Research Article |
Corresponding author: Ning Xiao ( armiger@163.com ) Corresponding author: Jiang Zhou ( zhoujiang@ioz.ac.cn ) Academic editor: Tihomir Stefanov
© 2023 Tao Luo, Ming-Le Mao, Chang-Ting Lan, Ling-Xing Song, Xin-Rui Zhao, Jing Yu, Xing-Liang Wang, Ning Xiao, Jia-Jun Zhou, Jiang Zhou.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Luo T, Mao M-L, Lan C-T, Song L-X, Zhao X-R, Yu J, Wang X-L, Xiao N, Zhou J-J, Zhou J (2023) Four new hypogean species of the genus Triplophysa (Osteichthyes, Cypriniformes, Nemacheilidae) from Guizhou Province, Southwest China, based on molecular and morphological data. ZooKeys 1185: 43-81. https://doi.org/10.3897/zookeys.1185.105499
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Recently described cave species of the genus Triplophysa have been discovered in southwestern China, suggesting that the diversity of the genus is severely underestimated and that there may be many undescribed species. In this work, four new species of the genus Triplophysa are described from southwestern Guizhou Province, China, namely Triplophysa cehengensis Luo, Mao, Zhao, Xiao & Zhou, sp. nov. and Triplophysa rongduensis Mao, Zhao, Yu, Xiao & Zhou, sp. nov. from Rongdu Town, Ceheng County, Guizhou, Triplophysa panzhouensis Yu, Luo, Lan, Xiao & Zhou, sp. nov. from Hongguo Town, Panzhou City, Guizhou, and Triplophysa anlongensis Song, Luo, Lan, Zhao, Xiao & Zhou, sp. nov. from Xinglong Town, Anlong County, Guizhou. These four new species can be distinguished from all recognized congeners by a combination of morphological characteristics and significant genetic divergences. The discovery of these species increases the number of known cave species within the genus Triplophysa to 39, making the genus the second most diverse group of cave fishes in China after the golden-line fish genus Sinocyclocheilus. Based on the non-monophyletic relationships of the different watershed systems in the phylogenetic tree, this study also discusses the use of cave species of the genus Triplophysa to determine the possible historical connectivity of river systems.
Diversity, karst cave, morphology, new species, taxonomy, Triplophysa
The Southwest China Karst (SCK) topography is widely distributed and varied, with its extremely high level of biodiversity making the region a priority area for biological conservation in China (
The genus Triplophysa Rendahl, 1933 is a large group of smaller loaches distributed on the Qinghai-Tibetan Plateau and adjacent regions and currently containing over 180 recognized valid species (
A list of 35 species of hypogean fishes of the genus Triplophysa distributed in the Southwest China Karst.
ID | Species | Province | Main drainage | Tributary | Reference |
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1 | T. aluensis Li & Zhu, 2000 | Yunnan | Pearl River | Nanpanjiang River |
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2 | T. anshuiensis Wu, Wei, Lan & Du, 2018 | Guangxi | Pearl River | Hongshui River |
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3 | T. baotianensis Li, Liu & Li, 2018 | Guizhou | Pearl River | Nanpanjiang River |
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4 | T. erythraea Liu & Huang, 2019 | Hunan | Yangtze River | Wujiang River |
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5 | T. fengshanensis Lan, 2013 | Guangxi | Pearl River | Hongshui River |
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6 | T. flavicorpus Yang, Chen & Lan, 2004 | Guangxi | Pearl River | Hongshui River |
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7 | T. gejiuensis (Chu & Chen, 1979) | Yunnan | Pearl River | Nanpanjiang River |
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8 | T. guizhouensis Wu, He & Yang, 2018 | Guizhou | Pearl River | Hongshui River |
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9 | T. huapingensis Zheng, Yang & Che, 2012 | Guangxi | Pearl River | Hongshui River |
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10 | T. langpingensis Yang, 2013 | Guangxi | Pearl River | Hongshui River |
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11 | T. longipectoralis Zheng, Du, Chen & Yang, 2009 | Guangxi | Pearl River | Liujiang River |
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12 | T. longliensis Qiu, Yang & Chen, 2012 | Guizhou | Pearl River | Hongshui River | Qiu et al. 2012 |
13 | T. luochengensis Li, Lan, Chen & Du, 2017 | Guangxi | Pearl River | Hongshui River |
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14 | T. macrocephala Yang, Wu & Yang, 2012 | Guangxi | Pearl River | Liujiang River |
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15 | T. nandanensis Lan, Yang & Chen, 1995 | Guangxi | Pearl River | Hongshui River |
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16 | T. nanpanjiangensis Zhu & Cao, 1988 | Yunnan | Pearl River | Nanpanjiang River |
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17 | T. nasobarbatula Wang & Li, 2001 | Guizhou | Pearl River | Liujiang River |
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18 | T. posterodorsalus (Li, Ran & Chen, 2006) | Guangxi | Pearl River | Liujiang River |
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19 | T. qingzhenensis Liu, Zen, & Gong, 2022 | Guizhou | Yangtze River | Wujiang River |
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20 | T. qini Deng, Wang & Zhang, 2022 | Chongqing | Yangtze River | Wujiang River |
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21 | T. qiubeiensis Li & Yang, 2008 | Yunnan | Pearl River | Nanpanjiang River |
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22 | T. rosa Chen & Yang, 2005 | Chongqing | Yangtze River | Wujiang River |
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23 | T. sanduensis Chen & Peng, 2019 | Guizhou | Pearl River | Duliujiang River |
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24 | T. shilinensis Chen,Yang & Xu, 1992 | Yunnan | Pearl River | Nanpangjiang River |
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25 | T. tianeensis Chen, Cui & Yang, 2004 | Guangxi | Pearl River | Hongshui River |
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26 | T. tianlinensis Li, Li, Lan & Du, 2017 | Yunnan | Pearl River | Hongshui River |
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27 | T. tianxingensis Yang, Li & Chen, 2016 | Yunnan | Pearl River | Nanpangjiang River |
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28 | T. wudangensis Liu, Zen & Gong, 2022 | Guizhou | Yangtze River | Wujiang River |
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29 | T. wulongensis Chen, Sheraliev, Shu & Peng, 2021 | Chongqing, Guizhou | Yangtze River | Wujiang River |
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30 | T. xiangshuingensis Li, 2004 | Yunnan | Pearl River | Nanpanjiang River |
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31 | T. xiangxiensis Yang, Yuan & Liao, 1986 | Hunan | Yangtze River | Yuanjiang River |
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32 | T. xichouensis Liu, Pan, Yang & Chen, 2017 | Yunnan | Red River | Red River |
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33 | T. xuanweiensis Lu, Li, Mao & Zhao, 2022 | Yunnan | Pearl River | Hongshui River |
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34 | T. yunnanensis Yang, 1990 | Yunnan | Pearl River | Nanpanjiang River |
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35 | T. zhenfengensis Wang & Li, 2001 | Guizhou | Pearl River | Beipanjiang River |
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Guizhou province is located in southwest China adjacent to Guangxi, Yunnan, Chongqing, and Sichuan provinces and is home to extensive karsts and well-developed underground rivers as well as being recognized as a cavefish biodiversity hotspot and a priority biodiversity reserve in China (
Sampling collection localities and distributions of the four new species and 35 hypogean species of the genus Triplophysa in southern China. For details concerning the ID numbers, please see Suppl. material
In January 2023, we collected several specimens of the genus Triplophysa, identified by the anterior and posterior nostrils being closely set, during a survey of cave fishes in southwestern Guizhou Province, China. Morphological examination and molecular phylogenetic analysis suggested that these specimens could be distinguished from the 35 hypogean species of the genus Triplophysa. Here, we formally describe the specimens as four new species, Triplophysa cehengensis sp. nov., Triplophysa rongduensis sp. nov., Triplophysa panzhouensis sp. nov., and Triplophysa anlongensis sp. nov.
In total, 73 samples from 20 species were collected for morphology comparison and genetic analysis (Fig.
Genomic DNA was extracted from muscle tissue using a DNA extraction kit from Tiangen Biotech (Beijing) Co. Ltd. In total, 12 tissue samples used for molecular analysis were amplified and sequenced for mitochondrial gene cytochrome b (Cyt b) using the primers L3975 (5’-CGCCTGTTTACCAAAAACAT-3’) and H4551 (5’-CCGGTCTGAACTCAGATCACGT-3’) following
Localities, voucher information, and GenBank numbers for all samples used. Numbers in bold were generated in this study.
ID | Species | Localities (* type localities) | Voucher ID | Cyt b |
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1 | T. panzhouensis sp. nov. | Hongguo Town, Panzhou City, Guizhou, China* |
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OQ754119 |
2 | T. panzhouensis sp. nov. | Hongguo Town, Panzhou City, Guizhou, China* |
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OQ754120 |
3 | T. panzhouensis sp. nov. | Hongguo Town, Panzhou City, Guizhou, China* |
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OQ754121 |
4 | T. cehengensis sp. nov. | Rongdu Town, Ceheng County, Guizhou, China* |
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OQ754132 |
5 | T. cehengensis sp. nov. | Rongdu Town, Ceheng County, Guizhou, China* |
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OQ754133 |
6 | T. cehengensis sp. nov. | Rongdu Town, Ceheng County, Guizhou, China* |
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OQ754134 |
7 | T. rongduensis sp. nov. | Rongdu Town, Ceheng County, Guizhou, China* |
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OQ754135 |
8 | T. rongduensis sp. nov. | Rongdu Town, Ceheng County, Guizhou, China* |
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OQ754136 |
9 | T. rongduensis sp. nov. | Rongdu Town, Ceheng County, Guizhou, China* |
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OQ754137 |
10 | T. anlongensis sp. nov. | Xinglong Town, Anlong County, Guizhou, China* |
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OQ754138 |
11 | T. anlongensis sp. nov. | Xinglong Town, Anlong County, Guizhou, China* |
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OQ754139 |
12 | T. anlongensis sp. nov. | Xinglong Town, Anlong County, Guizhou, China* |
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OQ754140 |
13 | T. baotianensis | Baotian Town, Panzhou City, Guizhou, China* |
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MT992550 |
14 | T. baotianensis | Baotian Town, Panzhou City, Guizhou, China |
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OQ241181 |
15 | T. erythraea | Dalong Cave, Huayuan County, Hunan, China* | / | MG967615 |
16 | T. huapingensis | / | F3917 | MG697589 |
17 | T. huapingensis | Huaping Town, Leye County, Guangxi, China* |
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OQ754125 |
18 | T. langpingensis | Longping Township, Tianlin County, Guangxi* |
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OQ754122 |
19 | T. longliensis | / | SWU2016090300 | MW582825 |
20 | T. macrocephala | Lihu Town, Nandan County, Guangxi, China* |
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OQ754123 |
21 | T. nasobarbatula | Dongtang Township, Libo County, Guizhou, China* |
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MH685911 |
22 | T. nasobarbatula | Dongtang Township, Libo County, Guizhou, China* |
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OQ241175 |
23 | T. nasobarbatula | Dongtang Township, Libo County, Guizhou, China* |
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OQ241176 |
24 | T. nandanensis | Hechi City, Guangxi,China | SWU20151123046 | MG697588 |
25 | T. nandanensis | Liuzhai Town, Nandan County, Guangxi, China* |
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OQ754126 |
26 | T. nandanensis | Liuzhai Town, Nandan County, Guangxi, China* |
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OQ754128 |
27 | T. qini | Houping Village, Wulong County, Chongqing, China* | ON528184 | |
28 | T. qiubeiensis | NijiaoVillage, Qiubei County, Yunnan, China* |
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OQ754127 |
29 | T. qingzhenensis | Qingzhen County, Guiyang City, Guizhou China* | IHB 201911150004 | MT700458 |
30 | T. rosa | Huolu Town, Wulong County, Chongqing, China | SWU10100503 | JF268621 |
31 | T. rosa | / | F3911 | MG697587 |
32 | T. rosa | Huolu Town, Wulong County, Chongqing City, China |
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OQ754130 |
33 | T. sanduensis | Zhonghe Town, Sandu County, Guizhou, China* | SWU20170613001 | MW582822 |
34 | T. tianeensis | / | / | MW582826 |
35 | T. tianeensis | Bala Township, Tian ‘e County, Guangxi, China* |
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OQ754124 |
36 | T. wudangensis | Wudang District, Guiyang City, Guizhou Province, China* | IHB 201908090003 | MT700460 |
37 | T. wudangensis | Wudang District, Guiyang City, Guizhou Province, China* |
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OQ754131 |
38 | T. wulongensis | Wulong County, Chongqing, China* | / | MW582823 |
39 | T. wulongensis | Huolu Town, Wulong County, Chongqing City, China |
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OQ754129 |
40 | T. xiangxiensis | Feihu Cave, Hunan, China* | / | JN696407 |
41 | T. xiangxiensis | / | IHB 2015010002 | KT751089 |
42 | T. xuanweiensis | Tangtang Town, Xuanwei City, Yunnan, China* | ASIZB223818 | OL675196 |
43 | T. xuanweiensis | Tangtang Town, Xuanwei City, Yunnan, China* | ASIZB223819 | OL675197 |
44 | T. xuanweiensis | Tangtang Town, Xuanwei City, Yunnan, China* | ASIZB223820 | OL675198 |
45 | T. zhenfengensis | Xinlongchang Town, Xingren City, Guizhou, China* |
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OQ241177 |
46 | T. zhenfengensis | Xinlongchang Town, Xingren City, Guizhou, China* |
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OQ241178 |
47 | T. zhenfengensis | Xinlongchang Town, Xingren City, Guizhou, China* |
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OQ241179 |
48 | T. zhenfengensis | Xinlongchang Town, Xingren City, Guizhou, China* |
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OQ241180 |
49 | T. nujiangensa | Fugong County, Yunnan, China | IHB201315814 | KT213598 |
50 | T. tibetana | Mafamu lake, Xinjiang, China | NWIPB1106069 | KT224364 |
51 | T. tenuis | Niutou river, Qingshui County, Gansu, China | IHB0917490 | KT224363 |
52 | T. wuweiensis | Yongchang County, Gansu, China | IHB201307124 | KT224365 |
53 | Barbatula barbatula | / | / | KP715096 |
54 | Barbatula labiata | Xinyuan County, Xinjiang, China | IHB201306569 | KT192057 |
55 | Homatula berezowskii | Qujing City, Yunnan, China | FS-2014-Y03 | NC_040302 |
In total, 55 mitochondrial Cyt b sequences were used for molecular analysis, including 22 that were newly sequenced and 33 that were downloaded from GenBank. We followed the phylogenetic study from
All sequences were assembled and aligned using the MUSCLE (
The analysis suggested the best partition scheme for each codon position of Cyt b. TRNEF+I+G, HKY+I, and TIM+I+G were selected for first, second, and third codons, respectively. Two independent runs were conducted in the BI analysis, each of which was performed for 2 × 107 generations and sampled every 1000 generations. The first 25% of the samples were discarded as a burn-in, resulting in a potential scale reduction factor of < 0.01. Nodes in the trees were considered well supported when Bayesian posterior probabilities (BPP) were ≥ 0.95 and the ML ultrafast bootstrap value (UBP) was ≥ 95%. Uncorrected p-distances (1000 replicates) based on Cyt b were estimated using MEGA v. 7.0.
Morphometric data were collected from 36 well-preserved specimens of the genus Triplophysa (Suppl. material
Comparative data for the 35 hypogean species of the genus Triplophysa were obtained from the literature and specimen examination (Suppl. material
Both ML and BI phylogenies were constructed based on mitochondrial Cyt b sequences, with the sequence length being 1140 base pairs. The BI and ML phylogenetic trees showed a highly consistent topology that strongly supported the monophyly of the genus Triplophysa and indicated that the genus could be divided into two major clades, the hypogean group and the epigean group (Fig.
Phylogenetic tree based on A mitochondrial Cyt b (1140 bp) and B the corresponding ranges of the three clades. In this phylogenetic tree, Bayesian posterior probabilities (BPP) from BI analysis/ultrafast bootstrap supports (UBP) from ML analysis are noted beside nodes. Scale bars represent 0.05 nucleotide substitutions per site. The numbers at the tips of branches correspond to the ID numbers listed in Table
The hypogean group contained 19 known species from the karsts of southwestern China (Guizhou, Yunnan, Guangxi, Chongqing, and Hubei) and four other lineages from southwestern Guizhou that could be further divided into three clades (Fig.
Within subclade B1, all samples from Rongdu Town, Ceheng County, Guizhou (samples 4–6 in Table
Uncorrected p-distance (%) between 23 species of the genus Triplophysa based on mitochondrial Cyt b.
ID | Species | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | 17 | 18 | 19 | 20 | 21 | 22 |
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1 | T. anlongensis sp. nov. | ||||||||||||||||||||||
2 | T. panzhouensis sp. nov. | 7.1 | |||||||||||||||||||||
3 | T. cehengensis sp. nov. | 10.2 | 10.1 | ||||||||||||||||||||
4 | T. rongduensis sp. nov. | 4.0 | 8.3 | 9.6 | |||||||||||||||||||
5 | T. baotianensis | 6.9 | 7.8 | 11.2 | 6.7 | ||||||||||||||||||
6 | T. erythraea | 14.7 | 13.7 | 15.4 | 14.7 | 14.2 | |||||||||||||||||
7 | T. huapingensis | 10.1 | 10.1 | 6.1 | 9.3 | 10.6 | 15.4 | ||||||||||||||||
8 | T. longliensis | 14.8 | 13.2 | 14.1 | 14.3 | 14.6 | 11.0 | 15.0 | |||||||||||||||
9 | T. langpingensis | 14.5 | 14.8 | 14.3 | 14.0 | 14.3 | 16.2 | 14.9 | 15.3 | ||||||||||||||
10 | T. macrocephala | 15.5 | 15.2 | 15.5 | 15.4 | 15.9 | 12.0 | 15.1 | 9.5 | 15.8 | |||||||||||||
11 | T. nandanensis | 16.8 | 16.1 | 16.3 | 16.2 | 16.8 | 12.4 | 16.2 | 10.7 | 16.6 | 5.1 | ||||||||||||
12 | T. nasobarbatula | 15.6 | 15.3 | 15.4 | 15.2 | 15.8 | 12.1 | 15.0 | 9.5 | 15.2 | 0.7 | 5.3 | |||||||||||
13 | T. qingzhenensis | 15.6 | 15.4 | 14.5 | 15.4 | 15.2 | 11.5 | 14.9 | 9.4 | 14.5 | 10.1 | 10.5 | 9.9 | ||||||||||
14 | T. qiubeiensis | 14.6 | 14.4 | 14.6 | 14.0 | 14.3 | 13.8 | 14.7 | 13.6 | 14.6 | 14.5 | 14.9 | 14.1 | 13.9 | |||||||||
15 | T. qini | 14.8 | 14.4 | 14.1 | 14.6 | 15.5 | 10.8 | 15.3 | 5.4 | 15.8 | 9.3 | 10.7 | 9.5 | 9.1 | 13.5 | ||||||||
16 | T. rosa | 15.6 | 15.8 | 14.8 | 15.6 | 15.2 | 11.8 | 15.6 | 9.7 | 14.6 | 10.4 | 10.9 | 10.1 | 1.5 | 14.1 | 9.4 | |||||||
17 | T. sanduensis | 15.0 | 13.6 | 14.1 | 14.5 | 14.8 | 11.2 | 15.0 | 0.5 | 15.3 | 9.4 | 10.8 | 9.4 | 9.4 | 13.8 | 5.5 | 9.7 | ||||||
18 | T. tianeensis | 16.9 | 16.0 | 16.7 | 16.7 | 16.8 | 11.9 | 16.4 | 10.9 | 16.5 | 5.2 | 1.9 | 5.3 | 10.4 | 14.8 | 10.4 | 11.0 | 11.0 | |||||
19 | T. wudangensis | 15.4 | 15.6 | 14.7 | 15.4 | 14.9 | 11.5 | 15.0 | 9.6 | 14.5 | 10.5 | 10.8 | 10.2 | 1.6 | 14.3 | 9.6 | 1.5 | 9.7 | 10.8 | ||||
20 | T. wulongensis | 17.1 | 16.5 | 16.7 | 16.7 | 16.8 | 15.6 | 18.0 | 13.8 | 15.6 | 15.2 | 14.9 | 15.2 | 13.9 | 15.6 | 13.5 | 14.0 | 13.6 | 14.6 | 14.0 | |||
21 | T. xiangxiensis | 14.5 | 14.3 | 14.6 | 14.3 | 15.1 | 11.7 | 15.6 | 7.7 | 14.9 | 8.5 | 10.0 | 8.4 | 9.0 | 14.1 | 6.0 | 9.0 | 8.0 | 9.3 | 9.2 | 14.7 | ||
22 | T. xuanweiensis | 14.9 | 14.1 | 15.0 | 14.8 | 14.5 | 12.1 | 14.9 | 11.8 | 14.1 | 11.5 | 11.9 | 11.4 | 11.6 | 12.1 | 11.4 | 11.7 | 11.8 | 12.1 | 11.3 | 14.2 | 11.4 | |
23 | T. zhenfengensis | 3.5 | 8.0 | 9.4 | 0.8 | 6.9 | 14.8 | 9.3 | 14.7 | 13.9 | 15.7 | 16.6 | 15.5 | 15.7 | 14.1 | 14.9 | 15.9 | 14.9 | 17.0 | 15.8 | 16.6 | 14.6 | 14.6 |
Within subclade B1, all samples from Rongdu Town, Ceheng County, Guizhou (samples 7–9 in Table
Within subclade B1, all samples from Xinglong Town, Anlong County, Guizhou (samples 10–12 in Table
Within subclade B1, all samples from Hongguo Town, Panzhou City, Guizhou (samples 1–3 in Table
Mann-Whitney U tests and boxplots revealed differences in several morphological characteristics among the four new species (T. cehengensis sp. nov., T. rongduensis sp. nov., T. panzhouensis sp. nov., and T. anlongensis sp. nov.), and between the new species and the closely related species T. baotianensis and T. zhenfengensis (Suppl. materials
Seven principal component factors with eigenvalues greater than one were extracted based on PCA of the morphological data. These factors accounted for 84.05% of the total variation (Suppl. material
The hypogean group of the genus Triplophysa in this study can be phylogenetically divided into two clades A and B, where Clade B can be further subdivided into subclades B1 and B2. These clades can be defined by the following morphology: (1) for Clade A, the dorsal-fin origin posterior to the pelvic-fin origin; (2) for Clade B, the dorsal-fin origin anterior/opposite to the pelvic-fin origin and subclades B1 and B2 can be defined by the number of unbranched pelvic fin rays (two vs one). The four new species are based on the following characteristics: the dorsal fin origin anterior to the pelvic fin origin and the two unbranched pelvic-fin rays and can be easily placed into subclade B1. Thus, the new species can be compared to the five species within the remaining subclade B1 and the 15 species in the undefined clade.
Holotype. GZNU20230214010 (Fig.
Paratypes. Two specimens from the same locality as the holotype: GZNU202302140011 and GZNU20230214012 collected by Tao Luo, Wei-Feng Wang, Xin-Rui Zhao, Jing Yu, and Chang-Ting Lan on January 7, 2023.
Triplophysa cehengensis sp. nov. can be distinguished from all other congeners by the following combination of characteristics: (1) body naked, scaleless, without skin pigmentation; (2) eyes reduced, diameter 2% of head length (HL), interorbital width 27–37% of HL; (3) dorsal fin distal margin emarginated; (4) pelvic-fin tip reaching to anus; (5) tip of pectoral fin not reaching to pelvic fin origin; (6) anterior and posterior nostrils closely set, with the anterior nostril elongated to a barbel-like tip; (7) tip of outrostral barbel extending backward, not reaching to anterior margin of the eye; (8) lateral line complete; (9) dorsal-fin rays iv-9, pectoral-fin rays i-10, pelvic-fin rays ii-8, anal-fin rays iii-5, 16 branched caudal-fin rays; and (10) total vertebrae 39.
Morphological data of the specimens of the Triplophysa cehengensis sp. nov. are provided in Suppl. materials
Dorsal-fin rays iv-9, pectoral-fin rays i-10, pelvic-fin rays ii-8, anal-fin rays iii-5, 16 branched caudal-fin rays. Dorsal fin short, length 22–25% of SL, distal margin emarginated, origin anterior to pelvic-fin insertion and situated slightly posterior to the midpoint between snout tip and caudal-fin base, first branched ray longest, shorter than head length, tip of dorsal fin vertical to the anus. Pectoral fin moderately developed, length 21–23% of SL, tip of pectoral fin extending backward almost to the midpoint between origin of pectoral and pelvic fin origins, not reaching to pelvic fin origin. Pelvic fin length 17–18% of SL, vertically aligned with third branched ray of dorsal fin, tips of pelvic fin reaching to anus. Anal fin long, length 19–21% of SL, distal margin truncated, origin close to anus, tips of anal fin not reaching caudal-fin base, distance between tips of anal fin and anus 8.5× the eye diameter. Caudal fin forked, upper lobe is equal in length to lower lobe, tips pointed, caudal peduncle length 6.9 mm, caudal peduncle depth 3.5 mm, with weak adipose crests along both dorsal and ventral sides. Total vertebrae: 39 (n = 1).
Cephalic lateral line system developed. Lateral line complete, exceeding tip of pectoral fin and reaching base of caudal fin. Two chambers of air bladder, anterior chamber dumbbell-shaped and membranous, open on both sides, slightly closed posteriorly (Fig.
In cave water bodies when alive, body semi-translucent and pale pink, without skin pigment, and all fins hyaline (Fig.
No secondary sex characteristics was observed based on the present specimens of Triplophysa cehengensis sp. nov.
Comparative data of Triplophysa cehengensis sp. nov. with the 34 recognized hypogean species within genus Triplophysa are given in Suppl. material
Triplophysa cehengensis sp. nov. can be distinguished from T. baotianensis, T. zhenfengensis, T. huapingensis, T. flavicorpus, T. guizhouensis, T. longipectoralis, T. luochengensis, T. nanpanjiangensis, T. tianxingensis, T. xiangshuingensis, T. xichouensis, and T. yunnanensis by body pigmentation absence (vs body pigmentation presence). Triplophysa cehengensis sp. nov. can be distinguished from T. qiubeiensis, T. anshuiensis, T. fengshanensis, T. gejiuensis, T. posterodorsalus, and T. shilinensis by eyes reduced, diameter 2% of HL (vs eyes absent).
For the remaining two species, the new species share the following morphological characteristics: body pigmentation absent, eyes reduced, and body scaleless. However, Triplophysa cehengensis sp. nov. differs from T. aluensis and T. langpingensis by the dorsal fin distal margin being emarginated (vs truncated); from T. aluensis by dorsal-fin rays (iv, 9 vs iii, 7), pelvic-fin rays (ii, 8 vs i, 6), 16 branched caudal-fin rays (vs 13), tip of pelvic fin reaching to anus (vs not reaching to anus); and from T. langpingensis by dorsal-fin rays (iv, 9 vs iii, 7–8), pelvic fin rays (ii, 8 vs i, 6), 16 branched caudal-fin rays (vs 14), lateral line complete (vs incomplete), and 16 branched caudal-fin rays (vs 14).
Currently, this new species Triplophysa cehengensis sp. nov. has only been found in a cave in Longjing Village, Rongdu Town, Ceheng County, Guizhou Province, China, at an elevation of 1228 m. The pool where the new species was found to live is more than 30 cm long, 25 cm wide, and 6 cm deep, with a slow flow of water, and is located ~ 30 m vertically down from the cave entrance. Inside the cave, bats (Ia io, six individuals) and crabs (Diyutamon cereum, ten individuals) were found. Outside the cave, maize and rice are being grown. The population of the new species is very small. From a total of eight sample collections, only specimen number three was obtained.
The specific epithet cehengensis is in reference to the type locality of the new species: Longjing Village, Rongdu Town, Ceheng County. We propose the common English name “Ceheng high-plateau loach” and the Chinese name “Cè Hēng Gāo Yuán Qīu (册亨高原鳅)”.
Holotype. GZNU20230106001 (Fig.
Paratypes. Five specimens from the same locality as the holotype: GZNU20230214001–214005 collected by Tao Luo, Chang-Ting Lan, Jing Yu, Xin-Rui Zhao, and Wei-Feng Wang on January 6, 2023.
Triplophysa rongduensis sp. nov. can be distinguished from species of the hypogean group of its congeners by the following combination of characteristics: (1) body naked, scaleless, with skin pigmentation; (2) eyes normal, diameter 7–15% of HL, interorbital width 24–32% of HL; (3) dorsal fin distal margin truncated; (4) pelvic-fin tip not reaching to anus; (5) tip of pectoral fin not reaching to pelvic fin origin; (6) anterior and posterior nostrils closely set, anterior nostril elongated to a barbel-like tip; (7) tip of outrostral barbel extending backward to posterior margin of the eye; (8) lateral line complete; (9) dorsal-fin rays iv-9, pectoral-fin rays i-10, pelvic-fin rays ii-7, anal-fin rays iii-5, 16 branched caudal-fin rays; and (10) total vertebrae 43.
Morphological data of the specimens of the Triplophysa rongduensis sp. nov. are provided in Suppl. materials
Dorsal-fin rays iv-9, pectoral-fin rays i-10, pelvic-fin rays ii-7, anal-fin rays iii-5, 16 branched caudal fin rays. Dorsal fin long, 19–25% of SL, distal margin emarginated, origin slightly anterior to pelvic-fin insertion, situated slightly posterior to the midpoint between snout tip and caudal-fin base; first branched ray longest, shorter than head length, tip of dorsal fin not extending to vertical of anus. Pectoral fin more developed, 18–20% of SL, tip of pectoral fin extends backwards not to the midpoint between origin of pectoral and pelvic fin origins, not reaching to pelvic fin origin. Pelvic fin length 7–17% of SL, vertically aligned with third branched ray of dorsal fin, tips of pelvic fin not reaching to anus, distance between tips of pelvic fin and anus 0.9× the eye diameter. Anal fin long, length 16–17% of SL, distal margin truncated, origin close to anus, spacing 2.1 mm, tips of anal fin not reaching caudal-fin base, distance between tip of anal fin and anus 1.5× the eye diameter. Caudal fin forked, upper lobe equal in length to lower lobe, tips pointed, caudal peduncle length 9.7 mm, caudal peduncle depth 6.0 mm, without adipose crests along both dorsal and ventral sides. Total vertebrae: 43 (n = 1).
Cephalic lateral line canals unclear. Lateral line complete and straight, exceeding tip of pectoral fin and reaching base of caudal fin. Two chambers of air bladder, anterior chamber dumbbell-shaped and membranous, open on both sides, slightly closed posteriorly (Fig.
In life, body coloration of Triplophysa rongduensis sp. nov. yellowish and heavily pigmented (Fig.
In one male holotype specimen (GZNU20230106001), microvillous microspinules were present on the anterior margin of the eye, outside of nostrils, upper lip, and outrostral barbel area.
Comparative data of Triplophysa rongduensis sp. nov. with T. cehengensis sp. nov. and the 34 recognized hypogean species within genus Triplophysa are given in Suppl. material
Triplophysa rongduensis sp. nov. differs from T. cehengensis sp. nov. by body pigmentation presence (vs absence), eye normal, diameter 7–15% of HL (vs eye reduced, diameter 2% of HL), dorsal fin distal margin emarginated (vs truncated), seven branched pelvic-fin rays (vs 8), tip of pectoral fin reaching to pelvic fin origin (vs not reaching to pelvic fin origin), tip of pelvic fin not reaching to anus (vs reaching to anus), and total vertebrae 43 (vs 39).
Triplophysa rongduensis sp. nov. can be distinguished from T. anshuiensis, T. langpingensis, T. qiubeiensis, T. aluensis, T. fengshanensis, T. gejiuensis, T. posterodorsalus, and T. shilinensis by body pigmentation presence (vs absence) and eye normal, diameter 7–15% of HL (vs eye reduced or absence); from T. huapingensis, T. flavicorpus, T. guizhouensis, T. longipectoralis, T. luochengensis, and T. yunnanensis by body scaleless (vs body covered by sparse scales).
Triplophysa rongduensis sp. nov. can be distinguished from T. baotianensis, T. nanpanjiangensis, T. tianxingensis, T. xiangshuingensis, and T. xichouensis by four unbranched dorsal-fin rays (vs 3) and two unbranched pectoral-fin rays (vs 1). Triplophysa rongduensis sp. nov. can be futher distinguished from T. baotianensis by four unbranched dorsal -fin rays (vs 3), four three unbranched anal -fin rays (vs 2), 16 branched caudal-fin rays (vs 11–13), and ten branched pectoral-fin rays (vs 9); from T. nanpanjiangensis, T. tianxingensis, and T. xichouensis by three unbranched anal-fin rays (vs 2); and from T. xiangshuingensis by small interorbital width, 24–29% of HL (vs 31–43% of HL), nine branched dorsal-fin rays (vs 6), ten branched pectoral-fin rays (vs 8 or 9), and 16 branched caudal-fin rays (vs 14).
Triplophysa rongduensis sp. nov. can be morphologically distinguished from its close relative T. zhenfengensis by dorsal fin distal margin emarginated (vs truncated), body scaleless (vs body covered by sparse scales), dorsal fin rays (iv, 9 vs iii, 7), ten branched pectoral-fin rays (vs 9), two unbranched pectoral-fin rays (vs 1), 16 branched caudal-fin rays (vs 14 or 15), tip of pectoral fin reaching to pelvic fin origin (vs not reaching to pelvic fin origin), and total vertebrae 43 (vs 40).
Currently, this new species Triplophysa rongduensis sp. nov. has only been found in a cave in Rongbei Village, Rongdu Town, Ceheng County, Guizhou Province, China, at an elevation of 1228 m. The pool where the new species was discovered is more than 4.5 m long, 6 m wide, and 5 m deep, with a slow flow of water, and is located ~ 30 m vertically down from the cave entrance. Inside the cave, crabs (Diyutamon cereum, 25 individuals), Sinocyclocheilus sp., and Oreolalax rhodostigmatus were found. Outside the caves, edible rape and rice are being grown. The caves in the habitat of this new species are the main source of drinking water for the local population.
The specific epithet rongduensis is in reference to the type locality of the new species: Rongdu Town, Ceheng County, Guizhou Province, China. We propose the common English name “Rongdu high-plateau loach” and the Chinese name “Rǒng Dù Gāo Yuán Qīu (冗渡高原鳅).”
Holotype. GZNU20230226002 (Fig.
Paratypes. Eleven specimens from the same locality as the holotype: GZNU20230215025–215029, GZNU20230216042–216044, GZNU20230226001–226003, and GZNU20230105001 collected by Tao Luo, Xing-Liang Wang, Ya-Li Wang, Jing Yu, and Wei-Feng Wang on February 15, 2023.
Triplophysa panzhouensis sp. nov. can be distinguished from species of the hypogean group of its congeners by the following combination of characteristics: (1) body naked, scaleless, with skin pigmentation; (2) eyes normal, diameter 7–11% of HL, interorbital width 22–31% of HL; (3) dorsal fin distal margin truncated; (4) pelvic-fin tip not reaching to anus; (5) tip of pectoral-fin not reaching to pelvic fin origin; (6) anterior and posterior nostrils closely set, anterior nostril elongated to a barbel-like tip; (7) tip of outrostral barbel extending backward to anterior margin of the eye; (8) lateral line complete; (9) dorsal-fin rays iv-7–8, pectoral-fin rays i-11, pelvic-fin rays ii-7, anal-fin rays iii-5, 16 branched caudal-fin rays; and (10) total vertebrae 39.
Morphological data of the specimens of the Triplophysa panzhouensis sp. nov. are provided in Suppl. materials
Dorsal-fin rays iv-7–8, pectoral-fin rays i-11, pelvic-fin rays ii-7, anal-fin rays iii-5, 16 branched caudal-fin rays. Dorsal fin long, length 17–21% of HL, distally margin truncated, origin slightly anterior to pelvic-fin insertion, situated slightly posterior to midpoint between snout tip and caudal-fin base, first branched ray longest, shorter than head length, tip of dorsal fin not extending to vertical limit of anus. Pectoral fin moderately developed, length 15–20% of SL, tip of pectoral fin extends backwards not to the midpoint between origins of pectoral and pelvic fins, not reaching origin of pelvic fin. Pelvic fin length 13–17% of SL, vertically aligned with first branched ray of dorsal fin, tips of pelvic fin not reaching to anus, distance between tips of pelvic fin and anus 3.7× the eye diameter. Anal fin long, length 13–17% of SL, distal margin truncated, origin close to anus, spacing 4.6 mm, tips of anal fin not reaching caudal-fin base, distance between tips of anal fin and anus 3.1× the eye diameter. Caudal fin forked, upper lobe is equal in length to lower lobe, tips pointed, caudal peduncle length 12.0 mm, caudal peduncle depth 5.4 mm, without adipose crests along both dorsal and ventral sides. Total vertebrae 39 (n = 1).
Cephalic lateral line canals unclear. Lateral line complete and straight, exceeding tip of pectoral fin and reaching base of caudal fin. Two chambers of air bladder, anterior chamber dumbbell-shaped and membranous, open on both sides, slightly larger pores posteriorly (Fig.
In cave water bodies when alive, Triplophysa panzhouensis sp. nov. has light yellow skin with irregular light brown patches on the body (Fig.
No secondary sex characteristics was observed based on the present specimens of Triplophysa panzhouensis sp. nov.
Comparative data of Triplophysa panzhouensis sp. nov. with T. cehengensis sp. nov., T. rongduensis sp. nov., and the 34 recognized hypogean species within genus Triplophysa are given in Suppl. material
Triplophysa panzhouensis sp. nov. differs from T. cehengensis sp. nov. by body pigmentation present (vs body pigmentation absent), eyes normal, diameter 7–11% of HL (vs eye reduced, diameter 2% of HL), dorsal fin distal margin truncated (vs emarginated), 7 branched pelvic-fin rays (vs 8), and tip of pelvic fin not reaching to anus (vs reaching to anus). Triplophysa panzhouensis sp. nov. differs from T. rongduensis sp. nov. by having nine branched dorsal-fin rays (vs 7 or 8), total vertebrae 39 (vs 43), and tip of outrostral barbel extending backward to anterior margin of the eye (vs extending backward to posterior margin of the eye).
Triplophysa panzhouensis sp. nov. can be distinguished from T. langpingensis, T. qiubeiensis, T. anshuiensis, T. aluensis, T. fengshanensis, T. gejiuensis, T. posterodorsalus, and T. shilinensis by body pigmentation presence (vs absence) and eye normal, diameter 7–11% HL(vs eye reduced or absence); from T. zhenfengensis, T. huapingensis, T. flavicorpus, T. guizhouensis, T. longipectoralis, T. luochengensis, and T. yunnanensis by body scaleless (vs body covered by sparse scales) and four unbranched dorsal-fin rays (vs 3).
Triplophysa panzhouensis sp. nov. can be distinguished from T. baotianensis, T. flavicorpus, T. nanpanjiangensis, T. tianxingensis, T. xiangshuingensis, and T. xichouensis by four unbranched dorsal-fin rays (vs 3) and two unbranched pectoral-fin rays (vs 1). Triplophysa panzhouensis sp. nov. can be futher distinguished from T. baotianensis by four unbranched dorsal-fin rays (vs 3), and 11 branched pectoral-fin rays (vs 9); from T. flavicorpus by interorbital width (vs 22.1–31.3% of HL vs 3.1–5.2% of HL), dorsal fin distal margin truncated (vs emarginated), and seven or eight branched dorsal-fin rays (vs 10); from T. nanpanjiangensis, T. tianxingensis, and T. xichouensis by three unbranched anal-fin rays (vs 2) and total vertebrae 39 (vs 40–42); and from T. xiangshuingensis by seven or eight branched dorsal-fin rays (vs 6), 11 branched pectoral-fin rays (vs 8 or 9), and 16 branched caudal-fin rays (vs 14).
Triplophysa panzhouensis sp. nov. is only known from the type locality, a vertical cave ~ 3 km from Hongguo Town, Panzhou city, Guizhou, China at an elevation of 2276 m. The cave was completely dark. Individuals of T. panzhouensis sp. nov. were located in a small pool ~ 25 m from the cave entrance. The pool was ~ 1.8 m wide and 80 cm deep, with a water temperature of ~ 16 °C at the time of collection and a water pH of 7.4. Within this cave, T. panzhouensis sp. nov. co-occurred with Sinocyclocheilus longicornus, Sinocyclocheilus sp., and Oreolalax rhodostigmatus. The arable land outside the cave was farmed to produce maize, wheat, and potatoes.
The specific epithet panzhouensis is in reference to the type locality of the new species: Hongguo Town, Panzhou City, Guizhou Province, China. We propose the common English name “Panzhou high-plateau loach” and the Chinese name “Pán Zhõu Gāo Yuán Qīu (盘州高原鳅).”
Holotype. GZNU20230215021 (Fig.
Paratypes. Six specimens from the same locality as the holotype: GZNU20230215021–215024, GZNU20230226004, and GZNU20230226005, collected by Tao Luo, Wei-Feng Wang, Jing Yu, Chang-Ting Lan, and Xin-Rui Zhao on February 15, 2023.
Triplophysa anlongensis sp. nov. can be distinguished from all other congeners by the following combination of characteristics: (1) body naked, scaleless, without skin pigmentation; (2) eyes normal, diameter 5–9% of HL, interorbital width 32–36% of HL; (3) dorsal fin distal margin truncated; (4) pelvic-fin tip not reaching to anus; (5) tip of pectoral fin not reaching to pelvic fin origin; (6) anterior and posterior nostrils closely set, anterior nostril elongated to a barbel-like tip; (7) tip of outrostral barbe backward extending to middle of the eye; (8) lateral line complete; (9) dorsal-fin rays iii-8, pectoral-fin rays i-11, pelvic-fin rays ii-8, anal-fin rays iii-5, and 16 branched caudal-fin rays; and (10) total vertebrae 41.
Morphological data of the six specimens of the Triplophysa anlongensis sp. nov. are provided in Suppl. materials
Dorsal-fin rays iii-8, pectoral-fin rays i-11, pelvic-fin rays ii-8, anal-fin rays iii-5, and 16 branched caudal-fin rays. Dorsal fin long, length 18–22% of HL, distally margin truncated, origin slight anterior to pelvic-fin insertion, situated slightly posterior to midpoint between snout tip and caudal-fin base, first branched ray longest, shorter than head length, tip of dorsal fin not extending to vertical of anus. Pectoral fin moderately developed, length 16–22% of SL, tip of pectoral fin extends backwards not reaching the midpoint between the pectoral and pelvic fin origins. Pelvic fin length 14–16% of SL, vertically aligned with third branched ray of dorsal fin, tips of pelvic fin not reaching to anus, distance between tips of pelvic fin and anus 1.5× the eye diameter. Anal fin long, length 13–17% of SL, distally margin truncated, origin close to anus, spacing 1.3 mm, tips of anal fin backwards not reaching caudal-fin base, distance between tips of anal fin and anus 1.8× eye diameter. Caudal fin forked, upper lobe equal in length to lower lobe, tips pointed, caudal peduncle length 7.7 mm, caudal peduncle depth 4.5 mm, without adipose crests along both dorsal and ventral sides. Total vertebrae 41 (n = 1).
Cephalic lateral line canals unclear. Lateral line complete and straight, exceeding tip of pectoral fin and reaching base of caudal fin. Two chambers of air bladder, anterior chamber dumbbell-shaped and membranous, open on both sides, slightly closed posteriorly (Fig.
In cave water bodies when alive, Triplophysa anlongensis sp. nov. has the ground color of body light yellow, slightly lighter ventrally. Dorsal and lateral parts of body and head gray and black. Dorsal, pectoral, pelvic, and anal fin rays brown, fin membrane hyaline. Blotches present on dorsal, pectoral, and caudal fins. Pelvic and anal fins without blotches (Fig.
No secondary sex characteristics was observed based on the present specimens of Triplophysa anlongensis sp. nov.
Comparative data of Triplophysa anlongensis sp. nov. with T. cehengensis sp. nov., T. rongduensis sp. nov., Triplophysa panzhouensis sp. nov. and the 34 recognized hypogean species within genus Triplophysa are given in Suppl. material
Triplophysa anlongensis sp. nov. differs from T. cehengensis sp. nov. By body pigmentation present (vs body pigmentation absent), eyes normal, diameter 5–9% of HL (vs eyes reduced, diameter 2% of HL), dorsal-fin rays (iii, 8 vs iv, 9), and tip of pelvic fin not reaching to anus (vs reaching to anus); from T. rongduensis sp. nov. by seven branched pelvic-fin rays (vs 6) and large interorbital width, 32–36% of HL (vs 24–29 of HL); and from Triplophysa panzhouensis sp. nov. by total vertebrae 41 (vs 39), dorsal fin rays (iii, 8 vs iv, 7–8), and eight branched pelvic fin rays (vs 7).
Triplophysa anlongensis sp. nov. can be distinguished from T. langpingensis, T. qiubeiensis, T. anshuiensis, T. fengshanensis, T. gejiuensis, T. posterodorsalus, T. shilinensis, and T. xichouensis body pigmentation presence (vs absence). Triplophysa anlongensis sp. nov. can be distinguished from T. flavicorpus, T. longipectoralis, T. xiangshuingensis, and T. yunnanensis by dorsal fin distal margin truncated (vs emarginated). Triplophysa anlongensis sp. nov. differs from T. zhenfengensis, T. huapingensis, T. guizhouensis, and T. luochengensis by body scaleless (vs body covered by sparse scales). Triplophysa anlongensis sp. nov. can be further distinguished from T. langpingensis, T. qiubeiensis, T. anshuiensis, T. flavicorpus, and T. longipectoralis by tip of pelvic fin not reaching to anus (vs reaching to anus).
Triplophysa anlongensis sp. nov. differs from T. baotianensis by slight small eyes, diameter 5–9% of HL (vs 9–13% of HL), large interorbital width, 32–36% of HL (vs 3–5% of HL), eight branched dorsal fin rays (vs 6 or 7), three unbranched anal-fin rays (vs 2), 11 branched pectoral-fin rays (vs 9), pelvic-fin rays (ii, 8 vs i, 6 or 7), and six branched caudal-fin rays (vs 11–13); from T. tianxingensis by anterior nostril elongated to a short barbel-like tip (vs not elongated to a short barbel-like tip), three unbranched anal-fin rays (vs 2), 11 branched pectoral-fin rays (vs 9), pelvic-fin rays (ii, 8 vs i, 5–7), and total vertebrae 41 (vs 42); and from T. xichouensis by anal-fin rays (iii, 5 vs ii, 6), 11 branched pectoral-fin rays (vs 9 or 10), pelvic-fin rays (ii, 8 vs i, 5 or 6), tip of pelvic fin not reaching to anus (vs reaching to anus), and total vertebrae 41 (vs 40).
Triplophysa anlongensis sp. nov. is only known from the type locality, a vertical cave ~ 3 km from NaNao Village, Xinglong Town, Anlong County, Guizhou Province, China at an elevation of 1387 m. There is no surface stream outside the cave. The cave is approximately 50 m long and has a small volume of water during dry periods, although it is a source of domestic water for the local population. Within this cave, Triplophysa anlongensis sp. nov. co-occurred with fish (Balitora anlongensis and Misgurnus anguillicaudatus), frogs (Odorrana sp.), the red-eared slider (Trachemys scripta), and crabs (Diyutamon cereum). Outside the cave, the arable land was farmed to produce maize, wheat, and potatoes.
The specific epithet anlongensis is in reference to the type locality of the new species: NaNao Village, Xinglong Town, Anlong County, Guizhou Province, China. We propose the common English name “Anlong high-plateau loach” and the Chinese name “ān lóng Gāo Yuán Qīu (安龙高原鳅).”
In this work, we have described four new species named T. cehengensis sp. nov., T. rongduensis sp. nov., T. panzhouensis sp. nov., and T. anlongensis sp. nov. based on morphological comparisons and mitochondrial DNA sequence differences (Fig.
Cave-dwelling species of the genus Triplophysa distributed in the karst region of southwest China, including Guangxi, Guizhou, Yunnan, Chongqing, and Hunan provinces (Fig.
The Yangtze River system (seven species) has a low number of species with highly specialized phenotypes (
We thank Wei-Feng Wang, Li Wu, Cui Fan, Zhi-Xia Chen, and others for help with specimen collections. We thank LetPub (www.letpub.com) for its linguistic assistance during manuscript preparation.
The authors have declared that no competing interests exist.
No ethical statement was reported.
This research was supported by the Guizhou Province Top Discipline Construction Program Project (Qianjiao Keyan Fa [2019] 125), the Guizhou Normal University Academic Emerging Talent Fund Project (Qianshi Xin Miao [2021] 20), the programs of the Strategic Priority Research Program B of the Chinese Academy of Sciences (CAS) (No. XDB31000000), and the Postgraduate Education Innovation Programme of Guizhou Province (No. Qianjiaohe YJSKYJJ [2021] 091).
Jiang Zhou conceived and supervised the project. Tao Luo, Ming-Le Mao, Xin-Rui Zhao, Jing Yu, Chang-Ting Lan, Jia-Jun Zhou, and Xing-Liang performed the sample collection and DNA preparation. Tao Luo and Ning Xiao performed the data analyses. Tao Luo, Ming-Le Mao, Xin-Rui Zhao, Ling-Xing Song, Jing Yu, and Chang-Ting Lan wrote the manuscript with input from Jiang Zhou.
Tao Luo https://orcid.org/0000-0003-4186-1192
Ming-Le Mao https://orcid.org/0000-0001-9764-4731
Chang-Ting Lan https://orcid.org/0009-0007-2381-3601
Ling-Xing Song https://orcid.org/0009-0007-2897-1852
Xin-Rui Zhao https://orcid.org/0000-0002-9125-6276
Jing Yu https://orcid.org/0009-0004-3629-3826
Xing-Liang Wang https://orcid.org/0009-0003-0654-8640
Ning Xiao https://orcid.org/0000-0002-7240-6726
Jia-Jun Zhou https://orcid.org/0000-0003-1038-1540
Jiang Zhou https://orcid.org/0000-0003-1560-8759
All of the data that support the findings of this study are available in the main text or Supplementary Information.
Specimens examined in this work
Triplophysa baotianensis
(n = 5): China: Guizhou: Panzhou City: Baotian Town (type locality):
Triplophysa erythraea (n = 2): China: Hunan: Huayuan County: Dalong Cave (type locality): GZNU20230216052 and GZNU20230216053.
Triplophysa nasobarbatula
(n = 8): China: Guizhou: Libo County: Dongtang Township (type locality):
Triplophysa guizhouensis
(n = 6): China: Guizhou: Huisui County: Baijin Town (type locality):
Triplophysa zhenfengensis (n = 5): China: Guizhou: Xingren City: Xinlongchang Town: GZNU20180419002, GZNU20190521001–052104.
Triplophysa langpingensis
(n = 2): China: Guangxi: Leye County: Huaping Township (type locality):
Triplophysa macrocephala
(n = 8): China: Guangxi: Nandan County: Lihu Township (type locality):
Triplophysa tianeensis
(n = 7): China: Guangxi: Tian’e County: Bala Township (type locality):
Triplophysa huapingensis
(n = 7): China: Guangxi: Leye County: Huaping Township (type locality):
Triplophysa nandanensis
(n = 2): China: Guangxi: Nandan County: Liuzhai Town (type locality):
Triplophysa qiubeiensis
(n = 2): China: Yunan: Qiubei County: Nijiao Town (type locality):
Triplophysa qingzhenensis (n = 8): China: Guizhou: Guiyang City: Qingzhen County (type locality): GZNU20220830001–0830008.
Triplophysa qini (n = 5): China: Chongqing: Wulong District: Fengdu County: Dudu Village (type locality): GZNU20230216045–0216045.
Triplophysa wudangensis (n = 4): China: Guizhou: Guiyang City: Wudang District (type locality): IHB 201908090001–090004.
Triplophysa rosa (n = 9): China: Chongqing: Wulong County: Huolu Town: GZNU20230216070–0216078.
Boxplots of morphometrics based on 35 morphometric measurements distinguishing T. anlongensis sp. nov., T. cehengensis sp. nov., T. panzhouensis sp. nov., T. rongduensis sp. nov., T. baotianensis, and T. zhenfengensis
Data type: png
GPS information on the geographical distribution of 39 hypogean species of the genus Triplophysa
Data type: xlsx
Morphological characters and measurement data
Data type: xlsx
Explanation note: Morphological characters and measurement data of the new species Triplophysa cehengensis sp. nov., Triplophysa rongduensis sp. nov., Triplophysa panzhouensis sp. nov., Triplophysa anlongensis sp. nov., T. zhenfengensis, and T. baotianensis. *Designates the holotype.
Comparison of the diagnostic features of the four new species described here with those selected for the 35 recognized hypogean species of the genus Triplophysa
Data type: xlsx
Explanation note: Modified from Wu et al. (2018) and
Morphological comparison of T. cehengensis sp. nov. (TC), T. rongduensis sp. nov. (TR), T. panzhouensis sp. nov. (TP), T. anlongensis sp. nov. (TA), T. baotianensis (TB), and T. zhenfengensis (TZ)
Data type: docx
Explanation note: All units in mm. P-values are at the 95% significance level.
Results and percentage of variance explained by principal component and discriminant function analyses
Data type: xlsx