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Nidirana chapaensis (Bourret, 1937), one additional anuran species for the amphibian fauna of China


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The China-Vietnam border region is a well-known biodiversity hotspot, harboring a striking diversity of species and endemism. However, the largest part of this region is so far understudied due to restrictions by political boundaries and difficulties in accessibility. Consequently, many species have only been reported from one side of the political boundary, despite the presence of continuous habitat on both sides. In this study, we present our discovery of the frog Nidirana chapaensis in southern Yunnan Province, China. This species was previously known only from Vietnam. All major morphometric characters of these Chinese specimens were found to match with the Vietnamese specimens. In addition, our mitochondrial phylogeny suggests that the Chinese population is monophyletic with respect to topotypic Nidirana chapaensis, with uncorrected pairwise distances of 0.2% at the COI gene fragment analyzed. Therefore, we report Nidirana chapaensis as a new member of amphibian fauna of China, describe the morphological variation of the Chinese population, and provide additional natural history data of this species based on our observation from China.
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Accepted by M. Vences: 21 Feb. 2019; published: 28 Mar. 2019
ISSN 1175-5326 (print edition)
(online edition)
Copyright © 2019 Magnolia Press
Zootaxa 4571 (4): 580
Nidirana chapaensis (Bourret, 1937), one additional anuran species for the
amphibian fauna of China
Key Laboratory for Conserving Wildlife with Small Populations in Yunnan, Southwest Forestry University, Kunming 650224, Yunnan,
Sam Noble Oklahoma Museum of Natural History & Department of Biology, University of Oklahoma, Norman, Oklahoma, USA
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming
650223, Yunnan, China
College of Life Science and Technology, Honghe University, Mengzi 661199, Yunnan, China
Imaging Biodiversity Expedition Institute, Beijing100107, Beijing, China
These authors contributed equally
Corresponding authors. E-mail:;
The China-Vietnam border region is a well-known biodiversity hotspot, harboring a striking diversity of species and en-
demism. However, the largest part of this region is so far understudied due to restrictions by political boundaries and dif-
ficulties in accessibility. Consequently, many species have only been reported from one side of the political boundary,
despite the presence of continuous habitat on both sides. In this study, we present our discovery of the frog Nidirana cha-
paensis in southern Yunnan Province, China. This species was previously known only from Vietnam. All major morpho-
metric characters of these Chinese specimens were found to match with the Vietnamese specimens. In addition, our
mitochondrial phylogeny suggests that the Chinese population is monophyletic with respect to topotypic Nidirana cha-
paensis, with uncorrected pairwise distances of 0.2% at the COI gene fragment analyzed. Therefore, we report Nidirana
chapaensis as a new member of amphibian fauna of China, describe the morphological variation of the Chinese popula-
tion, and provide additional natural history data of this species based on our observation from China.
Key words: new records, frog, China-Vietnam border, Yunnan Province
As a direct result of increasing attentions and efforts in herpetological researches in China for the past decade, the
Chinese amphibian fauna has grown dramatically, with new records and new species described almost every year
(Murphy 2016). However, many regions of the country, particularly along the southern borders in Tibet, Yunnan,
and Guangxi, have not been surveyed in detail. Among these is the subtropical region along the China-Vietnam
border in Wenshan Prefecture of south-eastern Yunnan Province.
Music Frogs of the genus Nidirana comprise eight recognized species, including Nidirana adenopleura
(Boulenger 1909), N. chapaensis (Bourret 1937), N. daunchina (Chang 1933), N. hainanensis (Fei et al. 2007), N.
lini (Chou 1999), N. nankunensis (Lyu et al. 2017), N. okinavana (Boettger 1895), and N. pleuraden (Boulenger
1904). Most of these taxa are found in southern China (Fei et al. 2009). Among the three species that have not been
recorded in China is N. chapaensis, which occurs in northern Vietnam, Laos and Thailand, close to the southern
border of Yunnan Province. Given the continuous habitat that stretches across political boundaries in this region,
and its under-surveyed nature, it was considered likely that N. chapaensis was also found in the territory of China.
During the herpetological surveys of south-eastern Yunnan Province in 2016, we collected three Nidirana
individuals from a hitherto unknown population in Wenshan Prefecture (Fig. 1). Toes of seventeen additional frogs
were sampled. Following detailed morphological comparisons and phylogenetic analyses, we here confirm that the
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newly discovered Nidirana population in Yunnan should be assigned to N. chapaensis. Therefore, we report N.
chapaensis as a new amphibian record of China, provide descriptions of additional specimens, and provide remarks
on the natural history of the species based on our field observations.
FIGURE 1. Distribution of Nidirana chapaensis near the China-Vietnam border. Red circle (Xichou, Wenshan, Yunnan,
China) represent the new distribution record of the species in China.
Material and methods
Field sampling. Field surveys were conducted at Xiaoqiaogou Nature Reserve (23.362 ºN, 104.839 ºE, 1312 m
asl.), Xichou, Wenshan, Yunnan Province, China in April, June and August of 2016. A total of three specimens
were collected (males: KIZ 028112, KIZ 028136-37). Toes of seventeen additional individuals were clipped and
collected before releasing the living frogs in the locality of capture. The three collected individuals were euthanized
using a chlorobutanol solution, and liver or muscle tissues were taken from the specimens before fixing them in
75% ethanol. All specimens were deposited at the Museum of the Kunming Institute of Zoology (KIZ), Chinese
Academy of Sciences, Kunming, China.
Morphology and morphometrics. Measurements were taken of the three preserved individuals (KIZ028112,
KIZ028136-37) with a digital calliper to the nearest 0.1 mm by Zhiyong Yuan. Morphological characteristics and
measuring methods follow Fei et al. (2007). Abbreviations used are as follows: SVL (snout-vent length); HL(head
length); HW (head width); SNL (snout length); IND (internarial distance); IOD (interorbital distance at narrowest
point); EYE (diameter of the exposed portion of the eyeball); UEW (width of upper eyelid);TMP (horizontal
diameter of tympanum); TEY (tympanum–eye distance); HND (hand length); LAD (diameter of lower arm); FEM
(femur length); TIB (tibia length); TW (tibia width); TFL (length of tarsus and foot); FTL (foot length).
For comparative material, morphological measurements were obtained from recent studies of N. chapaensis,
which frogs from the newly discovered locality resembled (Luu et al. 2013; Ziegler et al. 2014). In addition,
morphological data of congeners was obtained from the literatures as well (Fei et al. 2007; Le et al. 2014; Iwai &
Shoda-Kagaya 2012; Fei et al. 2007).
DNA sequencing and analysis of sequences. To build a phylogeny for the genus Nidirana, we started by
extracting DNA from ethanol-preserved tissues of two individuals originating from the newly discovered locality
in Yunnan Province, and two individuals of N. hainanensis from Hainan Province (Table 1) using standard phenol-
chloroform extraction (Sambrook et al. 1989). A fragment of the mitochondrial gene for cytochrome oxidase
subunit 1 (COI) was amplified by polymerase chain reaction (PCRs). The primers and PCR conditions followed
Che et al. (2012). Then, we downloaded twelve available COI sequences from GenBank for the all the currently
recognized species of Nidirana excepted N. nankunensis, which was described recently and proved to sister with
the clade including N. adenopleura and N. okinavana (Lyu et al. 2017). Odorrana graminea was used as outgroup
based on a current phylogenetic hypothesis (Che et al. 2007). Sequences were checked for ambiguities and aligned
with the default settings (Edgar 2004). The interspecific and within-species mean uncorrected pairwise distances
were calculated in MEGA v6.0.6 (Tamura et al. 2013).
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TABLE 1 . Specimens used in the study, including their voucher numbers, localities, and GenBank accession numbers
for the COI gene.
We reconstructed the phylogeny of the genus Nidirana using both Maximum Likelihood (ML) and Bayesian
Inference (BI) analyses based on the COI sequences. The best-fit partitioning scheme and the best substitution
models for the respective partitions were identified with PartitionFinder v.1.1.1 (Lanfear et al. 2012) using the
Bayesian Information Criterion. A GTR + G model for all the three codon positions was selected as the best mode
and partition which was used in the ML and BI analyses. Maximum Likelihood analysis was conducted with
RAxML version 8.2.4 (Stamatakis 2014). The robustness of nodes of the best ML tree was assessed with 1000
bootstrap replicates. Bayesian Inference was executed in MrBayes v. 3.2.2 (Ronquist et al. 2012). The partitioned
analysis was conducted with two independent runs, each with four Metropolis coupled Markov Chain Monte Carlo
chains of 10 million generations and a burn-in of 25%. Convergence of sampled parameters (effective sampling
size/ESS for all parameters >200) was assessed by Tracer 1.6 (Rambaut et al. 2014).
Results and discussion
Morphological measurements of all the specimens from Yunnan are summarized in Table 2. All of the major
diagnostic characters of the Chinese specimens (KIZ 028112, KIZ 028136-37) match with the data presented in the
original description of N. chapaensis. Measurements of a few additional characters slightly differed from the
Vietnamese specimens (Luu et al. 2013; Ziegler et al. 2014), but nevertheless fell within close range of the latter
(Table 2).
A total of 555 base-pairs of new COI sequences were obtained from four samples obtained during field works
in Yunnan and Hainan (GenBank numbers MK492282–MK492285, Table 1). For all the alignment sequences, 187
variable positions and 163 parsimony informative sites were identified. Interspecific uncorrected pairwise
distances between the new population from Yunnan, China and the other species of Nidirana varied from 4.3%
(between N. hainanensis) to 11.5% (between N. pleuraden) (Table 3). The uncorrected pairwise genetic distance
between the new populations of Nidirana from Yunnan and the topotypic N. chapaensis (Sapa, Lao Cai, Vietnam)
was very small (0.9%). Furthermore, both the ML and BI analyses showed with strong support that the Yunnan
population forms a monophyletic clade with topotypic N. chapaensis (Fig. 2). Thus, based on evidence from both
molecular and morphological data, we conclude that the Nidirana population from Wenshan, Yunnan, China is
conspecific with N. chapaensis, and we report N. chapaensis as a new amphibian species occurring in China.
Species Voucher Locality GenBank Acc. No. Source
Nidirana adenopleura A-A-WZ001 unknown NC018771 Yu et al. 2012
Nidirana chapaensis 2000.485 Sapa, Lao Cai, Viet Nam KR087625 Grosjean et al. 2015
Nidirana chapaensis KIZ028136 Wenshan, Yunnan, China MK492282 This study
Nidirana chapaensis KIZ028137 Wenshan, Yunnan, China MK492283 This study
Nidirana daunchina SYNU12050568 Hangzhou, Zhejiang, China KF020615 Zhou et al. 2015
Nidirana hainanensis Tissue ID:
Diaoluoshan, Hainan, China MK492284 This study
Nidirana hainanensis Tissue ID:
Diaoluoshan, Hainan, China MK492285 This study
Nidirana lini unknown unknown HQ395352 Li et al. 2015
Nidirana okinavana unknown Ryukyu islands, Japan NC022872 Kakehashi et al. 2013
Nidirana pleuraden unknown Longling, Yunnan, China HQ395340 Li et al. 2015
Babina holsti unknown Ryukyu islands, Japan NC022870 Kakehashi et al. 2013
Babina subaspera AB679645 Amami islands, Japan AB679645 Iwai and Shodakagaya 2012
Babina subaspera AB761265 Amami islands, Japan AB761265 Iwai and Shodakagaya 2012
Odorrana graminea 2005.021 Long Nai Khao, Phongsali, Laos KR087839 Grosjean et al. 2015
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TABLE 2. Morphometric comparisons of Nidirana chapaensis populations from Yunnan, China (this study) and
Vietnam (Luu et al. 2013; Ziegler et al. 2014). Abbreviations are provided in the materials and method section. All
measurements are in the unit of millimeter (mm). “-” indicates without data.
TABLE 3. The pairwise uncorrected p-distance (%) of COI gene sequences between species of Nidirana. “-” indicates
genetic distance less than 0.1%.
Taxonomic conclusion
Nidirana chapaensis (Bourret, 1937)
(Figs. 3 and 4)
Description. Males: SVL 43.0–47.3mm; head longer than width (HL14.9–15.4 mm, HW 14.2–14.9 mm); snout
obtusely pointed in dorsal view, projecting beyond lower jaw; canthus rostralis distinct; nostril dorsolaterally
oriented, close to tip of snout; loreal region concave, sloping; tympanum distinct, rounded, smaller than eye
diameter (TMP = 58.2–69.6% EYE); supratympanic fold absent; pineal body distinct; internarial distance
relatively greater than interorbital distance (IOD = 86.4–96.5% IND). Two short rows of vomerine teeth present;
tongue notched posteriorly; a pair of internal vocal sacs present in males. Skin smooth, except small granular
tubercles present on posterior dorsum, near cloacae and hindlimbs; rictal glands distinct; dorsolateral fold distinct.
Catalog No. KIZ028112 KIZ028136 KIZ028137 Males from Quang Binh, Vietnam Male from Ha Giang, Vietnam
(IEBR A.2013.98)
SVL 47.3 46.6 43.0 43.0–46.0 42.1
HL 15.4 15.0 14.9 17.3–20.4 19.7
HW 14.9 14.4 14.2 14.9–18.0 15.2
SNL 7.4 7.5 6.6. 6.9–7.8. 6.9.
IND 5.8 5.9 5.7 5.1–6.3 5.1–6.3
IOD 5.4 5.1 5.5 3.3–4.5 3.3–4.5
UEW 3.3 4.2 3.3 - -
EYE 6.0 5.5 5.6 4.7–5.6 5.7
TMP 3.6 3.2 3.9 3.9–4.2 4.6
TEY 1.1 1.0 1.0 0.3–1.4 -
HND 18.2 18.9 15.9 - -
LAD 3.6 3.8 4.5 - -
FEM 18.3 22.9 19.4 21.4–25.2 -
TIB 21.0 23.8 21.0 24.5–29.5 -
TW 6.9 6.9 6.3 - -
TFL 34.0 35.8 32.2 - -
FTL 24.7 26.5 21.2 24.1–29.2 -
1N. adenopleura -
2N. chapaensis 0.096 -
3N. daunchina 0.047 0.099 -
4N. hainanensis 0.096 0.043 0.095 -
5N. lini 0.099 0.093 0.101 0.101 -
6N. okinavana 0.045 0.094 0.048 0.096 0.101 -
7N. pleuraden 0.106 0.115 0.111 0.122 0.111 0.110 -
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Relative finger length II < I <IV<III; fingers webbing absent; lateral fringes absent; finger tips expanded with
circummarginal grooves without discs; subarticular tubercles distinct; supernumerary tubercle indistinct; three oval
palmar tubercles present. Nuptial pad absent in males. Hind limbs long, tibia longer than thigh (TIB =87.1–96.2%
TFL); toes long and thin, relative lengths I<II<III<V<IV (Fig. 4); webbing present on toes, webbing formula: I 1–2
II 1–2 III 1–2 IV 2–1 V; narrowly lateral fringes distinct on toes; subarticular tubercles distinct; oval inner
metatarsal tubercle distinct; outer metatarsal tubercle indistinct; supernumerary tubercles absent on toes.
Coloration. In life, dorsal coloration of the adults ranges from orange to dark brown, with a narrow white
stripe running from pine body to vent, and a pattern of irregular black spots covering the dorsum. The loreal region
is blackish, and rictal glands are white. The region around tympanum is usually black, but can be creamy yellow
instead. The upper flank is blackish, with spots close to the dorsolateral fold. The remaining parts of flank are
mostly creamy yellow, sometimes with black spots scattered across. The iris is bronze. Dark bars are present on the
dorsal surface of the limbs. Ventral throat and chest are gray, covered densely with black spots; the belly is
yellowish or white. Ventral surfaces of the limbs are creamy-yellow or white with gray spots. The fingers have dark
gray spots; subarticular tubercles on the fingers and toes are pale gray and dark brown respectively, and the inner
metatarsal tubercle is creamy-yellow.
Distribution. The currently known ranges include northern Vietnam, Laos, Thailand, and south-eastern
Yunnan Province of China (Nguyen et al. 2009; Stuart 2005).
Natural history notes. Specimens were collected in small ponds adjacent to a stream (Fig. 5) between 19:00
and 23:30. The surrounding habitat was coniferous mixed forest at an elevation of ca. 1300 m a.s.l. Call of males
were heard at an air temperature of 24°C–28°C and a relative humidity of 53%–85% during April, June and
August, 2016. Many individuals were fund in April, but very few were found in June and August. Other frog
species observed at the same locality included Microhyla heymonsi, Limnonectes bannaensis, Fejervarya
multistriata, Sylvirana maosonensis.
FIGURE 2. Bayesian phylogenetic tree of the genus Nidirana inferred from a fragment of COI gene. “-” denotes low support
by Bayesian posterior probabilities (BPP < 95%), and bootstrap support (BS < 70%). The scale bar represents 0.05 nucleotide
substitutions per site.
On the distribution of herpetofauna across the China-Vietnam border. Our new record presented here
adds to the list of amphibian species which have recently been found to occur on both sides of the China-Vietnam
border, such as Limnonectes bannaensis (Suwannapoom et al. 2016), Paramesotriton deloustali (Zhang et al.
2017), and Tylototriton ziegleri (Jiang et al. 2017). Given that the area surrounding the China-Vietnam border is
located in the same zoogeographic region (Holt et al. 2013), and since habitats on both sides of the border are
Zootaxa 4571 (4) © 2019 Magnolia Press
continuous without any known biogeographic barriers, it is highly possible that species many previously believed
endemic to either side of the border are found on the other side, and their distributions are much wider than
previously believed. Future collaborative research between herpetologists from both countries are needed in order
to obtain a better understanding of the biodiversity in this region.
FIGURE 3. Color variations of Nidirana chapaensis from the Yunnan population in China: (A–D) dorsolateral, dorsal (E), and
ventral (F) views of Nidirana chapaensis in life from China.
We thank Wouter Beukema, Miguel Vences and two anonymous reviewers for their help on English improve and
valuable suggestions during manuscript review. We thank Shaobin Hou, Baolin Zhang, Boyan Li, Jiansong Zhang
and the staff from Xiaoqiaogou nature reserve for helping with sample collections. This work was supported by
grants from the National Natural Science Foundation of China (31702008), Yunnan Applied Basic Research
Project (2018FD047), the First Class Discipline Construction Project for Forestry in Yunnan (No.51600625), and
the Biodiversity Conservation Program of Ministry of Environmental Protection of China to ZYY; Southeast Asia
Biodiversity Research Institute, CAS (Y4ZK111B01: 2017CASSEABRIQG002), China's Biodiversity
Observation Network (Sino-BON), and the Animal Branch of the Germplasm Bank of Wild Species, Chinese
Academy of Sciences (Large Research Infrastructure Funding) to JC.
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FIGURE 4. (A) Dorsal and (B) ventral views of the hand, and (C) dorsal and (D) ventral views of the foot of Nidirana
chapaensis in life from China.
FIGURE 5. (A) Habitat of Nidirana chapaensis in Xichou, Wenshan Prefecture, Yunnan Province, China.
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... Furthermore, our study further highlights a lack of existing knowledge on amphibian taxonomy and identifies an underestimation of the potential biodiversity along these national border areas. In recent years, many new recorded species have been found in border areas, such as Nidirana chapaensis ( Yuan et al. 2019), Gracixalus quangi (Lorphengsy et al. 2021) and Theloderma pyaukkya (Du et al. 2020), while a number of studies have called for enhanced surveys of species diversity in these border areas , Chen et al. 2018, Wu et al. 2020, Yuan et al. 2019. The findings of our research further support this view. ...
... Furthermore, our study further highlights a lack of existing knowledge on amphibian taxonomy and identifies an underestimation of the potential biodiversity along these national border areas. In recent years, many new recorded species have been found in border areas, such as Nidirana chapaensis ( Yuan et al. 2019), Gracixalus quangi (Lorphengsy et al. 2021) and Theloderma pyaukkya (Du et al. 2020), while a number of studies have called for enhanced surveys of species diversity in these border areas , Chen et al. 2018, Wu et al. 2020, Yuan et al. 2019. The findings of our research further support this view. ...
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Thailand is considered a global biodiversity hotspot that is known to harbour a striking diversity of endemic species. However, several research studies have determined that the level of amphibian diversity in the country has been significantly underestimated. The megophryid genus Leptobrachella Smith, 1925 is currently known to include 89 species that are primarily distributed throughout southern China and Southeast Asia; however, only seven species have been found in Thailand. Based on an integrative approach encompassing genetic and morphological analyses, we have concluded that the population identified from Chiang Rai Province of Thailand is conspecific with Leptobrachella ventripunctata (Fei, Ye, and Li, 1990). Importantly, this is the first confirmation record of this species, based on molecular and morphological evidence in Thailand. The discovery of this species reaffirms that the diversity within the genus has been underestimated with many species yet to be discovered. In addition, the findings of our study further highlight the lack of existing knowledge on amphibian taxonomy and an underestimation of the biodiversity that exists along these national border areas.
... The karstic ha bitats of southwestern China represent the world's largest karst habitat system, and harbor an extraordinary diverse endemic flora and vertebrate fauna, which are in danger to anthropogenic threats (National Bureau Statistics of China, 2011;Luo et al., 2016). Fieldwork in these areas since 2015 has discovered eight amphibians and reptiles as new species or new locality records (Böhme, 2003;Nicodemo and Bain, 2007;Wang et al., 2015;Chen et al., 2018;Ren et al., 2018;Wang et al., 2018;Zhang et al., 2018;Yuan et al., 2019). Although these recent discoveries provide a greater understanding of the reptilian biodiversity in this region, it also indicates that there is likely much biodiversity that has yet to be discovered. ...
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Cyrtodactylus geckos are one of the most speciose and diverse groups of extant lizards known, distributed throughout the Asian and Pacific realms. Using molecular phylogenetic methods and supporting morphological data, we describe a new species of Cyrtodactylus in Daweishan National Nature Reserve, Yunnan Province, China. Cyrtodactylus hekouensis sp. nov. can be morphologically distinguished from its nearby congeners by the following characters: maximum SVL 92.3 mm and TL 98.5 mm; 11-12 supralabials; 11-12 infralabials; 36-57 scale rows between the fifth supralabials; 10-13 dorsal tubercles rows; 3 postnasals on blunt and smooth front snout; precloacal-femoral pores in a continuous series of 33-39 (females with pitted scales) located under vent/cloaca and thighs in both sexes; precloacal groove absent; 3/3 postcloacal tubercles; subdigital lamellae under the fourth finger 21 or 22, under the fourth toe 20-23; smooth midbody with smooth venter and tuberculate dorsal scale rows, tubercles from head to tail base; dorsal transverse patterns are generally large, bilaterally symmetrical. The results of the phylogenetic analysis recover specimens of this new species as sister to a clade containing C. wayakonei and C. martini. Uncorrected pairwise intraspecific distances were < 1%, and distances between our new species and other Cyrtodactylus species from nearby countries ranged from 14.2% to 26.8%.
... ;Nguyen S. V. et al., 2009;Chuaynkern et al., 2010;Luu et al., 2013;Oliver et al., 2015;Lyu et al., 2017;Yuan et al., 2019]. ...
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The Indochinese Peninsula is recognized as one of the key global biodiversity hotspots. The amphibian fauna of Indochina (including Vietnam, Laos, Cambodia and Thailand) is one of the richest in the world. About 60% of the known species were described within the last 20 years. We review the literature and our data to assess all recent discoveries and taxonomic changes and compile the first annotated checklist of the amphibian fauna of Indochina since the middle of the 20th century, including updated faunal lists for Vietnam and Thailand. Amphibian checklists for Laos and Cambodia are published for the first time. For each species we provide the following information: scientific name; recommended common name in English; information on type specimens; type locality; data on its distribution within Indochina and beyond; IUCN conservation status; taxonomic comment and the most important references. We review the distribution of each species across the 23 biogeographic subregions of Indochina, estimate the similarity among the regional faunas and evaluate their species richness and endemism. In total we record 423 amphibian species belonging to three orders, 11 families and 71 genera; 199 species (47%) are endemic to Indochina. Comprising 270 known species, the amphibian fauna of Vietnam is the richest (98 endemics, 36.3%), followed by Thailand with 194 species (29 endemics, 14.9%), Laos with 121 species (13 endemics, 10.7%), and Cambodia with 72 species (6 endemics, 8.3%). A cluster analysis of faunal similarity between the subregions shows two major assemblages, divided by the Isthmus of Kra. Within the northern mainland portion of Indochina three clusters can be distinguished: (1) northeastern and northwestern uplands of Vietnam and northern Laos; (2) Northern, Central, and Southern Annamites, the Bolaven Plateau, and central-south Vietnam lowlands; (3) western Indochinese subregions. We identify the Northeast and Northwest Uplands of Vietnam, the Northern, Central and Southern Annamites, the Cardamom Mountains, the mountains of Northeast Thailand, Northern Tenasserim and southern Peninsular Thailand as nine major centers of diversity and endemism of Indochinese amphibians. The analysis of amphibian distribution patterns across Indochina suggests the presence of 14 chorotypes grouped in five major range types. Our results underline the role of Indochina as a key area for amphibian diversity and conservation. Among 423 species of Indochinese amphibians, 152 species (35.9%) were considered as data deficient (DD) or were not evaluated (NE) according to the IUCN Red List criteria; while 76 species (18.0%) were considered vulnerable (VU), endangered (EN) or critically endangered (CR), 20 species (4.7%) were considered to be near threatened (NT), and 175 species (41.4%) to be of the least concern (LC). Our study thus has implications for further conservation efforts on regional and global levels, as well as for understanding the biogeographic patterns of amphibian richness and endemism in Asia.
... A clear need therefore exists to conduct collaborative herpetological surveys involving both countries to obtain a better understanding of the biodiversity in this region. Similar efforts in the border area between China and Vietnam have previously demonstrated the value of such international surveys Yuan et al. 2019a). ...
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The Chinese-Myanmar border area forms part of a long-acknowledged biodiversity hotspot. This region is characterised by dramatic topography and diverse landscapes, which support a high degree of biodiversity and endemism that remains largely understudied. Based on recent survey efforts we here describe a new frog species of the genus Odorrana from this area. Found in Dulongjiang village, Yunnan, China, the new species differs from a morphological perspective from its congeners through the combination of a grass-green dorsum with black posterior spots, absence of dorsolateral folds, a distinct pineal body, presence of supratympanic folds, absence of outer metacarpal, metatarsal and supernumerary tubercles, and a pair of external subgular vocal sacs. From a molecular perspective, pairwise genetic distances of 4.8% to 11.6% for DNA sequences of the mitochondrial gene for 16S ribosomal RNA differentiate the new species from other Odorrana. Our mitochondrial phylogeny suggests that the new species is a deeply divergent genetic lineage clustering with the clade including Odorrana wuchuanensis, and the Odorrana andersonii and Odorrana margaretae groups. The discovery of this new species emphasizes the need for further herpetological studies in the China-Myanmar border region.
... This area forms the border of several countries, including Afghanistan, Bangladesh, Bhutan, China, India, Myanmar, Nepal, and Pakistan. Our findings highlight the importance of international cooperation to facilitate extensive sampling across political boundaries, in the interest of understanding biodiversity (e.g., Chen et al., 2017Chen et al., , 2018Suwannapoom et al., 2016;Yuan et al., 2019a). In addition, our analyses reduce the range sizes of some species. ...
The genus Amolops (“torrent frogs”) is one of the most species-rich genera in Ranidae, with 59 recognized species. This genus currently includes six species groups diagnosed mainly by morphology. Several recent molecular studies indicated that the classification of species groups within Amolops remains controversial, and key nodes in the phylogeny have been inadequately resolved. In addition, the diversity of Amolops remains poorly understood, especially for those from incompletely sampled regions. Herein, we investigate species-level diversity within the genus Amolops throughout southern China and Southeast Asia, and infer evolutionary relationships among the species using mtDNA data (16S, COI, and ND2). Molecular analyses indicate nine unnamed species, mostly distributed in the Himalayas. We then utilized anchored hybrid enrichment to generate a dataset representing the major mitochondrial lineages to resolve phylogenetic relationships, biogeography, and pattern of species diversification. Our resulting phylogeny strongly supports the monophyly of four previously identified species groups (the A. ricketti, A. daiyunensis, A. hainanensis, and A. monticola groups), but paraphyly for the A. mantzorum and A. marmoratus groups, as previously defined. We erect one new species group, the A. viridimaculatus group, and recognize Dubois (1992) ‘subgenus’ Amo as the A. larutensis species group. Biogeographic analysis suggests that Amolops originated on the Indo-Burma/Thai-Malay Peninsula at the Eocene/Oligocene boundary, and dispersed outward, exemplifying a common pattern observed for the origin of Asian biodiversity. The early divergence within Amolops coincides with the Himalayas uplift and the lateral extrusion of Indochina at the Oligocene/Miocene boundary. Our results show that paleoclimatic and geomorphological events have profoundly influenced the patterns of lineage diversification within Amolops.
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To date, 10 species of the genus Microhyla have been recorded in China, of which six were distributed in Yunnan Province. Microhyla hmongorum Hoang, Nguyen, Phan, Pham, Ninh, Wang, Jiang, Ziegler, and Nguyen, 2022 was also speculated to be distributed in Xishuangbana, Yunnan Province, China. However, there is no evidence of documentation of M. hmongorum . We report the first country record of Microhyla hmongorum , based on specimens collected from Yunnan border region. Morphologically, the specimen was consistent with the original descriptions of M. hmongorum . Phylogenetically, the sequences of the specimens from China clustered with the sequence of type specimens of M. hmongorum from Vietnam, with uncorrected pairwise distances of 0.9% at the 16S gene fragment analysed. Therefore, we report M. hmongorum as a new record species in China.
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Nine species of amphibians (Leptobrachella namdongensis, Amolops cremnobatus, Nidirana chapaensis, Odorrana graminea, Odorrana tiannanensis, Chirixalus doriae, Raorchestes parvulus, Rohanixalus vittatus, Paramesotriton deloustali) are reported for the first time from Son La Province, Vietnam. Our findings bring the species number of amphibians known from this province to 79. In addition, we provide morphological data and ecological notes for the aforementioned species.
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Amolops splendissimus Orlov and Ho, 2007 and A. caelumnoctis Rao and Wilkinson, 2007 were described almost simultaneously from either side of the China-Vietnam border. The two species share a strong morphological resemblance, and their taxonomic distinctiveness has been questioned, yet no one has confirmed the taxonomic relationship and status between the two taxa. To resolve this taxonomic issue, we collected additional topotypic and near-topotypic specimens of A. splendissimus and A. caelumnoctis from both China (A. caelumnoctis: Wenshan County, Yunnan Province; type locality Luchun County, Yunnan Province), and Vietnam (A. splendissimus: Tam Duong District, Lai Chau Province; type locality Mount Ky Quan San, Bat Xat, Lao Cai Province). Molecular analysis based on a 16S rRNA fragment revealed minimal genetic divergences between the two taxa (0.0%-0.4% uncorrected p-distance), and both species are closely related to A. viridimaculatus (2.1%-2.3%) and A. medogensis (3.5%-3.7%). Morphological comparisons between the newly collected specimens and the original descriptions of both species further support the lack of distinctiveness of the two species, hence, we conclude that A. caelumnoctis is a junior synonym of A. splendissimus.
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We updated the checklists of extant, native amphibians and reptiles of China based on the previously published checklist of reptiles in 2015, the online checklist of amphibians on the database AmphibiaChina, newly published data as of December 2019, and previously uncollected literature prior to 2015. In total, the amphibian fauna of China consists of 515 species in 62 genera, 13 families, and three orders (Anura: 431 species in 47 genera and nine families; Caudata: 82 species in 14 genera and four families; Gymnophiona: one species in one genus and one family), while the reptilian fauna of China consists of 511 species in 135 genera, 35 families, and three orders (Crocodylia: one species in one genus and one family; Testudines: 34 species in 18 genera and six families; Squamata 466 species in 116 genera and 28 families [Serpentes: 256 species in 73 genera, 18 families; Lacertilia: 211 species in 43 genera and 10 families]). Specifically, for amphibians between 2015 and 2019, one family was recorded from China for the first time, two new genera were described, a genus was resurrected, a genus was recorded from China for the first time, 74 new, valid species were either described or resurrected, 18 recognized species were recorded from China for the first time, and six genera and eight species were considered as junior synonyms. For reptiles between 2015 and 2019, five subfamilies were elevated to the full family status, one new subfamily and a new genus were described, three genera were resurrected, three recognized genera were recorded from China for the first time, 35 new species were described, two species were resurrected from synonyms, six subspecies were elevated to the full species status, 10 recognized species were recorded from China for the first time, four genera and four species were considered as junior synonyms, and distribution records of one genus and four recognized species were removed from China. Furthermore, by reviewing literature before 2015, we make additional changes on the previous reptile checklist, including adding new records of three genera, elevating three subspecies to full species status, adding new records of three recognized species, synonymizing three genera and two species as junior synonyms, and removing the distribution record of a single recognized species from China. Lastly, we revise the Chinese common names of some reptilian groups with recomanda-tions to maintain the stability of the Chinese common names. The number of new species and new national records for amphibians and reptiles between 2015 and 2019 in China accounts for 17.1% and 10.2% of the total number of species in each group, respectively. Because new species are described at considerable speed and given the constant changes in the taxonomy of China’s herpetofuna, it is crucial to update the checklists regularly and discuss the existing taxonomic problems, so that such information reflects the most current state of knowledge and are available for taxonomic researchers and conservation biologists alike.
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The taxonomy of Babina sensu lato was controversial in the past decades. In this study, the phylogeny of genus Babina sensu lato was reconstructed based on genetic analysis, morphological comparison and advertisement call analysis. We found that Babina sensu stricto and previous subgenus Nidirana should be two distinct genera in the family Ranidae. N. caldwelli is confirmed to be a synonym of N. adenopleura because of the small genetic divergence and the lack of distinct morphological differences. A new species, Nidirana nankunensis sp. nov. is described based on a series of specimens collected from Mt. Nankun, Guangdong Province, China, which can be distinguished from other known congeners by having a behavior of nest construction, distinctive advertisement calls, significant divergence in the mitochondrial genes, and a combination of morphological characters. Currently, the genus Babina contains two species and the genus Nidirana contains eight species.
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The Vietnam Warty Newt (Paramesotriton deloustali) is threatened by habitat loss and illegal capture for traditional medicines use and domestic pet trade. In this study, we described the complete mitochondrial genome (mitogenome) of P. deloustali. The entire mitogenome sequence of P. deloustali is 16,391 bp long, containing 13 protein coding genes, two ribosomal RNA, 22 transfer RNA genes, one control region and an additional 261 bp non-coding sequence between tRNAThr and tRNAPro genes. Our new data of the complete mitogenome of P. deloustali will offer solid genetic information for further conservation genetics research of this threatened species.
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Tylototriton ziegleri is distributed in northern Vietnam and is a new member of the genus Tylototriton. The conservation status of this species has not yet been assessed by the IUCN, but several other species of Tylototriton have been listed as near threatened or vulnerable due to habitat destruction and illegal hunting. We determined the complete sequence and structural characteristics of the mitochondrial genome of T. ziegleri. The entire mitogenome is 16,266 bp in length, and contains 13 protein-coding genes, 22 tRNA genes, two rRNA genes, a control region, and a non-coding region. The length, gene arrangement, and composition are similar to mitogenomes of other species of Tylototriton. A phylogenetic tree based on the mitogenome sequences of several Asian species within Salamandridae confirmed the genus Tylototriton as monophyletic, with T. ziegleri being the sister species of T. asperrimus and both constituting the sister group of T. wenxianensis. This work provides basic molecular data which are critical for further genetic research and conservation of the genus Tylototriton.
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Phylogenetic reconstructions derived from DNA sequence data play a central role in documenting the number of species in a complex. Such analyses are pointing to the existence of many cryptic species, especially in poorly understood groups such as the genus Limnonectes, and the L. kuhlii species complex in particular. To understand the Limnonectes frogs of China, we reconstruct the major matrilineal genealogy of Limnonectes from China and Southeast Asia based on 12S rRNA, tRNAVal and 16S rRNA gene sequences. Based on new data we recognize five species of Limnonectes in China including L. bannaensis, L. fujianensis, L. fragilis, L. taylori (new record), and a new species from southern China and Myanmar. Phylogenetically, the new species is more closely related to the clade comprising L. taylori, L. megastomias, L. isanensis, L. nguyenorum, and L. jarujini from Thailand than to other Chinese species. This study supports previous findings of sympatric members of a species complex that are not each other's closest relatives.
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A new species of the genus Rana (subgenus 'Nidirana'; Ranidae) is described from Yunnan, China. Rana lini is distinguished from the other species of 'Nidirana' by a combination of relatively large size in females, dilated tips of digits, prominent vocal sacs, extensive toe webbing, dermal folds on inner margins of 2nd and 3rd fingers wider than those of outer margins, and a distinct dermal flap along the lateral edge of the 5th toe. The new species is currently known from the subtropical forests of southern Yunnan, China, at an elevation of 1400-1650 m. A key to the species of the subgenus 'Nidirana' is provided.
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Two ranid species, Babina lini and Hylarana menglaensis, are recorded for the first time from Vietnam on the basis of a new amphibian collection from Dien Bien and Son La provinces. In addition, acoustic analyses of these species are also provided based on the advertisement calls recorded in Muong Nhe Nature Reserve of Dien Bien Province.
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We investigate the relationship between Rana coreana and Rana kunyuensis using morphological and molecular data. Morphological comparisons showed these two species to be identical in terms of body measurements and qualitative characteristics. Phylogenetic analyses found that R. kunyuensis and R. coreana are nested within a single clade and exhibited low divergence across several mitochondrial and nuclear genes. All evidence indicates that R. kunyuensis and R. coreana are the same species, making R. kunyuensis a junior synonym of R. coreana. This study stresses the importance of comprehensive taxon sampling, especially in neighboring countries.
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Amphibian populations are dramatically declining, while their inventory is far from being achieved. Tadpoles are usually overlooked from biodiversity survey, whereas their consideration will optimize species counts and knowledge of their ecological and developmental requirements is essential in conservation planning. Two mitochondrial markers, 16S (397 new sequences obtained) and COI (343 new sequences obtained), are used to test DNA barcoding on a set of larval and adult Asian amphibians represented by 83 recognized species from 65 sites. The advantages and drawbacks of each marker are assessed, COI barcoding being advocated for global DNA barcoding, whereas 16S suits for taxonomically or geographically restricted DNA barcoding. About half of the collected tadpoles were badly identified or incompletely named in the field. All tadpole sequences (except one case of probable introgressive hybridization) were correctly assigned to their respective species. Finally six clusters of tadpole sequences without conspecific adults were revealed, stressing the importance of collecting and taking into account tadpoles in biodiversity survey and conservation planning. Copyright © 2015 Académie des sciences. Published by Elsevier SAS. All rights reserved.