Research Article |
Corresponding author: Jiang Zhou ( zhoujiang@ioz.ac.cn ) Academic editor: Annemarie Ohler
© 2021 Tao Luo, Yali Wang, Siwei Wang, Xueli Lu, Weifeng Wang, Huaiqing Deng, 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, Wang Y, Wang S, Lu X, Wang W, Deng H, Zhou J (2021) A species of the genus Panophrys (Anura, Megophryidae) from southeastern Guizhou Province, China. ZooKeys 1047: 27-60. https://doi.org/10.3897/zookeys.1047.61097
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Herein, we describe Panophrys congjiangensis sp. nov. obtained from the Yueliangshan Nature Reserve, Congjiang County, Guizhou Province, China. Phylogenetic analyses based on the mitochondrial genes 16S rRNA and COI indicated that this new species represented an independent lineage, closely related to P. leishanensis. The uncorrected genetic distances between the new species and its closest congener, P. leishanensis, were 3.0% for 16S rRNA and 8.4% for COI. The new species is distinguished from its congeners by a combination of the following morphological characteristics (1) medium body size (SVL 28.6–33.4 mm in males and 38.4–40.2 mm in females); (2) a small horn-like tubercle at the edge of each upper eyelid; (3) the tympanum distinctly visible (TD/ED ratio 0.47–0.66); (4) vomerine teeth absent; (5) the tongue not notched behind; (6) a narrow and unobvious lateral fringe on toes; (7) relative finger lengths II < I < V < III; (8) rudimentary webs on toes; (9) hindlimbs slender, heels overlapping when thighs are positioned at right angles to the body; (10) two metacarpal tubercles on the palm, with the inner metatarsal tubercle long and oval-shaped; (11) the tibiotarsal articulation reaching the nostril when the leg is adpressed and stretched forward; (12) dorsal skin rough with numerous orange–red granules, ventral surface smooth; (13) a single internal subgular vocal sac present in males; and (14) in breeding males, weak gray-black nuptial pads with black nuptial spines present on the dorsal surface of the bases of the first and second fingers. To date, the new species is only known from the type locality.
Morphology, new species, Panophrys, southwest China, taxonomy
The Asian horned toad subfamily Megophryinae (Bonaparte, 1850) is widely distributed in southern China, the southern and eastern Himalayas, and across Indochina to the islands of the Sunda Shelf and the Philippines (
In the most recent revision, 59 species were assigned to the genus Panophrys (
During herpetological surveys conducted between 2019 and 2020 in Yueliangshan Nature Reserve, Congjiang County, Guizhou Province, China (Fig.
A total of 25 specimens were collected in this study: 22 were collected in Congjiang County, Guizhou Province, China, and were identified as an unknown species. The remaining 3 specimens, collected in Kuankuoshui National Nature Reserve, Suiyang County, Guizhou Province, China, were identified as P. jiangi. All specimens were fixed in 10% buffered formalin and later transferred to 75% ethanol for preservation. The muscle samples used for molecular analysis were preserved in 95% alcohol and stored at -20 °C. All specimens are housed at Guizhou Normal University (GZNU), Guiyang City, Guizhou Province, China.
Genomic DNA was extracted from the muscle tissue samples using DNA extraction kits (Tiangen Biotech (Beijing) Co., Ltd.). We amplified and sequenced two mitochondrial genes from each DNA sample: partial 16S ribosomal RNA (16S rRNA), using primers L3975 (5’-CGCCTGTTTACCAAAAACAT-3’) and H4551 (5’-CCGGTCTGAACTCAGATCACGT-3’) following
Localities, voucher information, and GenBank numbers for all samples used in this study.
ID | Species | Locality | Voucher number | Genbank accession No. | |
---|---|---|---|---|---|
16S rRNA | COI | ||||
1 | Panophrys obesa | Heishiding Nature Reserve, Guangdong, China | SYS a002271 | KJ579121 | MH406123 |
2 | Panophrys ombrophila | Wuyi Shan, Fujian, China | WUYI2015101 | KX856397 | – |
3 | Panophrys cheni | Taoyuandong Nature Reserve, Hunan, China | SYS a002123 | KJ560396 | MF667904 |
4 | Panophrys dongguanensis | Yinping Shan, Guangdong, China | SYS a002007 | MH406654 | MH406090 |
5 | Panophrys nankunensis | Nankun Shan, Guangdong, China | SYS a004498 | MK524108 | MK524139 |
6 | Panophrys wugongensis | Wugongshan Scenic Area, Jiangxi, China | SYS a002610 | MK524114 | MK524145 |
7 | Panophrys insularis | Nan’ao Island, Guangdong, China | SYS a002169 | MF667887 | MF667924 |
8 | Panophrys lini | Nanfengmian Nature Reserve, Jiangxi, China | SYS a002128 | KJ560416 | MF667907 |
9 | Panophrys nanlingensis | Nanling Nature Reserve, Guangdong, China | SYS a001959 | MK524111 | MK524142 |
10 | Panophrys xiangnanensis | Yangming Shan, Hunan, China | SYS a002875 | MH406714 | MH406166 |
11 | Panophrys baishanzuensis | Baishanzu National Park, Qingyuan, Zhejiang, China |
CIBQY20200719001 | MW001150 | MT998291 |
12 | Panophrys brachykolos | Hongkong, China | SYS a005563 | MK524122 | MK524153 |
13 | Panophrys kuatunensis | Wuyi Shan, Jiangxi, China | SYS a003449 | MF667881 | MF667916 |
14 | Panophrys lishuiensis | Lishui City, Zhejiang, China | CIBWYF00169 | KY021418 | – |
15 | Panophrys xianjuensis | Xianju, County, Zhejiang, China | CIBXJ20190801 | MN563754 | MN563770 |
16 | Panophrys jinggangensis | Jinggang Shan, Jiangxi, China | SYS a004028 | MH406780 | MH406239 |
17 | Panophrys liboensis | Libo Country, Guizhou, China | GZNU20150813001 | MF285253 | MW959767 |
18 | Libo Country, Guizhou, China | GZNU20160408007 | MF285258 | MW959768 | |
19 | Libo Country, Guizhou, China | GZNU20160408006 | MF285257 | MW959769 | |
20 | Libo Country, Guizhou, China | GZNU20160408004 | MF285256 | MW959770 | |
21 | Panophrys boettgeri | Longhu Forest Station, Fujian, China | SYS a004126 | MH406785 | MH406245 |
22 | Panophrys huangshanensis | Huang Shan, Anhui, China | SYS a002702 | MF667882 | MF667919 |
23 | Panophrys congjiangensis sp. nov. | Yueliangshan Nature Reserve, Congjiang, Guizhou, China | GZNU20200706003 | MW959773 | MW959761 |
24 | Yueliangshan Nature Reserve, Congjiang, Guizhou, China | GZNU20200706004 | MW959774 | MW959762 | |
25 | Yueliangshan Nature Reserve, Congjiang, Guizhou, China | GZNU20200706005 | MW959775 | MW959763 | |
26 | Yueliangshan Nature Reserve, Congjiang, Guizhou, China | GZNU20200706000 | MW959776 | MW959764 | |
27 | Panophrys leishanensis | Leigong Shan, Guizhou, China | CIBLS20141004003 | MK005308 | MK005304 |
28 | Leigong Shan, Guizhou, China | SYSa002213 | MH406673 | MH406113 | |
29 | Leigong Shan, Guizhou, China | CIBLS20160610005 | MK005309 | MK005305 | |
30 | Panophrys baolongensis | Baolong, Chongqing, China | KIZ019216 | KX811813 | KX812093 |
31 | Panophrys wushanensis | Shennongjia Nature Reserve, Hubei, China | SYS a003008 | MH406732 | MH406184 |
32 | Panophrys tuberogranulata | Badagong Shan, Hunan, China | SYS a004310 | MH406801 | MH406263 |
33 | Panophrys shimentaina | Shimentai Nature Reserve, Guangdong, China | SYS a002078 | MH406656 | MH406093 |
34 | Panophrys yangmingensis | Yangming Shan, Hunan, China | SYS a002889 | MH406720 | MH406172 |
35 | Panophrys jiulianensis | Nankun Shan, Guangdong, China | SYS a003623 | MK524103 | MK524134 |
36 | Panophrys mirabilis | Huaping Nature Reserve, Guangxi, China | SYS a002193 | MH406670 | MH406110 |
37 | Panophrys shunhuangensis | Nanshan National Forest Park, Hunan, China | HNNU18NS01 | MK836023 | MK977594 |
38 | Panophrys acuta | Heishiding Nature Reserve, Guangdong, China | SYS a002159 | MF667869 | MF667899 |
39 | Panophrys mufumontana | Mufu Shan, Hunan, China | SYS a006390/CIB110012 | MK524104 | MK524135 |
40 | Panophrys caudoprocta | Badagong Shan, Hunan, China | SYS a004281 | MH406795 | MH406257 |
41 | Panophrys sangzhiensis | Badagong Shan, Hunan, China | SYS a004307 | MH406798 | MH406260 |
42 | Panophrys spinata | Leigong Shan, Guizhou, China | SYS a002226 | MH406675 | MH406115 |
43 | Panophrys qianbeiensis | Huanglian Nature Reserve, Guizhou, China | CIBTZ20190608015 | MT651553 | MT654520 |
44 | Panophrys binlingensis | Wawu Shan, Sichuan, China | SYS a005313 | MH406892 | MH406354 |
45 | Panophrys binchuanensis | Jizu Shan, Yunnan, China | KIZ019441 | KX811849 | KX812112 |
46 | Panophrys angka | Kiew Mae Pan nature trail, Chiang Mai, Thailand |
KIZ040591 | MN508052 | – |
47 | Panophrys anlongensis | Anlong County, Guizhou, China | CIBAL20190531018 | MT823184 | MT823261 |
48 | Panophrys omeimontis | Laojun Shan, Sichuan, China | SYS a002741 | MH406710 | MH406162 |
49 | Panophrys palpebralespinosa | Pu Hu Nature Reserve, Thanh Hoa, Vietnam | KIZ011603 | KX811888 | KX812137 |
50 | Panophrys caobangensis | Nguyen Binh, Cao Bang,Vietnam | IEBR 4385 | LC483945 | – |
51 | Panophrys daweimontis | Dawei Shan, Yunnan, China | KIZ048997 | KX811867 | KX812125 |
52 | Panophrys jingdongensis | Wuliang Shan, Yunnan, China | SYS a003928 | MH406773 | MH406232 |
53 | Panophrys rubrimera | Lao Cai, Sa Pa, Vietnam | AMSR177676 | MF536419 | – |
54 | Panophrys wuliangshanensis | Wuliang Shan, Yunnan, China | SYS a003924 | MH406771 | MH406230 |
55 | Panophrys fansipanensis | Lao Cai, Sa Pa, Vietnam | VNMN 2018.01 | MH514886 | – |
56 | Panophrys hoanglienensis | Lao Cai, Sa Pa, Vietnam | VNMN 2018.02 | MH514889 | – |
57 | Panophrys jiangi | Huoqiuba Nature Reserve, Guizhou, China | GZNU20180606020 | MW959777 | MW959765 |
58 | Kuankuosui Nature Reserve, Guizhou, China | GZNU20070712001 | MW959778 | MW959766 | |
59 | Kuankuosui Nature Reserve, Guizhou, China | CIBKKS20180722006 | MN107743 | MN107748 | |
60 | Panophrys minor | Qingcheng Shan, Sichuan, China | SYS a003209 | MF667862 | MF667891 |
61 | Panophrys chishuiensis | Chishui, County, Guizhou, China | CIBCS20190518031 | MN954707 | MN928958 |
62 | Ophryophryne pachyproctus | Beibeng, Xizang, China | KIZ010978 | KX811908 | KX812153 |
63 | Panophrys yeae | Didong, Medog, Tibet, China | CIB201706MT01 | MN963217 | MN964312 |
64 | Panophrys zhoui | Renqinbeng, Medog, Tibet, China | CIBMT171053 | MN963207 | MN964322 |
65 | Xenophrys vegrandis | West Kameng district, Arunachal Pradesh, India | ZSI A 11605 | KY022305 | MH647530 |
66 | Ophryophryne elfina | Bidoup Mountain, Lam Dong, Vietnam | ZMMU ABV000454 | KY425379 | – |
67 | Ophryophryne gerti | Nui Chua National Park, Ninh Thuan, Vietnam | ITBCZ 1108 | KX811917 | KX812161 |
68 | Ophryophryne synoria | O’Reang, Mondolkiri, Cambodia | FMNH 262779 | MN629394 | – |
69 | Ophryophryne hansi | Phong Dien Nature Reserve, Thua Thien Hue, Vietnam | KIZ010360 | KX811913 | KX812155 |
70 | Ophryophryne microstoma | Wuhuang Shan,, Guangxi, China | SYS a003492 | MK524125 | MK524156 |
71 | Megophrys montana | Sukabumi, Java, Indonesia | LSUMZ 81916 | KX811927 | KX812163 |
72 | Megophrys parallela | – | RMAS 021 | KY679897 | – |
73 | Megophrys lancip | Ulubelu, Ngarip, Indonesia | MZB:Amp:22233 | KY679891 | – |
74 | Xenophrys medogensis | Medog County, Tibet, China | SYS a002932 | MH406725 | MH406177 |
75 | Xenophrys robusta | Darjeeling dist, West Bengal, India | SDBDU 2011.1057 | KY022314 | MH647535 |
76 | Xenophrys glandulosa | Gaoligong Shan,, Yunnan, China | SYS a003758 | MH406755 | MH406214 |
77 | Xenophrys himalayana | East Siang dist, Arunachal Pradesh, India | SDBDU2009.75 | KY022311 | |
78 | Xenophrys periosa | East Siang dist, Arunachal Pradesh, India | BNHS 6061 | KY022309 | MH647528 |
79 | Xenophrys monticola | Darjeeling dist, West Bengal, India | SDBDU 2011.1047 | KX894679 | |
80 | Xenophrys zhangi | Zhangmu, Xizang, China | KIZ014278 | KX811765 | KX812084 |
81 | Xenophrys flavipunctata | Hills dist, East Khasi, Meghalaya | SDBDU2009.297 | KY022307 | MH647536 |
82 | Xenophrys mangshanensis | Longtou glandulosa, Guangdong, China | SYS a002750 | MF667866 | MF667895 |
83 | Xenophrys maosonensis | Xiaoqiaogou Nature Reserve, Yunnan, China | KIZ016045 | KX811780 | KX812080 |
84 | Xenophrys oreocrypta | West Garo Hills dist, Meghalaya | BNHS 6046 | KY022306 | – |
85 | Xenophrys major | Zhushihe, Yunnan, China | SYSa002961 | MH406728 | MH406180 |
86 | Xenophrys awuh | – | SDBDU2007.161 | KY022319 | – |
87 | Xenophrys serchhipii | North dist, Tripura, India | SDBDU 2009.612 | KY022323 | MH647532 |
88 | Xenophrys zunhebotoensis | – | SDBDU 2009.110 | KY022321 | |
89 | Xenophrys ancrae | Changlang dist, Arunachal Pradesh, India | SDBDU 2009.727 | KY022318 | MH647531 |
90 | Xenophrys numhbumaeng | – | SDBDU 2007.041 | KY022316 | |
91 | Xenophrys oropedion | Mawphlang Sacred Forest, Meghalaya, India | SDBDU 2009.299 | KY022317 | MH647534 |
92 | Xenophrys megacephala | – | ZSI A 11213 | KY022315 | MH647533 |
93 | Xenophrys dzukou | – | SDBDU2007.106 | KY022324 | – |
94 | Xenophrys lekaguli | Pang Si Da National Park, Sa Kaeo, Thailand | FMNH 265955 | KY022214 | – |
95 | Xenophrys takensis | – | FMNH 261711 | KY022215 | – |
96 | Xenophrys auralensis | Aural, Kampong Speu, Cambodia | NCSM 79599 | KX811807 | – |
97 | Xenophrys parva | Zhushihe, Yunnan, China | SYSa003042 | MH406737 | MH406189 |
98 | Xenophrys aceras | Khao Nan National Park, Nakhon Si Thammarat, Thailand | KIZ025467 | KX811925 | KX812159 |
99 | Xenophryslongipes | Genting highland, Malaysia | IABHU 21101 | AB530656 | – |
100 | Atympanophrys gigantica | Ailao Shan, Yunnan, China | SYS a003883 | MH406766 | MH406225 |
101 | Atympanophrys shapingensis | Wawu Shan, Sichuan, China | SYS a005310 | MH406890 | MH406352 |
102 | Atympanophrys nankiangensis | Nanjiang, Sichuan, China | CIB ZYC517 | KX811900 | – |
103 | Atympanophrys wawuensis | Wawu Shan, Sichuan, China | KIZ025799 | KX811902 | KX812062 |
104 | Brachytarsophrys feae | Huangcaoling, Yunnan, China | KIZ046706 | KX811810 | KX812056 |
105 | Brachytarsophrys platyparietus | – | W01395 | AY526206 | – |
106 | Brachytarsophrys chuannanensis | Hejiang County, Sichuan, China | SYS a004926 | MH406901 | MH406364 |
107 | Brachytarsophrys carinense | Dayao Shan, Guangxi, China | Tissue ID: YPX20455 | KX811811 | KX812057 |
108 | Brachytarsophrys popei | Jinggang Shan, Jiangxi, China | SYS a004209 | MK524124 | MK524155 |
109 | Brachytarsophrys intermedia | Phong Nha0Ke Bang NP, U Bo, Vietnam | ZFMK 87596 | HQ588950 | – |
110 | “Megophrys”dringi | Mulu National Park, Sarawak Gunung, Malaysia | UNIMAS 8943 | KJ831317 | – |
111 | Pelobatrachus baluensis | Gunung Kinabalu National Park, Kogopan Trail, Malaysia | ZMH A13125 | KJ831310 | – |
112 | Pelobatrachus stejnegeri | Pasonanca Natural Park, Zamboanga, Philippines | KU 314303 | KX811922 | KX812052 |
113 | Pelobatrachus kobayashii | Gunung, Sabah, Meghalaya | UNIMAS 8148 | KJ831313 | – |
114 | Pelobatrachus ligayae | Palawan, Philippines | ZMMU NAP005015 | KX811919 | KX812051 |
115 | Pelobatrachus kalimantanensis | Kalimantan Selatan, Borneo, Indonesia | MZB. Amph 21482 | MG993554 | – |
116 | Pelobatrachus nasuta | Sabah, Lahad Datu District, Malaysia | FMNH 231281 | KY022186 | – |
117 | Pelobatrachus edwardinae | Bintulu, Sarawak, Malaysia | FMNH 273694 | KX811918 | KX812050 |
118 | Leptobrachium boringii | Emei Shan, Sichuan, China | Tissue ID: YPX37539 | KX811930 | KX812164 |
119 | Leptobrachella oshanensis | Emei Shan, Sichuan, China | KIZ025778 | KX811928 | KX812166 |
We used a total of 194 gene sequences (112 16S rRNA sequences and 82 COI sequences) for the molecular analyses, representing 102 species of subfamily Megophryinae. Two mitochondrial genes were sequenced in 10 muscle tissue samples from the specimens collected in this study, and 178 sequences were downloaded from GenBank. Samples included those from the undescribed species collected and named in this study (Fig.
Sampling collection localities and distribution of the Panophrys congjiangensis sp. nov., P. leishanensis, P. liboensis, and P. jiangi in Guizhou Province, China 1 Yueliangshan Nature Reserve, Congjiang County, Guizhou Province 2 Leigongshan National Nature Reserve, Leishan County, Guizhou Province 3 Maolan National Nature Reserve, Libo County, Guizhou Province 4, 5 Huoqiuba Nature Reserve, Suiyang County, Guizhou Province 6 Xingxiu Township, Dafang County, Guizhou Province 7 Fenghuang Township, Shuicheng County, Guizhou Province. The base maps are from Standard Map Service website (http://bzdt.ch.mnr.gov.cn/index.html).
All sequences were assembled and aligned using the MUSCLE (
To assess whether new species represent a valid species, two different methods were executed. We chose to include new species in the phylogenetic tree as well as several closely related species for species delimitation analysis. First, a Bayesian hypothesis-testing approach (Bayes Factor Delimitation, BFD) was implemented to statistically test alternate hypotheses of species delimitation (
In addition to the Bayesian methods tested, we also applied three tree-based species-delimitation methods, i.e., Bayesian implementation of the Poisson Tree Processes model (bPTP;
Measurements of the adult specimens of Panophrys congjiangensis sp. nov. All units in mm. See abbreviations for the morphological characters in the Materials and Methods section. (M = male, F = female, other abbreviations defined in text), * for the holotype.
Voucher number | Sex | SVL | HDL | HDW | SNT | ED | IOD | IND | TD | UEW | NED | TED | HND | LAHL | LW | FIL | FIIL | FIIIL | FIVL | TL | THL | FL | TFL | HLL | TW | IPTL | OPTL | IMTL |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
GZNU20200706010* | M | 33.4 | 11.3 | 11.1 | 4.2 | 3.8 | 3.1 | 3.7 | 2.4 | 2.9 | 2.3 | 2.0 | 8.6 | 14.7 | 2.3 | 4.5 | 4.1 | 6.3 | 5.5 | 17.5 | 17.8 | 15.7 | 24.9 | 60.2 | 3.5 | 1.8 | 1.6 | 2.2 |
GZNU20200706001 | M | 33.1 | 11.2 | 11.4 | 4.2 | 3.5 | 3.2 | 3.7 | 2.3 | 2.8 | 2.5 | 1.9 | 8.3 | 14.3 | 2.1 | 4.7 | 4.2 | 6.1 | 5.2 | 16.7 | 16.5 | 15.6 | 23.6 | 56.8 | 3.2 | 2.1 | 1.5 | 2.3 |
GZNU20200706002 | M | 30.1 | 11.2 | 10.8 | 3.9 | 3.5 | 3.5 | 3.4 | 2.1 | 2.4 | 1.9 | 1.9 | 7.9 | 13.6 | 2 | 4.6 | 4.2 | 5.8 | 5.2 | 14.9 | 14.6 | 14.7 | 20.8 | 50.3 | 3.1 | 1.9 | 1.3 | 2.3 |
GZNU20200706003 | M | 30.6 | 10.8 | 10.3 | 4.0 | 3.6 | 3.7 | 3.1 | 2.3 | 2.7 | 2.4 | 1.8 | 7.7 | 13.9 | 2.1 | 3.9 | 3.3 | 6.2 | 4.9 | 15.3 | 14.6 | 13.4 | 20.8 | 50.7 | 2.6 | 2.0 | 1.2 | 1.8 |
GZNU20200706004 | M | 32.3 | 11.2 | 11.1 | 4.3 | 3.3 | 4.3 | 3.4 | 2.0 | 2.8 | 2.2 | 2.0 | 8.3 | 14.5 | 2.3 | 4.3 | 4.5 | 6.1 | 4.7 | 16.5 | 15.4 | 15.1 | 22.5 | 54.4 | 3.3 | 2.1 | 1.2 | 2.0 |
GZNU20200706005 | M | 29.8 | 11.0 | 11.2 | 4.4 | 3.7 | 3.7 | 3.3 | 2.0 | 2.6 | 2.5 | 2.2 | 8.2 | 14.3 | 2 | 3.7 | 5.7 | 7.7 | 5.9 | 15.9 | 15.3 | 14.6 | 22.8 | 54.0 | 3.1 | 1.6 | 1.2 | 1.7 |
GZNU20200706006 | M | 30.4 | 11.2 | 10.6 | 4.2 | 3.3 | 3.7 | 3.3 | 1.7 | 2.5 | 2.7 | 1.7 | 7.3 | 13.2 | 1.9 | 4.3 | 4.0 | 5.6 | 4.4 | 15.6 | 15.1 | 13.7 | 21.1 | 51.8 | 3.2 | 2.1 | 1.7 | 1.9 |
GZNU20200706007 | M | 28.6 | 10.2 | 9.1 | 3.9 | 3.4 | 3.9 | 2.8 | 1.7 | 2.7 | 2.3 | 2.1 | 7.5 | 13.9 | 1.9 | 3.9 | 3.6 | 5.8 | 4.1 | 14.6 | 13.8 | 13.5 | 20.9 | 49.3 | 2.8 | 1.8 | 1.6 | 1.9 |
GZNU20200706008 | M | 31.4 | 10.8 | 10.4 | 4.9 | 3.6 | 3.2 | 3.3 | 2.1 | 2.9 | 2.2 | 2.0 | 8.23 | 14.7 | 1.7 | 4.4 | 4.3 | 6.3 | 4.5 | 16.2 | 16.3 | 13.8 | 22.1 | 54.6 | 2.9 | 2.3 | 1.4 | 1.8 |
GZNU20200706009 | M | 31.1 | 10.7 | 11.2 | 3.4 | 3.9 | 3.2 | 3.5 | 2.4 | 2.9 | 1.9 | 1.9 | 7.9 | 14.8 | 2.4 | 4.4 | 3.9 | 6.3 | 5.1 | 15.6 | 15.2 | 14.5 | 22.9 | 53.7 | 3.1 | 1.7 | 1.4 | 1.9 |
GZNU20200706012 | M | 33.3 | 11.5 | 10.9 | 4.3 | 3.6 | 3.8 | 3.3 | 2.2 | 2.6 | 1.9 | 2.1 | 8.1 | 14.6 | 2.1 | 4.3 | 4.9 | 6.1 | 4.8 | 16.2 | 16.1 | 15.3 | 23.5 | 55.8 | 3.4 | 1.8 | 1.2 | 2.1 |
GZNU20200706013 | M | 30.2 | 10.6 | 9.8 | 4.1 | 3.6 | 3.5 | 3.1 | 1.7 | 3.0 | 1.9 | 1.9 | 7.9 | 13.8 | 2.1 | 4.5 | 3.9 | 5.7 | 4.9 | 14.8 | 13.4 | 13.5 | 22.1 | 50.3 | 3.1 | 2.3 | 1.4 | 2.2 |
GZNU20200707001 | M | 31.2 | 11.4 | 10.4 | 4.1 | 3.9 | 3.9 | 3.4 | 2.3 | 2.9 | 2.4 | 1.9 | 8.5 | 15.2 | 2.0 | 4.5 | 4.5 | 6.9 | 4.9 | 16.9 | 15.5 | 15.1 | 23.1 | 55.5 | 3.4 | 1.7 | 1.4 | 2.0 |
GZNU20200707002 | M | 30.1 | 11.2 | 10.8 | 3.9 | 3.5 | 3.5 | 3.4 | 2.1 | 2.4 | 1.9 | 1.9 | 7.9 | 13.6 | 2 | 4.1 | 3.6 | 5.7 | 4.9 | 14.9 | 14.6 | 14.7 | 20.8 | 50.3 | 3.1 | 2.2 | 1.6 | 1.9 |
GZNU20200707003 | M | 31.8 | 11.4 | 11.1 | 4.5 | 3.5 | 3.7 | 3.3 | 2.3 | 2.9 | 2.3 | 1.9 | 8.5 | 14.9 | 2.4 | 4.7 | 3.8 | 6.5 | 4.8 | 16.6 | 15.6 | 15.1 | 22.4 | 54.6 | 3.2 | 2.5 | 1.5 | 2.6 |
GZNU20200706011 | F | 38.4 | 12.1 | 11.8 | 4.5 | 4.8 | 3.8 | 3.5 | 2.5 | 3.7 | 2.5 | 1.8 | 8.9 | 16.2 | 2.0 | 5.1 | 4.3 | 6.6 | 5.7 | 19.1 | 18.6 | 16.6 | 25.7 | 63.4 | 3.9 | 2.8 | 2.1 | 2.2 |
GZNU20200706004 | F | 39.2 | 13.2 | 11.7 | 4.3 | 4.3 | 3.7 | 3.9 | 2.5 | 3.9 | 2.3 | 1.9 | 9.5 | 16.3 | 1.9 | 4.8 | 5.9 | 7.5 | 6.4 | 19.2 | 19.5 | 17.8 | 26.3 | 65 | 3.6 | 2.9 | 2.3 | 2.4 |
GZNU20200706005 | F | 39.5 | 13.3 | 11.5 | 4.4 | 4.2 | 3.8 | 3.5 | 2.4 | 3.8 | 2.6 | 1.9 | 9.1 | 16.2 | 2.1 | 4.8 | 5.9 | 7.7 | 6.4 | 19.3 | 19.2 | 18.2 | 26.2 | 64.7 | 3.5 | 2.8 | 2.4 | 2.6 |
GZNU20200706006 | F | 40.2 | 14.5 | 13.2 | 5.1 | 5.2 | 4.5 | 4.8 | 3.4 | 3.9 | 2.5 | 2.2 | 9.8 | 17.4 | 2.0 | 4.8 | 6.0 | 7.7 | 6.8 | 20 | 19.9 | 18.9 | 26.8 | 66.7 | 3.8 | 2.9 | 2.9 | 2.9 |
Morphometric data were collected from 19 well-preserved adult specimens (voucher information given in Table
ED eye diameter (diameter of exposed portion of eyeball);
FIL first finger length;
FIIL second finger length;
FIIIL third finger length;
FIVL fourth finger length;
FL foot length (distance from distal end of tibia to the tip of the distal phalanx of toe IV);
HDL head length (from tip of snout to the articulation of the jaw);
HDW head width (head width at the commissure of the jaws);
HLL hindlimb length (distance from tip of fourth toe to vent);
HND hand length (from the proximal border of the outer palmar tubercle to the tip of digit III);
IMTL inner metatarsal tubercle length (taken as maximal length of inner metatarsal tubercle);
IND internasal distance (distance between nares);
IOD interorbital distance (minimum distance between upper eyelids);
IPTL inner palmar tubercle length (measured as maximal distance from proximal to distal ends of the inner palmar tubercle);
LAHL length of lower arm and hand (distance from the elbow to the distal end of finger IV);
LW lower arm width (maximum width of the lower arm);
NED nasal to eye distance (distance between the nasal and the anterior corner of the eye);
OPTL outer metacarpal tubercle length (measured as maximal diameter of outer metacarpal tubercle);
SNT snout length (from tip of snout to the anterior corner of the eye);
SVL snout-vent length (from tip of snout to posterior margin of vent);
TD tympanum diameter (horizontal diameter of tympanum);
TED tympanum to eye distance (distance from anterior edge of tympanum to posterior corner of eye);
TFL length of foot and tarsus (distance from the tibiotarsal articulation to the distal end of toe IV);
THL thigh length (distance from vent to knee);
TL tibia length (distance from knee to heel);
TW tibia width (maximum width of the tibia);
UEW upper eyelid width (greatest width of the upper eyelid margins measured perpendicular to the anterior-posterior axis).
To reduce allometric effects, all measurements were size-corrected with respect to SVL prior to morphometric analysis. Principal component analyses (PCAs) of size-corrected measurements and simple bivariate scatterplots were used to explore and characterize the morphometric differences between the new species and P. leishanensis. Mann–Whitney U tests were conducted to determine the significance of differences in morphometric characters between the new species and P. leishanensis. Mann–Whitney U tests also were used to test the significance of morphometric differences between males and females of the new species. All statistical analyses were performed using SPSS 21.0 (SPSS, Inc., Chicago, IL, USA), and differences were considered statistically significant at P < 0.05. Sex was determined based on male secondary sexual characters: the presence of a vocal sac and nuptial pads/spines (
We compared the morphological characters of the new species with literature data for 59 other species in the Panophrys (Table
References for morphological characters for congeners of the genus Panophrys.
ID | Species | Literature consulted |
---|---|---|
1 | Panophrys acuta (Wang, Li & Jin, 2014) |
|
2 | Panophrys angka (Wu, Suwannapoom, Poyarkov, Chen, Pawangkhanant, Xu, Jin, Murphy & Che, 2019) |
|
3 | Panophrys anlongensis (Li, Lu, Liu & Wang, 2020) |
|
4 | Panophrys baishanzuensis (Wu, Li, Liu, Wang & Wu, 2020) |
|
5 | Panophrys baolongensis (Ye, Fei & Xie, 2007) |
|
6 | Panophrys binchuanensis (Ye & Fei, 1995) |
|
7 | Panophrys binlingensis (Jiang, Fei & Ye, 2009) |
|
8 | Panophrys boettgeri (Boulenger, 1899) |
|
9 | Panophrys brachykolos (Inger & Romer, 1961) |
|
10 | Panophrys caobangensis (Nguyen, Pham, Nguyen, Luong & Ziegler, 2020) |
|
11 | Panophrys caudoprocta (Shen, 1994) |
|
12 | Panophrys cheni (Wang & Liu, 2014) |
|
13 | Panophrys chishuiensis (Xu, Li, Liu, Wei & Wang, 2020) |
|
14 | Panophrys daiyunensis Lyu, Wang & Wang, 2021 |
|
15 | Panophrys daoji Lyu, Zeng, Wang & Wang, 2021 |
|
16 | Panophrys daweimontis (Rao & Yang, 1997) |
|
17 | Panophrys dongguanensis (Wang & Wang, 2019) |
|
18 | Panophrys fansipanensis (Tapley, Cutajar, Mahony, Nguyen, Dau, Luong, Le, Nguyen, Nguyen, Portway, Luong & Rowley, 2018) |
|
19 | Panophrys frigida (Tapley, Cutajar, Nguyen, Portway, Mahony, Nguyen, Harding, Luong & Rowley, 2021) |
|
20 | Panophrys hoanglienensis (Tapley, Cutajar, Mahony, Nguyen, Dau, Luong, Le, Nguyen, Nguyen, Portway, Luong & Rowley, 2018) |
|
21 | Panophrys huangshanensis (Fei & Ye, 2005) |
|
22 | Panophrys insularis (Wang, Liu, Lyu, Zeng & Wang, 2017) |
|
23 | Panophrys jiangi (Liu, Li, Wei, Xu, Cheng, Wang & Wu, 2020) |
|
24 | Panophrys jingdongensis (Fei & Ye, 1983) |
|
25 | Panophrys jinggangensis (Wang, 2012) |
|
26 | Panophrys jiulianensis (Wang, Zeng, Lyu & Wang, 2019) |
|
27 | Panophrys kuatunensis (Pope, 1929) |
|
28 | Panophrys leishanensis (Li, Xu, Liu, Jiang, Wei & Wang, 2019 «2018») |
|
29 | Panophrys liboensis (Zhang, Li, Xiao, Li, Pan, Wang, Zhang & Zhou, 2017) |
|
30 | Panophrys lini (Wang & Yang, 2014) |
|
31 | Panophrys lishuiensis (Wang, Liu & Jiang, 2017) |
|
32 | Panophrys lushuiensis (Shi, Li, Zhu, Jiang, Jiang & Wang, 2021) |
|
33 | Panophrys minor (Stejneger, 1926) |
|
34 | Panophrys mirabilis (Lyu, Wang & Zhao, 2020) |
|
35 | Panophrys mufumontana (J. Wang, Lyu & Y.Y. Wang, 2019) |
|
36 | Panophrys nankunensis (Wang, Zeng &. Wang, 2019) |
|
37 | Panophrys nanlingensis (Lyu, J. Wang, Liu & Y.Y. Wang, 2019) |
|
38 | Panophrys obesa (Wang, Li & Zhao, 2014) |
|
39 | Panophrys ombrophila (Messenger & Dahn, 2019) |
|
40 | Panophrys omeimontis (Liu, 1950) |
|
41 | Panophrys palpebralespinosa (Bourret, 1937) |
|
42 | Panophrys qianbeiensis (Su, Shi, Wu, Li, Yao, Wang & Li, 2020) |
|
43 | Panophrys rubrimera (Tapley, Cutajar, Mahony, Chung, Dau, Nguyen, Luong & Rowley, 2017) |
|
44 | Panophrys sangzhiensis (Jiang, Ye & Fei, 2008) |
|
45 | Panophrys sanmingensis Lyu & Wang, 2021 |
|
46 | Panophrys shimentaina (Lyu, Liu & Wang, 2020) |
|
47 | Panophrys shuichengensis (Tian & Sun, 1995) |
|
48 | Panophrys shunhuangensis (Wang, Deng, Liu, Wu & Liu, 2019) |
|
49 | Panophrys spinata (Liu & Hu, 1973) |
|
50 | Panophrys tongboensis Wang & Lyu, 2021 |
|
51 | Panophrys tuberogranulata (Shen, Mo & Li, 2010) |
|
52 | Panophrys wugongensis (J. Wang, Lyu & Y.Y. Wang, 2019) |
|
53 | Panophrys wuliangshanensis (Ye & Fei, 1995) |
|
54 | Panophrys wushanensis (Ye & Fei, 1995) |
|
55 | Panophrys xianjuensis (Wang, Wu, Peng, Shi, Lu & Wu, 2020) |
|
56 | Panophrys xiangnanensis (Lyu, Zeng & Wang, 2020) |
|
57 | Panophrys yangmingensis (Lyu, Zeng & Wang, 2020) |
|
58 | Panophrys yeae (Shi, Zhang, Xie, Jiang, Liu, Ding, Luan & Wang, 2020) |
|
59 | Panophrys zhoui (Shi, Zhang, Xie, Jiang, Liu, Ding, Luan & Wang, 2020) |
|
The advertisement calls of the new species were recorded from the holotype specimen (voucher number GZNU20200706010) in the field on 5 July 2020 at the Yueliangshan Nature Reserve, Congjiang County, Guizhou Province, China. The advertisement calls were recorded in a stream, using a digital sound recorder (TASCAM DR-40) at an ambient air temperature of 25 °C and 92% humidity. Sounds were recorded within 5 cm of the calling individual. The wave-format sound files were sampled at 44 kHz, with sampling depth 24 bits. Praat 6.1.16 (
ML and BI phylogenies were constructed based on two concatenated mitochondrial gene sequences: 16S rRNA (548 bp) and COI (672 bp). The ML and BI topologies were largely identical (Fig.
Phylogenetic tree based on mitochondrial 16S+COI genes. In this phylogenetic tree, ultrafast bootstrap supports (UFB) from ML analyses/Bayesian posterior probabilities (BPP) from BI analyses were noted beside nodes. The symbol “-” represents value below 0.60/60. Photos of new collections and 11 of 4 Panophrys species in Guizhou Province. The scale bar represents 0.08 nucleotide substitutions per site. The numbers at the tip of branches corresponds to the ID numbers in Table
The smallest p-distance between this lineage and any other species of Panophrys was 1.2% in 16S rRNA (with P. huangshanensis) and 6.5% in COI (with P. wushanensis). These levels of divergence were similar to those between other pairs of recognized congeners. For example, the 16S rRNA p-distance was 1.2% between P. leishanensis and P. huangshanensis, 1.2% between P. jingdongensis and P. binchuanensis, while the COI p-distance was 5.9% between P. lini and P. nanlingensis, 3.6% between P. spinata and P. sangzhiensis, and 4.5% between Brachytarsophrys carinense and B. popei (Suppl. material
The results of the *BEAST analysis for the alternative species model are provided in Table
The species delimitation results of new species and several closely related species in BF method.
Model | Species delimitation | MLE Path Sampling (PS) | MLE Stepping Stone (SS) | Rank | BF (PS) | BF (SS) |
---|---|---|---|---|---|---|
M1 | 11 species: | -4011.49 | -4011.48 | 1 | 14.14 | 14.02 |
AC+BA+JU+MI+SI+SU+TU+WU+YA+LE+CO | ||||||
M2 | 10 species: | -4018.56 | -4018.49 | 2 | – | – |
AC+BA+JU+MI+SI+SU+TU+WU+YA+{LE+CO} |
The results of the Mann–Whitney U tests indicated that males of the new species differed significantly from P. leishanensis males based on several morphometric characters (all p-values < 0.05; Table
Morphological comparison of Panophrys congjiangensis sp. nov. (PC) and P. leishanensis (PL). All units in mm. P-values are at 95% significance. Morphometric characters are explained in the methods section. CM and CF are the abbreviations of male and female from Panophrys congjiangensis sp. nov.
Measurements | Panophrys congjiangensis sp. nov. | Panophrys leishanensis# | P-value from Mann-Whitney U test | |||||||
Male (N=15) | Female (N=4) | Male (N=10) | Female (N=2) | Male | Female | CM vs.CF | ||||
Range | Mean ± SD | Range | Mean ± SD | Range | Mean ± SD | Range | PC vs. PL | PC vs. PL | ||
SVL | 28.6–33.4 | 31.2 ± 1.4 | 38.4–40.2 | 39.3 ± 0.7 | 30.4–38.7 | 34.3 ± 2.7 | 42.3–42.3 | 0.000 | 0.133 | 0.003 |
HDL | 10.2–11.5 | 11.0 ± 0.4 | 12.1–14.5 | 13.3 ± 1.0 | 9.1–11.0 | 10.1 ± 0.7 | 11.3–11.7 | 0.000 | 0.267 | 0.002 |
HDW | 9.1–11.4 | 10.7 ± 0.6 | 11.5–13.2 | 12.1 ± 0.8 | 10.5–12.0 | 11.4 ± 0.5 | 12.1–12.4 | 0.014 | 0.133 | 0.003 |
SNT | 3.4–4.9 | 4.2 ± 0.3 | 4.3–5.1 | 4.6 ± 0.4 | 3.6–4.5 | 4.2 ± 0.3 | 4.5–5.0 | 0.007 | 0.267 | 0.027 |
ED | 3.3–3.9 | 3.6 ± 0.2 | 4.2–5.2 | 4.6 ± 0.5 | 3.3–4.3 | 3.9 ± 0.3 | 4.1–4.8 | 0.643 | 1.000 | 0.002 |
IOD | 3.1–4.3 | 3.6 ± 0.3 | 3.7–4.5 | 4.0 ± 0.4 | 3.3–4.3 | 3.7 ± 0.3 | 3.9–4.2 | 0.062 | 0.267 | 0.084 |
IND | 2.8–3.7 | 3.3 ± 0.2 | 3.5–4.8 | 3.9 ± 0.6 | 3.5–4.7 | 4.0 ± 0.4 | 4.1–4.3 | 0.392 | 0.267 | 0.011 |
TD | 1.7–2.4 | 2.1 ± 0.2 | 2.4–3.4 | 2.7 ± 0.5 | 2.0–2.6 | 2.3 ± 0.2 | 2.5–2.8 | 0.461 | 1.000 | 0.003 |
UEW | 2.4–3.0 | 2.7 ± 0.2 | 3.7–3.9 | 3.8 ± 0.1 | / | / | / | / | / | 0.002 |
NED | 1.9–2.7 | 2.2 ± 0.3 | 2.3–2.6 | 2.5 ± 0.1 | / | / | / | / | / | 0.059 |
TED | 1.7–2.2 | 1.9 ± 0.1 | 1.8–2.2 | 2.0 ± 0.2 | / | / | / | / | / | 0.750 |
HND | 7.3–8.6 | 8.1 ± 0.4 | 8.9–9.8 | 9.3 ± 0.4 | / | / | / | / | / | 0.003 |
LAHL | 13.2–15.2 | 14.3 ± 0.6 | 16.2–17.4 | 16.5 ± 0.6 | 14.4–16.3 | 15.3 ± 0.6 | 18.1–18.4 | 0.036 | 0.133 | 0.003 |
LW | 1.7–2.4 | 2.1 ± 0.2 | 1.9–2.1 | 2.0 ± 0.1 | 2.7–3.9 | 3.2 ± 0.5 | 2.8–2.9 | 0.001 | 1.000 | 0.355 |
TL | 14.6–17.5 | 15.9 ± 0.9 | 19.1–20 | 19.4 ± 0.4 | 16.2–18.6 | 17.5 ± 0.9 | 19.2–19.2 | 0.129 | 0.800 | 0.003 |
THL | 13.4–17.8 | 15.3 ± 1.1 | 18.6–19.9 | 19.3 ± 0.5 | 14.4–16.8 | 15.4 ± 0.8 | 17.6–17.7 | 0.000 | 1.000 | 0.003 |
FL | 13.4–15.7 | 14.6 ± 0.8 | 16.6–18.9 | 17.9 ± 1.0 | 14.9–17.3 | 15.9 ± 1.0 | 18.1–19.0 | 0.129 | 0.800 | 0.003 |
TFL | 20.8–24.9 | 22.3 ± 1.2 | 25.7–26.8 | 26.3 ± 0.5 | 21.1–25.9 | 23.5 ± 0.5 | 27.5–27.9 | 0.004 | 1.000 | 0.003 |
HLL | 49.3–60.2 | 53.5 ± 3.0 | 63.4–66.7 | 65.0 ± 1.4 | 50.3–60.2 | 54.2 ± 3.0 | 49.3–50.3 | 0.023 | 0.533 | 0.003 |
TW | 2.6–3.5 | 3.1 ± 0.2 | 3.5–3.9 | 3.7 ± 0.2 | 3.6–4.7 | 4.2 ± 0.3 | 4.8–5.1 | 0.000 | 0.133 | 0.003 |
IPTL | 1.6–2.5 | 2.0 ± 0.3 | 2.8–2.9 | 2.9 ± 0.1 | / | / | / | / | / | 0.003 |
OPTL | 1.2–1.7 | 1.4 ± 0.2 | 2.1–2.9 | 2.4 ± 0.3 | / | / | / | / | / | 0.002 |
IMTL | 1.7–2.6 | 2.0 ± 0.2 | 2.2–2.9 | 2.5 ± 0.3 | / | / | / | / | / | 0.014 |
FIL | 3.7–4.7 | 4.3 ± 0.3 | 4.8–5.1 | 4.9 ± 0.2 | 3.2–3.9 | 3.5 ± 0.2 | 4.0–4.3 | 0.000 | 0.133 | 0.003 |
FIIL | 3.3–5.7 | 4.2 ± 0.6 | 4.3–6 | 5.5 ± 0.8 | 2.8–3.5 | 3.2 ± 0.3 | 3.8–4.1 | 0.000 | 0.133 | 0.011 |
FIIIL | 5.6–7.7 | 6.2 ± 0.5 | 6.6–7.7 | 7.4 ± 0.5 | 4.2–5.4 | 4.8 ± 0.4 | 5.4–5.8 | 0.000 | 0.133 | 0.009 |
FIVL | 4.1–5.9 | 4.9 ± 0.4 | 5.7–6.8 | 6.3 ± 0.5 | 3.4–4.1 | 3.7 ± 0.2 | 4.2–4.3 | 0.000 | 0.133 | 0.004 |
Holotype. GZNU20200706010 (Figs
Morphological features of the live adult male holotype GZNU20200706010 of Panophrys congjiangensis sp. nov. A single subgular vocal sac B dorsal view C dorsolateral view D ventral view E ventral view of hand F ventral view of foot G dorsal view of hand (1 indicates villiform gray-black nuptial spines) H iris. A was photographed at about 9 p.m., and B to H during the day, respectively.
Paratypes. Nineteen adult specimens (15 males and 4 females) from the same locality. Eleven males (GZNU20200706001–06009, GZNU20200706012–06013) collected with the holotype on 6 July 2020 by Tao Luo, Xueli Lu, and Weifeng Wang. One female (GZNU20200706011) collected with the holotype by Tao Luo. Three males (GZNU200707001–07003) collected on 7 July 2020 by Tao Luo. Three females (GZNU20200706004, GZNU20200706005, and GZNU20200706006) collected on 7 July 2020 by Tao Luo.
The specific epithet “congjiangensis” refers to the holotype locality, which is Yueliangshan Nature Reserve, located in Congjiang County, Guizhou Province, China. We propose the English common name “Congjiang Horned Toad” and the Chinese common name “Cong Jiang Jiao Chan (从江角蟾)”.
Panophrys congjiangensis sp. nov. is assigned to the genus Panophrys based on molecular phylogenetic analyses and the following characteristics, which are diagnostic for this genus: (1) snout shield-like; (2) snout projecting beyond the lower jaw; (3) tympanum distinct (4) canthus rostralis distinct; (5) chest gland small and round, closer to axilla than to midventral line; (6) femoral gland on rear of thigh; (7) vertical pupils (Fei et al. 2006;
Panophrys congjiangensis sp. nov. is distinguished from its congeners by a combination of the following characters: (1) medium body size (SVL: 28.6–33.4 mm in males and 38.4–40.2 mm in females); (2) single small horn-like tubercle at edge of each upper eyelid; (3) tympanum distinctly visible (TD/ED ratio 0.47–0.66); (4) vomerine teeth absent; (5) tongue not notched behind; (6) a narrow and unobvious lateral fringe on toes; (7) relative finger lengths II < I < V < III; (8) rudimentary webs on toes; (9) hindlimbs slender, heels overlapping when thighs are positioned at right angles to body; (10) two metacarpal tubercles on palm, with inner metatarsal tubercle long and oval-shaped; (11) tibiotarsal articulation reaching nostril when leg is stretched forward; (12) dorsal skin rough, with numerous orange-red granules, ventral surface smooth; (13) single internal subgular vocal sac present in males; (14) in breeding males, weak gray-black nuptial pads with black nuptial spines present on dorsal surfaces of bases of first and second fingers.
GZNU20200706010 (Figs
Forelimbs slender and comparatively short, the length of lower arm and hand 44.01% of SVL; fingers slender, relative finger lengths: II < I < IV < III; tips of fingers slightly dilated, round, without lateral fringes; one distinct subarticular tubercle at the base of each finger; two metacarpal tubercles on the palm; prominent, the outer one long and thin, the inner one oval-shaped, inner metacarpal tubercles longer than outer metacarpal tubercles (IPTL/OPT ratio 1.13).
Hindlimbs slender (HLL/SVL ratio 1.80); heels slightly overlapping when thighs are positioned at right angles to the body; tibiotarsal articulation reaching the nostril when leg stretched forward; foot length less than tibia length (FL/TL ratio 0.90); relative toe lengths I < II < V < III < IV; tips of toes round and slightly dilated; toes with narrow and unobvious lateral fringes and rudiment webs; one subarticular tubercle at the base of each toe; inner metatarsal tubercle long oval-shaped and the outer one absent.
Dorsal skin rough with numerous orange-red granules; several large warts scattered on flanks and dorsal limbs; several tubercles on upper eyelid, including a small horn-like prominent tubercle on the edge (Fig.
(Fig.
(Fig.
Measurements of the type series are shown in Tables
The call description is based on recordings of the holotype GZNU 20200706010 (Fig.
Adult males (SVL 28.6–33.4 mm) smaller than adult females (SVL 38.4–40.2 mm). Adult males with single internal subgular vocal sac (Fig.
Comparative data of Panophrys congjiangensis sp. nov. with 59 recognized congeners of Panophrys are given in Suppl. material
By having small body size, SVL 30.4–34.1 mm in males, Panophrys congjiangensis sp. nov. differs from P. baolongensis (42.0–45.0 in males), P. binlingensis (45.1–51.0 in males), P. boettgeri (34.5–37.8 in males), P. caobangensis (34.9–38.9 in males), P. caudoprocta (81.3 in single male), P. hoanglienensis (37.4–47.6 in males), P. huangshanensis (36.0–41.6 in males), P. insularis (36.8–41.2 in males), P. jingdongensis (53.0–56.5 in males), P. mirabilis (55.8–61.4 in males), P. obesa (35.6 in single male), P. palpebralespinosa (36.2–38.0 in males), P. sangzhiensis (54.7 in single male), and P. xiangnanensis (38.6–42.0 in males). By having larger body size, SVL 30.4–34.1 mm in males, Panophrys congjiangensis sp. nov. differs from P. cheni (26.2–29.5 in males), P. daiyunensis (27.6–28.7 in males), P. kuatunensis (26.2–29.6 in males), P. sanmingensis (27.0–29.5 in males), P. yeae (23 in single male), and P. zhoui (23.8–29.1 in males). By having small body size, SVL 38.9–40.2 mm in females, Panophrys congjiangensis sp. nov. differs from P. fansipanensis (41.7–42.5 in females), P. minor (42.0–48.2 in females), P. tuberogranulata (50.5 in single female), P. wuliangshanensis (41.3 in single female), P. xianjuensis (41.6 in single female), and P. yangmingensis (45.2 in single female).
Nine Panophrys species were previously recorded from the Guizhou Province, namely P. anlongensis, P. chishuiensis, P. jiangi, P. leishanensis, P. liboensis, P. omeimontis, P. shuichengensis, P. spinata, and P. qianbeiensis. Panophrys congjiangensis sp. nov. differs from P. anlongensis by having small body size, SVL 30.4–34.1 mm in males and 38.9–40.2 mm in females (vs. 40.0–45.5 mm in males and 48.9–51.2 in females), tibiotarsal articulation reaching the nostril when leg stretched forward (vs. reaching to the level of mid-eye). Panophrys congjiangensis sp. nov. differs from P. chishuiensis by having small body size, SVL 30.4–34.1 mm in males and 38.9–40.2 mm in females (vs. 43.4–44.1 mm in males and 44.8–49.8 in females), rudimentary webs on toes (vs. lacking webs), subarticular tubercles present on each toes (vs. absent), tibiotarsal articulation reaching the nostril when leg stretched forward (vs. reaching the level between tympanum and eye). Panophrys congjiangensis sp. nov. differs from P. jiangi by having slightly small body size, SVL 30.4–34.1 mm in males (vs. 34.4–39.2 mm in males), relative finger lengths are II < I < V < III (vs. I < II < V < III), tibiotarsal articulation reaching the nostril when leg stretched forward (vs. reaching forward to the region between tympanum and eye). Panophrys congjiangensis sp. nov. differs from P. liboensis, P. omeimontis, P. qianbeiensis, P. shuichengensis, and P. spinata by having small body size, SVL 30.4–34.1 mm in males and 38.9–40.2 mm in females (vs. SVL>40 mm in males in P. liboensis, P. omeimontis, P. qianbeiensis, P. shuichengensis, and P. spinata; vs. SVL>50 mm in females in P. liboensis, P. omeimontis, P. shuichengensis, and P. spinata), small horn-like tubercle at the edge of each upper eyelid (vs. slightly large in P. liboensis and P. shuichengensis; absence in P. qianbeiensis), absence of vomerine teeth (vs. present in P. liboensis, P. omeimontis, and P. qianbeiensis), tongue not notched behind (vs. notched in P. liboensis, P. omeimontis, P. qianbeiensis, P. shuichengensis, and P. spinata), lateral fringes on toes narrow and unobvious (vs. wide in P. liboensis, P. qianbeiensis, P. shuichengensis, and P. spinata), rudimentary webs on toes (vs. more than one-fourth webs in P. qianbeiensis, P. shuichengensis, and P. spinata), subarticular tubercles present on each toes (vs. absent in P. liboensis and P. shuichengensis), tibiotarsal articulation reaching the nostril when leg stretched forward (vs. reaching to ocular region in P. liboensis, P. omeimontis, P. shuichengensis, and P. spinata; reaching to the level between tympanum and eye in P. qianbeiensis). Panophrys congjiangensis sp. nov. differs from P. leishanensis by having slightly small body size, SVL 38.9–40.2 mm in females (vs. 42.3 in single female), having narrow and unobvious lateral fringes on toes (vs. lacking), tibiotarsal articulation reaching the nostril when leg stretched forward (vs. reaching between tympanum to eye). The mean SVL of male Panophrys congjiangensis sp. nov. was significantly greater than that of P. leishanensis. In addition, the ratios of HDL, HDW, SNT, LAHL, LW, THL, TFL, HLL, and TW to SVL were all significantly greater in male Panophrys congjiangensis than in male P. leishanensis (all p-values < 0.05; Table
From the remaining 24 species occurring in Panophrys, Panophrys congjiangensis sp. nov. can be distinguished by the absence of vomerine teeth (vs. present in P. daweimontis, P. dongguanensis, P. frigida, P. jinggangensis, P. jiulianensis, P. nankunensis, P. nanlingensis, P. rubrimera, P. shimentaina, and P. tongboensis), by the unnotched tongue (vs. tongue notched in P. binchuanensis, P. cheni, P. kuatunensis, and P. lushuiensis), by the small horn-like tubercle at edge of upper eyelid (vs. slightly large in P. acuta), by the absence of lateral fringes on toes (vs. lacking in P. angka, P. brachykolos, P. lishuiensis, P. ombrophila, P. shunhuangensis, and P. wugongensis; vs. wide in P. lini; vs. lacking in males in P. wushanensis, wide in females in P. wushanensis), by the subarticular tubercles present (vs. absent in P. baishanzuensis and P. mufumontana), tibiotarsal articulation reaching the nostril when leg stretched forward (vs. reaching to ocular region in P. acuta, P. baishanzuensis, P. binchuanensis, P. jiulianensis, P. lini, P. nanlingensis, P. ombrophila, and P. wushanensis; vs. reaching to the level between tympanum and eye in P. angka, P. dongguanensis, P. kuatunensis, P. lishuiensis, and P. nankunensis; vs. reaching to the level between eye and snout in P. cheni, P. daweimontis, and P. shunhuangensis; vs. reaching to the level behind the eye in P. brachykolos, P. mufumontana, P. shimentaina, and P. wugongensis; vs. reaching to the level at center of tympanum P. daoji).
Panophrys congjiangensis sp. nov. is only known from the type locality, Yueliangshan Nature Reserve, Congjiang County, Guizhou Province, China at elevations of 1142–1206 m. Individuals of the new species were frequently found in bamboo forests, grasses, and shrubberies near streams. Plants in the type locality predominantly fall into the families Urticaceae, Gramineae, Cyperaceae, Rosaceae, Dryopteridaceae, Polygonaceae, Aquifoliaceae, and Fagaceae. In the Yueliang Mountains, Panophrys congjiangensis sp. nov. is sympatric with Pachytriton inexpectatus Nishikawa, Jiang, Matsui & Mo, 2010; Amolops sinensis Lyu, Wang & Wang, 2019; Nidirana leishanensis Li, Wei, Xu, Cui, Fei, Jiang, Liu & Wang, 2019; Hylarana latouchii (Boulenger, 1899); Quasipaa boulengeri (Günther, 1889); Hyla annectans (Jerdon, 1870); Opisthotropis zhaoermii Ren, Wang, Jiang, Guo & Li, 2017; Trimeresurus stejnegeri (Schmidt, 1925); and Rhabdophis tigrinus (Boie, 1826). These species were often found in the same streams as Panophrys congjiangensis sp. nov.
Phylogenetic analyses based on two mitochondrial genes suggested that the specimens collected in this study fell into the Panophrys, but were distinct from all previously described species in this genus. Genetic distances between Panophrys congjiangensis sp. nov. and its sister species P. leishanensis were 3.0% for 16S rRNA and 8.4% for COI, within the ranges expected for interspecific divergences in amphibians (
Climatic fluctuations, habitat heterogeneity, habitat diversity, and the dynamics of montane forests may play important roles in driving diversification in the Panophrys (
Biodiversity conservation in southwestern China is a priority of the Chinese government (
In the past three years alone, 11 new amphibian species have been described from Guizhou Province, China (
This work was supported by the programs of the Ministry of Science and Technology of the People’s Republic of China major project “Protect technology and endangered mechanism research on rare animals” (No. 2016YFC0503200), the Strategic Priority Research Program B of the Chinese Academy of Sciences(CAS) (No. XDB31000000), the National Animal Collection Resource Center, China (Grant No. 2005DKA21402), the Application of Amphibian Natural Antioxidant Peptides as Cosmetic Raw Material Antioxidants (QKZYD [2020]4002). We thank Professor Paul A. Garber for editing assistance during the preparation of this manuscript. We thank LetPub (www.letpub.com) for its linguistic assistance during the preparation of this manuscript.
Specimens examined
Panophrys jiangi (N=3): China: Guizhou Province: Suiyang County: Kuankuosui National Nature Reserve (type locality): GZNU20070712001. China: Guizhou: Suiyang: Huoqiuba Nature Reserve (topotype locality): GZNU20180606020–606022.
Panophrys liboensis (N=5): China: Guizhou Province: Libo County (type locality): GNUG20150813001, 408002, 408004, 408006–408008.
Panophrys shuichengensis (N=7): China: Guizhou Province: Shuicheng County (type locality): 944001, 98001, 98002, 945005, 2007030, 2007031, 2007032.
Panophrys spinata (N=6): China: Guizhou Province: Dafang County (topotype locality): GZNU201707015011–606016.
Table S1. Uncorrected p-distance between Megophryinae species based on 16s rRNA sequences
Data type: molecular data
Table S2. Uncorrected p-distance between Megophryinae species based on COI gene sequences
Data type: molecular data
Table S3. Variable loadings for principal components with Eigenvalues greater than 2, from morphometric characters corrected by SVL
Data type: statistical data
Table S4. Diagnostic characters separating all 59 species of the Panophrys
Data type: species data
Explanation note: Data modified from