2 urn:lsid:arphahub.com:pub:45048D35-BB1D-5CE8-9668-537E44BD4C7E urn:lsid:zoobank.org:pub:91BD42D4-90F1-4B45-9350-EEF175B1727A ZooKeys ZK 1313-2989 1313-2970 Pensoft Publishers 10.3897/zookeys.1208.126092 126092 Research Article Salamandridae Taxonomy Asia A new species of the newt genus Hypselotriton (Amphibia, Urodela, Salamandridae) from Jiangxi Province, southeastern China Jiang Zhihao https://orcid.org/0009-0004-1492-0665 1 Writing - original draft Writing - review and editing Formal analysis Investigation Huang Song snakeman@sinoophis.com https://orcid.org/0000-0001-6786-8523 1 Writing - review and editing Fan Qiangyong https://orcid.org/0009-0002-5515-2143 2 Funding acquisition Cheng Lin https://orcid.org/0009-0004-4674-4213 2 Funding acquisition Gong Yanan https://orcid.org/0000-0002-8155-7024 1 Funding acquisition Cui Zhangbo https://orcid.org/0009-0000-1501-0402 1 Investigation Zhang Tierui https://orcid.org/0009-0000-1869-9313 1 Investigation Lan Wenjun https://orcid.org/0009-0005-0690-1082 2 Formal analysis Zou Zhian https://orcid.org/0009-0001-2556-2590 2 Formal analysis Huang Xuanzhi https://orcid.org/0009-0008-2873-3640 1 Formal analysis Raffaëlli Jean https://orcid.org/0000-0003-0567-3661 3 Writing - review and editing Chen Jinmin chenjinminkiz@126.com https://orcid.org/0000-0001-6432-7721 1 Writing - review and editing The Anhui Provincial Key Laboratory of Biodiversity Conservation and Ecological Security in the Yangtze River Basin, College of Life Sciences, Anhui Normal University, Wuhu 241000, China Anhui Normal University Wuhu China Jiangxi Wuyishan National Nature Reserve Administration Bureau, Wuyishan National Nature Reserve, Shangrao, Jiangxi, China Wuyishan National Nature Reserve Shangrao China Penclen, Plumelec, 56420, France Unaffiliated Plumelec France

Corresponding authors: Song Huang (snakeman@ahnu.edu.cn);Jinmin Chen (chenjinminkiz@126.com)

Academic editor: Anthony Herrel

 2024 05 08 2024 1208 331 346 8483E148-B973-5124-AA61-7DB5B851A079 9B405A9D-B91D-43D3-89EE-CDC597A873EC 04 05 2024 02 07 2024 Zhihao Jiang, Song Huang, Qiangyong Fan, Lin Cheng, Yanan Gong, Zhangbo Cui, Tierui Zhang, Wenjun Lan, Zhian Zou, Xuanzhi Huang, Jean Raffaëlli, Jinmin Chen 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. http://zoobank.org/9B405A9D-B91D-43D3-89EE-CDC597A873EC

A new newt species, Hypselotriton huanggangensissp. nov., is described based on nine specimens collected from Huanggangshan Mountains, Yanshan County, Jiangxi, China. Morphologically, the new species is characterized by the combination of nine external characters: (1) obvious black patches with clear boundaries on the whole body; (2) ground color of the dorsal body tan; (3) ground color of venter bright orange; (4) skin rough; (5) vertebral ridge weak; (6) fingers and toes overlapping when forelimb and hindlimb adpressed towards each other along body; (7) postocular orange spot absent; (8) small white warty glands around the eye; (9) two discontinuous longitudinal lines formed by white warty glands from neck to lateral parts of tail. Molecularly, the new species forms an independent clade with strong support in the phylogenetic trees of the genus based on the mitochondrial locus of NADH dehydrogenase subunit 2 (ND2) gene fragments. The new species distinctly differs from H. fudingensis by differences in its body measurements, vertebral ridge, dorsal black patches, and ventral black patches. Furthermore, the new species and H. fudingensis are geographically isolated by a series of high mountain ranges, including the Wuyishan and Jiufengshan Mountains. The number of Hypselotriton species is now 11.

 Black patches fire-bellied newts geographical isolation morphology phylogenetics taxonomy Doctoral Research Starting Foundation of Anhui Normal University (752017), National Natural Science Foundation of China (NSFC 31471968; 32001222; 31900323). Citation

Jiang Z, Huang S, Fan Q, Cheng L, Gong Y, Cui Z, Zhang T, Lan W, Zou Z, Huang X, Raffaëlli J, Chen J (2024) A new species of the newt genus Hypselotriton (Amphibia, Urodela, Salamandridae) from Jiangxi Province, southeastern China. ZooKeys 1208: 331–346. https://doi.org/10.3897/zookeys.1208.126092

 Introduction

The newt genus Hypselotriton Wolterstorff, 1934 (Urodela, Salamandridae) is distributed in China, including Anhui, Jiangsu, Zhejiang, Fujian, Jiangxi, Guangdong, Hubei, Henan, Hunan, Yunnan, and Guizhou Provinces (AmphibiaChina 2024; AmphibiaWeb 2024; Frost 2024). The generic classifications between Hypselotriton and Cynops has a lengthy history of taxonomic debate (Tschudi and Johann 1838; Wolterstorff 1934; Chang 1935; Zhao and Hu 1984; Zhao et al. 1988; Chan et al. 2001; Weisrock et al. 2006; Zhang et al. 2008; Dubois and Raffaëlli 2011, 2012; Raffaëlli 2013, 2022; Tominaga et al. 2013; Fei and Ye 2016). Previously, some arrangements suggested that Hypselotriton is a junior synonym of Cynops (Ye et al. 1993; Wu et al. 2010a; Fei and Ye 2016). However, recent phylogenetic studies have presented evidence that Cynops sensu lato (including Hypselotriton) is paraphyletic with respect to Pachytriton and Paramesotriton (Rancilhac et al. 2021; Zhong et al. 2021; Yuan et al. 2022). Following the latest taxonomic arrangements (Dubois and Raffaëlli 2009; Dubois et al. 2021; Frost 2024) and the premise of monophyly, the genus Cynops Tschudi, 1838 is restricted to the Japanese species and all Chinese species are placed in the genus Hypselotriton.

Currently, the following 10 species of Hypselotriton have been recorded: H. cyanurus Liu, Hu & Yang, 1962; H. yunnanensis Yang, 1983; H. chenggongensis Kou & Xing, 1983; H. wolterstorffi Boulenger, 1905; H. orientalis David, 1873; H. orphicus Risch, 1983; H. fudingensis Wu, Wang, Jiang & Hanken, 2010; H. maguae Lyu, Qi & Wang, 2023; H. jiaoren Lyu, Qi & Wang, 2023; H. glaucus Yuan, Jiang, Ding, Zhang & Che, 2013. Recent studies suggest that overall species richness of Hypselotriton is underestimated (Yuan et al. 2022; Lyu et al. 2023). The reevaluations of the “widespread” species (H. orientalis and H. yunnanensis) and the survey of unexplored areas are likely to reveal overlooked diversity.

Huanggangshan Mountains (about 10 km long) is located on the northwestern side of the Wuyishan Mountains (about 550 km long), China. The highest peak (2161 m a.s.l) of Huanggangshan Mountains is known as the “roof of Eastern Mainland China” and “the first peak in the southeast of the mainland” (Lin and Ye 1985). During the recent surveys, on the northwestern side of the Huanggangshan Mountains, nine fire-bellied newts of unidentified Hypselotriton population were sampled in a small waterhole. After examination, they were found to differ from other congeneric members in both morphological and molecular characteristics. As a result, we herein describe it as a new species of Hypselotriton.

 Materials and methods Sampling

Nine specimens were collected in a small waterhole (28.15°N, 117.53°E; elevation 84 m) from Huanggangshan Mountains, Yanshan County, Shangrao City, Jiangxi (Fig. 1). The tiptoes of the specimens (the first toe of each specimen) were cut off and immediately preserved in 75% ethanol. These samples were then used for DNA analysis. After identifying that it is a new species, all fire-bellied newts were humanly euthanized by the injection of 0.7% tricaine methanesulfonate (MS222) solution (Yang et al. 2023), and fresh liver tissue was extracted and immediately preserved in 95% ethanol. The specimens were fixed in 10% formalin for one day, subsequently preserved in 75% ethanol and deposited in Anhui Normal University Museum (voucher numbers: HSA23097–23103, HSA23075–23076). Collections of all animals used for this present study obey the Wildlife Protection Act of China, following the guidelines and regulations approved by the internal review board of AHNU (approval no. AHNU-ET2023110), and with the permissions of local government authorities.

10.3897/zookeys.1208.126092.figure1 8D4C659D-1FB1-57DD-AD53-563E5A979572

Geographic distribution of Hypselotriton huanggangensis sp. nov. (red star) and H. fudingensis (green triangles) in southeastern China. They are separated by the Wuyishan and Jiufengshan Mountains.

https://binary.pensoft.net/fig/1108189 Morphological examination

External measurements were made for the seven specimens of H. huanggangensis sp. nov. and nine specimens of H. fudingensis with digital calipers to the nearest 0.1 mm. Only adult specimens were measured (Lyu et al. 2023). These 14 measurements are as follows: total length (TOL) from tip of snout to tip of tail; snout–vent length (SVL) from tip of snout to posterior edge of vent; tail length (TAL) from posterior edge of vent to tip of tail; maximum tail depth (TAD); head length (HL) from tip of snout to the posterior edge of the parotoid gland; maximum head width (HW); snout length (SL) from tip of snout to the anterior corner of eye; eye diameter (ED) from the anterior corner to the posterior corner of the eye; interorbital distance (IOD) between the anterior corner of each eye; eye–nostril length (EN) from the anterior corner of the eye to the nostril; internasal distance (IND) between the external nares; axilla–groin length (AG) between the axilla and the groin along the body; forelimb length (FLL) from elbow to tip of finger III; and hindlimb length (HLL) from knee to tip of toe III.

Statistical analyses on the morphometric measurements were performed in IBM SPSS Statistics 27.0. Males and females were analyzed separately, due to obvious sexual size dimorphism (Fei et al. 1990, 2006, 2012; Fei and Ye 2016). All measurements were made to normalize and reduce the variance (most P values > 0.05 in the Levene’s test). Univariate Analysis of Covariance (ANCOVA) with SVL as the covariate were used to test for differences between H. huanggangensis sp. nov. and H. fudingensis (Lai & Lue, 2008).

 Molecular phylogeny

Total genomic DNA was extracted from ethanol-preserved liver tissues, using the Qiagen DNEasy blood and tissue extraction kit (Qiagen Inc., Valencia, CA, USA). The phylogenetic relationships within Hypselotriton were derived from an analysis of the mtDNA fragment that codes for subunit two of NADH dehydrogenase (ND2) and its flanking tRNAs. A 1026-bp fragment was amplified using primers KIZL4437 (Yuan et al. 2011) and 5081R (Wu et al. 2010b). Two internal primers ND2–38R (5′–TATTCAYCCTAARTGTGCR–3′) and 4416F (Wu et al. 2010b) were applied for sequencing (Yuan et al. 2013). Standard polymerase chain reactions (PCR) were performed in a final volume of 15 ul with the following procedures: initial denaturation at 94 °C for 5 min, 35 amplification cycles at 94 °C for 1 min, annealing for 1 min at 52 °C, extension for 1 min at 72 °C. Final extension at 72 °C was conducted for 10 min. The successfully amplified products were purified using ExoSAP-IT purification kit according to the manufacturer’s instruction (Yuan et al. 2013). Purified PCR products were directly sequenced in both directions using a BigDye Terminator Cycle Sequencing Kit (v. 2.0, Applied Biosystems, Foster City, California, USA) and an ABI PRISM 3730 automated DNA sequencer (Yuan et al. 2013). For the phylogenetic analyses, 38 sequences from additional Chinese Hypselotriton congeners and two sequences of outgroup species of the genera Pachytriton Boulenger, 1878 and Paramesotriton Chang, 1935, were obtained from GenBank and incorporated into our dataset. Detailed information is provided in Table 1 (ID means the ordinal numbers of the species). DNA sequences were aligned using MEGA v. 6.0.6 (Kumar et al. 2018) with default parameters and manually checked.

Localities, voucher information (Holotype: HSA23097), and GenBank accession numbers for all samples of ND2 used in this study.

ID Species Localities Voucher ND2
1 Hypselotriton huanggangensis sp. nov. China: Jiangxi: Shangrao: Yanshan HSA23075 PP590780
2 Hypselotriton huanggangensis sp. nov. China: Jiangxi: Shangrao: Yanshan HSA23076 PP590788
3 Hypselotriton huanggangensis sp. nov. China: Jiangxi: Shangrao: Yanshan HSA23097 PP590781
4 Hypselotriton huanggangensis sp. nov. China: Jiangxi: Shangrao: Yanshan HSA23098 PP590782
5 Hypselotriton huanggangensis sp. nov. China: Jiangxi: Shangrao: Yanshan HSA23099 PP590783
6 Hypselotriton huanggangensis sp. nov. China: Jiangxi: Shangrao: Yanshan HSA23100 PP590784
7 Hypselotriton huanggangensis sp. nov. China: Jiangxi: Shangrao: Yanshan HSA23101 PP590785
8 Hypselotriton huanggangensis sp. nov. China: Jiangxi: Shangrao: Yanshan HSA23102 PP590786
9 Hypselotriton huanggangensis sp. nov. China: Jiangxi: Shangrao: Yanshan HSA23103 PP590787
10 Hypselotriton orientalis China: Anhui: Huangshan: Furonggu SYS a002711 OQ116690
11 Hypselotriton orientalis China: Anhui: Huoshan: Shangtushi KIZ 021844 ON793742
12 Hypselotriton orientalis China: Anhui: Xiuning: Dafu KIZ 021962 ON793737
13 Hypselotriton orientalis China: Henan: Xinyang: Mt Jigong KIZ 013021 ON793736
14 Hypselotriton orientalis China: Jiangxi: Jiujiang KIZ 020539 ON793739
15 Hypselotriton orientalis China: Jiangxi: Shangrao: Wannian CIB 97867 GU301788
16 Hypselotriton orientalis China: Jiangxi: Shangrao: Wuyuan KIZ YPX25002 ON793740
17 Hypselotriton orientalis China: Zhejiang: Jinhua KIZ 06358 ON793718
18 Hypselotriton orientalis China: Zhejiang: Quzhou CIB 97919 GU301790
19 Hypselotriton orientalis China: Zhejiang: Taizhou: Tiantai KIZ 012941 ON793732
20 Hypselotriton fudingensis China: Fujian: Ningde: Mt Taimu CIB 97874 GU301785
21 Hypselotriton fudingensis China: Fujian: Ningde: Jiulongjing SYS a008487 OQ116688
22 Hypselotriton fudingensis China: Fujian: Ningde: Jiulongjing SYS a008488 OQ116689
23 Hypselotriton fudingensis China: Fujian: Ningde: Qingyu KIZ 012214 ON793743
24 Hypselotriton glaucus China: Guangdong: Meizhou: Mianyang KIZ 09793 ON793715
25 Hypselotriton glaucus China: Guangdong: Meizhou: Mianyang KIZ 09799 ON793716
26 Hypselotriton glaucus China: Guangdong: Meizhou: Mianyang KIZ 09800 ON793717
27 Hypselotriton orphicus China: Fujian: Fuzhou: Yongtai KIZ 09905 ON793728
28 Hypselotriton orphicus China: Fujian: Quanzhou: Mt Daiyun KIZ 09839 ON793723
29 Hypselotriton orphicus China: Guangdong: Chaozou KIZ 09816 ON793719
30 Hypselotriton jiaoren China: Guangdong: Qingyuan: Yingde SYS a008786 OQ116679
31 Hypselotriton jiaoren China: Guangdong: Qingyuan: Yingde SYS a008787 OQ116680
32 Hypselotriton jiaoren China: Guangdong: Qingyuan: Yingde SYS a008788 OQ116681
33 Hypselotriton jiaoren China: Guangdong: Qingyuan: Yingde SYS a008789 OQ116682
34 Hypselotriton jiaoren China: Guangdong: Qingyuan: Yingde CIB 118534 OQ116683
35 Hypselotriton jiaoren China: Guangdong: Qingyuan: Yingde SYS a008791 OQ116684
36 Hypselotriton maguae China: Jiangxi: Fuzhou: Mt Magu CIB 118535 OQ116685
37 Hypselotriton maguae China: Jiangxi: Fuzhou: Mt Magu SYS a007032 OQ116686
38 Hypselotriton cyanurus China: Guizhou: Liupanshui: Shuicheng CIB 95897 GU301784
39 Hypselotriton cyanurus China: Guizhou: Liupanshui: Shuicheng KIZ 02331 ON793754
40 Hypselotriton cyanurus China: Guizhou: Liupanshui: Shuicheng KIZ 02332 ON793755
41 Hypselotriton yunnanensis China: Yunnan: Chuxiong: Zijing KIZ 021922 ON793749
42 Hypselotriton yunnanensis China: Yunnan: Chuxiong: Zijing KIZ 021923 ON793750
43 Hypselotriton yunnanensis China: Yunnan: Kunming: Gulu KIZ 022160 ON793752
44 Hypselotriton yunnanensis China: Yunnan: Kunming: Huahongdong KIZ 022157 ON793751
45 Hypselotriton yunnanensis China: Yunnan: Pu’er: Ning’er KIZ 01445 ON793756
46 Hypselotriton yunnanensis China: Yunnan: Pu’er: Ning’er KIZ 03900 ON793747
47 Hypselotriton yunnanensis China: Yunnan: Pu’er: Ning’er KIZ 03901 ON793748
48 Pachytriton archospotus China: Hunan: Guidong KIZ 04563 KU375007
49 Paramesotriton chinensis China: Zhejiang: Jinhua:Panan KIZ 06335 KU375034

The matrilineal genealogy was reconstructed using Bayesian-inference (BI) and maximum-likelihood (ML) methods based on ND2 gene. PartitionFinder2 was used to test the best partitioning scheme and jModelTest v. 2.1.2 was used to test the best fitting nucleotide substitution model. The data were analyzed using BI in MrBayes v. 3.2.4 (Ronquist et al. 2012), and ML in RaxmlGUI v. 1.3 (Silvestro and Michalak 2012). Two independent runs were conducted in a BI analysis, each of which was performed for 10 million generations and sampled every 1000 generations with the first 25% samples discarded as burn-in, resulting in a potential scale reduction factor (PSRF) of <0.005. The analyses used the proportion of invariable sites estimated from the data and 1000 bootstrap pseudoreplicates under the GTR+gamma model (Chen et al. 2021). Nodes in the trees were considered well supported when Bayesian posterior probabilities (BPP) were ≥ 0.95 and ML bootstrap support (BS) was ≥ 70% (Chen et al. 2021). Mean genetic distances between and within species were calculated in MEGA v. 6.0.6 using the uncorrected genetic distance (p-distances) model.

 Results

Morphologically, our newly collected specimens can be distinguished from all known congeners (details in the taxonomic account below), which can be reliably identified by the obvious black patches with clear boundaries on the whole body and weak vertebral ridge. Statistical analyses on the morphometric measurements were performed on the specimens from northeastern Jiangxi and its sister species H. fudingensis from northeastern Fujian (Table 2). The results of T-test on morphometrics showed that individuals of the northeastern Jiangxi population and H. fudingensis are obviously different in HL, ED, and IND for males (p-values < 0.05), and in TAD for females (p-values < 0.05). Furthermore, once differences attributable to SVL were accounted for (Table 4), there were significant differences between H. huanggangensis sp. nov. and H. fudingensis for TAD, HL, ED, and AG in males, and significant differences for TOL, TAD, HL, HW, AG, FLL and HLL in females.

Morphometric comparisons based on the morphometric measurements (in mm) of Hypselotriton huanggangensis sp. nov. and H. fudingensis. * p-values < 0.05, ** p-values < 0.01.

Holotype H. huanggangensis sp. nov. H. fudingensis P-values
HSR23097 Male (n = 5) Female (n = 2) Male (n = 2) Female (n = 7) Males Females
TOL 75.14 73.0–79.1 (74.8±2.5) 79.6–89.4 69.4–77.7 81.1–101.0 (90.8±6.6) 0.691 0.277
SVL 45.62 42.9–47.3 (44.9±1.7) 44.4–51.1 42.9–45.8 46.5–54.5 (51.0±3.3) 0.754 0.314
TAL 30.35 29.0–33.0 (30.8±1.5) 36.1–38.7 28.8–31.6 36.1–49.0 (40.7±4.4) 0.669 0.263
TAD 7.08 6.7–7.4 (7.0±0.3) 7.4–8.4 5.1–6.2 5.5–7.4 (6.4±0.6) 0.224 0.045*
HL 13.6 12.6–13.6 (13.0±0.4) 12.0–14.4 11.7–12.5 13.2–15.5 (14.1±0.7) 0.040* 0.298
HW 9.16 8.7–9.5 (9.1±0.3) 9.0–10.4 8.3–8.8 8.9–10.5 (9.8±0.5) 0.084 0.878
SL 3.98 4.0–4.4 (4.2±0.2) 3.5–4.6 4.2–4.4 4.1–4.9 (4.4±0.3) 0.472 0.645
ED 3.63 3.5–3.7 (3.6±0.1) 3.7–4.3 3.2–3.3 3.1–4.0 (3.5±0.3) 0.003** 0.202
IOD 5.37 5.0–5.4 (5.3±0.2) 4.9–5.2 4.4–5.3 4.8–5.8 (5.4±0.3) 0.507 0.333
EN 2.86 2.9–3.2 (3.0±0.1) 2.6–3.5 2.9–3.1 2.6–3.3 (3.1±0.2) 0.809 0.914
IND 2.77 2.4–2.8 (2.5±0.2) 2.6–2.9 1.9–2.1 1.9–3.8 (2.5±0.6) 0.021* 0.668
AG 20.19 18.4–21.4 (19.8±1.1) 21.5–24.5 18.2–19.7 20.8–26.1 (24.2±1.9) 0.386 0.659
FLL 12.28 12.3–14.5 (13.4±1.0) 13.6–15.2 12.9–14.2 13.3–15.2 (14.2±0.8) 0.807 0.780
HLL 13.95 14.0–16.2 (15.0±1.0) 13.8–17.2 14.0–14.4 13.5–16.3 (14.9±1.1) 0.188 0.878

BI and ML analyses resulted in similar identical topologies (Fig. 2). As shown in the tree (Fig. 2), three major clades with strong support were revealed for the samples of Hypselotriton, while the relationship among these clades are not resolved. The first clade is composed of samples of H. cyanurus (BPP 1.00, BS 100) and H. yunnanensis (BPP 1.00, BS 97). The second clade consists of H. glaucus from eastern Guangdong (BPP 1.00, BS 100) and H. jiaoren from northern Guangdong (BPP 1.00, BS 100). In the third clade, the new specimens from northeastern Jiangxi form a distinct lineage (BPP 1.00, BS 100), which is sister to H. fudingensis with support values (BPP 1.00, BS 99). The genetic distances based on the ND2 gene among species of Hypselotriton are presented in Table 3. The putative new species from Huanggangshan Mountains showed obvious genetic divergence from other congeners. When compared with closely related recognized congeners, the minimum uncorrected genetic distance was 2.2% between the clade from Huanggangshan Mountains and H. fudingensis (Table 3).

Uncorrected p-distances (%) based on the ND2 gene among Hypselotriton species (in 0.1%).

ID Species 1 2 3 4 5 6 7 8 9
1 Hypselotriton huanggangensis sp. nov. 0
2 Hypselotriton orientalis 7.2 2.7
3 Hypselotriton fudingensis 2.2 8.0 0.5
4 Hypselotriton glaucus 18.8 19.6 18.7 0.5
5 Hypselotriton orphicus 11.8 14.0 12.3 19.9 2.0
6 Hypselotriton jiaoren 18.3 19.5 17.8 6.6 19.3 0.6
7 Hypselotriton maguae 13.9 13.8 13.3 22.5 13.2 21.3 0
8 Hypselotriton cyanurus 21.1 21.3 20.6 20.8 20.8 19.5 23.4 0
9 Hypselotriton yunnanensis 19.6 20.4 20.0 20.9 20.8 20.9 22.8 11.5 2.6
10.3897/zookeys.1208.126092.figure2 5648A503-194F-5308-9F4F-FB38E389C280

Bayesian-inference tree and maximum-likelihood phylogenies based on mitochondrial ND2 gene. Bayesian posterior probabilities and the bootstrap supports (BPP/BS) are shown near the notes. Number at the ends of the lineages correspond to the IDs in Table 1.

https://binary.pensoft.net/fig/1108190

Accordingly, combining the results of the morphological examination presented below and the phylogenetic analysis, the specimens from northeastern Jiangxi are regarded as a new species that is described herein.

Taxonomic account Animalia Urodela Salamandridae 158DB4EF-DDF4-5E74-8F65-F1437DC0BE85 Hypselotriton huanggangensis https://zoobank.org/18334E5C-BED9-47AE-A42D-FA916E7B2CC2 Jiang, Huang, Fan, Cheng, Raffaëlli & Chen sp. nov. Figs 3 , 4 , 5 Type material.

Holotype. HSA 23097, adult male from Huanggangshan Mountains (28.72°N, 117.33°E; elevation 84 m), Yanshan County, Shangrao City, Jiangxi Province, China, collected by Zhihao JIANG on 22 July 2023.

10.3897/zookeys.1208.126092.figure3 346C488F-2C05-56D3-A2DA-24A8DE875DF2

Paratypes of Hypselotriton huanggangensis sp. nov. (HSA23075) A dorsalateral view in life B ventral view in life C small waterhole around mountain shrubs without direct sunlight at the type locality.

https://binary.pensoft.net/fig/1108191

Paratypes. Six adult males: HSA23075–23076, HSA23098–23099, HSA23102–23103, and two adult females: HSA23100–23101. Same collection date and locality as the holotype.

 Etymology.

The specific name huanggangensis refers to the type locality in the Huanggangshan Mountains. For the English common name, we suggest “Huanggangshan Fire-bellied Newt” and for the Chinese name, 黄岗山蝾螈 (huáng gǎng shān róng yuán).

 Diagnosis.

(1) Small body size, TOL 73.0–79.1 mm in adult males, TOL 79.6–89.4 mm in adult females; (2) obvious black patches with clear boundaries on the whole body; (3) ground color of the dorsal body tan; (4) skin rough; (5) ground color of venter bright orange; (6) vertebral ridge weak; (7) fingers and toes overlapping when forelimb and hindlimb adpressed towards each other along body; (8) parotoid gland inconspicuous; (9) postocular orange spot absent; (10) white warty glands around the eye; (11) two discontinuous longitudinal lines which consist of white warty glands from neck to lateral parts of tail (Fig. 6).

 Description of the holotype.

HSA 23097 (Figs 4, 5), adult male with a small, slender body (TOL 79.1 mm, SVL 47.3 mm). Head oval in dorsal view; snout truncate, projecting slightly beyond mandible; nostril small but conspicuous; tongue elongate, enlarged anteriorly, with free lateral margin; vomerine tooth patch ∧–shaped; parotoid gland inconspicuous, gill remnants absent; gular fold present; skin with fine granules, covering most parts of dorsum, venter, chin and tail; vertebral ridge weak; cloacal opening oval, slightly protruding; limbs slender, fingers and toes overlapping when forelimb and hindlimb adpressed towards each other along the body; four fingers and five toes, slender and elongated, lacking webbing; relative length of fingers I < IV < II < III; relative length of toes I < V < II < IV < III. Tail laterally compressed, tapers posteriorly; caudal fin distinct; tail tip bluntly pointed.

10.3897/zookeys.1208.126092.figure4 0AED1545-B6BF-51E5-9855-9329DEFFCA5F

Comparison of holotype of Hypselotriton huanggangensis sp. nov. (left, HSA23097) and Hypselotriton fudingensis (right, HSA23108) in life A dorsal view B ventral view, 1, 5 obvious black patches, 2 weak vertebral ridge, 3 small spots, 4 conspicuous vertebral ridge, 6 bright orange venter without dark blotches.

https://binary.pensoft.net/fig/1108192 10.3897/zookeys.1208.126092.figure5 B1F905B5-DD0C-5C30-AA88-2FA9FA522F32

Comparison of holotype of Hypselotriton huanggangensis sp. nov. (left, HSA23097) and Hypselotriton fudingensis (right, HSA23104) in preservative A dorsal views B ventral views, 1–6 same as in Fig. 4.

https://binary.pensoft.net/fig/1108193 10.3897/zookeys.1208.126092.figure6 DD5BC876-314A-58EE-BF3B-B8A72FF50E3B

Two discontinuous longitudinal lines which consist of white warty glands from neck to lateral parts of tail (Hypselotriton huanggangensis sp. nov., HSA23099) in life.

https://binary.pensoft.net/fig/1108194 Coloration of the holotype.

In life, obvious black patches with clear boundaries on the whole body; ground color of the dorsal body tan; ground color of venter bright orange; white warty glands from the lateral part of head to tail; axilla, cloacal opening, and venter of tail bright orange. In preservative after six months (Fig. 5), dorsum, flanks, and limbs slightly darker. All orange coloration of venter fades to creamy white.

Test for differences between H. huanggangensis sp. nov. and H. fudingensis using ANCOVA (SVL as covariate).

Variable Males Females
Total length (TOL)
F 6.78 12.17
P 0.052 0.008
Tail length (TAL)
F 1.09 2.27
P 0.419 0.185
Maximum tail depth (TAD)
F 12.34 15.48
P 0.019 0.004
Head length (HL)
F 8.76 9.66
P 0.035 0.013
Maximum head width (HW)
F 3.77 16.71
P O.120 0.004
Snout length (SL)
F 1.18 1.23
P 0.395 0.357
Eye diameter (ED)
F 12.00 1.91
P 0.020 0.228
Interorbital distance (IOD)
F 1.20 1.48
P 0.390 0.301
Eye–nostril length (EN)
F 0.03 0.38
P 0.967 0.700
Internasal distance (IND)
F 5.91 0.189
P 0.064 0.833
Axilla–groin length (AG)
F 76.89 9.38
P 0.010 0.014
Forelimb length (FLL)
F 0.07 8.86
P 0.938 0.016
Hindlimb length (HLL)
F 1.48 5.65
P 0.330 0.042
Variation.

Linear measurements are summarized in Table 2. Females (TOL 79.6–89.4 mm) are distinctly larger than males (TOL 73.0–79.1 mm). All paratypes resemble the holotype except that the cloaca is wider and more swollen in males than in females, the irregular bright-orange patches on ventral surface occupy more surface in females than in males, and the gular fold absent in some individuals.

 Comparisons.

Hypselotriton huanggangensis sp. nov. is phylogenetically close to H. fudingensis, which is distributed in northeastern Fujian. However, H. huanggangensis sp. nov. differs from H. fudingensis by its weak vertebral ridge (vs vertebral ridge conspicuous), dorsal black patches (vs small or no spots on dorsum), and ventral black patches (vs venter bright orange without dark blotches).

In addition, H. huanggangensis sp. nov. further differs from the remaining congeners.

Hypselotriton huanggangensis sp. nov. differs from H. orphicus by its weak vertebral ridge (vs slightly bulged) and obvious black patches with clear boundaries on the whole body (vs small or moderate dorsal blackish dots in H. orphicus).

Hypselotriton huanggangensis sp. nov. differs from H. orientalis by its parotoid gland inconspicuous (vs conspicuous) and the presence of obvious black patches with clear boundaries on the dorsum (vs absent in H. orientalis).

Hypselotriton huanggangensis sp. nov. differs from H. glaucus by its obvious black patches with clear boundaries on the whole body (vs dorsum, flanks, limbs, and upper side of tail with irregular obscure greyish blue patches in H. glaucus).

Hypselotriton huanggangensis sp. nov. differs from H. jiaoren by its rough skin (vs smooth), and obvious black patches with clear boundaries on the whole body (vs dorsum, flanks, limbs, and upper side of tail uniformly dark brown in H. jiaoren).

Hypselotriton huanggangensis sp. nov. differs from H. maguae by having its fingers and toes overlapping when forelimbs and hindlimbs are adpressed (vs forelimbs and hindlimbs not meeting when adpressed towards each other along body), and obvious black patches with clear boundaries on the whole body (vs dorsum, flanks, limbs, and upper side of tail uniformly dark brown in H. maguae).

Hypselotriton huanggangensis sp. nov. can be distinctly distinguished from H. wolterstorffi, H. cyanurus, H. chenggongensis and H. yunnanensis by its absent postocular orange spot (vs present).

 Distribution and habitat.

Hypselotriton huanggangensis sp. nov. is currently known only from the type locality on the western side of the Wuyishan Mountains in northeastern Jiangxi. Newts were found in a small waterhole around mountain shrubs without direct sunlight, at 84 m a.s.l. All individuals were observed in July, September, and February.

 Discussion

Despite more than a century of effort, taxonomists have yet to reach a consensus on the concept of species and methods of all species delimitation (Mayden 1997; de Queiroz 1998; Fu and Zeng 2008; Yang and Rannala 2010; Chen et al. 2013; Peng et al. 2014). Trying to solve “the notorious problem of taxonomic uncertainty (Uetz et al. 2024)”, Huang et al. (2021) proposed the principle of “species subdivision” with recommendable “subdivision” at the species level. It should help to understand species natural history more effectively and facilitate consistent actions in taxonomy and practices of conservation biology.

In the present study, although the genetic distance based on the ND2 gene fragment between H. huanggangensis sp. nov. and its sister species H. fudingensis is not very large (uncorrected p-distance = 2.2%), morphological differences between them are distinct (Fig. 4; Table 2). In addition, H. huanggangensis sp. nov. and H. fudingensis are isolated by a series of high mountain ranges, including the Wuyishan and Jiufengshan Mountains. These barriers often isolate different amphibian species (Chen et al. 2020).

With the addition of the new species described here, the genus Hypselotriton now includes 11 species. Until now, seven species of Hypselotriton were found in the southeastern Chinese hilly area (H. huanggangensis, H. jiaoren, H. glaucus, H. maguae, H. orphicus, H. fudingensis and H. orientalis), and another four species of Hypselotriton are distributed in southwestern China. Due to unresolved relationships, a more extensive sampling of taxa and molecular data are necessary for reliable conclusions on the evolution and taxonomy of Hypselotriton.

 Acknowledgements

We thank Cong Liu for assistance in the field. We also thank Ben Liu for discussion of the analysis of phylogenetic data.

 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 the Second Tibetan Plateau Scientific Expedition and Research Program (STEP), Grant No. 2019QZKK0501, Doctoral Research Starting Foundation of Anhui Normal University (752017), National Natural Science Foundation of China (NSFC 31471968; 32001222; 31900323).

 Author contributions

Writing–original draft: ZJ. Writing–reviewing and editing: ZJ, SH, JC, JR. Funding acquisition & Supervision: QF, LC, YG. Investigation–sample collection: ZJ, ZC, TZ. Formal analysis–phylogenetic & morphological analysis: ZJ, WL, ZZ, XH.

 Author ORCIDs

Zhihao Jiang https://orcid.org/0009-0004-1492-0665

Song Huang https://orcid.org/0000-0001-6786-8523

Qiangyong Fan https://orcid.org/0009-0002-5515-2143

Lin Cheng https://orcid.org/0009-0004-4674-4213

Yanan Gong https://orcid.org/0000-0002-8155-7024

Zhangbo Cui https://orcid.org/0009-0000-1501-0402

Tierui Zhang https://orcid.org/0009-0000-1869-9313

Wenjun Lan https://orcid.org/0009-0005-0690-1082

Zhian Zou https://orcid.org/0009-0001-2556-2590

Xuanzhi Huang https://orcid.org/0009-0008-2873-3640

Jean Raffaëlli https://orcid.org/0000-0003-0567-3661

Jinmin Chen https://orcid.org/0000-0001-6432-7721

 Data availability

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

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