Research Article |
Corresponding author: Jiang Zhou ( zhoujiang@ioz.ac.cn ) Academic editor: Angelica Crottini
© 2020 Tao Luo, Ning Xiao, Kai Gao, 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, Xiao N, Gao K, Zhou J (2020) A new species of Leptobrachella (Anura, Megophryidae) from Guizhou Province, China. ZooKeys 923: 115-140. https://doi.org/10.3897/zookeys.923.47172
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This study describes a new species of the genus Leptobrachella, Leptobrachella suiyangensis sp. nov. from the Huoqiuba Nature Reserve, Suiyang County, Guizhou Province, China, based on morphological data and phylogenetic analyses (16S rRNA mtDNA). The new species can be distinguished from other congeners by the molecular divergence and by a combination of morphological characters, including body size, dorsal and ventral patterns, dorsal skin texture, size of the pectoral and femoral glands, degree of webbing and fringing on the toes and fingers, dorsum coloration, and iris coloration in life. Currently, the genus Leptobrachella contains 75 species, 21 of which are found in China, including seven species reported from Guizhou Province. The uncorrected sequence divergence percentage between Leptobrachella suiyangensis sp. nov. and all homologous DNA sequences available for the 16S rRNA gene was found to be >4.7%. The new record of the species and its relationships with others in the same genus imply that species distribution, habitat variation, environmental adaptation, and diversity of the genus Leptobrachella in southwest China need to be further investigated.
Leptobrachella suiyangensis sp. nov., mitochondrial DNA, morphology, Southwest China
The genus Leptolalax Dubois, 1983 in the family Megophryidae Bonaparte, 1850 is regarded to be closely associated with the genus Leptobrachella Smith, 1925 and has been assigned as a synonym of the genus Leptobrachella based on a large-scale molecular analysis (
During a field survey in June 2018 in a montane evergreen forest, Suiyang County, Guizhou Province (Fig.
Eight specimens collected from the aforementioned area (Fig.
DNA samples were extracted from muscular tissues with a DNA extraction kit (Tiangen Biotech (Beijing) Co. Ltd). The mitochondrial gene and 16S ribosomal RNA gene (16S rRNA) were sequenced (951bp). The fragmented genes were amplified with primer pairs L3975 (5'-CGCCTGTTTACCAAAAACAT-3') and H4551 (5'-CCGGTCTGAACTCAGATCACGT-3') for 16S rRNA (
ID | Species | Locality | Voucher no. | GenBank no. |
---|---|---|---|---|
1 | Leptobrachella suiyangensis sp.nov. | Suiyang County, Guizhou, China | GZNU20180606002 | MK829648 |
2 | Leptobrachella suiyangensis sp.nov. | Suiyang County, Guizhou, China | GZNU20180606005 | MK829649 |
3 | Leptobrachella suiyangensis sp.nov. | Suiyang County, Guizhou, China | GZNU20180606006 | MK829650 |
4 | Leptobrachella aerea | Vilabuly, Savannakhet, Laos | NCSM 76038 | MH055809 |
5 | Leptobrachella aerea | Phong Nha-Ke Bang, Quang Binh, Vietnam | RH60165 | JN848437 |
6 | Leptobrachella alpina | Huangcaoling, Yunnan, China | KIZ046816 | MH055866 |
7 | Leptobrachella applebyi | Song Thanh Nature Reserve, Quang Nam, Vietnam | AMS R171704 | HM133598 |
8 | Leptobrachella baluensis | Tambunan, Sabah, Borneo, Malaysia | SP 21604 | LC056792 |
9 | Leptobrachella bidoupensis | Bidoup, Lam Dong, Vietnam | NCSM 77321 | HQ902883 |
10 | Leptobrachella bijie | Zhaozishan Nature Reserve, Bijie City, Guizhou, China | SYS a007313/CIB110002 | MK414532 |
11 | Leptobrachella bijie | Zhaozishan Nature Reserve, Bijie City, Guizhou, China | SYS a007314 | MK414533 |
12 | Leptobrachella botsfordi | Fansipan, Lao Cai, Vietnam | AMS R 176540 | MH055952 |
13 | Leptobrachella bourreti | Sapa, Lao Cai, Vietnam | 1999.566 | KR827860 |
14 | Leptobrachella brevicrus | Gunung Mulu National Park, Sarawak, Malaysia | UNIMAS 8957 | KJ831303 |
15 | Leptobrachella dringi | Gunung Mulu, Malaysia | KUHE:55610 | AB847553 |
16 | Leptobrachella eos | Boun Tay, Phongsaly, Laos | NCSM 80551 | MH055887 |
17 | Leptobrachella eos | Zhushihe, Yunnan, China | SYS a003959 | MH055888 |
18 | Leptobrachella firthi | Ngoc Linh Nature Reserve, Kon Tum, Vietnam | AMS: R 176506 | JQ739207 |
19 | Leptobrachella fritinniens | Gunung Mulu, Malaysia | KUHE55371 | AB847557 |
20 | Leptobrachella gracilis | Gunung Mulu, Malaysia | KUHE55624 | AB847560 |
21 | Leptobrachella hamidi | Bukit Lanjan, Selangor, Malaysia | KUHE17545 | AB969286 |
22 | Leptobrachella heteropus | Larut, Perak, Malaysia | KUHE15487 | AB530453 |
23 | Leptobrachella isos | Gia Lai, Vietnam | AMS R 176469 | KT824767 |
24 | Leptobrachella itiokai | Mulu NP, Sarawak, Borneo, Malaysia | KUHE 55845 | LC137802 |
25 | Leptobrachella juliandringi | Mulu NP, Sarawak, Borneo, Malaysia | KUHE 55333 | LC056780 |
26 | Leptobrachella kajangensis | Tioman, Malaysia | LSUHC 4431 | LC202001 |
27 | Leptobrachella kecil | Cameron, Malaysia | KUHE 52440 | LC202004 |
28 | Leptobrachella khasiorum | Khasi Hills, Meghalaya, India | SDBDU 2009.329 | KY022303 |
29 | Leptobrachella liui | Wuyi Shan, Fujian, China | SYS a001597 | KM014547 |
30 | Leptobrachella liui | Wuyi Shan, Fujian, China | ZYCA907 | MH055908 |
31 | Leptobrachella laui | Shenzhen, Guangdong, China | SYS a002450 | MH055904 |
32 | Leptobrachella laui | Shenzhen, Guangdong, China | SYS a001515 | KM014545 |
33 | Leptobrachella macrops | Phu Yen, Vietnam | ZMMU-A5823 | MG787993 |
34 | Leptobrachella mangshanensis | Mangshan, Hunan, China | MSZTC201701 | MG132196 |
35 | Leptobrachella mangshanensis | Mangshan, Hunan, China | MSZTC201702 | MG132197 |
36 | Leptobrachella maoershanensis | Mao’er Shan, Guangxi, China | KIZ07614 | MH055927 |
37 | Leptobrachella maoershanensis | Mao’er Shan, Guangxi, China | KIZ027236 | MH055928 |
38 | Leptobrachella marmorata | Borneo, Malaysia | KUHE53227 | AB969289 |
39 | Leptobrachella maura | Borneo, Malaysia | SP21450 | AB847559 |
40 | Leptobrachella melanoleucus | Surat Thani, Thailand | KUHE:23845 | LC201999 |
41 | Leptobrachella melica | Cambodia, Ratanakiri | MVZ258198 | HM133600 |
42 | Leptobrachella minimus | Doi Chiang Dao, Chiangmai, Thailand | THNHM07418 | JN848402 |
43 | Leptobrachella minimus | Doi Suthep, Thailand | KUHE:19201 | LC201981 |
44 | Leptobrachella mjobergi | Gading NP, Sarawak, Borneo, Malaysia | KUHE:47872 | LC056787 |
45 | Leptobrachella nahangensis | Na Hang Nature Reserve, Tuyen Quang, Vietnam | ROM 7035 | MH055853 |
46 | Leptobrachella nahangensis | Na Hang, Tuyen Quang, Vietnam | ZMMU-NAP-02259 | MH055854 |
47 | Leptobrachella nyx | Ha Giang, Vietnam | ROM 36692 | MH055816 |
48 | Leptobrachella oshanensis | Emei Shan, Sichuan, China | KIZ025776 | MH055895 |
49 | Leptobrachella oshanensis | Emei Shan, Sichuan, China | Tissue ID: YPX37492 | MH055896 |
50 | Leptobrachella pallida | Vietnam: Lam Dong | UNS00511 | KU530190 |
51 | Leptobrachella parva | Mulu National Park, Sarawak, Malaysia | KUHE:55308 | LC056791 |
52 | Leptobrachella petrops | Cham Chu Nature Reserve, Tuyen Quang, Vietnam | VNMN:2016 A.06 | KY459998 |
53 | Leptobrachella picta | Borneo, Malaysia | UNIMAS 8705 | KJ831295 |
54 | Leptobrachella pluvialis | Fansipan, Lao Cai, Vietnam | ROM 30685 | MH055843 |
55 | Leptobrachella pluvialis | Sapa, Lao Cai, Vietnam | ZMMU-A-5222-02262 | MH055844 |
56 | Leptobrachella puhoatensis | Pu Hu, Thanh Hoa, Vietnam | VNMN:2016 A.23 | KY849587 |
57 | Leptobrachella purpura | Yingjiang, Yunnan Province, China | SYS a006530 | MG520354 |
58 | Leptobrachella purpura | Yingjiang, Yunnan Province, China | SYS a006531 | MG520355 |
59 | Leptobrachella purpuraventra | Wujing Nature Reserve, Bijie City, Guizhou, China | SYS a007081 | MK414517 |
60 | Leptobrachella purpuraventra | Wujing Nature Reserve, Bijie City, Guizhou, China | SYS a007277/CIB110003 | MK414518 |
61 | Leptobrachella pyrrhops | Lam Dong, Vietnam | ZMMU A-5208 | KP017575 |
62 | Leptobrachella sabahmontana | Borneo, Malaysia | BORNEENSIS 12632 | AB847551 |
63 | Leptobrachella shangsiensis | Guangxi, China | NHMG1401032 | MK095460 |
64 | Leptobrachella shangsiensis | Guangxi, China | NHMG1401033 | MK095461 |
65 | Leptobrachella solus | Hala-Bala, Thailand | KUHE:23261 | LC202007 |
66 | Leptobrachella solus | Tam Dao, Vinh Phuc, Vietnam | ROM 20236 | MH055858 |
67 | Leptobrachella tengchongensis | Gaoligong Shan, Yunnan, China | SYS a004598 | KU589209 |
68 | Leptobrachella tengchongensis | Gaoligong Shan, Yunnan, China | SYS a003766 | MH055897 |
69 | Leptobrachella ventripunctatus | Zhushihe, Yunnan, China | SYS a004536 | MH055831 |
70 | Leptobrachella wuhuangmontis | Mt. Wuhuang, Pubei County, Guangxi, China | SYS a003485 | MH605577 |
71 | Leptobrachella wuhuangmontis | Mt. Wuhuang, Pubei County, Guangxi, China | SYS a003486 | MH605578 |
72 | Leptobrachella yingjiangensis | Yingjiang, Yunnan, China | SYS a006533 | MG520350 |
73 | Leptobrachella yingjiangensis | Yingjiang, Yunnan, China | SYS a006532 | MG520351 |
74 | Leptobrachella yunkaiensis | Dawuling Forest Station, Maoming City, Guangdong, China | SYS a004663 | MH605584 |
75 | Leptobrachella yunkaiensis | Dawuling Forest Station, Maoming City, Guangdong, China | SYS a004664 / CIB107272 | MH605585 |
76 | Leptobrachella zhangyapingi | Chiang Mai, Thailand | KIZ07258 | MH055864 |
77 | Leptobrachella zhangyapingi | Pang Num Poo, Chiang Mai Province,Thailand | JK-2013 | JX069979 |
78 | Leptobrachium huashen | Yunnan, China | KIZ049025 | KX811931 |
79 | Leptobrachium cf. chapaense | Sapa, Lao Cai, Vietnam | AMS R 171623 | KR018126 |
80 | Megophrys major | Kon Tum, Vietnam | AMS R 173870 | KY476333 |
All sequences were aligned by MUSCLE v. 3.6 with the default settings (
Phylogenetic trees were constructed with both Maximum Likelihood (ML) and Bayesian Inference (BI). The ML was conducted in IQ-TREE (
Morphometric data were taken from eight of most well-preserved adult specimens. Measurements were recorded to the nearest 0.1 mm (
SVL snout-vent length (from tip of snout to vent)
HDL head length (from tip of snout to rear of jaws)
HDW head width (head width at commissure of jaws)
SNT snout length (from tip of snout to the anterior corner of the eye)
EYE eye diameter (diameter of the exposed portion of the eyeballs)
IOD interorbital distance (minimum distance between upper eyelids)
IND internasal distance (distance between nares)
UEW upper eyelid width (measured as the greatest width of the upper eyelid)
NEL nostril-eyelid length (distance from nostril to eyelid)
NSL nostril-snout length (distance from nostril to snout)
TMP tympanum diameter (horizontal diameter of tympanum)
TEY tympanum-eye distance (distance from anterior edge of tympanum to posterior corner of eye)
TIB tibia length (distance from knee to heel)
ML manus length (distance from tip of third digit to proximal edge of inner palmar tubercle)
LAHL length of the lower arm and hand (distance from tip of the third finger to elbow)
HLL hindlimb length (distance from tip of fourth toe to vent)
FOT foot length (from proximal edge of the inner metatarsal tubercle to the tip of the fourth toe)
Sex was determined by direct observation of calls in life, the presence of internal vocal sac openings, and the presence of eggs in the abdomen through external inspection. Comparative morphological data of Leptobrachella species were obtained from the references listed in Table
Obtained references of 74 known congeners of the genus Leptobrachella, respectively.
ID | Leptobrachella species | Literature obtained |
---|---|---|
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. baluensis Smith, 1931 |
|
7 | L. bidoupensis (Rowley, Le, Tran & Hoang, 2011) |
|
8 | L. bijie Wang, Li, Li, Chen & Wang, 2019 |
|
9 | L. bondangensis Eto, Matsui, Hamidy, Munir & Iskandar, 2018 |
|
10 | L. botsfordi (Rowley, Dau & Nguyen, 2013) |
|
11 | L. bourreti (Dubois, 1983) |
|
12 | L. brevicrus Dring, 1983 |
|
13 | L. crocea (Rowley, Hoang, Le, Dau & Cao, 2010) |
|
14 | L. dringi (Dubois, 1987) |
|
15 | L. eos (Ohler, Wollenberg, Grosjean, Hendrix, Vences, Ziegler & Dubois, 2011) |
|
16 | L. firthi (Rowley, Hoang, Dau, Le & Cao, 2012) |
|
17 | L. fritinniens (Dehling & Matsui, 2013) |
|
18 | L. fuliginosa (Matsui, 2006) |
|
19 | L. fusca Eto, Matsui, Hamidy, Munir & Iskandar, 2018 |
|
20 | L. gracilis (Günther, 1872) |
|
21 | L. hamidi (Matsui, 1997) |
|
22 | L. heteropus (Boulenger, 1900) |
|
23 | L. isos (Rowley, Stuart, Neang, Hoang, Dau, Nguyen & Emmett, 2015) |
|
24 | L. itiokai Eto, Matsui & Nishikawa, 2016 |
|
25 | L. juliandringi Eto, Matsui & Nishikawa, 2015 |
|
26 | L. kajangensis (Grismer, Grismer & Youmans, 2004) |
|
27 | L. kalonensis (Rowley, Tran, Le, Dau, Peloso, Nguyen, Hoang, Nguyen & Ziegler, 2016) |
|
28 | L. kecil (Matsui, Belabut, Ahmad & Yong, 2009) |
|
29 | L. khasiorum (Das, Tron, Rangad & Hooroo, 2010) |
|
30 | L. lateralis (Anderson, 1871) |
|
31 | L. laui (Sung, Yang & Wang, 2014) |
|
32 | L. liui (Fei & Ye, 1990) |
|
33 | L. macrops (Duong, Do, Ngo, Nguyen & Poyarkov, 2018) |
|
34 | L. maculosa (Rowley, Tran, Le, Dau, Peloso, Nguyen, Hoang, Nguyen & Ziegler, 2016) |
|
35 | L. mangshanensis (Hou, Zhang, Hu, Li, Shi, Chen, Mo & Wang, 2018) |
|
36 | L. maoershanensis (Yuan, Sun, Chen, Rowley & Che, 2017) |
|
37 | L. marmorata (Matsui, Zainudin & Nishikawa, 2014) |
|
38 | L. maura (Inger, Lakim, Biun & Yambun, 1997) | Inger et al. 1997 |
39 | L. melanoleuca (Matsui, 2006) |
|
40 | L. melica (Rowley, Stuart, Neang & Emmett, 2010) |
|
41 | L. minima (Taylor, 1962) |
|
42 | L. mjobergi Smith, 1925 |
|
43 | L. nahangensis (Lathrop, Murphy, Orlov & Ho, 1998) |
|
44 | L. natunae (Günther, 1895) |
|
45 | L. nokrekensis (Mathew & Sen, 2010) |
|
46 | L. nyx (Ohler, Wollenberg, Grosjean, Hendrix, Vences, Ziegler & Dubois, 2011) |
|
47 | L. oshanensis (Liu, 1950) |
|
48 | L. pallida (Rowley, Tran, Le, Dau, Peloso, Nguyen, Hoang, Nguyen & Ziegler, 2016) |
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49 | L. palmata Inger & Stuebing, 1992 |
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50 | L. parva Dring, 1983 |
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51 | L. pelodytoides (Boulenger, 1893) |
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52 | L. petrops (Rowley, Dau, Hoang, Le, Cutajar & Nguyen, 2017) |
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53 | L. picta (Malkmus, 1992) |
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54 | L. platycephala (Dehling, 2012) |
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55 | L. pluvialis (Ohler, Marquis, Swan & Grosjean, 2000) |
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56 | L. puhoatensis (Rowley, Dau & Cao, 2017) |
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57 | L. purpuraventra Wang, Li, Li, Chen & Wang, 2019 |
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58 | L. purpurus (Yang, Zeng & Wang, 2018) |
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59 | L. pyrrhops (Poyarkov, Rowley, Gogoleva, Vassilieva, Galoyan & Orlov, 2015) |
|
60 | L. rowleyae (Nguyen, Poyarkov, Le, Vo, Ninh, Duong, Murphy & Sang, 2018) |
|
61 | L. sabahmontana (Matsui, Nishikawa & Yambun, 2014) |
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62 | L. serasanae Dring, 1983 |
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63 | L. shangsiensis Chen, Liao, Zhou & Mo, 2019 |
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64 | L. sola (Matsui, 2006) |
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65 | L. sungi (Lathrop, Murphy, Orlov & Ho, 1998) |
|
66 | L. tadungensis (Rowley, Tran, Le, Dau, Peloso, Nguyen, Hoang, Nguyen & Ziegler, 2016) |
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67 | L. tamdil (Sengupta, Sailo, Lalremsanga, Das & Das, 2010) |
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68 | L. tengchongensis (Yang, Wang, Chen & Rao, 2016) |
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69 | L. tuberosa (Inger, Orlov & Darevsky, 1999) |
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70 | L. ventripunctata (Fei, Ye & Li, 1990) |
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71 | L. wuhuangmontis Wang, Yang & Wang, 2018 |
|
72 | L. yingjiangensis (Yang, Zeng & Wang, 2018) |
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73 | L. yunkaiensis Wang, Li, Lyu & Wang, 2018 |
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74 | L. zhangyapingi (Jiang, Yan, Suwannapoom, Chomdej & Che, 2013) |
|
Phylogenetic trees from Maximum likelihood (ML) and Bayesian inference (BI) were constructed based on DNA sequences of the mitochondrial 16S rRNA gene with a length of 500 bp. The trees present identical topologies (Fig.
Bayesian inference tree derived from partial DNA sequences of the mitochondrial 16S r RNA gene. Numbers before slashes indicate Bayesian posterior probabilities (displayed >0.60 values), and numbers after slashes are ultrafast bootstrap support for maximum likelihood (2000 replicates) analyses (>60 retained). The symbol “–” represents value below 0.60/60. The scale bar represents 0.05 nucleotide substitutions per site.
Holotype.
GZNU20180606007, adult male, collected by Tao Luo (TL hereafter) on 7 June 2018 from the Huoqiuba Nature Reserve (28.4805°N, 107.0764°E, 1501 m. a.s.l.; Fig.
Paratypes. Five adult males (GZNU20180606002, GZNU20180606005, GZNU20180606006, GZNU20180606008), and three adult females (GZNU20180606001, GZNU20180606003, GZNU20180606004). They were collected from the holotype locality on 6 June 2018.
The specific epithet “suiyangensis” refers to the name of the holotype locality, Taibai Town in Suiyang County of Guizohu Province, China. We suggest as its English name “Suiyang Leaf-litter Toad,” and its Chinese name as “Sui Yang Zhang Tu Chan (绥阳掌突蟾)”.
Measurements (in mm) of the type series of Leptobrachella suiyangensis sp.nov. (H = holotype, P = paratype,M= male, F= female, another abbreviations defined in text).
Specimen | Type status | Sex | SVL | HDL | HDW | SNT | EYE | IOD | IND | UEW | NEL | NSL | TMP | TEY | TIB | HND | LAHL | HLL | FOT |
GZNU20180606007 | H | M | 28.7 | 9.9 | 9.3 | 4.1 | 2.4 | 2.9 | 2.8 | 1.6 | 2.8 | 1.1 | 2.1 | 1.1 | 13.1 | 7.2 | 13.4 | 43.3 | 12.5 |
GZNU20180606008 | P | M | 29.2 | 10.5 | 9.8 | 4.6 | 2.8 | 2.8 | 2.9 | 2.1 | 2.3 | 1.7 | 1.2 | 1.9 | 13.4 | 7.0 | 13.2 | 43.4 | 12.9 |
GZNU20180606002 | P | M | 29.7 | 12.1 | 10.1 | 5.0 | 3.9 | 3.2 | 3.7 | 3.1 | 2.3 | 1.3 | 2.3 | 1.4 | 13.8 | 7.1 | 13.3 | 44.4 | 12.3 |
GZNU20180606005 | P | M | 29.0 | 11.8 | 10.3 | 4.5 | 3.3 | 3.4 | 3.1 | 2.0 | 2.7 | 1.1 | 1.9 | 1.6 | 13.5 | 6.5 | 13.4 | 41.8 | 12.9 |
GZNU20180606006 | P | M | 29.2 | 11.4 | 10.4 | 4.0 | 3.8 | 3.2 | 3.2 | 2.6 | 2.6 | 2.2 | 1.8 | 1.3 | 13.6 | 7.4 | 13.3 | 42.8 | 12.6 |
GZNU20180606001 | P | F | 32.0 | 12.6 | 10.7 | 4.7 | 3.7 | 3.5 | 3.5 | 3.0 | 2.4 | 1.3 | 2.6 | 1.7 | 15.2 | 7.1 | 13.4 | 44.7 | 13.9 |
GZNU20180606003 | P | F | 30.5 | 10.3 | 10.9 | 4.7 | 3.7 | 3.1 | 3.1 | 2.2 | 2.3 | 1.5 | 3.5 | 1.4 | 15.2 | 7.4 | 13.8 | 45.3 | 16.6 |
GZNU20180606004 | P | F | 33.5 | 13.1 | 12.1 | 4.9 | 3.6 | 3.1 | 3.6 | 2.8 | 3.1 | 1.8 | 3.8 | 1.7 | 17.4 | 8.1 | 16.7 | 53.8 | 14.4 |
The specimens were assigned to the genus Leptobrachella on the basis of the following characters: (1) small body size; (2) having an elevated inner metacarpal tubercle; (3) having macro-glands on body (including supra-axillary, femoral and ventrolateral glands); (4) lacking vomerine teeth; (5) having small tubercles on eyelids; (6) anterior tip of snout with whitish vertical bar (
Description of the holotype. GZNU20180606007 (adult male), small body size (SVL 28.7 mm); the head length is slightly larger than the head width (HDL/HDW ratio 1.06); the snout is slightly protruding, projecting beyond the margin of the lower jaw; the nostril is between the snout and the eye (NSL/NEL ratio 0.39); the canthus rostralis is gently rounded; the loreal region is slightly concave; the interorbital space is flat; larger (IOD 2.9 mm) than the upper eyelid (1.6 mm in width), and the internarial distance is 2.8 mm; with vertical pupil; snout length is slightly larger than eye diameter (SNT/EYE ratio 1.71); tympanum is distinct and rounded, its diameter (TMP 2.1 mm) is smaller than that of the eye diameter (EYE 2.4 mm) and longer than the tympanum-eye distance (TMP/TEY ratio 1.91); deep black supratympanic line is present; weakly black supratympanic line exists (Fig.
The tips of the fingers are rounded, slightly swollen; relative finger lengths are presented as: I <II < IV < III; nuptial pad is absent; absent subarticular tubercles (Fig.
Dorsal skin is shagreened and scattered with fine and rounded granules, some of the granules forming short longitudinal folds; ventral skin smooth; large pectoral gland, elongated oval, 1.5 mm in length; small femoral gland, rounded, 0.7 mm in diameter, situated on the posteroventral surface of the thigh, closer to tibiotarsal articulation than to the vent; risen supra-axillary gland, 1.3 mm in diameter; ventrolateral gland is distinct as small white dots forming an incomplete line (Fig.
Measurements of holotype (in mm). Holotype: SVL 28.7, HDL 9.9, HDW 9.3, SNT 4.1, EYE 2.4, IOD 2.9, INT 2.8, UEW 1.6, NEL 2.8, NSL 1.1, TMP 2.1, TEY 1.1, TIB 13.1, HND 7.2, LAHL 13.4, HLL 43.3, FOT 12.5.
Coloration of holotype in life. Dorsal skin purple-brown; brown-purplish with dark-brown marks between the eyes and the scapular region, which are scattered with some deep yellow-orange granules more concentrated on the upper eyelid (Fig.
In preservation, there are dark brown marks on the dorsum and flanks; dorsum of the body and hindlimbs are dark brown, while dorsum of the forelimbs is yellowish brown; transverse bars on the limbs become more distinct, and dark-brown patterns, marks and spots on the back are indistinct; ventral surface of the body is yellowish brown with brown marbling on the sides and chest; orange supra-axillary, femoral, pectoral and ventrolateral glands fade to greyish white.
Measurements of the type series are shown in Table
Currently, Leptobrachella suiyangensis sp. nov. is known only from its holotype locality, Huoqiuba Nature Reserve, Suiyang County, Guizhou Province, China (Fig.
Measurements (in mm), and body proportions of Leptobrachella suiyangensis sp.nov. from Suiyang County, Guizhou Province, China.
Measurements | Males Range (mean ± SD), n = 5 | Females Range (mean ± SD), n = 3 |
SVL | 28.7–29.7 (29.2 ± 0.4) | 30.5–33.5 (32.0 ± 1.5) |
HDL | 9.9–12.1 (11.1 ± 0.9) | 10.3–13.1 (12.0 ± 1.5) |
HDW | 9.3–10.4 (10.0 ± 0.4) | 10.7–12.1 (11.2 ± 0.8) |
SNT | 4.0–5.0 (4.4 ± 0.4) | 4.7–4.9 (4.8 ± 0.1) |
EYE | 2.4–3.9 (3.2 ± 0.6) | 3.6–3.7 (3.7 ± 0.1) |
IOD | 2.8–3.4 (3.1 ± 0.2) | 3.1–3.5 (3.2 ± 0.2) |
INT | 2.8–3.7 (3.1 ± 0.4) | 3.1–3.6 (3.4 ± 0.3) |
UEW | 1.6–3.1 (2.3 ± 0.6) | 2.2–3.0 (2.7 ± 0.4) |
NEL | 2.3–2.8 (2.5 ± 0.2) | 2.3–3.1 (2.6 ± 0.4) |
NSL | 1.1–2.2 (1.5 ± 0.5) | 1.3–1.8(1.5 ± 0.3) |
TMP | 1.2–2.3 (1.9 ± 0.4) | 2.6–3.8 (3.3 ± 0.6) |
TEY | 1.1–1.9 (1.5 ± 0.3) | 1.4–1.7 (1.6 ± 0.2) |
TIB | 13.1–13.8 (13.5 ± 0.3) | 15.2–17.4 (15.9 ± 1.3) |
HND | 6.5–7.4 (7.0 ± 0.3) | 7.1–8.1 (7.5 ± 0.5) |
LAHL | 13.2–13.4 (13.3 ± 0.1) | 13.4–16.7 (14.6 ± 1.8) |
HLL | 41.8–44.4 (43.1 ± 0.9) | 44.7–53.8 (47.9 ± 5.1) |
FOT | 12.3–12.9 (12.6 ± 0.3) | 13.9–16.6 (15.0 ± 1.4) |
HDL/HDW | 1.06–1.20 (1.12 ± 0.06) | 0.90–1.20 (1.07 ± 0.15) |
HDL/SVL | 0.34–0.41 (0.38 ± 0.03) | 0.30–0.40 (0.37 ± 0.06) |
SNT/HDL | 0.35–0.44 (0.40 ± 0.03) | 0.40–0.50 (0.43 ± 0.06) |
SNT/EYE | 1.05–1.71 (1.41 ± 0.27) | 1.30–1.40 (1.33 ± 0.06) |
EYE/TMP | 1.14–2.33 (1.80 ± 0.45) | 1.00–1.40 (1.17 ± 0.21) |
EYE/SNT | 0.59–0.95 (0.73 ± 0.15) | 0.70–0.80 (0.77 ± 0.06) |
TMP/EYE | 0.43–0.88 (0.59 ± 0.18) | 0.70–1.10 (0.93 ± 0.21) |
TIB/SVL | 0.46–0.47 (0.46 ± 0.01) | 0.50 |
LAHL/SVL | 0.45–0.47 (0.46 ± 0.01) | 0.40–0.50 (0.47 ± 0.06) |
HLL/SVL | 1.44–1.51 (1.48 ± 0.03) | 1.40–1.60 (1.50 ± 0.10) |
TIB/HLL | 0.30–0.32 (0.31 ± 0.01) | 0.30 |
Leptobrachella suiyangensis sp. nov. differs from all other species of Leptobrachella based on morphological and molecular evidence. Phylogenetically, L. suiyangensis sp. nov., L. alpina and L. purpurus form a clade. Genetically, among this clade, the smallest genetic distance, at 5.49%, is between L. suiyangensis sp. nov. and L. alpina, and the largest genetic distance is 6.27% (L. suiyangensis sp. nov. and L. purpurus). Morphologically, the new species can be distinguished from L. alpina by having a larger body size of males (28.7–29.7 mm vs 24.0–26.4 mm); having narrower lateral fringes on the toes of the male (vs wide in males); dorsum purple-brown to dark purple-brown or grey-purple ground colour; ventral yellowish creamy-white with marbled texture on the chest and belly or with irregular light-brown speckling (vs almost uniformly gray-brown on dorsal part, ventral nearly immaculately creamy white, brown specking on margins); ventrolateral glands are characterized by small white dots forming an incomplete line (vs small white dots forming a complete line longitudinally); shoulder-gland is orange-yellow (vs white, around gland); head length greater than head width, HDL/HDW ratio 1.12 (vs head length equal to head width, HDL/HDW ratio 1.00). The new species can be distinguished from L. purpurus by body size of males (28.7–29.7 mm vs 25.0–27.5 mm); having narrow lateral fringes on the toes of males (vs wide in males); dorsum purple-brown to dark purple-brown or grey-purple ground color, ventral yellowish creamy-white with marbled texture on the chest and belly or with irregular light-brown speckling (vs dorsum coloration purplish brown, ventral side dull white with an indistinct grey dusting); throat immaculate gray (vs throat immaculate pinkish; almost dark orange-yellow on the upper arm (vs upper arms with distinct coppery orange coloration); dark bars on dorsal surface of tibia and tarsus very narrow, especially those on dorsal skin of tarsus (vs relatively broader dark bars on dorsal surface of tibia and tarsus); tibiotarsal articulation reaches to the anterior eye (vs tibiotarsal articulation reaches to posterior corner of the eye); relative length of fingers I <II < IV < III (vs I = II = IV < III).
Compared with the 26 known congeners in the genus Leptobrachella found south of the Isthmus of Kra, referring to the presence or absence of supra-axillary and ventrolateral glands, L. suiyangensis sp. nov. can be easily distinguished 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, all of which are lack of supra-axillary and ventrolateral glands (
From the remaining 48 known congeners in the genus Leptobrachella found north of the Isthmus of Kra (Table
Selected diagnostic characters for species described herein and species in the genus Leptobrachella occurring north of the Isthmus of Kra (modified from Rowley et al. 2017;
ID | Species | Males SVL (mm) | Black spots on flanks | Toes webbing | Fringes on toes | Ventral coloration | Dorsal skin texture |
1 | L. suiyangensis sp. nov. | 28.7–29.7 | Yes | Rudimentary | Narrow | Yellowish creamy-white with marble texture chest and belly or with irregular light brown speckling | Shagreen with small granules |
2 | L. aerea | 25.1–28.9 | No | Rudimentary | Wide | Near immaculate creamy white, brown specking on margins | Finely tuberculate |
3 | L. alpinus | 24.0–26.4 | Yes | Rudimentary | Wide in males | Creamy-white with dark spots | Relatively smooth, some with small warts |
4 | L. applebyi | 19.6–22.3 | Yes | Rudimentary | No | Reddish brown with white speckling | Smooth |
5 | L. ardens | 21.3–24.7 | Yes | No | No | Reddish brown with white speckling | Smooth- finely shagreened |
6 | L. bidoupensis | 18.5–25.4 | Yes | Rudimentary | Weak | Reddish brown with white speckling | Smooth |
7 | L. botsfordi | 29.1–32.6 | No | Rudimentary | Narrow | Reddish brown with white speckling | Shagreened |
8 | L. bourreti | 28.0–36.2 | Yes | Rudimentary | Weak | Creamy white | Relatively smooth, some with small warts |
9 | L. crocea | 22.2–27.3 | No | Rudimentary | No | Bright orange | Highly tuberculate |
10 | L. eos | 33.1–34.7 | No | Rudimentary | Wide | Creamy white | Shagreened |
11 | L. firthi | 26.4–29.2 | No | Rudimentary | Wide in males | Creamy white | Shagreened with fine tubercles |
12 | L. fuliginosa | 28.2–30.0 | Yes | Rudimentary | Weak | White with brown dusting | Nearly smooth, few tubercles |
13 | L. isos | 23.7–27.9 | No | Rudimentary | Wide in males | Creamy white with white dusting on margins | Mostly smooth, females more tuberculate |
14 | L. kalonensis | 25.8–30.6 | Yes | No | No | Pale, speckled brown | Smooth |
15 | L. khasiorum | 24.5–27.3 | Yes | Rudimentary | Wide | Creamy white | Isolated, scattered tubercles |
16 | L. lateralis | 26.9–28.3 | Yes | Rudimentary | No | Creamy white | Roughly granular |
17 | L. laui | 24.8–26.7 | Yes | Rudimentary | Wide | Creamy white with dark brown dusting on margins | Round granular tubercles |
18 | L. liui | 23.0–28.7 | Yes | Rudimentary | Wide | Creamy white with dark brown spots on chest and margins | Round granular tubercles with glandular folds |
19 | L. macrops | 28.0–29.3 | Yes | Rudimentary | No | Greyish-violet with white speckling | Roughly granular with larger tubercles |
20 | L. maculosa | 24.2–26.6 | Yes | No | No | Brown, less white speckling | Dorsum mostly smooth with numerous tiny tubercles |
21 | L. mangshanensis | 22.2–27.8 | Yes | Rudimentary | Weak | White speckles on throat and belly | Nearly smooth |
22 | L. maoershanensis | 25.2–30.4 | Yes | Rudimentary | Narrow | Creamy white chest and belly with irregular black spots | Longitudinal folds |
23 | L. melica | 19.5–22.7 | Yes | Rudimentary | No | Reddish brown with white speckling | Smooth |
24 | L. minima | 25.7–31.4 | Yes | Rudimentary | No | Creamy white | Smooth |
25 | L. nahangensis | 40.8 | Yes | Rudimentary | No | Creamy white with light specking on throat and chest | Smooth |
26 | L. nokrekensis | 26.0–33.0 | Yes | Rudimentary | unknown | Creamy white | Tubercles and longitudinal folds |
27 | L. nyx | 26.7–32.6 | Yes | Rudimentary | No | Creamy white with white with brown margins | Rounded tubercles |
28 | L. oshanensis | 26.6–30.7 | Yes | No | No | Whitish with no markings or only small, light grey spots | Smooth with few glandular ridges |
29 | L. pallida | 24.5–27.7 | No | No | No | Reddish brown with white speckling | Tuberculate |
30 | L. pelodytoides | 27.5–32.3 | Yes | Wide | Narrow | Whitish | Small, smooth warts |
31 | L. petrops | 23.6–27.6 | No | No | Narrow | Immaculate creamy white | Highly tuberculate |
32 | L. pluvialis | 21.3–22.3 | Yes | Rudimentary | No | Dirty white with dark brown marbling | Smooth, flattened tubercles on flanks |
33 | L. puhoatensis | 24.2–28.1 | Yes | Rudimentary | Narrow | Reddish brown with white dusting | Longitudinal skin ridges |
34 | L. purpurus | 25.0–27.5 | Yes | Rudimentary | Wide | Dull white with indistinct grey dusting | Shagreen with small tubercles |
35 | L. pyrrhops | 30.8–34.3 | Yes | Rudimentary | No | Reddish brown with white speckling | Slightly shagreened |
36 | L. rowleyae | 23.4–25.4 | Yes | No | No | Pinkish milk-white to light brown chest and belly with numerous white speckles | Smooth with numerous tiny tubercles |
37 | L. sungi | 48.3–52.7 | No or small | Wide | Weak | White | Granular |
38 | L. shangsiensis | 24.9–29.4 | Yes | Narrow | Narrow | Yellowish creamy-white with marble texture | Smooth |
39 | L. tadungensis | 23.3–28.2 | Yes | No | No | Reddish brown with white speckling | Smooth |
40 | L. tamdil | 32.3 | Yes | Wide | Wide | White | Weakly tuberculate |
41 | L. tengchongensis | 23.9–26.0 | Yes | Rudimentary | Narrow | White with dark brown blotches | Shagreened with small tubercles |
42 | L. tuberosa | 24.4–29.5 | No | Rudimentary | No | White with small grey spots/streaks | Highly tuberculate |
43 | L. ventripunctata | 25.5–28.0 | Yes | Rudimentary | No | Chest and belly with dark brown spots | Longitudinal skin ridges |
44 | L. wuhuangmontis | 25.6–30.0 | Yes | Rudimentary | Narrow | Greyish white mixed by tiny white and black dots | Rough, scattered with dense conical tubercles |
45 | L. yingjiangensis | 25.7–27.6 | Yes | Rudimentary | Wide | Creamy white with dark brown flecks on chest and margins | Shagreened with small tubercles |
46 | L. yunkaiensis | 25.9–29.3 | Yes | Rudimentary | Wide | Belly pink with distinct or indistinct speckling | Shagreened with short skin ridges and raised warts |
47 | L. zhangyapingi | 45.8–52.5 | No | Rudimentary | Wide | Creamy-white with white with brown margins | Mostly smooth with distinct tubercles |
48 | L. bijie | 29.0–30.4 | Yes | Rudimentary | Narrow | White with distinct nebulous greyish speckling on chest and ventrolateral flanks | Shagreened and granular |
49 | L. purpuraventra | 27.3–29.8 | Yes | Rudimentary | Narrow | Grey purple with distinct nebulous greyish speckling on chest and ventrolateral flanks | Shagreened and granular |
In having irregular, light-brown speckling on the flanks, the new species differs from L. aerea, L. botsfordi, L. crocea, L. firthi, L. isos, L. pallida, L. petrops and L. tuberosa, all of which lack distinct irregular, light-brown speckling on the flanks. By having rudimentary webbing on the toes, the new species differs from L. kalonensis, L. oshanensis, L. pallida, L. petrops, and L. tadungensis, all of which lack webbing on the toes; and from L. pelodytoides, which has wide webbing on the toes. By having narrow lateral fringes on toes, the new species differs from L. ardens, L. eos, L. firthi, L. isos, L. khasiorum, L. laui, L. liui, L. purpurus, L. tamdil, L. yingjiangensis and L. yunkaiensis, all of which have wide lateral fringes on the toes; from L. bidoupensis, L. bourreti, L. fuliginosa and L. mangshanensis, all of which have weak lateral fringes on the toes; and from L. crocea, L. kalonensis, L. lateralis, L. macrops, L. minima, L. nyx, L. oshanensis, L. pallida, L. pyrrhops, L. tadungensis, L. tuberosa, and L. ventripunctata, all of which lack lateral fringes on the toes. By having dorsal surface shagreened with small granules, and in lacking enlarge tubercles or warts, the new species 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. alpina (dorsum smooth, some with small warts), L. fuliginosa (dorsum smooth with fine tubercles), L. laui (dorsum with round granular tubercle, lacking skin ridges), L. liui (dorsum with round tubercles), L. macrops (dorsum roughly granular with large tubercles), L. maoershanensis (dorsum smooth with small warts), L. nokrekensis (dorsum tubercles and longitudinal folds), L. pelodytoides (dorsum with small, smooth warts), L. puhoatensis (dorsum longitudinal skin ridges), L. tuberosa (dorsum highly tuberculate), L. yunkaiensis (dorsum with raised warts), L. wuhuangmontis (dorsum rough with conical tubercles), and L. bijie and L. purpuraventra (dorsum shagreened and granular). By the yellowish creamy-white with marbled chest and belly or with irregular light-brown speckling, the new species differs from L. alpinus, L. applebyi, L. ardens, L. bidoupensis, L. botsfordi and L. pyrrhops (ventral reddish brown with white speckling), L. aerea (ventral nearly immaculate creamy-white with brown specking on margins), L. bijie (ventral white with distinct nebulous greyish speckling on chest and ventrolateral flanks), L. crocea (ventral bright orange), L. khasiorum, L. nokrekensis and L. yingjiangensis (ventral creamy white), L. macrops (ventral greyish-violet with white speckling), L. puhoatensis (ventral reddish-brown with white dusting), L. purpurus (ventral dull white with indistinct grey dusting), L. purpuraventra (ventral grey-purple with distinct nebulous greyish speckling on the chest and ventrolateral flanks), L. tuberosa (ventral white with small grey spots and streaks), L. ventripunctata (chest and belly with large dark brown spots), L. wuhuangmontis (ventral greyish white), and L. yunkaiensis (belly pink with speckling). A comparative morphological data (selection) of Leptobrachella suiyangensis sp. nov. and 48 recognized Leptobrachella species occurring north of the Isthmus of Kra are listed in Table
Phylogenetic analyses based on mitochondrial DNA and nuclear DNA all suggested that the new species belongs to Leptobrachella but is separate from its congeners. Genetic distance of the 16S rRNA gene between the new species and its closely related species (L. bijie, L. purpuraventra, L. alpina and L. purpurus) was 4.71–6.27%, within the expected range of interspecific divergences in amphibians (
The new species described in this study increases the number of species of Leptobrachella to 75, with 21 recorded from China (
Studies on the taxonomy and phylogeny of the genus Leptobrachella were difficult to perform because of the morphological conservativeness of the species; in the field, many species appear to be very similar morphologically, and there exist sympatric species. This likely hinders our understanding of these cryptic species (Ohler et al. 2010;
Currently, to our knowledge, L. suiyangensis sp. nov. is restricted to rocky streams in bamboo forests. However, the type locality of L. suiyangensis sp. nov. has faced habitat loss and human disturbance, such as artificial grazing and herb collection, which could possibly threaten this species. Leptobrachella suiyangensis sp. nov. is range-restricted to Kuankuoshui National Nature Reserve, which borders the nearby Huoqiuba Nature Reserve and is in the eastern Ta-lou Mountains. These areas feature subtropical evergreen broad-leaved forest and evergreen deciduous broad-leaved mixed forest. Thus, it is likely that other populations of L. suiyangensis sp. nov. may be discovered in the Kuankuoshui Nature Reserve in the near future.
This project was supported by the key project of science-technology of basic condition platform granted by The Ministry of Science and Technology of the People’s Republic of China (Grant No. 2005DKA21402); The National Top Discipline Construction Project of Guizhou Province; Geography in Guizhou Normal University (85 2017 Qianjiao Keyan Fa); and The Project of Science and Technology Program of Guizhou Province, “Study of biological and ecological vales for Fanjingshan World Natural Heritage Nomination of Tongren City (3052 2015 Qiankehe SY)”. We thank Bo Song and Hongtao Cui for their help in sample collection. We thank Professor Ruliang Pan and LetPub (http://www.letpub.com) for its linguistic assistance during the preparation of this manuscript.