Coptis austrogaoligongensis (Ranunculaceae), a new species from West Yunnan, China

Zhuo Cheng; Jiahua Li; Congli Xu; Lixiang Zhang; Qiangbang Gong; Chunlin Long

doi:10.3897/phytokeys.244.127978

Abstract

Based on morphological and plastid data, we have described and confirmed that Coptis austrogaoligongensis distributed in Tongbiguan Provincial Nature Reserve, Yingjiang County, Yunnan Province, is a new species of Coptis. It is distinctly different from C. teeta subsp. teeta and C. teeta subsp. lohitensis with differences mainly reflected in the following features: former leaf segment lobes contiguous to each other, and lateral segments equal to central one; plants without developed stolons; inflorescences with only 1–3 flowers; petals have short claws. Phylogenetic analysis indicated that C. austrogaoligongensis is a sister to C. teeta subsp. teeta and C. teeta subsp. lohitensis.

Key words

Coptis, Coptis austrogaoligongensis, taxonomy, Yunnan Province

Introduction

Coptis Salisb. is a small genus of Ranunculaceae, consisting of perennial herbs with yellowish rhizomes and numerous fibrous roots (Wang 1979, 2001). The taxonomy of Coptis has traditionally been based on vegetative (leaf type and shape) and reproductive (flower number, color and shape of sepals and petals, and beak length) characters (Xiang et al. 2016, 2018). The genus Coptis comprises 17 species worldwide, which is mainly distributed in the warm temperate to the cold coniferous forests of eastern Asia and North America (Xiang et al. 2016, 2018; Wang et al. 2020). Representatives of the genus Coptis are considered essential medicinal plants in Eastern Asia. They are rich in isoquinoline type alkaloids, such as berberine, epiberberine, coptisine, palmatine, jatrorrhizine, and columbamine, and their dried rhizomes are used in traditional Chinese medicine as Rhizoma Coptidis, which is famous for clearing heat, removing dampness, and reducing fire (Wang et al. 2019; Yang et al. 2021).

According to the morphology, especially flower and leaf, genus representatives distributed in China had been classified into seven species, one variant and one subspecies, i. e., C. chinensis Franch. (endemic to SW China), C. chinensis var. brevisepala W. T. Wang & P. K. Hsiao (endemic to SE China), C. deltoidea C. Y. Cheng & P. K. Hsiao (endemic to Sichuan, China), C. omeiensis (Chen) C. Y. Cheng (endemic to Sichuan, China), C. quinquefolia Miq. (distributed in Taiwan Province and Japan), C. quinquesecta W. T. Wang (endemic to Yunnan, China), C. teeta Wall. (distributed in East Himalaya), C. teeta subsp. lohitensis Pandit & Babu (distributed in East Himalaya), and C. huanjiangensis L.Q. Huang, Q.J. Yuan & Y.H. Wang. C. huanjiangensis as a new species was described during the survey of traditional Chinese medicine resources in Huanjiang, Guangxi in 2022 (Wang et al. 2022).

Coptis teeta complex is an important local medicinal plant in China. The C. teeta complex is divided into two subspecies, C. teeta subsp. teeta and C. teeta subsp. lohitensis, based on the presence of stolons and reproduction methods (Pandit and Babu 1993, 2000, 2003). Coptis teeta subsp. teeta is mainly distributed in Changdu and Linzhi of Xizang, China. It used as a medicinal plant by the Monba and Lhoba ethnic groups in southeastern Xizang. C. teeta subsp. lohitensis is mainly distributed in the Gaoligong Mountains in the northwest of Yunnan Province. It was used as a currency equivalent exchange in the past. C. teeta subsp. lohitensis is used in the treatment of diarrhea, dysentery and eye diseases by the Lisu, Dulong and Nu ethnic minorities in Gaoligong Mountain of Yunnan Province (Cheng et al. 2022, 2024).

In March 2023, during the investigation of C. teeta resources in southeast Tibet and northwest Yunnan, we found one population in Yingjiang County of Dehong Dai and Jinpo Autonomous Prefecture, which was different from the previously observed materials of C. teeta complex. The difference was mainly reflected in the fact that leaf segment lobes were contiguous to each other, and lateral segments equal to central one. Plants had no developed stolons. But there was no option to evaluate the morphological features of flowers at that time. In March 2024, we obtained flowers material. It was found that inflorescence consists of 1–3 flowers. The petals have short claws. This was obviously different from C. teeta subsp. teeta and C. teeta subsp. lohitensis characteristics. At the same time, we also collected materials for the molecular studies, and the results of consequent phylogenetic analysis proved that this population is the sister group of C. teeta subsp. teeta and C. teeta subsp. lohitensis.

Methods

Material sampling and DNA extraction

Samples of the new species were collected from Yingjiang County, Dehong Dai and Jinpo Autonomous Prefecture. The plastome sequences of 8 related Coptis species (a total of sixteen accessions) and an outgroup species were obtained from GenBank (http://www.ncbi.nlm.nih.gov). The total genomic DNA was extracted from the fresh leaves using the modified CTAB method (Doyle and Doyle 1987), and libraries were prepared using the TruePrep DNA Library Prep Kit (Vazyme Biotech Co., Ltd, Nanjing, CN). All the DNA and molecular materials were deposited in the herbarium of Minzu University of China (MUC). Sample information is listed in Suppl. material 1: table S1. For principal component analysis (PCA), we measured more than 20 individuals with complete traits. In our examination, we focused on 19 morphological characters, which also encompassed both vegetative and reproductive characteristics (Suppl. material 1: table S2). These characters were chosen based on their relevance in species identification and establishment, as described by Sun and Zhang (1995).

Plastome sequencing and assembly

Genomic paired-end sequencing was conducted using the Illumina Novaseq 6000 platform. The chloroplast genome was assembled and analyzed using the program NOVOPlasty v. 4.3.1 (Dierckxsens et al. 2017). Annotation was performed with CPGView to determine the initial location of the chloroplast genome and the IR region (Liu et al. 2023), with the chloroplast genome of C. teeta (NC 054331) serving as a reference. The annotations were manually checked for errors using Zhou et al. (2021) as reference. The final chloroplast genome of new species was deposited in the NCBI GenBank under accession numbers: PP786562 and PP786563.

Phylogenetic reconstruction

Fifty-six single copy protein-coding genes (PCGs) were extracted from 19 chloroplast sequences using the PhyloSuite v. 1.2.3 software (Zhang et al. 2020; Xiang et al. 2023). They were aligned using the MAFFT v. 7.149b algorithm (Katoh et al. 2019). All these single gene alignments were concatenated to create a document for phylogenetic analyses. The best-fit model was determined using the Akaike information criterion (AIC) in ModelFinder program (Kalyaanamoorthy et al. 2017). To determine its phylogenetic position, a maximum likelihood (ML) tree was constructed by IQ-TREE v. 1.6.10. Bayesian inference (BI) analysis was performed with MrBayes based on 56 PCGs of 8 other Coptis species through PhyloSuite v. 1.2.3 software. Phylogenetic trees were visualized, rooted with Asteropyrum peltatum, and edited using the iTOL v. 5 (Ivica and Peer 2021). R v.4.3.2 was employed for data analysis, the ggplot2 package was used for statistical chart visualization, and the factoextra package was used for presenting the PCA plot charts.

Results

Phylogenetic and morphological analysis

Consensus phylogenetic tree reconstructed by ML and BI analyze based on 56 PCGs of 10 species, with Asteropyrum peltatum as outgroups is represented in the Fig. 1. The topologies of the ML and BI trees were identical with all the branches strongly-supported (ML BS≥90% and BI PP = 1). All the accessions of Coptis formed a monophyletic group with high support. The two samples of the new species (C. austrogaoligongensis C. L. Long & Z. Cheng, sp. nov.) were clustered into one clade and sister to the C. teeta subsp. teeta and C. teeta subsp. lohitensis clade (Fig. 1). Morphological PCA results show that the three species are obviously divided into three clusters, with an interpretation of 39.9% for PCA1 and 23.6% for PCA2. The long distance between the three species indicates obvious differences between them (Fig. 2).

Figure 1.

Consensus phylogenetic tree reconstructed by ML and BI analysis based on 56 protein-coding sequences (CDS) of 10 species, with Asteropyrum peltatum as outgroups. Asterisks near the branches indicate bootstrap support (BS) percentages obtained from maximum likelihood inference and posterior probabilities (PP) obtained from Bayesian analysis (BS/PP). Those nodes with BS≥90%, PP =1.00 were shown with asterisks.

Figure 2.

Morphological principal component analysis (PCA) of three species based on some individuals (20 NJ individuals, 20 YJ individuals, and 27 MT individuals) and 19 morphological traits. MT: C. teeta subsp. teeta, NJ: C. teeta subsp. lohitensis, YJ: C. austrogaoligongensis.

Taxonomic treatment

Coptis austrogaoligongensis C. L. Long & Z. Cheng, sp. nov.

urn:lsid:ipni.org:names:77345372-1

Figs 3, 4 “南高黎贡黄连”(Nan Gao Li Gong Huang Lian)

Type

China, Yunnan Province, Dehong Dai and Jingpo Autonomous Prefecture, Yingjiang County, Zhina Township, 2444 m a.s.l., 25°15'55"N, 98°4'11"E, 20 March 2023, Zhuo Cheng YNHL021 (holotype: KUN!; isotype: KUN!).

Description

Herbs perennial, rhizomes branched, without stolons. Leaves basal, petioles 24–35 cm, glabrous. Leaf blade ovate, 7–12 × 6–12 cm, three-segmented, margin with sparsely upturned spiny hairs; central segment petiolulate (petiole 0.5–1 cm), ovate-rhombic, 6–12 × 3.5–6 cm, four-ten-lobed, lobes remote, ultimate lobes margin acute serrate, apex acute or obtuse. Scapes one to several, erect, longer or shorter than the leaves, 25–40 cm wide, glabrous, sulcate. Inflorescences terminal, often monochasial, three-five-flowered; flowers small, actinomorphic, bisexual; bracts lanceolate, palmately divided. Sepals five, greenish, long ellipsoid or lanceolate, 0.5–0.6 × 0.15–0.2 cm, sparsely puberulous. Petals spatulate, 0.15–0.25 cm long, glabrous, apex rounded to obtuse, 1/4–1/3 as long as sepals. Stamens numerous, glabrous, 2–4 mm long, outer ones slightly shorter than petals. Pistils 8–14, 3–5 mm long; follicles 4.5–9.0 mm long, stipitate; seeds ellipsoid, ca. 2–3 mm long, brown.

Figure 3.

Coptis austrogaoligongensis C. L. Long & Z. Cheng, sp. nov. A species habitat B leaf C plant in florescence stage D whole plant E petiole F margin with sparsely upturned spiny hairs G rhizome and fibrous roots H calyx I petal J, K bract L fruit. Photos by Zhuo Cheng & Jiahua Li.

Distribution and habitat

The only known locality of this taxon is in Zhidong Village, Zhina Township, Yingjiang County, Dehong Dai and Jingpo Autonomous Prefecture, Yunnan Province. The site is located in an open area in a primeval forest dominated by Fagaceae and Magnoliaceae. The observed population is very small, with about 100 plants growing in the bamboo forest along the roadside, accompanied by some pteridophytes. The elevation is 2400–2500 m above sea level.

Figure 4.

Coptis austrogaoligongensis C. L. Long & Z. Cheng, sp. nov. A species habitat B Fruit C Flower D margin with sparsely upturned spiny hairs E calyx F petal G bract H bract. Drawn by Xinchen Qu.

Etymology

The specific epithet is derived from the type locality, Gaoligongshan, Yunnan.

Phenology

The species was observed flowering in February – March and fruiting in April–June.

Conservation status

This species has not been recorded or described so far, and there is only one known site in Yingjiang County, which is relatively unknown to botanists. In addition, C. austrogaoligongensis is very small and has a short flowering period, making it easily overlooked. At the moment, without further biogeographical investigations, we can suggest that this species satisfies the IUCN 3.1 Red List CR (Critically Endangered) Criteria B1ab(ii,iii)+2ab(i,ii,iii) (IUCN 2012), which has an EOO (Extent of occurrence) < 100 km² and AOO (Area of occupancy) < 10 km², it may be classified as “critically endangered” (CR). The distribution site of C. austrogaoligongensis is next to the road, which causes a great risk of human disturbance and extreme weather, such as tourist activities, road building, grazing and landslides. Additionally, regional management in pursuit of economic development is likely to pose a threat through trampling and pollution of soil and water, probably causing negative impacts to the small and fragile habitat.

Taxonomic notes

The new species belongs to Coptis section Chrysocoptis based on the following characters: leaves tri-lobed, leaflets sessile or short petiolate (Cheng et al. 1965; Wang 2001; Wang et al. 2022). There were seven species, one variant and one subspecies belonging to Coptis Section Chrysocoptis in China prior to the discovery of C. austrogaoligongensis. From the perspective of geographical distribution, C. teeta subsp. lohitensis and C. quinquesecta are both distributed in Yunnan, C. teeta subsp. lohitensis is mainly distributed in north Gaoligong Mountains and C. quinquesecta is mainly found in Jinping County, whereas C. austrogaoligongensis is mainly distributed in south Gaoligong Mountains. According to the key to the species of Coptis occurring in China (Wang 2001), the morphology of C. austrogaoligongensis is similar to C. teeta subsp. teeta and C. teeta subsp. lohitensis. However, C. austrogaoligongensis can be clearly distinguished by the following features: leaves with deep pinnate cleavages on whole lobes adjacent to each other, lateral segments equal to central one; plants without developed stolons; inflorescences with only 1–3 flowers; petals have short claws. A comparative summary of the characters that differentiate these three taxa is presented in Table 1.

Table 1.

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Distinguishing features of C. austrogaoligongensis in comparison with C. teeta subsp. lohitensis and C. teeta subsp. teeta.

Characters	C. austrogaoligongensis	C. teeta subsp. lohitensis	C. teeta subsp. teeta
Leaf blade	Ovate, 7–12 × 6–12 cm	Ovate-triangular, 6–12 × 5–9 cm	Ovate-triangular, 12–17.5 ×7.7–14.5cm
Leaves shape	lateral segments equal to central one, the petiole length of the middle lobe is 0.3–0.8 cm; leaves with deep pinnate cleavages on whole lobes adjacent to each other	lateral segments subsessile, shorter than central one, the petiole length of the middle lobe is 1–2.1 cm; obliquely ovate, unequally parted	lateral segments equal to central one, the petiole length of the middle lobe is 1.5–2.4 cm; obliquely ovate, unequally parted
Inflorescences	1–3 flowers	3–5 flowers	3–5 flowers
Petiole length	24–35 cm	8–19 cm	19–31 cm
Scape length	25–40 cm	15–25 cm	20–30 cm
Sepal number	5 or 6	5	5
Sepal shape	long ellipsoid or lanceolate, 0.5–0.6 × 0.15–0.2 cm, sparsely puberulous	elliptic, 0.75–0.8 × 0.25–0.3 cm, glabrous.	long ellipsoid or lanceolate, 0.5–0.6 × 0.2–0.25 cm, sparsely puberulous
Petal length	spatulate, 0.2–0.3 cm, glabrous, apex rounded to obtuse	spatulate, 0.54–0.59 cm, glabrous, apex rounded to obtuse	0.3–0.35 cm, glabrous, apex rounded to obtuse
Petal shape	petal with short claws	petals have long claws	petals have long claws
The length ratio of sepal vs petal	ca. 3 times	ca. 2 times	ca. 2 times
Are there any stolons	No	Yes	No

Additional C. austrogaoligongensis specimens examined

China. Yunnan: Dehong Dai and Jingpo Autonomous Prefecture, Yingjiang County, Zhina Township, 2444 m a.s.l., 25°15'55"N, 98°4'11"E, 20 March 2023, Zhuo Cheng YNHL021, Zhuo Cheng YNHL022, Zhuo Cheng YNHL023, Zhuo Cheng YNHL024, Zhuo Cheng YNHL025, Zhuo Cheng YNHL026 (KUN!).

Specimens of C. teeta subsp. lohitensis examined

China. Yunnan: Lushui County, 29 September 2009, L. Xie 83-0381(KUN); Lushui County, 20 November 2007, H. Li 24283 (PE); Longyang District, 23 April 2014, H.J. Dong et al. 935 (KUN); Fugong County, 15 March 2008, X.H. Jin & T. Zhang 071 (PE); Fugong County, 12 November 2007, H. Li 20256 (PE); Gongshan County, 27 September 1984, Qingzang team 9763 (PE).

Key to the species of Coptis in China

There are eight species and one variant of Coptis distributed in China. An identification key is presented below.

1	Leaves five-sectioned	2
–	Leaves three-sectioned	3
2	Rhizome robust; leaf blade 5.5–14 cm wide, central segment pinnately divided, apex Attenuate	*C. quinquesecta*
–	Rhizome slender; leaf blade 2–6 cm wide, central segment three-lobed, apex acute	*C. quinquefolia*
3	Leaf blade lanceolate to narrowly ovate; lateral segments 3–3.5× shorter than central segment; sepals linear-lanceolate	*C. omeiensis*
–	Leaf blade ovate to ovate-triangular; lateral segments slightly shorter than central segment; sepals lanceolate, elliptic, or narrowly ovate	4
4	Petals spatulate	5
–	Petals lanceolate to linear-lanceolate	8
5	Inflorescences three–five-flowered	6
–	Inflorescences more than five-flowered	*C. huanjiangensis*
6	Sparse lobes of leaf, long petals clawed	7
–	Close lobes of leaf; without stolons; short claws in petals	*C. austrogaoligongensis*
7	stolons developed	C. teeta subsp. lohitensis
–	stolons absent	C. teeta subsp. teeta
8	Leaf segment lobes ± contiguous to each other; stamens ca. 1/2 as long as petals	*C. deltoidea*
–	Leaf segment lobes remote; outer stamens slightly shorter than petals	9
9	Sepals 9–13 mm, ca. 2× as long as petals	C. chinensis var. chinensis
–	Sepals ca. 6.5 mm, slightly longer than petals	C. chinensis var. brevisepala

Acknowledgements

We are very grateful to the local people of Yingjiang County, Yunnan, who guided us to the habitats of Coptis species and continued to send us information in different seasons. Many thanks to Xinlei Zhao, a respected taxonomist and expert of Coptis from the Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, for his confirmation of this new species. We also thank Jun Yang from the Kunming Institute of Botany, Chinese Academy of Sciences for their helpful suggestions.

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This work was supported by grants from Yunnan Province for the Baoshan Administrative of Gaoligongshan National Nature Reserve (GBP-2022-01 and 202305AF150121), the National Natural Science Foundation of China (32370407, 31761143001 and 31870316), and the Minzu University of China (2020MDJC03, 2023GJAQ09 and 2022ZDPY10).

Author contributions

Conceptualization: CL, ZC. Data curation: LZ, ZC. Formal analysis: ZC. Funding acquisition: CL. Investigation: CX, ZC, QG, JL, LZ. Resources: JL. Supervision: CX. Writing - original draft: ZC. Writing - review and editing: CL.

Author ORCIDs

Zhuo Cheng https://orcid.org/0000-0001-7807-2571

Chunlin Long https://orcid.org/0000-0002-6573-6049

Data availability

All of the data that support the findings of this study are available in the main text or Supplementary Information.

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Supplementary material

Supplementary material 1

Supplementary information

Zhuo Cheng, Jiahua Li, Congli Xu, Lixiang Zhang, Qiangbang Gong, Chunlin Long

Data type: xlsx

Explanation note: table S1. Accession numbers of 21 sequenced or downloaded chloroplast genomes in this study. table S2. List of analyzed morphological traits. Data type: table (excel file).

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.

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﻿Abstract