genus was subdivided into six well-supported clades, the earliest diverging clade is clade A (G. sect. Pseudodistichi), successively followed by clade B (G. sect. Brachycaules) (BP = 79, PP = 0.7916), clade C (G. sect. Gastrochilus) (BP = 100, PP = 0.9993) and clade D (G. sect. Acinacifolii) (BP = 99, PP = 1). Clades E and F are sister to each other and they together comprise a clade sisiter to clade D. Our two new species, G. lihengiae (see below) and G. bernhardtianus (see below) are resolved as distinct species in clade E (G. sect. Microphylli) in all data sets. Gastrochilus distichus (Lindl.) Kuntze + G. prionophyllus H. Jiang, D. P. Ye & Q. Liu is sister to G. lihengiae while G. heminii M. Liao, B. Xu & Yue.H. Cheng is sister to G. bernhardtianus and they consisting a sister group to G. alatus. Within clade F, two samples of G. gongshanensis Z.H.Tsi and two unidentified species from the Dali, Yunnan form a distinct subclade, which is sister to G. yunlongensis W. H. Rao, L. J. Chen & Z. J. Liu.

Figure 3. 

a Phylogenetic relationships in genus Gastrochilus based on nrDNA ITS and plastid matK, trnL-trnF, psbA-trnH, psbM-trnD sequences. ML and BI trees have the same topology and BP and PP are given beside the branches b phylogenetic relationships of Gastrochilus based on the complete plastomic sequences. All nodes are supported with a posterior probability (pp) of 1.0. “*” indicates 100% bootstrap support c phylogenetic tree based on combined nrITS and plastid DNA markers and conflicting topologies (clade B and clade F) are highlighted.

Table 2.
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Best-fit models and parameters for each genus.

Genus Region AIC select model Base frequencies Base frequencies p-inv (I) Gamma shape (G)
A C G T A-C A-G A-T C-G C-T G-T
Neottia ITS, matK, rbcL GTR+I+G 0.2907 0.1881 0.1987 0.3226 1.1924 1.7728 0.3011 0.2107 2.0924 1.0000 0.4320 0.9250
Papilionanthe ITS, matK, trnL-F, psbA-trnH, trnS-trnfM GTR+G 0.3083 0.1866 0.1675 0.3376 1.1439 1.3935 0.4909 0.4617 1.3426 1.0000 0.3190
Gastrochilus Plastome GTR+I+G 0.3129 0.1844 0.1785 0.3242 1.0779 1.2742 0.3062 0.2105 1.1734 1.0000 0.7880 0.8410
Gastrochilus ITS, matK, trnL-F, psbA-trnH, psbM-trnD GTR+I+G 0.3207 0.1879 0.18 0.3114 0.8602 1.6575 0.3075 0.5360 1.8524 1.0000 0.1610 0.0210

In the present study, analysis of 20 complete chloroplast genomes of Gastrochilus specimens provide a wealth of information to determine phylogenetic relationships within this genus, including a fully resolved phylogenetic tree with almost 100% bootstrap values and 1.00 posterior probabilities, and are better supported than in the studies of Liu et al. (2019) and Zhang et al. (2023) based on nrITS and plastid sequences (Fig. 3b). However, these phylogenetic relationships based on whole plastomes and chloroplast sequences suggest different topologies, particularly among the relationship between Clade B and subclade including G. fargesii (Fig. 3c). The analysis by Zhang et al. (2023) suggested that G. obliquus diverged early in the clade B with G. formosanus falling into clade F (Fig. 3a). In contrast, our plastome-based topology showed that species of clade B recovered as sister to clade F with high statistic support (Fig. 3c). One explanation is the difference in information sites, and the other possibility is sampling size. More samples with plastomic data should be used in future study to resolve the difference.

Conservation status

We preliminarly assesed the conservation status of the five new species using the IUCN Red List Categories and Criteria (IUCN 2022). Neottia lihengiae is known from two sites with a population of >2,000 individuals in an area of ca. 80 square kilometers scattered under a protected, evergreen broadleaved forest and a mossy dwarf forest in northwestern Yunnan, respectively. Based on population size and healthy habitats, the conservation status is proposed as Least Concern (LC). In contrast, the remaining four species, are known only from type localities and adjacent areas. For each of these species, only one or two populations with few individuals were detected during our two to three field surveys. More extensive fieldwork is needed to objectively assess their conservation status.. Therefore, the status of all remaining species are temporarily rated as Data Deficient (DD).

Taxonomic treatments

Neottia Guett

Neottia lihengiae J.D.Ya, H.Jiang & D.Z.Li, sp. nov.

urn:lsid:ipni.org:names:77331171-1
(李恒对叶兰 Li Heng Dui Ye Lan) Fig. 4

Diagnosis

Neottia lihengiae is morphologically similar to N. biflora (Schltr.) Szlach., but can be distinguished by its smaller plant size, ca. 5.5–9.0 cm tall (vs. 10–13 cm tall), its lax rachis of 2–5–flowered (vs. 1- or 2-flowered), floral bracts and sepals longer than their pedicel (vs. shorter than pedicel), smaller flowers with sepals and petals connivant and ca. 3.0 mm long (vs. spreading and ca. 6.0–7.0 mm long). The outer surfaces of the sepals are not carinate (vs. carinate). The labeullum is ligulate and its midvein is not thickened (vs. cuneate and midvein slightly thickened). The rostellum is almost equal to the anther (vs. distinctly shorter than the anther).

Figure 4. 

Neottia lihengiae J.D.Ya, H.Jiang & D.Z.Li, sp. nov. A habitat B plant C inflorescence D leaves E flower (front view) F flower (dorsal view) G column and labellum H column I anther cap. Photographed by J.-D. Ya.

Type

China. Yunnan Province, Diqing Prefecture, Shangri-La County, Tianbao mountain, 3800 m, under shrubs of a scree slope, 4 July 2020, J.-D. Ya et al. 20CS19095 (Holotype: KUN! isotype: KUN!)

Description

Terrestrial, autotrophic herbs, 5.5–9.0 cm tall. Rhizome with many elongate, filiform roots. Stem erect, slender, usually with 1 or 2 membranous ca. 8.0 mm long tubular sheaths at its base. Leaves 2, opposite, borne above the middle of the plant, 7 veined from the base, subsessile, broadly ovate or broadly ovate-triangular, unequal in size, the larger leaf ca. 1.2 × 1.2 cm, the smaller one ca. 1.0 × 1.0 cm, with bases rounded and apices acute. Peduncle 0.7–1.2 cm, puberulous, rachis 1.2–1.8 cm, laxly 2–5-flowered; floral bracts ovate-lanceolate, concave, longer than the pedicel, 3–4 × ca. 0.8 mm, apex acute to acuminate. Flowers resupinate, uniformly green; pedicel and ovary 2.0–3.0 mm long, glabrous; sepals and petals connivent. Dorsal sepal ovate-lanceolate, ca. 3.2 × 1.1 mm, 1-veined, apex subacute; lateral sepals lanceolate, slightly oblique, ca. 3.5 × 0.8 mm, 1-veined, apex acute. Lateral petals linear-lanceolate, ca. 3.0 × 0.6 mm, 1-veined, apices subacute; labellum ligulate, ca. 4.0 × 1.6 mm, entire to shallowly notched or emarginate at apex, usually with a minute tooth in the notch. Column slightly arcuate, ca. 1.7 mm long, anther inclined toward rostellum, ca. 0.9 mm; rostellum spreading forward, nearly as long as the anther.

Phenology

Flowers from June to July.

Etymology

Named in honor of late Prof. Li Heng, a Chinese botanist who made significant contributions to our understanding of plant diversity and phytogeography of the Gaoligong Mountains at the border between China and Myanmar (Guo et al. 2023).

Distribution and habitat

It is known from Northwest Yunnan including Lijiang and Diqing. It grows under shrubs colonizing scree slopes at elevations of 3700–3800 m.

Additional specimen examined

China. Yunnan Province, Lijiang City, Gucheng District, Dadong Xiang, 3192 m, in the scree slope area under the forest dominated by Pinus densata Mast. 17 June 2017, H. Jiang and W.P. Zhang 08835 (paratypes: YAF!); Yunnan Province, Diqing Prefecture, Shangri-La County, Tianbao mountain, 3719 m, under the shrub of scree slope, 15 Aug. 2018, C. Liu et al. 18CS17401 (paratypes: KUN!). N. biflora: China. Sichuan, Dongrergo, K. A. H. Smith 3656 (isotypes, PE00027184!). N. tianschanica: China. Xinjiang Uygur Autonomous Region, Tian-Shan, 18 July 1957, K.-Z. Guan 172 (holotype, LE 01012234!); China. Xinjiang Uygur Autonomous Region, Urumqi, Houxia Zhen, 2161 m, J.D. Ya et al. 17CS16209 (KUN1437961!).

Neottia chawalongensis J.D.Ya & D.Z.Li, sp. nov.

urn:lsid:ipni.org:names:77331172-1
(察瓦龙对叶兰 Cha Wa Long Dui Ye Lan) Fig. 5

Diagnosis

Neottia chawalongensis is similar to N. pinetorum (Lindl.) Szlach., but differs in having floral bracts longer than its pedicel (vs. shorter or as long as pedicel), a reduced pedicel ca. 1.9 mm (vs. 4–6 mm), and a shorter but pubescent ovary, ca. 2.8 mm (vs. glabrous, 3–4.5 mm). The labellum is lanceolate (vs. obovate-cuneate, oblong-cuneate, sublinear-cuneate, or oblanceolate), densely papillate (vs. slightly papillate), with labellum lobes narrowly lanceolate and apices acuminate (oblong-ovate and apices obtuse-rounded) while its sinus usually lacks a short tooth between the lobes.

Figure 5. 

Neottia chawalongensis J.D.Ya & D.Z.Li, sp. nov. A, B plant C leaves D inflorescence E flower (front view) F adaxial sepals, petals and lip G abaxial sepals, petals and lip H column and lip I column J anther cap. Photographed by J.-D. Ya.

Type

China. Tibetan Autonomous Region, Linzhi City, Chayu County, Chawalong Township, 3757 m, under the shrub of scree slope, 21 July 2022, J.-D. Ya et al. 22CS22851 (KUN!)

Description

Terrestrial, autotrophic plants, 9.0–13.5 cm tall. Rhizome with many elongate, filiform roots. Stem 5.0–7.0 cm, erect, slender, ridged, usually with 1 or 2 membranous, long, tubular sheaths at its base. Leaves 2, opposite, borne in the middle of the plant, 5 veined from the base, subsessile, broadly ovate or broadly ovate-triangular, ca. 2.0 × 1.8 cm in diameter. Peduncle 1.8–2.4 cm, puberulous, rachis 2.7–4.3 cm, held laxly bearing 7–9-flowered; floral bracts ovate, concave, longer than the pedicel, 3–4 mm long with acute apices. Flowers resupinate, uniformly green; pedicel ca. 1.9 mm, glabrous to sparsely pubescent; ovary ca. 2.8 mm, pubescent with sepals and petals widely spreading. Dorsal sepal narrowly elliptical, ca. 3.5 × 1.3 mm, 1-veined, apex obtuse; lateral sepals narrowly elliptic-falcate, ca. 3.5 × 1.5 mm, with an obtuse apex. Lateral petals linear, ca. 3.0 × 0.4 mm, apices subacute; labellum pendulous, lanceolate, ca. 8.0 × 2.2 mm, margins densely papillate, apex deeply 2-lobed with lobes parallel, narrowly lanceolate, ca. 2.6 × 0.7 mm, and apices acuminate; disk with a thickened longitudinal channel extending from the base of the labellum almost to the sinus. Column slightly arcuate above the middle, 3.5 mm long; anther inclined towards the rostellum, ca. 0.8 mm; rostellum spreading forward.

Phenology

Flowers from July to August.

Etymology

The specific epithet “chawalongensis” refers to the type locality Chawalong (Cawarong) Township.

Distribution and habitat

At present, this new species is only found in Chawalong, Chayu, Tibet (Xizang), China. It is a predominantly terrestrial species growing on the scree slopes under the forest of Abies and Picea at an elevation of 3757 m a.s.l. It appears to be locally abundant with other orchid species including Ponerorchis chusua (D. Don) Soó, Galearis spathulate (Lindl.) P. F. Hunt, Cypripedium wardii Rolfe, C. bardolphianum W.W.Sm. & Farrer and C. flavum P. F. Hunt & Summerh.

Additional specimen examined

N. pinetorum: India. Sikkim, 10–11000 feet., J. D. Hooker 355 (holotype, K000974204!, isotype, AMES 00101020!); China. Yunnan, upper Kiukiang valley, 2500 m, T.T.Yu 19644 (PE00027188!). N. bambusetorum: China, Yunnan, Prope fines Tibeto-Birmanicas inter fluvios Lu-djiang (Salween) et Djiou-djiang (Irrawadi or. sup.), in jugi Tschiangschel, 27°52', lateris orientalis regione (frigide) temperata in bambusetis, 3275–3350 m, Hand.-Mazz.9238 (holorypus, WU0061594!)

Papilionanthe Schltr

Papilionanthe motuoensis J.D.Ya & D.Z.Li, sp. nov.

urn:lsid:ipni.org:names:77331173-1
(墨脱凤蝶兰 Mo Tuo Feng Die Lan) Figs 6, 7A–C

Diagnosis

Papilionanthe motuoensis is similar to P. uniflora (Lindl.) Garay but differs in having a glabrous pedicel and ovary (vs. glandular-pubescent). Its lateral petals are oblong-ovate (vs. oblong) with irregularly denticulate margins (vs. with undulating margins), truncate apices (vs. obtuse apices). Its labellum is white tinged with yellow (vs. uniformly white), with a subflabellate mid-lobe and a labellum base with an apically dilate to reniform claw, its apex is emarginated with an irregularly denticulate margin (vs. mid-lobe simple, oblong, apex widely cuneate).

Figure 6. 

Papilionanthe motuoensis J.D.Ya & D.Z.Li, sp. nov. A plant B flower (dorsal view) C flower (front view) D flower (lateral view) E abaxial sepals and petals F adaxial sepals and petals G column and lip (lateral view) H column and lip (front view) I lip (front view) J lip (rip cutting) K column (lateral view) L column (front view) M anther cap N pollinarium. Photographed by J.-D. Ya.

Type

China. Yunnan, Kunming, voucher from cultivated plants at Kunming Institute of Botany, CAS, 20 Oct. 2020 (flowering), J.-D. Ya BC201015 (holotype: KUN!), plants originally collected from Tibet (Xizang), Linzhi City, Motuo County, 1625 m, at the edge of a subtropical, evergreen, broadleaved forest.

Description

Stems pendulous, terete, to 50 cm, 2.0 mm in diam. branched, enclosed in leaf sheaths. Leaves laxly alternate, terete, 9–16 × 0.2 cm, base with amplexicaul-sheathing, apex apiculate; sheaths tubular, 2.0–2.8 cm long, glabrous. Inflorescence ca. 1.5 cm, usually 1–2-flowered; peduncle slender, ca. 1.3 cm; floral bracts ovate-triangular, 1.5 × 1.2 mm. Flowers 2.5 cm in diam. sepals and lateral petals white, mid-vein pink, labellum white tinged with yellow, its spur with a whitish and/or pink tinge, apex yellowish green. Pedicel and ovary, ca. 1.2 cm long, glabrous. Dorsal sepal ovate, ca. 1.0 × 0.5 cm, acute, 5-veined; lateral sepals oblong, slightly falcate, ca. 1.1 × 0.4 cm, acuminate, 5-veined; lateral petals oblong-ovate, 1.1 × 0.6 cm, margin irregularly denticulate, apices truncate, 7-veined; labellum adnate to column foot, 3-lobed; lateral lobes deeply bifid, unequal, linear, acute, long lobule ca. 7.0 × 2.0 mm, short lobule ca. 4.0 × 1.0 mm; mid-lobe spreading, subflabellate, ca. 4.8 × 5.0 cm, base with a claw ca. 2.2 × 2.0 mm, apical dilate to reniform, apex emarginate, margin irregularly denticulate; spur slightly curved forward, cylindrical, ca. 22.0 × 3.5 mm, narrowing towards the terminus, its interior pubescent. Column 7.0 × 3.0 mm, foot ca 5.2 mm, with narrowly and entire wings decurrent to foot; anther cap galeate with a acuminate apex, 2.0 × 2.5 mm; pollinia 2, subglobose, ca. 1.0 mm in diameter, waxy, porate, attached by a stipe to a broad cellular viscidium.

Figure 7. 

A–C Papilionanthe motuoensis D–F Papilionanthe uniflora (Nepal). Photographed by: A–C J.-D. Ya D–F Bhakta Bahadur Raskoti.

Phenology

Observed flowering in October.

Etymology

The specific epithet “motuoensis” refers to the type locality Motuo (Medog) County.

Additional specimens examined

Tibet (Xizang), Linzhi City, Motuo County, Bangxin Xiang, 1330 m, from subtropical, evergreen, broadleaved forest. Oct. 2019, M.-K. Li and W. Wang 2019343 (paratypes, Herbarium of Tibet Agricultural and Animal Husbandry University, No. 8 Xueyuan Road, Bayi District, Nyingchi, Tibet). P. uniflora: Nepal. Gosain Than, N. Wallich no. 1993 (K001114863!); India, Mao, C.B. Clarke 41790 (K000891405!)

Distribution and habitat.

The new epiphytic species was found only in Motuo County, Tibet (Xizang), China, growing on limbs in a subtropical, evergreen, broadleaved forest at elevations of 1300–1650 m.

Gastrochilus D.Don

Gastrochilus lihengiae J.D.Ya, Ting Zhang & Z.D.Han, sp. nov.

urn:lsid:ipni.org:names:77331174-1
(纤细盆距兰 Xian Xi Pen Ju Lan) Figs 8, 9

Diagnosis

The floral morphology of Gastrochilus lihengiae is similar to G. distichus (Lindl.) O. Kuntze and G. prionophyllus H. Jiang, D. P.Ye & Q. Liu, but can be distinguished from the former by its narrower leaves, blades 0.25–0.35 cm wide (vs. 0.4–0.6 cm), and distinctly serrate leaf margins (vs. entire), with acuminate and mucronate apices (vs. apex acute bearing 2 or 3 awns). The lateral petals are narrowly oblong (vs. subobovate). The labellum with a hypochile, ca 7.0 mm (vs. 4.0 mm). The outside of the hypochile with three ridges (vs. glabrous), and from the latter by its falcate-lanceolate (vs. ovate) leaves with mucronate apices (vs. apex with 2 unequally awns), the lateral petals are narrowly oblong (vs. subobovate), the central cushion on the epichile of the labellum is not thickened (vs. thickened), while the outer surface of its hypochile has three ridges (vs. glabrous).

Figure 8. 

Gastrochilus lihengiae J.D.Ya, Ting Zhang & Z.D.Han, sp. nov. A flowering plant B fruiting plant; Stem C leaf D inflorescence E–I flower (different view) J adaxial sepals and petals K abaxial sepals and petals L column and lip M, N lip (rip cutting) O, P column Q pollinarium R anther cap. Photographed by J.-D. Ya & Z.-D. Han.

Type

China. Yunnan Province, Nujiang Prefecture, Gongshan County, Cikai Township, 1935 m, in the montane moist evergreen broad-leaved forest, 24 Apr. 2020, J.-D. Ya et al. 22CS21828 (Holotype: KUN! isotype: KUN!)

Description

Epiphytic herbs, stem pendulous, to 20 cm long, ca. 1.0–1.5 mm in diameter, slender, with 0.5–0.6 cm internodes, often branched with tiny red-purple spots. Leaves alternate, distichous, falcate-lanceolate, ca. 1.6–1.8 × 0.25–0.35 cm, the margin significantly serrate with an acuminate and mucronate apex. Inflorescences several, held opposite to nearly opposite the leaves, subumbellate, 1–3-flowered; peduncle 0.7–1.0 cm, slender, upper part enlarged, lower part with 2 cupular sheaths; floral bracts ovate, ca. 1.0 mm; pedicel and ovary 1.0–1.1 cm. Flowers yellow-green, with reddish brown spots. Dorsal sepal concave, oblong-ovate, ca. 4.0 × 2.0 mm, apex obtuse; lateral sepals concave, narrowly oblong, ca. 5.5 × 1.8 mm, apex obtuse; lateral petals narrowly oblong, 4.0 × 1.8 mm, apices subtruncate. Labellum subdivided into an epichile and a saccate hypochile; the epichile subovate triangular, ca. 5.0 × 2.5 mm, adaxially glabrous, with a central cushion and 2 conic calli near its base, the margin entire to irregularly denticulate, apex rounded; the hypochile subcupular, ca. 7.0 mm tall and ca. 5.2 mm in diam. outside with three ridges from the base of the column to its apex. Column stout, ca. 2.0 mm long, with rounded-auriculate wings at the base; anther cap narrowed into a beak towards its apex; the rostellum bilobed with an acuminate tip, and a horn-like awn arising from the center of each lobe; pollinarium ca 2.1 mm long; pollinia 2, yellow, 0.8 × 0.5 mm, almost hemispheric with a depression at the center; stipe elongate, obovate, ca.1.5 mm long; cellular viscidium elliptic, 1.0 × 0.5 mm. Capsules cylindrical, ca. 1.5 × 0.6 cm.

Figure 9. 

Gastrochilus lihengiae J.D.Ya, Ting Zhang & Z.D.Han, sp. nov. A plant B flower C sepals and petals D column and lip E epichile F pollinarium G anther cap H leaf. Drawn by Z.-D. Han.

Phenology

Flowering from March to April, while the fruits matured in March in the following year.

Etymology

Named in honor of late Prof. Li Heng for her contributions to the orchid flora of Yunnan (Guo et al. 2023).

Distribution and habitat

At present, two populations of this new species were found in Gongshan County, Yunnan, China. It is epiphytic on tree trunks in the mixed evergreen broad-leaved forest or montane moist evergreen broad-leaved forest at an elevation of 1900–2100 m.

Additional specimens examined

China. Yunnan Province, Nujiang Prefecture, Gongshan County, Dulongjiang Xiang, 2051 m, in the mixed evergreen broad-leaved forest, 4 Mar. 2023, Ting Zhang et al. 23CS24145 (paratypes, KUN!). G. distichus: India. Skimm, J.D. Hooker 206. (holotype: K000873754!). G. prionophyllus: China. Yunnan, Malipo County, Xia jinchang town, limestone forest, 1550–1650 m a.s.l., epiphytic on tree trunks or on rocks, 15 Mar. 2016, Qiang Liu 359 (holotype, HITBC!). G. fargesii: China. Sichuan, Tschen-keou-tin, P.G. Farges 1236 (type, K00083803! isotype, AMES00271835!).

Gastrochilus bernhardtianus J.D.Ya & D.Z.Li, sp. nov.

urn:lsid:ipni.org:names:77331175-1
(丽江盆距兰 Li Jiang Pen Ju Lan) Figs 10, 11, 12 A–C

Diagnosis

Gastrochilus bernhardtianus is similar to G. affinis (King & Pantl.) Schltr. in floral morphology, but can be distinguished by its shorter peduncle, ca. 0.3 cm (vs. 1.5–2.0 cm), pedicel and ovary ca. 4.5 mm (vs.0.6–1.3 cm). Sepals and lateral petals dark yellowish-green with densely purplish-red marks or spots flushed brown to purplish brown (vs. green flushed with brown to purplish brown). The dorsal sepal elliptic, ca 3.4 mm wide (vs. elliptic-oblong, 1.0–1.3 cm wide), lateral sepals narrowly ovate, ca. 5.5 × 2.8 mm (vs. elliptic-ovate, 3.5–4.0 × 0.7–1.3 mm). Lateral petals narrowly oblong, ca. 5.2 × 2.7 mm (vs. ovate-elliptic to elliptic, 3.0–4.0 × 1.0–1.3 cm). Labellum with purplish-red spots (vs. yellowish to greenish-yellow marks) and yellowish-green calli (vs. brown to purplish brown) with a transversely oblong epichile (vs. broadly subtriangular) and a green center (vs. deep purple to purplish-brown).

Figure 10. 

Gastrochilus bernhardtianus J.D.Ya & D.Z.Li, sp. nov. A plant B inflorescence C–E flower F adaxial sepals and petals G abaxial sepals and petals H column and lip I column J–M lip N lip (rip cutting) O pollinarium P anther cap. Photographed by J.-D. Ya.

Type

China. Yunnan Province: Lijiang Prefecture, Yulong County, Yunshanping, 3308 m, in cold-temperate, evergreen conifer forest, 20 May 2020, J.-D. Ya et al. 20CS19022 (Holotype: KUN!)

Description

Epiphytic herb, stem pendulous, with purplish spots, ca. 5.0 cm long, 1.5–2.0 mm in diameter. Leaves distichous, blade oblong-lanceolate, with purple-red spots on the abaxial leaf surface, 1.8–2.5 × 0.4–0.7 cm, base sheathing, apex acute and slightly trilobate. Racemes axillary, sub-umbellate, 1–2 flowered; peduncle ca. 0.3 cm, with purple-red spots; floral bracts ovate-triangular, ca. 1.0 mm; pedicel and ovary yellow-green with purple-red spots, ca. 4.5 mm. Flower densely marked with purplish-red spots flushed with brown to purplish brown, sepals and lateral petals dark yellowish-green. Dorsal sepal elliptical, ca. 5.2 × 3.4 mm, apex obtuse; lateral sepals narrowly ovate, ca. 5.5 × 2.8 mm, apices obtuse; lateral petals narrowly oblong, ca. 5.2 × 2.7 mm, apices obtuse. Labellum epichile with a green center and yellowish green margins, transversely oblong, ca. 8.0 × 2.8 mm, adaxially glabrous, with a central cushion and 2 conic calli near its base, margins erose, apex rounded; hypochile saccate, light yellowish green, subconical, ca. 5.1 mm tall and ca. 3.8 mm in diam. dorsally compressed, slightly bent outward, subacute to obtuse and shortly bifid at apex, with one internal ridge at the bottom. Column stout, ca. 2.0 mm, with rounded-auriculate wings at the base, anther cap galeate with recurved–acuminate apex, 1.2 × 0.9 mm; rostellum bilobed with an acuminated terminus; pollinarium ca 2.0 mm long; pollinia 2, yellow, 0.6 × 0.5 mm, almost hemispheric with a depression at the centre; stipe elongate, obovate, ca.1.0 mm long; cellular viscidium elliptic, 0.8 × 0.3 mm.

Figure 11. 

Gastrochilus bernhardtianus J.D.Ya & D.Z.Li, sp. nov. A plant B flower C sepals and petals D column and lip E lip F anther cap G pollinarium. Drawn by Z.-D. Han.

Figure 12. 

Comparison of two taxa of Gastrochilus A–C G. bernhardtianus with various colours in different areas D, E G. affinis. Photographed by J.-D. Ya.

Phenology

Flowering from May to June.

Etymology

The species is named after Peter Bernhardt, pollination biologist and orchidologist, for his contributions to pollination ecology of Chinese orchids in collaboration with botanists of China. Previously Professor of Biology at St. Louis University, USA,Peter Bernhardt was the 2022 recipient of the Peter H. Raven Scientific Outreach Award (Raven 2023). Currently he works closely with colleagues in Yunnan as a research associate of the Missouri Botanical Garden, USA and as an adjunct professor at Curtin University, Perth, Western Australia.

Distribution and habitat

The new species is found only in Yulong County, Yunnan, China, and epiphytic on trees of the cold-temperate, evergreen needleleaved forest dominated by Picea likiangensis (Franch.) E.Pritz. and Abies forrestii Coltm.-Rog. at an elevation of 3300 m a.s.l.

Additional specimens examined

G. affinis: India. Sikkim, Lachong Valey, R. Pantling 444 (K000891609!); China. Yunnan, Fugong, Jiakedi, east slope of Gaoligongshan, epiphyticon trunk, alt., 2555 m,16 May 2005, X. H. Jin6984 (PE!); Yunnan, Tengchong, 2828 m, 31 Mar 2007, X.H. Jin 8936 (PE!). G. alatus: China: Yunnan, Fugong, Zhuminglin, 2758 m, 16 May 2005, H.X. Jin 6998 (Holotype, PE!).

Acknowledgments

We are grateful to Prof. Xiao-hua Jin for his valuable discussions and suggestions, to Dr. Yan-Hui Zhao, Mr. Wei Zhang, Mr. Cheng Liu, Mr. Sheng-Ping Ming, Mr. Jin Li, and Mr. Chang-Hong Li for their kind assistance in the field, to Dr. Bhakta Bahadur Raskoti for his outstanding image. We convey our special thanks to the Gongshan Branch of the Gaoligong Mountains National Nature Reserve for their kind help in the field. This study was financially supported by the Science and Technology Basic Resources Investigation Program of China (grant No. 2021FY100200), the Key Basic Research Program of Yunnan Province, China (grant 202101BC070003), the National Wild Plant Germplasm Resource Center and a CAS Technology Talent Program to J. Cai, and National Forestry and Grassland Administration (grants 2019073017, 2019073019).

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This study was financially supported by the Science and Technology Basic Resources Investigation Program of China (grant No. 2021FY100200), the Key Basic Research Program of Yunnan Province, China (grant 202101BC070003), the National Wild Plant Germplasm Resource Center and a CAS Technology Talent Program to J. Cai, and National Forestry and Grassland Administration (grants 2019073017, 2019073019).

Author contributions

J.D.Y. and D.Z.L. conceived of and designed the study. J.D.Y. J.H. Z.D.H. T.Z. and H.H. contributed plant materials. W.T.W. and Y.L.L. performed the plastome assembly, annotation, and phylogenetic analyses. J.D.Y. W.T.W. Y.L.L. and D.Z.L. wrote the manuscript. J.C. and D.Z.L. supported funding acquisition. All authors read and approved the final version of manuscript.

Author ORCIDs

Ji-Dong Ya https://orcid.org/0000-0003-3389-1412

Wan-Ting Wang https://orcid.org/0009-0007-9822-4983

Yun-Long Liu https://orcid.org/0000-0003-4650-3466

Hong Jiang https://orcid.org/0000-0001-6613-8588

Zhou-Dong Han https://orcid.org/0009-0001-9431-1349

Ting Zhang https://orcid.org/0000-0003-0939-8468

Hua Huang https://orcid.org/0000-0002-7400-5651

Jie Cai https://orcid.org/0000-0003-1627-3700

De-Zhu Li https://orcid.org/0000-0002-4990-724X

Data availability

All relevant data are within the manuscript and its Additional files. The data that support the findings of this study are openly available in the Science Data Bank at https://www.doi.org/10.57760/sciencedb.09591 or http://resolve.pid21.cn/31253.11.sciencedb.09591.

References

  • Cai J, Yu WB, Zhang T, Wang H, Li DZ (2019) China’s biodiversity hotspots revisited: A treasure chest for plants. In: Cai J, Yu W-B, Zhang T, Li D-Z (Eds) Revealing of the plant diversity in China’s biodiversity hotspots. PhytoKeys 130: 1–24. https://doi.org/10.3897/phytokeys.130.38417
  • Chase MW, Cameron KM, Freudenstein JV, Pridgeon AM, Salazar G, Berg C, Schuiteman A (2015) An updated classification of Orchidaceae. Botanical Journal of the Linnean Society 177(2): 151–174. https://doi.org/10.1111/boj.12234
  • Chen SC (1999) Neottia and Listera. In: Lang KY (Ed.) Flora Republicae Popularis Sinicae, Vol. 17. Science Press, Beijing, 97–121.
  • Chen SC, Gale SW, Cribb PJ (2009) Neottia, Papilionanthe, Gastrochilus. In: Wu ZY, Raven PH, Hong DY (Eds) Flora of China, vol. 25. Sciences Press, Beijing, in association with Missouri Botanical Garden Press, St. Louis, 184–195, 477–478, 491–498.
  • Darriba D, Taboada GL, Doallo R, Posada D (2012) jModelTest 2: More models, new heuristics and parallel computing. Nature Methods 9(8): 772. https://doi.org/10.1038/nmeth.2109
  • Feng YL, Wicke S, Li JW, Han Y, Lin CS, Li DZ, Zhou TT, Huang WC, Huang LQ, Jin XH (2016) Lineage-Specific Reductions of Plastid Genomes in an Orchid Tribe with Partially and Fully Mycoheterotrophic Species. Genome Biology and Evolution 8(7): 2164–2175. https://doi.org/10.1093/gbe/evw144
  • Guo HJ, Huang JL, Ji YH, Li R, Long CL, Yang QE, Yang YP, Yi TS (2023) Honoring a legend: Celebrating the life and legacy of Professor Heng Li. Plant Diversity 45(3): 239–240. https://doi.org/10.1016/j.pld.2023.03.003
  • Jin XH, Li JW, Ye DP (2019) Atlas of Native Orchids in China. Henan Science and Tecnology Press, Zhenzhou.
  • Jin JJ, Yu WB, Yang JB, Song Y, DePamphilis CW, Yi TS, Li DZ (2020) GetOrganelle: A fast and versatile toolkit for accurate de novo assembly of organelle genomes. Genome Biology 21(1): 241. https://doi.org/10.1186/s13059-020-02154-5
  • Katoh K, Standley DM (2013) MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Molecular Biology and Evolution 30(4): 772–780. https://doi.org/10.1093/molbev/mst010
  • Li ZH, Ma X, Wang DY, Li YX, Wang CW, Jin XH (2019) Evolution of plastid genomes of Holcoglossum (Orchidaceae) with recent radiation. BMC Evolutionary Biology 19(1): 63. https://doi.org/10.1186/s12862-019-1384-5
  • Li MK, Pang SS, Deng JP, Wang W, Chen XD, Luo Y, Xing Z (2023) New Records of tribe Vandeae (Orchidaceae) in Tibet. Redai Zuowu Xuebao 44(2): 282–288.
  • Liao M, Cheng YH, Zhang JY, Feng Y, Liu GY, Ye P, Jin SL, Lin HQ, Xu B (2022) Gastrochilus heminii (Orchidaceae, Epidendroideae), a new species from Sichuan, China, based on molecular and morphological data. PhytoKeys 215: 95–106. https://doi.org/10.3897/phytokeys.215.91061
  • Liu Q, Song Y, Jin XH, Ga JY (2019) Phylogenetic relationships of Gastrochilus (Orchidaceae) based on nuclear and plastid DNA data. Botanical Journal of the Linnean Society 189(3): 228–243. https://doi.org/10.1093/botlinnean/boy084
  • Liu DK, Tu XD, Zhao Z, Zeng MY, Zhang S, Ma L, Zhang GQ, Wang MM, Liu ZJ, Lan SR, Li MH, Chen SP (2020b) Plastid phylogenomic data yield new and robust insights into the phylogeny of Cleisostoma–Gastrochilus clades (Orchidaceae, Aeridinae). Molecular Phylogenetics and Evolution 145: 106729. https://doi.org/10.1016/j.ympev.2019.106729
  • Liu J, Milne RI, Zhu GF, Spicer RA, Wambulwa MC, Wu ZY, Boufford DE, Gao LM, Li DZ (2022) Name and scale matters: Clarifying the geography of Tibetan Plateau and adjacent mountain regions. Global and Planetary Change 215: 103893. https://doi.org/10.1016/j.gloplacha.2022.103893
  • Pearce NR, Cribb PJ (2002) The Orchids of Bhutan. Royal Botanic Garden Edinburgh, Edinburgh.
  • Pridgeon AM, Cribb PJ, Chase MW, Rasmussen FN (2014) Genera Orchidacearum, 6. Oxford University Press, Oxford, 188–191, 218–220.
  • Raskoti BB (2009) The Orchids of Nepal. Quality Printers, Kathmandu, Nepal.
  • Ronquist F, Teslenko M, Van Der Mark P, Ayres DL, Darling A, Höhna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP (2012) MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61(3): 539–542. https://doi.org/10.1093/sysbio/sys029
  • Wang W, Li MK, Xing Z (2023) The Wild Orchids in Tibet. Tongji University Press, Shanghai.
  • Wu ZY [Ed.] (1987) Flora Xizangia (Orchidaceae), Vol. 5. Science Press, Beijing, 648–870.
  • Wu XF, Ye DP, Pan B, Lin XQ, Jiang H, Liu Q (2019) Validation of Gastrochilus prionophyllus (Vandeae, Orchidaceae), a new species from Yunnan Province, China. PhytoKeys 130: 161–169. https://doi.org/10.3897/phytokeys.130.34555
  • Zhang GQ, Liu KW, Chen LJ, Xiao XJ, Zhai JW, Li LQ, Cai J, Hsiao YY, Rao WH, Huang J, Ma XY, Chuang SW, Huang LQ, Tsai WC, Liu ZJ (2013) A new molecular phylogeny and a new genus, Pendulorchis, of the Aerides-Vanda alliance (Orchidaceae: Epidendroideae). PLoS ONE 8(4): e60097. [Epub2013] https://doi.org/10.1371/journal.pone.0060097
  • Zhang YB, Du HD, Jin XH, Ma KP (2015) Species diversity and geographic distribution of wild Orchidaceae in China. Kexue Tongbao 60(2): 179–188. https://doi.org/10.1360/N972014-00480 [In Chinese]
  • Zhang JY, Cheng YH, Liao M, Feng Y, Jin SL, He TM, He H, Xu B (2023) A new infrageneric classification of Gastrochilus (Orchidaceae: Epidendroideae) based on molecular and morphological data. Plant Diversity. https://doi.org/10.1016/j.pld.2023.08.001

Supplementary materials

Supplementary material 1 

Voucher and GenBank accession numbers of Neottia samples.

Ji-Dong Ya, Wan-Ting Wang, Yun-Long Liu, Hong Jiang, Zhou-Dong Han, Ting Zhang, Hua Huang, Jie Cai, De-Zhu Li

Data type: xlsx

Explanation note: List of taxa and plant materials. “XXXX” indicates plastid sequences used in analysis without Genbank accession numbers. Alignment of sequences are available under the Science data bank.

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (12.17 kb)
Supplementary material 2 

List of taxa, vouchers and GenBank accession numbers of Papilionanthe samples downloaded from NCBI (with newly sampled sequences added, below)

Ji-Dong Ya, Wan-Ting Wang, Yun-Long Liu, Hong Jiang, Zhou-Dong Han, Ting Zhang, Hua Huang, Jie Cai, De-Zhu Li

Data type: xlsx

Explanation note: List of taxa and plant materials. “XXXX” indicates plastid sequences used in analysis without Genbank accession numbers. Alignment of sequences are available under the Science data bank.

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (11.51 kb)
Supplementary material 3 

Summary of publicly available Gastrochilus plastomes sequences in this study

Ji-Dong Ya, Wan-Ting Wang, Yun-Long Liu, Hong Jiang, Zhou-Dong Han, Ting Zhang, Hua Huang, Jie Cai, De-Zhu Li

Data type: xlsx

Explanation note: List of taxa and plant materials. “XXXX” indicates plastid sequences used in analysis without Genbank accession numbers. Alignment of sequences are available under the Science data bank.

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (10.28 kb)
Supplementary material 4 

Summary of publicly available Gastrochilus plastid sequences in this study

Ji-Dong Ya, Wan-Ting Wang, Yun-Long Liu, Hong Jiang, Zhou-Dong Han, Ting Zhang, Hua Huang, Jie Cai, De-Zhu Li

Data type: xlsx

Explanation note: List of taxa and plant materials. “XXXX” indicates plastid sequences used in analysis without Genbank accession numbers. Alignment of sequences are available under the Science data bank.

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (13.57 kb)
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