Corresponding author: Jun Wu (
Academic editor: A. Ohler
A new species of the Asian horned toad genus
Wu Y, Li S, Liu W, Wang B, Wu J (2020) Description of a new horned toad of
The Asian horned toad
Wuyi Mountain region, located in northern Fujian, southeastern Jiangxi and south Zhejiang provinces of China, is a biodiversity hotspot. In this region, four
During field surveys in Qingyuan County, Zhejiang Province, China, we collected
A total of 15 specimens were sampled in this study: six adult males and one tadpole of the undescribed species and two adult males of
Information for samples used in molecular phylogenetic analyses in this study.
ID | Species | Voucher number | Locality | GenBank accession number | |
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1 | CIBQY20200719001 | Baishanzu National Park, Qingyuan, Zhejiang, China |
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2 | CIBQY20200719002 |
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3 | CIBQY20200719003 |
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4 | CIBQY20200719004 |
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5 | CIBQY20200719006 |
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6 | CIBQY20200726001 |
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7 | CIBQY20200726002 |
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8 |
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CIBWY18082407 | Wuyi Shan, Fujian, China |
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9 |
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CIBWY18082408 |
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10 |
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SYS a001579 |
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– | |
11 |
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SYS a002370 | Suichuan, Jiangxi, China |
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– |
12 |
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SYS a002874 | Yangming Shan, Hunan, China |
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13 |
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SYS a001959 | Nanling Nature Reserve, Guangdong, China |
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14 |
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SYS a001972 | Yinping Shan, Guangdong, China |
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15 |
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SYS a004498 | Nankun Shan, Guangdong, China |
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16 |
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SYS a001427 | Jinggang Shan, Jiangxi, China |
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– |
17 |
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SYS a002610 | Wugongshan Scenic Area, Jiangxi, China |
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18 |
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KRM18 | Wuyishan, Fujian, China |
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– |
19 |
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CIBWY18082308 |
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20 |
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SYS a002272 | Heishiding Nature Reserve, Guangdong, China |
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– |
21 |
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WYF00169 | Lishui, zhejiang, China |
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– |
22 |
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CIBXJ190505 | Xianju, zhejiang, China |
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23 |
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KIZ07132 | Chashan Forest Farm, Jiangxi, China |
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24 |
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CIB20200718001 | Baishanzu National Park, Qingyuan, Zhejiang, China |
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25 |
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CIB20200718002 | Baishanzu National Park, Qingyuan, Zhejiang, China |
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26 |
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Tissue ID: YPXJK033 | Wuyi Shan, Fujian, China |
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27 |
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KIZ022004 | Huang Shan, Anhui, China |
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28 |
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GNUG:20160408003 | Libo, Guizhou, China |
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– |
29 |
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SYS a006391 | Mufu Shan, Hunan, China |
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30 |
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KIZ045469 | Guangwu Shan, Sichuan, China |
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31 |
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KIZ019216 | Baolong, Chongqing, China |
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32 |
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Tissue ID: YPX10987 | Badagongshan Nature Reserve, Hunan, China |
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33 |
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SYS a002877 | Yangming Shan, Hunan, China |
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34 |
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SYS a002077 | Shimentai Nature Reserve Guangdong, China |
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35 |
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SYS a002107 | Jiulian Shan, Jiangxi, China |
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36 |
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HNNU16SH02 | Shunhuang Mountains, Hunan, China |
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– |
37 |
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SYS a002192 | Huaping Nature Reserve, Guangxi, China |
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38 |
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CIBLS20171101001 | Leigong Shan, Guizhou, China |
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39 |
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KIZ025765 | Emei Shan, Sichuan, China |
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40 |
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KIZ040591 | Kiew Mae Pan nature trail, Chiang Mai, Thailand |
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– |
41 |
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KIZ019441 | Jizu Shan, Yunnan, China |
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42 |
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KIZ011603 | Pu Hu Nature Reserve, Thanh Hoa, Vietnam |
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43 |
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SYSa002227 | Leigong Shan, Guizhou, China |
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44 |
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SYSa004307 | Zhangjiajie, Hunan, China |
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45 |
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SYSa005313 | Wawu Shan, Sichuan, China |
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46 |
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KIZ046812 | Huangcaoling, Yunnan, China |
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47 |
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KIZ048997 | Dawei Shan, Yunnan, China |
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48 |
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KIZ-LC0805067 | Huanglianshan National Nature Reserve, Yunnan, China |
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49 |
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VNMN 2018.01 | Lao Cai, Sa Pa, Vietnam |
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– |
50 |
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VNMN 2018.02 | Lao Cai, Sa Pa, Vietnam |
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– |
51 |
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KIZ01939 | Qingcheng Shan, Sichuan, China |
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52 |
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CIBKKS20180722006 | Kuankuosui Nature Reserve, Guizhou, China |
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53 |
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CIBCS20190518031 | Chishui Nature Reserve, Guizhou, China |
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54 |
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ROM 16634 | Hong Kong, China |
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55 |
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SYS a001957 | Heishiding Nature Reserve, Guangdong, China |
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– |
56 |
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ITBCZ 1108 | Nui Chua National Park, Ninh Thuan, Vietnam |
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57 |
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ZMMU ABV-00454 | Bidoup Mountain, Lam Dong, Vietnam |
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– |
58 |
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FMNH 262778 | O’Reang, Mondolkiri, Cambodia |
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– |
59 |
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KIZ010360 | Phong Dien Nature Reserve, Thua Thien Hue, Vietnam |
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60 |
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KIZ048799 | Xiaoqiaogou Nature Reserve, Yunnan, China |
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61 |
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KIZ010978 | Beibeng, Xizang, China |
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62 |
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ZMH A13125 | Gunung Kinabalu National Park, Kogopan Trail, Malaysia |
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63 |
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KU 314303 | Pasonanca Natural Park, Zamboanga, Philippines |
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64 |
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ZMMU NAP-05015 | Palawan, Philippines |
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65 |
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UNIMAS 8148 | Gunung Kinabalu National Park, Sabah, Malaysia |
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66 |
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KIZ019419 | Malaysia |
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67 |
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FMNH 273694 | Bintulu, Sarawak, Malaysia |
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68 |
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KIZ025467 | Khao Nan National Park, Nakhon Si Thammarat, Thailand |
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69 |
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KIZ016045 | Xiaoqiaogou Nature Reserve, Yunnan, China |
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70 |
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KIZ021786 | Nanling National Forest Park, Guangdong, China |
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71 |
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SDBDU2009.297 | East Khasi Hills dist., Meghalaya |
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72 |
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KIZ048439 | Husa, Yunnan, China |
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73 |
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KIZ06621 | Beibeng, Xizang, China |
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74 |
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BNHS 6061 | West Kameng dist., Arunachal Pradesh, IN |
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75 |
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SDBDU2009.75 | East Siang dist., Arunachal Pradesh, IN |
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– |
76 |
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K5198/ZSI11393 | – |
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– |
77 |
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KIZ014278 | Zhangmu, Xizang, China |
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78 |
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ZSIA11799 | – |
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– |
79 |
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SYSa002961 | Zhushihe, Yunnan, China |
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80 |
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BNHS 6046 | West Garo Hills dist., Meghalaya |
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– |
81 |
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NCSM 79599 | Aural, Kampong Speu, Cambodia |
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– |
82 |
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SYSa003042 | Zhushihe, Yunnan, China |
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83 |
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UNIMAS 8943 | Gunung Mulu National Park, Sarawak, Malaysia |
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– |
84 |
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Nanjiang, Sichuan, China |
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– | |
85 |
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KIZ025799 | Wawu Shan, Sichuan, China |
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86 |
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SYSa003933 | Wuliang shan, Yunnan, China |
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87 |
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KIZ014512 | Liziping Nature Reserve, Sichuan, China |
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88 |
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KIZ046706 | Huangcaoling, Yunnan, China |
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89 |
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CIB20050081 | Hejiang, Sichuan, China |
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– |
90 |
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Tissue ID: YPX20455 | Dayao Shan, Guangxi, China |
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91 |
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SYS a000589 | Naling Nature Reserve, Guangdong, China |
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– |
92 |
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ZFMK 87596 | U Bo, Phong Nha-Ke Bang NP, Vietnam |
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93 |
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LSUMZ 81916 | Sukabumi, Java, Indonesia |
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94 |
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MZB: Amp:22233 | – |
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95 |
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Tissue ID: YPX37539 | Emei Shan, Sichuan, China |
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96 |
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KIZ025778 | Emei Shan, Sichuan, China |
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Sampling localities of
Six adult males and one tadpole of the undescribed species, two
For molecular analyses, the available sequences for congeners of
Six adult males and one tadpole of the undescribed species were measured (Table
For the single tadpole of the undescribed species, eleven morphometric characters were measured:
To reduce the impact of allometry, the correct value from the ratio of each character to
The new species was also compared with all other
References for morphological characters for congeners of the genus
Species | Literature obtained |
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Mahony et al. 2020 | |
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Mahony et al. 2020 | |
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Li et al. 2018 | |
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Mahony et al. 2020 | |
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Mo et al. 2012 | |
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The advertisement calls of the undescribed species were recorded from the holotype specimen CIBQY20200726001 in the field on 26 July 2020 from Qingyuan County, Zhejiang Province, China. When registrating the male in the stream the ambient air temperature was 21.5 °C and there was air humidity of 87%. For comparisons, the advertisement calls of
Aligned sequence matrix of
Maximum likelihood (
Genetic distances based on
In
Morphometric comparisons between the adult specimens of
Character |
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Mann-Whitney U value | P-value | |||
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Male (N = 6) | Male (N = 6) | |||||
Ranging | Mean ± SD | Ranging | Mean ± SD | |||
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28.4–32.4 | 30.5 ± 1.8 | 28.4–32.4 | 30.5 ± 1.8 | 13.000 | 0.423 |
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8.0–9.1 | 8.6 ± 0.4 | 8.0–9.1 | 8.6 ± 0.4 | 6.000 | 0.055 |
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9.3–10.5 | 10.2 ± 0.4 | 9.3–10.5 | 10.2 ± 0.4 | 8.000 | 0.109 |
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3.4–4.1 | 3.8 ± 0.3 | 3.4–4.1 | 3.8 ± 0.3 | 16.000 | 0.749 |
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1.5–2.6 | 2.0 ± 0.4 | 1.5–2.6 | 2.0 ± 0.4 | 18.000 | 1.000 |
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3.1–3.7 | 3.4 ± 0.3 | 3.1–3.7 | 3.48 ± 0.3 | 16.000 | 0.749 |
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2.8–3.3 | 3.0 ± 0.2 | 2.8–3.3 | 3.08 ± 0.2 | 6.000 | 0.055 |
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2.3–3.0 | 2.6 ± 0.2 | 2.3–3.0 | 2.6 ± 0.2 | 2.000 |
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3.7–4.0 | 3.8 ± 0.1 | 3.7–4.0 | 3.8 ± 0.1 | 15.000 | 0.631 |
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1.5–2.1 | 1.8 ± 0.2 | 1.5–2.1 | 1.8 ± 0.2 | 16.000 | 0.749 |
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13.4–14.6 | 14.1 ± 0.5 | 13.4–14.6 | 14.2 ± 0.5 | 9.000 | 0.150 |
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6.6–7.9 | 7.1 ± 0.5 | 6.6–7.9 | 7.1 ± 0.5 | 6.000 | 0.055 |
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2.2–2.7 | 2.4 ± 0.2 | 2.2–2.7 | 2.4 ± 0.2 | 10.000 | 0.200 |
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2.2–2.8 | 2.5 ± 0.2 | 2.2–2.8 | 2.5 ± 0.2 | 17.000 | 0.873 |
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2.4–3.0 | 2.7 ± 0.2 | 2.4–3.0 | 2.7 ± 0.2 | 12.000 | 0.200 |
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4.3–5.1 | 4.6 ± 0.3 | 4.3–5.1 | 4.6 ± 0.3 | 10.000 | 0.200 |
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2.6–3.6 | 3.0 ± 0.4 | 2.6–3.6 | 3.0 ± 0.4 | 15.000 | 0.631 |
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12.2–13.5 | 12.9 ± 0.5 | 12.2–13.5 | 12.9 ± 0.5 | 10.000 | 0.200 |
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12.8–14.9 | 13.9 ± 0.9 | 12.8–14.9 | 13.9 ± 0.9 | 13.000 | 0.423 |
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2.7–4.2 | 3.3 ± 0.5 | 2.7–4.2 | 3.3 ± 0.5 | 13.000 | 0.423 |
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17.8–20.4 | 19.4 ± 1.0 | 17.8–20.4 | 19.4 ± 1.0 | 1.000 |
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11.2–12.3 | 11.8 ± 0.4 | 11.2–12.3 | 11.8 ± 0.4 | 13.000 | 0.423 |
Plots of the first principal component (PC1) versus the second (PC2) for
There were two differences in sonograms and waveforms of calls between the undescribed species and
Comparisons of characteristics of advertisement calls of
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CIBQY20200726001 | CIBWY2018082410 | CIBWY2018082412 | WY2018082411 | |
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11 | 30 | 30 | 20 |
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22 | 30 | 30 | 40 |
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151.0–170.0 (162.4 ± 5.7) | 131.0–163.0 (147.2 ± 7.1) | 131.0–163.0 (147.2 ± 7.1) | 130.0–159.0 (120.9 ± 5.9) |
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0.79 | 1.18 | 1.13 | 1.3 |
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682.0–1869.0 (936.8 ±349.0) | 404–1548.0 (687.3 ± 206.8) | 404–1548.0 (687.3 ± 206.8) | 350.0–733.0 (458.4 ± 87.1) |
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23.0–30.0 (26.0 ± 2.4) | 25.0–36.0 (30.0 ± 2.3) | 25.0–36.0 (30.0 ± 2.3) | 32.0–40.4 (35.7 ± 2.3) |
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3.19–3.38 (3.36 ± 0.06) | 3.38–3.75 (3.46 ± 0.16) | 3.38–3.75 (3.46 ± 0.16) | 3.38–3.38 (3.38±0.01) |
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3.0–6.0 (4.9 ± 0.6) | 3.0–6.0 (4.4 ± 0.7) | 3.0–6.0 (4.4 ± 0.7) | 3.0–6.0 (4.5 ± 0.6) |
Visualization of advertisement calls of
Based on the molecular phylogenetic analyses, morphological comparisons (Supp. material 4), and bioacoustics differences, the specimens from Qiangyuan County, Zhejiang Province, China represent a new species which is described as follows.
CIBQY20200726001 (Figs
Five adult males collected from the same place as holotype collected by Bin Wang. CIBQY20200719001-CIBQY20200719004 collected on 19 July 2020 by Bin Wang, and CIBQY20200726002 collected by Zhonghao Luo on 26 July 2020.
One tadpole (CIBQY20200719005; Fig.
(Figs
Forelimbs slender, the length of lower arm and hand 47.0% of
Hindlimbs slender, tibia length 46.5% times of
Dorsal skin rough, several large warts scattered on flanks; a small horn-like tubercle at the edge of each upper eyelid; tubercles on the dorsum forming a X-shaped ridge, two dorsolateral parallel ridges on either side of the X-shaped ridges; an inverted triangular brown speckle between two upper eyelidsseveral tubercles scattered on dorsal, flanks and dorsal surface of thighs and tibias; supratympanic fold distinct.
Numerous granules scattered on ventrum; pectoral and femoral glands distinct; numerous white granules on outer thighs.
Photos of the holotype specimen CIBQY20200726001 of
Photos of the holotype CIBQY20200726001 of
Color variation in
Amplitude modulation within note was apparent, beginning with moderately high energy pulses, increasing to the maximum by approximately quarter, and then decreasing towards the end. The average dominant frequency was 3.36 ± 0.06 (3.19–3.38 kHz).
Supp. material 4. By having small body size,
By vomerine teeth absent,
By a small horn-like tubercle present at the edge of each upper eyelid,
By tongue not notched behind,
By toes with narrow lateral fringes,
By toes without webbing,
By heels overlapping when thighs are positioned at right angles to the body,
By tibiotarsal articulation reaching to the level to the middle of eye when leg stretched forward,
By having an internal single subgular vocal sac in male,
The congeners
Photos of the tadpole CIBQY20200719006 of
Habitats of
The specific name
Although
During our several and extensive surveys, we only found fewer than 15 adult males of
We thank Zhonghao Luo, Yang Li and Shengchao Shi for their help with collecting samples and data. This work was supported by the Biodiversity Survey Project of Lishui, Zhejiang Province and the Project supported by the Biodiversity investigation, Observation and Assessment Program (2019–2023) of Ministry of Ecology and Environment of China.
Table S1
morphological measurements
Measurements of the adult specimens of
Table S2
genetic distance
Uncorrected
Table S3
genetic distance
Uncorrected
Table S4
morphological comparisons
Diagnostic characters separating