Biodiversity Data Journal :
Taxonomy & Inventories
|
Corresponding author: Bin Wang (wangbin@cib.ac.cn)
Academic editor: Franco Andreone
Received: 28 Sep 2023 | Accepted: 29 Dec 2023 | Published: 09 Jan 2024
© 2024 Shize Li, Wei Li, Yanlin Cheng, Jing Liu, Gang Wei, Bin Wang
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:
Li S, Li W, Cheng Y, Liu J, Wei G, Wang B (2024) Description of a new Asian Leaf Litter Toad of the genus Leptobrachella Smith, 1925 (Anura, Megophryidae) from southern Guizhou Province, China. Biodiversity Data Journal 12: e113427. https://doi.org/10.3897/BDJ.12.e113427
|
The Asian leaf litter toads of the genus Leptobrachella Smith, 1925 (Anura, Megophryidae) inhabit the forest floor and rocky streams in hilly evergreen forests and are widely distributed from southern China, west to north-eastern India and Myanmar, through mainland Indochina to Peninsular Malaysia and the Island of Borneo.
A new species of the Asian leaf litter toad genus Leptobrachella from Guizhou Province, China is described. Molecular phylogenetic analyses, based on mitochondrial 16S rRNA and COI genes and nuclear RAG1 gene sequences indicated that the new species is genetically divergent from its congeners. The new species could be distinguished from its congeners by a combination of the following characters: (1) body of medium size in males (SVL 31.9 – 32.9 mm); (2) distinct black spots present on flanks; (3) toes rudimentarily webbed, with wide lateral fringes; (4) skin on dorsum shagreened with fine tiny granules and short ridges; (5) heels overlapped when thighs are positioned at right angles to the body; (6) tibia-tarsal articulation reaching interior corner of the eye.
A new species of the Asian leaf litter toad genus Leptobrachella from Guizhou Province, China is described. Molecular phylogenetic analyses, based on mitochondrial 16S rRNA and COI genes and nuclear RAG1 gene sequences indicated that the new species is genetically divergent from its congeners. The new species could be distinguished from its congeners by a combination of the following characters: (1) body of medium size in males (SVL 31.9 – 32.9 mm); (2) distinct black spots present on flanks; (3) toes rudimentarily webbed, with wide lateral fringes; (4) skin on dorsum shagreened with fine tiny granules and short ridges; (5) heels overlapped when thighs are positioned at right angles to the body; (6) tibia-tarsal articulation reaching interior corner of the eye.
Taxonomy, molecular phylogenetic analyses, morphology, new species
The Asian leaf litter toads of the genus Leptobrachella Smith, 1925 (Anura, Megophryidae) inhabit the forest floor and rocky streams in hilly evergreen forests and are widely distributed from southern China, west to north-eastern India and Myanmar, through mainland Indochina to Peninsular Malaysia and the Island of Borneo (
From 2022 to 2023, four specimens of Leptobrachella were collected from Dushan County, Guizhou Province, China. Morphologically, these specimens most closely resemble L. dong, but differ from L. dong and all other Leptobrachella from China and adjoining countries. To distinguish these specimens, we conducted phylogenetic analyses, based on mitochondrial DNA, nuclear DNA and morphological comparisons. All of the analyses consistently indicated that the specimens from Dushan County are a new taxon. Herein, we describe this taxon as a new species.
Four specimens of the undescribed species (Table
Information for samples used in molecular phylogenetic analyses in this study (/ = not available)
ID | Species | Locality | Voucher number | GenBank accession number | ||
16S | COI | RAG1 | ||||
1 | Leptobrachella dushanensis sp. nov. | Dushan County, Guizhou Province, China | CIB DS20220409002 | PP061389 | PP061403 | PP067958 |
2 | Leptobrachella dushanensis sp. nov. | Dushan County, Guizhou Province, China | CIB DS20230310002 | PP061391 | PP061405 | / |
3 | Leptobrachella dushanensis sp. nov. | Dushan County, Guizhou Province, China | CIB DS20230310001 | PP061390 | PP061404 | / |
4 | Leptobrachella dushanensis sp. nov. | Dushan County, Guizhou Province, China | CIB DS20220409001 | PP061388 | PP061402 | PP067957 |
5 | L. graminicola | Mount Pu Ta Leng, Lao Cai, Vietnam | VNMN 010905 | MZ224648 | / | / |
6 | L. graminicola | Mount Pu Ta Leng, Lao Cai, Vietnam | VNMN 010908 | MZ224653 | / | / |
7 | L. graminicola | Mount Pu Ta Leng, Lao Cai, Vietnam | VNMN 010909 | MZ224649 | / | / |
8 | L. graminicola | Mount Pu Ta Leng, Lao Cai, Vietnam | TOLO2022.518 | PP061392 | PP061406 | / |
9 | L. graminicola | Mount Pu Ta Leng, Lao Cai, Vietnam | VNMN 010904 | MZ224651 | / | / |
10 | L. graminicola | Mount Pu Ta Leng, Lao Cai, Vietnam | VNMN 010912 | MZ224647 | / | / |
11 | L. graminicola | Mount Pu Ta Leng, Lao Cai, Vietnam | VNMN 010910 | MZ224655 | / | / |
12 | L. yeae | Changshouqiao, Mount Emei, Sichuan, China | CIBEM1867 | MT957005 | / | MT975978 |
13 | L. yeae | Linggongli, Mount Emei, Sichuan, China | CIBEMLGL19052104 | MT957006 | / | MT975979 |
14 | L. yeae | Mount Emei, Sichuan, China | KIZ025778 | KX811928 | KX812166 | / |
15 | L. dong | Congjiang County, Guizhou Province, China | CIB LB20220305005 | OP764533 | PP061396 | / |
16 | L. dong | Congjiang County, Guizhou Province, China | CIB LB20220311002 | OP764535 | PP061398 | OP776441 |
17 | L. dong | Congjiang County, Guizhou Province, China | CIB LB20220306008 | OP764534 | PP061397 | OP776439 |
18 | L. dong | Suining County, Hunan Province, China | CIB ZNY2022001 | OP764538 | PP061399 | OP776442 |
19 | L. dong | Suining County, Hunan Province, China | CIB ZNY2022002 | OP764539 | PP061400 | OP776443 |
20 | L. dong | Tongdao County, Hunan Province, China | CIB WB2020277 | OP764531 | PP061401 | OP776448 |
21 | L. bourreti | Sapa, Lao Cai, Vietnam | 1999.566 | KR827860 | / | / |
22 | L. bourreti | Sapa, Lao Cai Province, Vietnam | AMS R 177673 | KR018124 | / | / |
23 | L. bourreti | Ky Quan San, Lao Cai, Vietnam | AMS R.188515 | MZ208835 | / | / |
24 | L. bourreti | Mount Pu Ta Leng, Lao Cai, Vietnam | VNMN 010916 | MZ209167 | / | / |
25 | L. bourreti | Bat Xat District, Lao Cai, Vietnam | VNMN 010073 | PP061393 | PP061407 | / |
26 | L. bourreti | Bat Xat District, Lao Cai, Vietnam | ZMMU-A5636-02280 | MH055872 | / | / |
27 | L. jinyunensis | Mt.Jinyun, Beibei District, Chongqing, China | CIB 119058 | OQ024797 | OQ024396 | OQ031199 |
28 | L. jinyunensis | Mt.Jinyun, Beibei District, Chongqing, China | CIB 119061 | OQ024800 | OQ024399 | OQ031202 |
29 | L. chishuiensis | Chishui National Nature Reserve, Chishui, Guizhou, China | CIBCS20190518047 | MT117055 | OQ024422 | OQ031225 |
30 | L. chishuiensis | Chishui National Nature Reserve, Chishui, Guizhou, China | CIBCS20190518049 | MT117053 | OQ024424 | OQ031227 |
31 | L. bijie | Mt. Baima, Qixingguan, Bijie, Guizhou, China | CIB 119068 | OQ024807 | OQ024406 | OQ031208 |
32 | L. bijie | Mt. Baima, Qixingguan, Bijie, Guizhou, China | CIB 119069 | OQ024808 | OQ024407 | OQ031209 |
33 | L. jinshaensis | Lengshuihe Nature Reserve, Jinsha County, Guizhou, China | CIBJS20200516005 | OQ024816 | OQ024417 | OQ031220 |
34 | L. jinshaensis | Lengshuihe Nature Reserve, Jinsha County, Guizhou, China | CIBJS20200516002 | OQ024815 | OQ024419 | OQ031222 |
35 | L. suiyangensis | Huoqiuba Nature Reserve, Suiyang County, Guizhou, China | GZNU20180606005 | MK829649 | / | OL800396 |
36 | L. suiyangensis | Huoqiuba Nature Reserve, Suiyang County, Guizhou, China | GZNU20180606006 | MK829650 | / | OL800397 |
37 | L. purpuraventra | Mt. Baima, Qixingguan, Bijie, Guizhou, China | CIB 119072 | OQ024811 | OQ024412 | OQ031215 |
38 | L. purpuraventra | Mt. Baima, Qixingguan, Bijie, Guizhou, China | CIB 119073 | OQ024812 | OQ024413 | OQ031216 |
39 | L. niveimontis | Daxueshan Nature Reserve, Yunnan Province, China | KIZ015734 | MT302618 | / | OL800394 |
40 | L. yunyangensis | Qiyaoshan Nature Reserve, Yunyang County, Chongqing, China | GZNU20210622001 | OL800364 | / | OL800393 |
41 | L. eos | Long Nai Khao, Phongsali, Laos | MNHN: 2004.0276 | KR827862 | KR087758 | / |
42 | L. eos | Boun Tay, Phongsaly, Laos | NCSM 80551 | MH055887 | / | / |
43 | L. purpura | Yingjiang, Yunnan Province, China | SYS a006530 | MG520354 | / | / |
44 | L. alpina | Caiyanghe, Yunnan Province, China | KIZ049024 | MH055867 | / | / |
45 | L. wulingensis | Tianquanshan Forest Park, Zhangjiajie, Hunan Province, China | CSUFT 177 | MT530315 | / | / |
46 | L. dorsospina | Yushe Forest Park, Shuicheng County, Guizhou Province, China | SYS a004961 | MW046194 | / | / |
47 | L. oshanensis | Baoguosi, Mount Emei, Sichuan Province, China | CIBEMS20190421BGS1 | MT957023 | / | / |
48 | L. oshanensis | Shengshuige, Mount Emei, Sichuan, China | CIBEMS20190422SSG1-4 | MT957025 | / | / |
49 | L. oshanensis | Heilongjiang, Mount Emei, Sichuan, China | CIBEMS20190422HLJ1-4 | MT957027 | / | / |
50 | L. oshanensis | Shengshuige, Mount Emei, Sichuan, China | CIBEMS20190421SSG1-10 | MT957031 | / | / |
51 | L. oshanensis | Heilongjiang, Mount Emei, Sichuan, China | CIBEMS20190422HLJ1-2 | MT957033 | / | / |
52 | L. korifi | Doi Inthanon,Thailand | KUHE 19134 | LC741033 | / | / |
53 | L. sinorensis | Mae Hong Son,Thailand | KUHE 19809 | LC741034 | / | / |
54 | L. murphyi | Doi Inthanon, Chiang Mai, Thailand | KIZ034039 | MZ710519 | / | / |
55 | L. tengchongensis | Gaoligong Shan, Yunnan Province, China | SYS a004598 | KU589209 | / | / |
56 | L. khasiorum | Khasi Hills, Meghalaya, India | SDBDU 2009.329 | KY022303 | / | / |
57 | L. tamdil | Mizoram, India | MZMU2224 | MW665130 | / | / |
58 | L. yingjiangensis | Yingjiang County, Yunnan Province, China | SYS a006533 | MG520350 | / | / |
59 | L. puhoatensis | Pu Hu, Thanh Hoa, Vietnam | VNMN:2016 A.23 | KY849587 | / | / |
60 | L. petrops | Ba Vi National Park, Ha Tay, Vietnam | ROM 13483 | MH055901 | / | / |
61 | L. namdongensis | Thanh Hoa Province, Vietnam | VNUF A.2017.37 | MK965389 | / | / |
62 | L. liui | Wuyi Shan City, Fujian Province, China | SYS a001597 | KM014547 | / | / |
63 | L. mangshanensis | Mangshan, Hunan Province, China | MSZTC201701 | MG132196 | / | / |
64 | L. verrucosa | Lianshan Bijiashan Nature Reserve, Guangdong, China | GEP a059 | OP279589 | / | / |
65 | L. shimentaina | Shimentai Nature Reserve, Guangdong, China | SYS a004712 | MH055926 | / | / |
66 | L. flaviglandulosa | Xiaoqiaogou Nature Reserve, Yunnan Province, China | KIZ016072 | MH055934 | / | / |
67 | L. yunkaiensis | Dawuling Forest Station, Maoming City, Guangdong Province, China | SYS a004663 | MH605584 | / | / |
68 | L. maoershanensis | Mao’er Shan, Guangxi Province, China | KIZ07614 | MH055927 | / | / |
69 | L. bashaensis | Basha Nature Reserve, Congjiang County, Guizhou Province, China | GIB196403 | MW136294 | / | / |
70 | L. laui | Shenzhen City, Guangdong Province, China | SYS a002450 | MH055904 | / | / |
71 | L. firthi | Quang Nam Province, Vietnam | AMS R 171714 | JQ739203 | / | / |
72 | L. isos | Gia Lai, Vietnam | AMS R 176469 | KT824767 | / | / |
73 | L. sungi | Tam Dao, Vinh Phuc, Vietnam | ROM 20236 | MH055858 | / | / |
74 | L. zhangyapingi | Chiang Mai, Thailand | KIZ07258 | MH055864 | / | / |
75 | L. pelodytoides | Tam Dao, Vinh Phu, Vietnam | ROM18282 | EF397244 | / | / |
76 | L. ventripunctata | Wenlong, Yunnan Province, China | KIZ013621 | MH055824 | / | / |
77 | L. feii | Xiaoqiaogou Nature Reserve, Yunnan Province, China | KIZ048894 | MT302634 | / | / |
78 | L. aspera | Huanglianshan Nature Reserve, Lyuchun, Yunnan, China | SYS a007743 | MW046199 | / | / |
79 | L. aerea | Vilabuly, Savannakhet, Laos | NCSM 76038 | MH055809 | / | / |
80 | L. minima | Doi Phu Fa, Nan, Thailand | KIZ024317 | MH055852 | / | / |
81 | L. nyx | Ha Giang, Vietnam | ROM 36692 | MH055816 | / | / |
82 | L. wuhuangmontis | Pubei County, Guangxi Province, China | SYS a003485 | MH605577 | / | / |
83 | L. shiwandashanensis | Fangcheng City, Guangxi Province, China | NNU202103146 | MZ326691 | / | / |
84 | L. wumingensis | Wuming County, Guangxi, China | NNU 00283 | OM935575 | / | / |
85 | L. pluvialis | Fansipan, Lao Cai, Vietnam | ROM 30685 | MH055843 | / | / |
86 | L. shangsiensis | Guangxi Province, China | NHMG1401032 | MK095460 | / | / |
87 | L. nahangensis | Na Hang Nature Reserve, Tuyen Quang, Vietnam | ROM 7035 | MH055853 | / | / |
88 | L. damingshanensis | Guangxi Province., China | NNU202103281 | MZ145229 | / | / |
89 | L. kalonensis | Binh Thuan Province, Vietnam | AMNH A191762 | KR018115 | / | / |
90 | L. bidoupensis | Bidoup-Nui Ba National Park, Lam Dong, Vietnam | ZMMU-A-4797-01454 | MH055945 | / | / |
91 | L. pallida | Lam Dong, Vietnam | UNS00511 | KU530190 | / | / |
92 | L. tadungensis | Dak Nong Province, Vietnam | UNS00515 | KR018121 | / | / |
93 | L. maculosa | Ninh Thuan Province, Vietnam | AMS R 177660 | KR018119 | / | / |
94 | L. pyrrhops | Loc Bac, Lam Dong, Vietnam | ZMMU-A-4873-00158 | MH055950 | / | / |
95 | L. macrops | Phu Yen, Vietnam | ZMMU-A5823 | MG787993 | / | / |
96 | L. rowleyae | Da Nang City, Vietnam | ITBCZ2783 | MG682552 | / | / |
97 | L. applebyi | Phong Dien Nature Reserve, Thua Thien-Hue, Vietnam | KIZ010701 | MH055947 | / | / |
98 | L. melica | Cambodia, Ratanakiri | MVZ258198 | HM133600 | / | / |
99 | L. ardens | Kon Ka Kinh National Park, Gia Lai, Vietnam | ZMMU-NAP-06099 | MH055949 | / | / |
100 | L. crocea | Thua Thien-Hue, Vietnam | ZMMU-NAP-02274 | MH055955 | / | / |
101 | L. tuberosa | Kon Ka Kinh National Park, Gia Lai, Vietnam | ZMMU-NAP-02275 | MH055959 | / | / |
102 | L. botsfordi | Fansipan, Lao Cai, Vietnam | AMS R 176540 | MH055952 | / | / |
103 | L. fuliginosa | Phetchaburi, Thailand | KUHE 20197 | LC201988 | / | / |
104 | L. melanoleuca | Kapoe, Ranong, Thailand | KIZ018031 | MH055967 | / | / |
105 | L. neangi | Veal Veng District, Pursat Province, Cambodia | CBC 1624 | MT644613 | / | / |
106 | L. brevicrus | Gunung Mulu National Park, Sarawak, Malaysia | UNIMAS 8957 | KJ831303 | / | / |
107 | L. itiokai | Mulu National Park, Sarawak, Malaysia | KUHE:55897 | LC137805 | / | / |
108 | L. parva | Mulu National Park, Sarawak, Malaysia | KUHE:55308 | LC056791 | / | / |
109 | L. baluensis | Tambunan, Sabah, Borneo, Malaysia | SP 21604 | LC056792 | / | / |
110 | L. mjobergi | Gading NP, Sarawak, Borneo, Malaysia | KUHE:47872 | LC056787 | / | / |
111 | L. juliandringi | Mulu NP, Sarawak, Borneo, Malaysia | KUHE 55333 | LC056780 | / | / |
112 | L. sabahmontana | Borneo, Malaysia | BORNEENSIS 12632 | AB847551 | / | / |
113 | L. dringi | Gunung Mulu, Malaysia | KUHE:55610 | AB847553 | / | / |
114 | L. picta | Borneo, Malaysia | UNIMAS 8705 | KJ831295 | / | / |
115 | L. fritinniens | Danum Valley Field Center, Sabah, Malaysia | FMNH 244800 | MH055971 | / | / |
116 | L. arayai | Liwagu, Kinabalu, Malaysia | BORNEEISIS 22931 | AB847558 | / | / |
117 | L. hamidi | Bukit Lanjan, Selangor, Malaysia | KUHE17545 | AB969286 | / | / |
118 | L. marmorata | Borneo, Malaysia | KUHE53227 | AB969289 | / | / |
119 | L. gracilis | Bukit Kana, Sarawak, Malaysia | FMNH 273682 | MH055972 | / | / |
120 | L. maura | Borneo, Malaysia | SP21450 | AB847559 | / | / |
121 | L. heteropus | Larut, Perak, Malaysia | KUHE15487 | AB530453 | / | / |
122 | L. sola | Gunung Stong, Kelantan, Malaysia | KU RMB20973 | MH055973 | / | / |
123 | L. kecil | Cameron, Malaysia | KUHE 52440 | LC202004 | / | / |
124 | L. kajangensis | Tioman, Malaysia | LSUHC 4431 | LC202001 | / | / |
125 | Megophrys glandulosa | Yunnan Province, China | KIZ048439 | KX811762 | KX812165 | / |
126 | Leptobrachium huashen | Yunnan Province, China | KIZ049025 | KX811931 | KX812075 | / |
Sampling localities of Leptobrachella dushanensis sp. nov., L. bourreti, L. dong and L. graminicola. 1 Dushan County, the type locality of Leptobrachella dushanensis sp. nov.; 2-4 represent the sampling localities of L. dong; 2 Congjiang County, Guizhou Province; 3 Tongdao County, Hunan Province; 4 Suining County, Hunan Province, China; 5 represent the sampling locality of L. bourreti from Bat Xat District, Lao Cai, Vietnam; 6 the sampling locality of L. graminicola from Mount Pu Ta Leng, Lao Cai, Vietnam.
Total DNA was extracted using a standard phenol-chloroform extraction protocol (
For phylogenetic analyses, we downloaded the corresponding gene sequences for all related species in the genus Leptobrachella from GenBank, based on previous studies (
Sequences were assembled and aligned using the Clustalw module in BioEdit v. 7.0.9.0 (
Morphologically, these specimens most closely resemble L. dong. To explore the morphological differences between the new taxon and L. dong, four specimens of the new taxon and 15 specimens of L. dong containing four specimens from Guizhou Province and 11 specimens from Hunan Province were measured. The terminology and methods followed
In order to reduce the impact of allometry, the correct value from the ratio of each measurement to SVL was calculated and then log-transformed for the following morphometric analyses. One-way analysis of variance (ANOVA) was used to test the significance of differences on morphometric characters between the undescribed species and L. dong from different populations in males. The significance level was set at 0.05. To show the spatial distribution of different species on the morphometric characters, principal component analyses (PCA) were performed. All statistical analyses were performed using SPSS 21.0 (SPSS, Inc., Chicago, IL, USA).
The undescribed taxon was also compared with all other congeners of Leptobrachella, based on morphological characters. Comparative morphological data were obtained from literature (Table
References for morphological characters for congeners of the genus Leptobrachella.
ID | Species | References |
1 | L. aerea (Rowley, Stuart, Richards, Phimmachak & Sivongxay, 2010) |
|
2 | L. alpina (Fei, Ye & Li, 1990) |
|
3 | L. applebyi (Rowley & Cao, 2009) |
|
4 | L. arayai (Matsui, 1997) |
|
5 | L. ardens (Rowley, Tran, Le, Dau, Peloso, Nguyen, Hoang, Nguyen & Ziegler, 2016) |
|
6 | L. aspera Wang, Lyu, Qi & Wang, 2020 |
|
7 | L. baluensis Smith, 1931 |
|
8 | L. bashaensis Lyu, Dai, Wei, He, Yuan, Shi, Zhou, Ran, Kuang, Guo, Wei & Yuan, 2020 |
|
9 | L. bidoupensis (Rowley, Le, Tran & Hoang, 2011) |
|
10 | L. bijie Wang, Li, Li, Chen & Wang, 2019 |
|
11 | L. bondangensis Eto, Matsui, Hamidy, Munir & Isk &ar, 2018 |
|
12 | L. botsfordi (Rowley, Dau & Nguyen, 2013) |
|
13 | L. bourreti (Dubois, 1983) |
|
14 | L. brevicrus Dring, 1983 |
|
15 | L. chishuiensis Li, Liu, Wei & Wang, 2020 |
|
16 | L. crocea (Rowley, Hoang, Le, Dau & Cao, 2010) |
|
17 | L. damingshanensis Chen, Yu, Cheng, Meng, Wei, Zhou, Lu, 2021 |
|
18 | L. dorsospina Wang, Lyu, Qi & Wang, 2020 |
|
19 | L. dringi (Dubois, 1987) |
|
20 | L. dong Liu, Shi, Li, Zhang, Xiang, Wei, and Wang, 2023 |
|
21 | L. eos (Ohler, Wollenberg, Grosjean, Hendrix, Vences, Ziegler & Dubois, 2011) |
|
22 | L. feii Chen, Yuan & Che, 2020 |
|
23 | L. firthi (Rowley, Hoang, Dau, Le & Cao, 2012) |
|
24 | L. flaviglandulosa Chen, Wang & Che, 2020 |
|
25 | L. fritinniens (Dehling & Matsui, 2013) |
|
26 | L. fuliginosa (Matsui, 2006) |
|
27 | L. fusca Eto, Matsui, Hamidy, Munir & Isk &ar, 2018 |
|
28 | L. gracilis (Günther, 1872) |
|
29 | L. graminicola Nguyen, Tapley, Nguyen, Luong & Rowley, 2021 |
|
30 | L. hamidi (Matsui, 1997) |
|
31 | L. heteropus (Boulenger, 1900) |
|
32 | L. isos (Rowley, Stuart, Neang, Hoang, Dau, Nguyen & Emmett, 2015) |
|
33 | L. itiokai Eto, Matsui & Nishikawa, 2016 |
|
34 | L. jinshaensis Cheng, Shi, Li, Liu, Li & Wang, 2021 |
|
35 | L. jinyunensis Shi, Shen, Wang, Jiang & Wang, 2023 |
|
36 | L. juliandringi Eto, Matsui & Nishikawa, 2015 |
|
37 | L. kajangensis (Grismer, Grismer & Youmans, 2004) |
|
38 | L. kalonensis (Rowley, Tran, Le, Dau, Peloso, Nguyen, Hoang, Nguyen & Ziegler, 2016) |
|
39 | L. kecil (Matsui, Belabut, Ahmad & Yong, 2009) |
|
40 | L. khasiorum (Das, Tron, Rangad & Hooroo, 2010) |
|
41 | L. korifi Matsui, Panha & Eto, 2023 |
|
42 | L. lateralis (Anderson, 1871) |
|
43 | L. laui (Sung, Yang & Wang, 2014) |
|
44 | L. liui (Fei & Ye, 1990) |
|
45 | L. macrops (Duong, Do, Ngo, Nguyen & Poyarkov, 2018) |
|
46 | L. maculosa (Rowley, Tran, Le, Dau, Peloso, Nguyen, Hoang, Nguyen & Ziegler, 2016) |
|
47 | L. mangshanensis (Hou, Zhang, Hu, Li, Shi, Chen, Mo & Wang, 2018) |
|
48 | L. maoershanensis (Yuan, Sun, Chen, Rowley & Che, 2017) |
|
49 | L. marmorata (Matsui, Zainudin & Nishikawa, 2014) |
|
50 | L. maura (Inger, Lakim, Biun & Yambun, 1997) |
|
51 | L. melanoleuca (Matsui, 2006) |
|
52 | L. melica (Rowley, Stuart, Neang & Emmett, 2010) |
|
53 | L. minima (Taylor, 1962) |
|
54 | L. mjobergi Smith, 1925 |
|
55 | L. murphyi Chen, Suwannapoom, Wu, Poyarkov, Xu, Pawangkhanant & Che, 2021 |
|
56 | L. nahangensis (Lathrop, Murphy, Orlov & Ho, 1998) |
|
57 | L. namdongensis Hoang, Nguyen, Luu, Nguyen & Jiang, 2019 |
|
58 | L. natunae (Günther, 1895) |
|
59 | L. neangi Stuart & Rowley, 2020 |
|
60 | L. niveimontis Chen, Poyarkov, Yuan & Che, 2020 |
|
61 | L. nokrekensis (Mathew & Sen, 2010) |
|
62 | L. nyx (Ohler, Wollenberg, Grosjean, Hendrix, Vences, Ziegler & Dubois, 2011) |
|
63 | L. oshanensis (Liu, 1950) |
|
64 | L. pallida (Rowley, Tran, Le, Dau, Peloso, Nguyen, Hoang, Nguyen & |
|
65 | L. palmata Inger & Stuebing, 1992 |
|
66 | L. parva Dring, 1983 |
|
67 | L. pelodytoides (Boulenger, 1893) |
|
68 | L. petrops (Rowley, Dau, Hoang, Le, Cutajar & Nguyen, 2017) |
|
69 |
L. phiadenensis Luong, Hoang, Pham, Ziegler, and Nguyen, 2023 |
|
70 |
L. phiaoacensisLuong, Hoang, Pham, Ziegler, and Nguyen, 2023 |
|
71 | L. picta (Malkmus, 1992) |
|
72 | L. pingbianensis (Rao, Hui, Zhu & Ma, 2022 "2020") |
|
73 | L. platycephala (Dehling, 2012) |
|
74 | L. pluvialis (Ohler, Marquis, Swan & Grosjean, 2000) |
|
75 | L. puhoatensis (Rowley, Dau & Cao, 2017) |
|
76 | L. purpuraventra Wang, Li, Li, Chen & Wang, 2019 |
|
77 | L. purpurus (Yang, Zeng & Wang, 2018) |
|
78 | L. pyrrhops (Poyarkov, Rowley, Gogoleva, Vassilieva, Galoyan & Orlov, 2015) |
|
79 | L. rowleyae (Nguyen, Poyarkov, Le, Vo, Ninh, Duong, Murphy & Sang, 2018) |
|
80 | L. sabahmontana (Matsui, Nishikawa & Yambun, 2014) |
|
81 | L. serasanae Dring, 1983 |
|
82 | L. shangsiensis Chen, Liao, Zhou & Mo, 2019 |
|
83 | L. shiwanshanensis Chen, Peng, Pan, Liao, Liu & Huang, 2021 |
|
84 | L. shimentaina Wang, Lyu & Wang, 2022 |
|
85 | L. sinorensis Matsui, Panha & Eto, 2023 |
|
86 | L. sola (Matsui, 2006) |
|
87 | L. suiyangensis Luo, Xiao, Gao & Zhou, 2020 |
|
88 | L. sungi (Lathrop, Murphy, Orlov & Ho, 1998) |
|
89 | L. tadungensis (Rowley, Tran, Le, Dau, Peloso, Nguyen, Hoang, Nguyen & Ziegler, 2016) |
|
90 | L. tamdil (Sengupta, Sailo, Lalremsanga, Das & Das, 2010) |
|
91 | L. tengchongensis (Yang, Wang, Chen & Rao, 2016) |
|
92 | L. tuberosa (Inger, Orlov & Darevsky, 1999) |
|
93 | L. ventripunctata (Fei, Ye & Li, 1990) |
|
94 | L. verrucosa Wang, Zeng, Lin & Li, 2022 |
|
95 | L. wuhuangmontis Wang, Yang & Wang, 2018 |
|
96 | L. wulingensis Qian, Xiao, Cao, Xiao & Yang, 2020 |
|
97 | L. wumingensis Chen, Peng, Li, and Yu, 2023 |
|
98 | L. yeae Shi, Hou, Song, Jiang & Wang, 2021 |
|
99 | L. yingjiangensis (Yang, Zeng & Wang, 2018) |
|
100 | L. yunyangensis Luo, Deng & Zhou, 2022 |
|
101 | L. yunkaiensis Wang, Li, Lyu & Wang, 2018 |
|
102 | L. zhangyapingi (Jiang, Yan, Suwannapoom, Chomdej & Che, 2013) |
|
All the sequences in this study were retrieved from GenBank and the accession numbers of the newly-determined sequences in this study are shown in Table
Leptobrachella dushanensis sp. nov. is assigned to the genus Leptobrachella, based on molecular data and the following morphological characters: medium size, rounded finger tips, the presence of two elevated inner palmar tubercle not continuous to the thumb, presence of macroglands on body (including supra-axillary, pectoral and femoral glands), vomerine teeth absent, tubercles on eyelids and anterior tip of snout with vertical white bar (Dubois 1983; Fei et al. 2009).
Leptobrachella dushanensis sp. nov. could be distinguished from its congeners by a combination of the following characters: body of medium size in males (SVL 31.9 – 32.9 mm); distinct black spots present on flanks; toes rudimentarily webbed, with wide lateral fringes; skin on dorsum shagreened with fine tiny granules and short ridges; heels overlapped when thighs are positioned at right angles to the body; tibia-tarsal articulation reaching interior corner of the eye.
Adult male. SVL in 33.2 mm. Head length slightly wider than head width (HDL/HDW = 1.04); snout sharply rounded in dorsal view, projecting slightly beyond margin of the lower jaw; nostril closer to snout than eye; loreal region oblique; canthus rostralis indistinct; eyes large and convex (ED/HDL = 0.37), slightly shorter than snout length (ED/SL = 0.93), pupil vertical; tympanum distinct, rounded, tympanum diameter smaller than eye (TD/ED = 0.43), upper margin of tympanum in contact with supratympanic ridge; vomerine teeth absent; tongue notched behind; supratympanic ridge distinct, extending from posterior corner of eye to supra-axillary gland.
Fore-limb relatively long (LAL / SVL = 0.45), fingers long and slender (ML/SVL = 0.25), without webbing, lateral fringes on fingers narrow; relative finger lengths II < I < IV < III; tips of fingers rounded and slightly swollen; subarticular tubercles absent on fingers, inner metacarpal tubercle large and rounded, separated from the smaller, round outer metacarpal; supra-axillary glands oval.
Hind-limb relatively long (HLL/SVL = 1.56), heels overlapping when the tibiae perpendicular to the body axis; tibio-tarsal articulation of adpressed limb reaching interior corner of the eye, tibia length about half of snout-vent length (TL/SVL = 0.49); relative toe length: I < II < III < V < IV; toe tips rounded and slightly swollen; rudimentary webbing present between all five toes; wide lateral fringes present on all toes; subarticular tubercles indistinct on the base of toe; inner metatarsal tubercle oval and distinct, outer metatarsal tubercle absent.
Skin on dorsum shagreened with fine tiny granules and short ridges; supra-axillary glands long, oval, close to the armpit; pectoral gland indistinct; round femoral glands present and protuberant on rear of thigh, closer to knee than to vent; femoral adipose glands distinct, attached to inner side of skin on posterior ventral surface of thigh; ventral skin smooth; ventrolateral glands distinctly visible and raised, forming an incomplete line.
In life, dorsal surface of head and trunk earth brown, with a distinct reverse-triangle taupe markings between eyes connecting to a taupe W-shaped marking between axillae that are fringed with greyish-white colour; very distinct, light brown markings between the nostrils; bicoloured iris, with the upper 1/3 of the iris being copper-orange and the lower 2/3 a light silvery-grey and a dark blotch under the eye; sparse, small, light brown granules and small dark brown patches present on the dorsum of the limbs; elbow to upper arm distinctly greyish-brown in colour on the dorsum; three transverse black bars present on dorsal surface of lower arm; distinct dark blotches on flanks from groin to axilla, longitudinally in two rows; ventral surfaces light coloured, throat and ventral arms pinkish, chest and belly cream-white and flanks of ventral with several granules and brown spots; ventral hind-limbs pinkish with sparse white glands (Fig.
Dorsum of body and limbs fade to brown; transverse bars on limbs become more distinct. Ventral surface of body and limbs fade to cream-white. Supra-axillary, femoral and pectoral glands fade to cream-yellow (Fig.
Measurements of adult specimens are presented in Suppl. material
Measurements of Leptobrachella dushanensis sp. nov and L. dong from Guizhou and Hunan populations. Units in mm. In addition, the results of one-way ANOVA with p-values for morphometric comparisons between Leptobrachella dushanensis sp. nov. and L. dong from Guizhou and Hunan populations (* p < 0.05, ** p < 0.01) are given. See abbreviations for characters in the Materials and Methods section.
Character | Leptobrachella dushanensis sp.nov. (n = 4) | L. dong (n = 4) Guizhou | L. dong (n = 11) Hunan | p-value | ||||
Ranging | Mean ± SD | Ranging | Mean ± SD | Ranging | Mean ± SD | Guizhou | Hunan | |
SVL | 31.9 – 32.9 | 32.3 ± 0.4 | 29.5 – 31.2 | 30.4 ± 0.86 | 29.2 – 34.2 | 31.1 ± 1.4 | 0.006** | 0.169 |
HDL | 10.5 – 10.8 | 10.6 ± 0.1 | 9.6 – 10.8 | 10.1 ± 0.5 | 10.0 – 11.7 | 10.7 ± 0.5 | 0.694 | 0.025* |
HDW | 10.0 – 10.4 | 10.1 ± 0.2 | 9.8 – 10.9 | 10.5 ± 0.5 | 10.5 – 12.2 | 11.2 ± 0.6 | 0.001** | 0.000** |
SL | 4.2 – 4.4 | 4.2 ± 0.1 | 4.2 – 4.9 | 4.6 ± 0.3 | 4.0 – 5.0 | 4.4 ± 0.3 | 0.027* | 0.044* |
IND | 2.6 – 2.7 | 2.6 ± 0.1 | 3.5 – 4.4 | 3.8 ± 0.4 | 2.8 – 3.6 | 3.1 ± 0.3 | 0.000** | 0.000** |
IOD | 2.8 – 3.1 | 2.9 ± 0.1 | 2.3 – 3.2 | 2.8 ± 0.4 | 2.8 – 3.9 | 3.4 ± 0.4 | 0.788 | 0.001** |
UEW | 2.9 – 3.1 | 3.0 ± 0.1 | 2.8 – 3.4 | 3.1 ± 0.3 | 2.6 – 3.1 | 2.9 ± 0.2 | 0.094 | 0.946 |
ED | 3.8 – 4.0 | 3.9 ± 0.1 | 3.9 – 4.4 | 4.1 ± 0.2 | 3.6 – 4.4 | 4.2 ± 0.2 | 0.039* | 0.006** |
TYD | 1.6 – 1.8 | 1.7 ± 0.1 | 1.9 – 2.2 | 2.1 ± 0.1 | 1.5 – 1.7 | 1.6 ± 0.1 | 0.001** | 0.705 |
LAL | 14.4 – 14.5 | 14.5 ± 0.1 | 13.5 – 14.2 | 13.9 ± 0.3 | 14.3 – 15.5 | 14.8 ± 0.4 | 0.127 | 0.011* |
ML | 8.0 – 8.5 | 8.1 ± 0.2 | 7.3 – 8.3 | 7.9 ± 0.4 | 7.2 – 8.6 | 7.9 ± 0.4 | 0.216 | 0.750 |
TL | 15.9 – 16.0 | 15.9 ± 0.1 | 14.5 – 14.7 | 14.6 ± 0.1 | 14.5 – 16.4 | 15.5 ± 0.6 | 0.079 | 0.749 |
FL | 14.8 – 15.0 | 14.9 ± 0.1 | 13.5 – 15 | 14.2 ± 0.6 | 13.5 – 15.5 | 15.0 ± 0.6 | 0.625 | 0.408 |
HLL | 50.0 – 50.5 | 50.3 ± 0.3 | 41.3 – 44.2 | 43.3 ± 1.3 | 46.2 – 51.6 | 49.1 ± 1.6 | 0.018* | 0.564 |
Compared with the 26 known congeners occurring south of the Isthmus of Kra, Leptobrachella dushanensis sp. nov. could be distinguished from them by several characters. By having supra-axillary and ventrolateral glands, the new species differs from L. arayai, L. dringi, L. fritinniens, L. gracilis, L. hamidi, L. heteropus, L. kajangensis, L. kecil, L. marmorata, L. maura, L. melanoleuca, L. picta, L. platycephala, L. sabahmontana and L. sola (vs. absent in the latter); by having rounded fingertips and moderate body size (31.9 – 32.9 mm in four adult males), the new species differs from the following species with pointed fingertips and smaller body size: L. baluensis (14.9–15.9 mm in males), L. bondangensis (17.8 mm in male), L. brevicrus (17.1–17.8 mm in males), L. fusca (16.3 mm in male), L. itiokai (15.2–16.7 mm in males), L. juliandringi (17.0–17.2 mm in males), L. mjobergi (15.7–19.0 mm in males), L. natunae (17.6 mm in one adult male), L. palmata (14.4–16.8 mm in males), L. parva (15.0–16.9 mm in males) and L. serasanae (16.9 mm in female).
Leptobrachella dushanensis sp. nov. could also be identified from 76 known Leptobrachella species occurring north of the Isthmus of Kra by some characters (Suppl. material
By having medium size of body (SVL 31.9–32.9 mm in males), Leptobrachella dushanensis sp. nov. differs from the the smaller in males L. aerea (25.1–28.9 mm), L. alpina (24.0–26.4 mm), L. applebyi (19.6–22.3 mm), L. ardens (21.3–24.7 mm), L. aspera (22.4 mm), L. bashaensis (22.9–25.6 mm), L. bidoupensis (23.6–24.6), L. bijie (29.0–30.4), L. crocea (22.2–27.3 mm), L. dorsospina (28.7–30.5 mm), L. feii (21.5–22.8 mm), L. firthi (26.4–29.2 mm), L. flaviglandulosa (23.0–27.0 mm), L. fuliginosa (28.2–30.0 mm), L. graminicola (23.1–24.6 mm) , L. isos (23.7–27.9 mm), L. khasiorum (24.5–27.3 mm), L. lateralis (26.9–28.3 mm), L. laui (24.8–26.7 mm), L. liui (24.8–26.7 mm), L. macrops (28.0–29.3 mm), L. maculosa (24.2–26.6 mm), L. mangshanensis (22.22–27.76 mm), L. maura (26.1 mm), L. melica (19.5–22.8 mm), L. murphyi (23.2–24.9 mm), L. niveimontis (22.5–23.6 mm), L. pallida (24.5–27.7 mm, L. petrops (23.6–27.6 mm), L. pingbianensis (28 mm), L. pluvialis (21.3–22.3 mm), L. puhoatensis (24.2–28.1 mm), L. purpuraventra (27.3–29.8 mm), L. purpurus (25.0–27.5 mm), L. rowleyae (23.4–25.4 mm), L. shangsiensis (24.9–29.4 mm), L. shiwandashanensis (26.8–29.7 mm), L. shimentaina (26.4–28.9 mm), L. sinorensis (26.6–27.1 mm), L. suiyangensis (28.7–29.7 mm), L. tadungensis (23.3–28.2 mm), L. tengchongensis (23.9–26.0 mm), L. tuberosa (24.4–29.5 mm), L. ventripunctata (23.7–27.7 mm), L. verrucosa (23.2–25.9 mm), L. wuhuangmontis (25.6–30.0 mm), L. wumingensis (26.0–26.7 mm), L. yingjiangensis (25.7–27.6 mm) and L. yunkaiensis (25.9–29.3 mm); differs from the larger in males L. nahangensis (40.8 mm), L. platycephala (35.1 mm), L. sungi (48.3–52.7 mm in males) and L. zhangyapingi (45.8–52.5 mm).
By having black spots on flanks, Leptobrachella dushanensis sp. nov. differs from L. aerea, L. botsfordi, L. crocea, L. eos, L. firthi, L. isos, L. pallida, L. petrops, L. tuberosa and L. zhangyapingi (vs. lacking distinct black spots on the flanks in the latter).
By having rudimentary webbing, Leptobrachella dushanensis sp. nov. differs from L. ardens, L. jinshaensis, L. kalonensis, L. maculosa, L. oshanensis, L. pallida, L. petrops, L. rowleyae, L. shiwandashanensis and L. tadungensis (vs. absent webbing in the latter).
By having wide fringes on toes, Leptobrachella dushanensis sp. nov. differs from L. applebyi, L. ardens, L. aspera, L. bashaensis, L. bidoupensis, L. bijie, L. botsfordi, L. bourreti, L. chishuiensis, L. crocea, L. damingshanensis, L. dorsospina, L. feii, L. flaviglandulosa, L. fuliginosa, L. jinshaensis, L. kalonensis, L. korifi, L. lateralis, L. macrops, L. maculosa, L. mangshanensis, L. melica, L. minima, L. nahangensis, L. namdongensis, L. niveimontis, L. nyx, L. oshanensis, L. pallida, L. pelodytoides, L. petrops, L. phiaoacensis, L. phiadenensis,
L. pluvialis, L. puhoatensis, L. purpuraventra, L. pyrrhops, L. rowleyae, L. shangsiensis, L. sinorensis, L. shiwandashanensis, L. sungi, L. tengchongensis, L. tuberosa, L. ventripunctata, L. wuhuangmontis, L. wulingensis, L. wumingensis, L. yeae and L. yunyangensis (vs. fringes on toes narrow or absent in the latter).
By having dorsal surface shagreened with fine tubercles, Leptobrachella dushanensis sp. nov. differs from L. applebyi, L. bidoupensis, L. kalonensis, L. melica, L. minima, L. nahangensis, L. shangsiensis and L. tadungensis, all of which have the dorsum smooth and L. bourreti (dorsum smooth with small warts), L. fuliginosa (dorsum smooth with fine tubercles), L. liui (dorsum with round tubercles), L. macrops (dorsum roughly granular with large tubercles), L. maoershanensis (dorsum shagreened with tubercles), L. minima (dorsum smooth), L. neangi (dorsum with small, irregular bumps and ridges), L. nyx (dorsum with round tubercles), L. nokrekensis (dorsum tubercles and longitudinal folds), L. pelodytoides (dorsum with small, smooth warts), L. tamdil (dorsum weakly tuberculate, with low, oval tubercles), L. tuberosa (dorsum highly tuberculate), L. yunkaiensis (dorsum with raised warts) and L. wuhuangmontis (dorsum rough with conical tubercles).
In mitochondrial DNA trees, Leptobrachella dushanensis sp. nov. and L. dong clustered into one clade, being sisters. The new species differs from L. dong by the following characters: head length slightly wider than head width (vs. head width slightly wider than head length); males with internal single subgular vocal sac (vs. a pair of subgular internal vocal sacs); tibiotarsal articulation reaching to anterior edge of eye (vs. reaching to middle of eye).
Adult males with internal single subgular vocal sac, femoral adipose glands present on posterior surface of thigh and tiny transparent spines on chest during breeding season. Nuptial pads and spines absent on males.
This specifc name “Dushan” refers to the distribution of this species in Dushan County, Guizhou Province, China. We suggest its English common name “Dushan leaf litter toad” and Chinese name “Dushan Zhang Tu Chan (独山掌突蟾)”.
Leptobrachella dushanensis sp. nov. was only found in Dushan County, Guizhou Province, China. Elevations recorded range from 1000 m to 1200 m.
Leptobrachella dushanensis sp. nov. was found under stones in fast-flowing mountain streams surrounded by evergreen broadleaf forest (Fig.
Aligned sequence matrix of 16S, COI and RAG1 genes contained 519, 615 and 831 bps respectively. ML and BI analyses, based on 16S and COI, resulted in essentially identical topologies. All samples of the undescribed species were clustered into one clade being deeply clustered into the Leptobrachella clade and being sister to L. dong (Fig.
Phylogenetic trees of the genus Leptobrachella and a haplotype network constructed, based on RAG1 gene sequences. A Bayesian inference (BI) tree, based on mitochondrial COI gene; B Bayesian Inference (BI) tree, based on mitochondrial 16S gene; C the haplotype network constructed, based on RAG1 gene sequences. In this phylogenetic tree, Bayesian posterior probabilities (BPP) from BI analyses/bootstrap supports (BS) from ML analyses are listed beside the nodes. The symbol “-” represents a value below 0.50/50. For information of samples 1–126, refer to Table
The results of one-way ANOVA showed that males of the undescribed species differed significantly from L. dong from both Guizhou and Hunan populations, based on several morphometric characters. From Guizhou population, the undescribed species was larger in SVL, HDW and HLL and shorter in SL, IND, ED and TYD; from Hunan population, the undescribed species was larger in HDL and shorter in HDW, SL, IND, IOD, ED and LAL (all p-values < 0.05; Table
Plots of principal component analyses of Leptobrachella dushanensis sp. nov. and L. dong in males. PC1, the first principal component; PC2, the second principal component. Different species were denoted as different shapes, the red triangle represents Leptobrachella dushanensis sp. nov., black square represents L. dong from Guizhou Province and blue square represents L. dong from Hunan Province.
More detailed descriptions of results from morphological comparisons between the undescribed species and its congeners were presented in the Taxon Treatment section for describing the new species.
As a result, molecular and morphological results supported that our specimens from Guizhou Province, China were a new taxon.
The Asian leaf litter toads of Leptobrachella have low vagility and an exclusive association with montane forests and their populations are often highly structured and underestimation of species diversity occurs in the genus, which suggests a high degree of localised diversification and micro-endemism (
In this study, two mitochondrial genes and one nuclear gene were amplified. On the 16S gene, the genetic distance between Leptobrachella dushanensis sp. nov. and its closely-related species L. dong was 1.6% which is at the same level with L. bijie and L. jinyunensis, but on COI gene, the genetic distance between them is 7.3%, much larger than that between other solid species (Suppl. materials
Measurements of adult specimen of Leptobrachella dushanensis sp. nov. and L. dong. Units in mm. See abbreviations for characters in the Materials and Methods section.
Diagnosis characters on morphology of Leptobrachella dong sp. nov. from other congeners.
Uncorrected p-distance between Leptobrachella species on the 16S rRNA gene. Mean value of genetic distance is given in the lower half of the table.
Uncorrected p-distance between Leptobrachella species on the COI. Mean value of genetic distance is given in the lower half of the table.