Research Article |
Corresponding author: Wei-Cai Chen ( chenweicai2003@126.com ) Academic editor: Bin Wang
© 2023 Wei-Cai Chen, Wan-Xiao Peng, Peng Li, Gui-Dong Yu.
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:
Chen W-C, Peng W-X, Li P, Yu G-D (2023) A new species of the genus Leptobrachella Smith 1925 (Amphibia, Anura, Megophryidae) from Guangxi, China. ZooKeys 1178: 1-16. https://doi.org/10.3897/zookeys.1178.106038
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A new species of Leptobrachella, L. wumingensis sp. nov., was described from the Damingshan National Nature Reserve, Wuming District, Nanning City, Guangxi, China based on morphological, molecular and bioacoustic data. Phylogenetic analysis of 16S mtDNA fragments revealed that the new species is closely related to L. damingshanensis. Uncorrected p-distances between the new species and all homologous DNA sequences available for the 16S gene of Leptobrachella are greater than 7.1%. Morphologically, L. wumingensis sp. nov. differs from its congeners in several ways, including a medium body size (SVL 26.0–26.7 mm in males, 30.6–34.8 mm in females), lack of toe webbing and lateral fringes, shagreened and granular dorsal surface, pale brown dorsum with darker brown markings, iris bicolored, with the upper half copper and fading to silver in the lower half, and the presence of small irregular black spots and tangerine tubercles on the flanks. Furthermore, we found the new species to have two types of advertisement calls and relatively high dominant frequencies, making it distinct from its congeners.
Bioacoustics, molecular analyses, morphological characters, sympatric species, taxonomy
The genus Leptobrachella Smith, 1925 is distributed in northeastern India, Southeast Asia, Vietnam, Thailand, Myanmar, Malaya, Borneo and Natuna Island (
Between 2019 and 2023, we conducted surveys at the Damingshan National Nature Reserve (DMS), Wuming District, Nanning City, Guangxi, China. We collected nine specimens (hereafter DMS specimens) of Leptobrachella that significantly differed from the sympatric species, L. damingshanensis Chen, Yu, Cheng, Meng, Wei, Zhou & Lu, 2021 and other congeners in morphological characters, including body size and color pattern as well as molecular data. The uncorrected p-distances between 16S mtDNA fragments of DMS specimens and all homologous sequences of Leptobrachella available in GenBank were greater than 7.1%. Additionally, the bioacoustic data of the DMS specimens distinguished them from L. damingshanensis and other advertisement calls available in the genus Leptobrachella. Given the unique morphological characters, the relatively high degree of mtDNA divergence, and the distinct bioacoustics data, we describe the DMS specimens as a new species of Leptobrachella.
Nine Leptobrachella specimens were collected from the Damingshan National Nature Reserve, Wuming District, Nanning City, Guangxi, China (Fig.
Genomic DNA was isolated from muscle tissue using Tiangen Biotech Co. Ltd. tissue extraction kits (Beijing, China). The primer pairs 16Sar_L 5’-CGCCTGTTTAC CAAAAACAT-3’ and 16Sbr_H 5’-CCGGTCTGAACTCAGATCACGT-3’ (
Phylogenetic relationships were inferred using 16S mtDNA fragments through maximum likelihood (ML) and Bayesian inference (BI) analyses. Table
ID | Species | Locality | Voucher no. | GenBank No. 16S rRNA |
---|---|---|---|---|
1 | L. wumingensis sp. nov. | Wuming County, Guangxi, China* | NNU 00283 | OM935575 |
2 | L. wumingensis sp. nov. | Wuming County, Guangxi, China* | NNU 00284 | OM935576 |
3 | L. wumingensis sp. nov. | Wuming County, Guangxi, China* | NNU 00285 | OM935577 |
4 | L. wumingensis sp. nov. | Wuming County, Guangxi, China* | NNU 00286 | OM935578 |
5 | L. wumingensis sp. nov. | Wuming County, Guangxi, China* | NNU 01058 | OR194551 |
6 | L. wumingensis sp. nov. | Wuming County, Guangxi, China* | NNU 01059 | OR194552 |
7 | L. wumingensis sp. nov. | Wuming County, Guangxi, China* | NNU 01086 | OR194553 |
8 | L. aerea | Quang Binh, Vietnam | ZFMK 86362 | JN848409 |
9 | L. alpina | Caiyanghe, Yunnan, China | KIZ 049024 | MH055867 |
10 | L. applebyi | Phong Dien Nature Reserve, Thua Thien-Hue, Vietnam | KIZ 010701 | MH055947 |
11 | L. arayai | Borneo, Malaysia* | AE 100/S9 | DQ642119 |
12 | L. ardens | Kon Ka Kinh National Park, Gia Lai, Vietnam* | ZMMU-NAP-06099 | MH055949 |
13 | L. aspera | Huanglianshan Nature Reserve, Lyuchun, Yunnan, China* | SYS a007743 | MW046199 |
14 | L. baluensis | Sabah, Borneo, Malaysia* | SP 21604 | LC056792 |
15 | L. bashaensis | Basha Nature Reserve, Guizhou, China* | GIB 196404 | MW136295 |
16 | L. bidoupensis | Bidoup-Nui Ba National Park, Lam Dong, Vietnam* | ZMMU-A-4797-01454 | MH055945 |
17 | L. bijie | Bijie City, Guizhou, China* | SYS a007313 | MK414532 |
18 | L. botsfordi | Lao Cai, Vietnam* | AMS R 176540 | MH055952 |
19 | L. bourreti | Mao’er Shan, Guangxi, China | KIZ 019389 | MH055869 |
20 | L. brevicrus | Sarawak, Borneo, Malaysia* | ZMH A09365 | KJ831302 |
21 | L. chishuiensis | Guizhou, China* | CIB CS20190518047 | MT117053 |
22 | L. crocea | Thua Thien-Hue, Vietnam | ZMMU-NAP-02274 | MH055955 |
23 | L. damingshanensis | Wuming County, Guangxi, China* | NNU 202103281 | MZ145229 |
24 | L. damingshanensis | Wuming County, Guangxi, China* | NNU 202103282 | MZ145230 |
25 | L. damingshanensis | Wuming County, Guangxi, China* | NNU 202103283 | MZ145231 |
26 | L. dong | Tongdao County, Hunan Province, China* | CIB SSC1758 | OP764529 |
27 | L. dorsospina | Yushe Forest Park, Shuicheng, Guizhou, China* | SYS a004961 | MW046194 |
28 | L. dringi | Borneo, Malaysia* | KUHE:55610 | AB847553 |
29 | L. eos | Phongsaly, Laos* | MNHN 2004.0274 | JN848452 |
30 | L. feii | Yunnan, China* | KIZ048894 | MT302634 |
31 | L. firthi | Kon Tum, Vietnam* | AMS: R 176524 | JQ739206 |
32 | L. flaviglandulosa | Xiaoqiaogou Nature Reserve, Yunnan, China* | KIZ 016072 | MH055934 |
33 | L. fritinniens | Danum Valley Field Center, Sabah, Malaysia | FMNH 244800 | MH055971 |
34 | L. fuliginosa | Phetchaburi, Thailand | KUHE:20197 | LC201988 |
35 | L. gracilis | Bukit Kana, Sarawak, Malaysia | FMNH 273682 | MH055972 |
36 | L. graminicola | Mount Pu Ta Leng, Lao Cai, Vietnam* | VNMN 010909 | MZ224649 |
37 | L. hamidi | Borneo, Malaysia* | KUHE 17545 | AB969286 |
38 | L. heteropus | Peninsular, Malaysia | KUHE 15487 | AB530453 |
39 | L. isos | Gia Lai, Vietnam* | AMS R 176480 | KT824769 |
40 | L. itiokai | Gunung Mulu National Park, Sarawak, Malaysia* | KUHE:55897 | LC137805 |
41 | L. jinshaensis | Lengshuihe Nature Reserve, Jinsha County, Guizhou, China* | CIB JS20200516001 | MT814014 |
42 | L. juliandringi | Sarawak, Borneo, Malaysia* | KUHE 17557 | LC056784 |
43 | L. kajangensis | Tioman, Malaysia* | LSUHC:4439 | LC202002 |
44 | L. kalonensis | Binh Thuan, Vietnam* | IEBR A.2014.15 | KR018114 |
45 | L. kecil | Cameron, Malaysia* | KUHE:52439 | LC202003 |
46 | L. khasiorum | Meghalaya, India* | SDBDU 2009.329 | KY022303 |
47 | L. korifi | Doi Inthanon, Thailand* | KUHE 19134 | LC741033 |
48 | L. laui | Wutongshan, Shenzhen city, China* | SYS a001507 | KM014544 |
49 | L. liui | Wuyi Shan, Fujian, China* | ZYCA907 | MH055908 |
50 | L. macrops | Dak Lak, Vietnam* | AMS R177663 | KR018118 |
51 | L. maculosa | Ninh Thuan, Vietnam* | AMS: R 177660 | KR018119 |
52 | L. mangshanensis | Manghan, Hunan, China* | MSZTC201703 | MG132198 |
53 | L. maoershanensis | Mao’er Shan, Guangxi, China | KIZ 07614 | MH055927 |
54 | L. marmorata | Borneo, Malaysia* | KUHE 53227 | AB969289 |
55 | L. maura | Borneo, Malaysia | SP 21450 | AB847559 |
56 | L. melanoleuca | Kapoe, Ranong, Thailand | KIZ 018031 | MH055967 |
57 | L. melica | Ratanakiri, Cambodia* | MVZ 258198 | HM133600 |
58 | L. minima | Doi Phu Fa, Nan, Thailand | KIZ 024317 | MH055852 |
59 | L. mjobergi | Sarawak, Borneo, Malaysia* | KUHE 47872 | LC056787 |
60 | L. murphyi | Doi Inthanon, Chiang Mai, Thailand* | KIZ 031199 | MZ710523 |
61 | L. nahangensis | Tuyen Quang, Vietnam* | ROM 7035 | MH055853 |
62 | L. namdongensis | Thanh Hoa, Vietnam* | VNUF A.2017.95 | MK965390 |
63 | L. neangi | Veal Veng District, Pursat, Cambodia* | CBC 1609 | MT644612 |
64 | L. niveimontis | Yongde County, Yunnan, China* | KIZ 028276 | MT302620 |
65 | L. nyx | Ha GiangProv., Vietnam* | AMNH A 163810 | DQ283381 |
66 | L. oshanensis | Emei Shan, Sichuan, China* | Tissue ID: YPX37492 | MH055896 |
67 | L. pallida | Lam Dong, Vietnam* | UNS 00510 | KR018112 |
68 | L. parva | Mulu National Park, Sarawak, Malaysia* | KUHE:55308 | LC056791 |
69 | L. pelodytoides | NA | TZ 819 | AF285192 |
70 | L. petrops | Ba Vi National Park, Ha Tay, Vietnam | ROM 13483 | MH055901 |
71 | L. picta | Borneo, Malaysia | UNIMAS 8705 | KJ831295 |
72 | L. pluvialis | Lao Cai, Vietnam* | MNHN:1999.5675 | JN848391 |
73 | L. puhoatensis | Nghe An, Vietnam* | VNMN 2016 A.22 | KY849586 |
74 | L. purpurus | Yunnan, China* | SYS a006530 | MG520354 |
75 | L. purpuraventra | Guizhou, China* | SYS a007281 | MK414517 |
76 | L. pyrrhops | Loc Bac, Lam Dong, Vietnam* | ZMMU-A-4873-00158 | MH055950 |
77 | L. rowleyae | Da Nang City, Vietnam* | ITBCZ2783 | MG682552 |
78 | L. sabahmontanus | Borneo, Malaysia* | BORNEENSIS 12632 | AB847551 |
79 | L. shangsiensis | Shangsi County, Guangxi, China* | NHMG1401032 | MK095460 |
80 | L. shimentaina | Shimentai Nature Reserve, Guangdong, China* | SYS a004712 | MH055926 |
81 | L. shiwandashanensis | Fangcheng City, Guangxi, China* | NNU202103146 | MZ326691 |
82 | L. sinorensis | Mae Hong Son, Thailand* | KUHE 19809 | LC741034 |
83 | L. sola | Gunung Stong, Kelantan, Malaysia | KU RMB20973 | MH055973 |
84 | L. suiyangensis | Guizhou, China* | GZNU 20180606005 | MK829649 |
85 | L. sungi | Vinh Phuc, Vietnam* | ROM 20236 | MH055858 |
86 | L. tadungensis | Dak Nong, Vietnam* | UNS 00515 | KR018121 |
87 | L. tengchongensis | Yunnan, China* | SYS a004598 | KU589209 |
88 | L. tuberosa | Kon Ka Kinh National Park, Gia Lai, Vietnam* | ZMMU-NAP-02275 | MH055959 |
89 | L. ventripunctata | Wenlong, Yunnan, China | KIZ 013621 | MH055824 |
90 | L. wuhuangmontis | Pubei County, Guangxi, China* | SYS a003486 | MH605578 |
91 | L. wulingensis | Hunan, China* | CSUFT194 | MT530316 |
92 | L. yeae | Mount Emei, Sichuan, China* | CIBEMS20190422HLJ1-6 | MT957019 |
93 | L. yingjiangensis | Yunnan, China* | SYS a006532 | MG520351 |
94 | L. yunkaiensis | Guangdong, China* | SYS a004663 | MH605584 |
95 | L. yunyangensis | Yunyang County, Chongqing, China* | GZNU 20210629001 | OL800366 |
96 | L. zhangyapingi | Chiang Mai, Thailand* | KIZ 07258 | MH055864 |
97 | Leptobrachium huashen | Yunnan, China | KIZ 049025 | KX811931 |
98 | Xenophrys glandulosa | Yunnan, China | KIZ 048439 | KX811762 |
Two male advertisement calls were recorded using a SONY ICD-TX50 recorder at an ambient temperature of approximately 21 °C. Acoustic analysis was performed using Raven Pro 1.6 (Cornell Laboratory of Ornithology, Ithaca, NY, USA) following the method outlined by
The molecular analyses conducted using BI and ML methods produced similar topologies, as depicted in Fig.
The calls of two individuals were recorded in the field (NNU 01058 and NNU 01086). Two types of advertisement calls (Type A and Type B; Fig.
The DMS specimens can be readily differentiated from other species in the same phylogenetic clade. For instance, adult males of L. damingshanensis have a significantly larger body size (SVL 33.6–34.4 mm) and possess a pair of distinct tangerine glands on the proximal thigh area, as well as rudimentary webbing and narrow lateral fringes on their toes. In contrast, the DMS specimens have indistinct tangerine glands on the proximal thigh area, and no toe webbing or lateral fringes.
Taken together, molecular data, acoustic analyses and morphological characters support the conclusion that the DMS specimens represent a distinct and previously unrecognized species of Leptobrachella, which is described below.
Holotype. NNU 01058, adult male, collected at the Damingshan National Nature Reserve, Wuming District, Nanning City, Guangxi, China (23.507°N, 108.395°E; elevation 1214 m), collected by Wei-Cai Chen on 12 April 2023. Paratypes. NNU 201907009, NNU 01086, two adult males, collected at the same locality as the holotype; NNU 201907009 collected by Gui-Dong Yu on 23 May 2019, NNU 01086 collected by Wei-Cai Chen on 14 April 2023; NNU 00283–6, four adult females collected at the same locality as the holotype on 16 June 2021 by Wei-Cai Chen; NNU 01059–60, two adult females collected at the same locality as the holotype on 12 April 2023 by Wei-Cai Chen.
The specific name ‘wumingensis’ is derived from the type locality, Wuming District, Nanning City, Guangxi, China. The proposed common name in English is Wuming Leaf Litter Toad, and in Chinese, it is called Wu Ming Zhang Tu Chan (武鸣掌突蟾).
Leptobrachella wumingensis sp. nov. is classified under Leptobrachella based on specific morphological features, including its relatively small body size, presence of an inner metacarpal tubercle, macro-glands on the supra-axillary and femoral glands, lack of vomerine teeth, and a whitish vertical bar on the anterior tip of the snout, according to previous studies (
Head length almost equal to width (HW/HL = 1.02); snout bluntly rounded in profile and dorsal view, projecting slightly over lower jaw; nostril oval-shaped, closer to tip of snout than eye; canthus rostralis distinct; loreal region distinctly sloping, slightly concave; pupil vertical; eye diameter less than snout length (ED/SNT = 0.96); tympanum distinct and rounded, diameter about 49% that of eye; vomerine teeth absent; tongue with a deep notch at posterior tip; supratympanic fold distinctly raised from corner of eye to the posterior of tympanum (Fig.
Tips of fingers slightly swollen; relative finger lengths I < II < IV < III; subarticular tubercles absent; prominent inner palmar tubercle, small outer palmar tubercle; finger webbing and dermal fringes absent; nuptial pad on fingers absent. Tips of toes rounded, slightly swollen; relative toe lengths I < II < V = III < IV; subarticular tubercles absent, replaced by dermal ridges; prominent and elongated inner metatarsal tubercle; outer metatarsal tubercle absent; toe webbing and lateral fringes absent. Tibia 48% of SVL; tibiotarsal articulation reaching middle of eye; heels meeting when hindlimbs flexed at right angles with respect to body (Fig.
Dorsal surface shagreened and granular; upper eyelid with small tubercles; ventral surface without tubercles; flanks with sparse tubercles; pectoral glands elongated, approximately 2.1 mm in diameter; small femoral glands oval, approximately 0.6 mm in diameter, closer to knee than to vent; supra-axillary glands oval, approximately 0.8 mm in diameter; ventrolateral glandular line discrete; ventral surface of thigh with some tubercles (Fig.
Dorsum light brown with brown markings, an inverted triangle marking between eyes, an irregular brown marking on scapular region, a brown ‘Λ’ marking on rear of dorsum; tympanum pale brown; supratympanic fold black from the posterior corner of eye to the anterior of supra-axillary glands; two wide black bars on upper lip; five irregular black spots and several small light tangerine tubercles on flanks; transverse brown bars on dorsal surface of hindlimbs; upper arms light orange; belly with tiny creamy white spots; throat creamy white with tiny light brown spots; lower jaw with creamy white tubercles; ventral surfaces of limbs with sparse creamy white tubercles; pectoral and femoral glands creamy white, supra-axillary glands light orange; pupil black; iris bicolored, upper half copper, fading to silver in lower half (Fig.
The dorsum and limb surfaces are faded to uniform light brown. Irregular black spots on flanks and bars on limbs are darkish brown. The throat, chest, and belly are creamy white, and pectoral, femoral, supra-axillary, and ventrolateral glands are also creamy white.
The measurements of the type series are shown in Table
Measurements of voucher specimens of L. wumingensis sp. nov. (mm). Abbreviations defined in text.
NNU 01058 | NNU 01086 | NNU 201907009 | NNU 00283 | NNU 00284 | NNU 00285 | NNU 00286 | NNU 01059 | NNU 01060 | |
---|---|---|---|---|---|---|---|---|---|
Sex | Male | Male | Male | Female | Female | Female | Female | Female | Female |
SVL | 26.7 | 26.6 | 26.0 | 31.2 | 31.5 | 30.6 | 31.9 | 34.8 | 31.6 |
HL | 8.7 | 8.7 | 8.9 | 10.2 | 10.0 | 10.2 | 10.8 | 10.4 | 11.2 |
HW | 8.9 | 9.0 | 9.3 | 10.5 | 10.6 | 10.8 | 10.8 | 11.0 | 11.2 |
SNT | 3.4 | 3.8 | 3.7 | 4.3 | 4.2 | 4.1 | 4.0 | 4.5 | 4.3 |
ED | 3.2 | 3.6 | 3.4 | 4.3 | 4.8 | 4.6 | 4.1 | 3.7 | 3.6 |
IOD | 3.0 | 2.7 | 2.8 | 3.1 | 3.3 | 3.6 | 3.3 | 3.4 | 2.9 |
IN | 2.7 | 2.6 | 2.2 | 3.4 | 3.5 | 3.6 | 3.4 | 3.2 | 3.0 |
TD | 1.6 | 1.8 | 1.8 | 2.4 | 2.4 | 1.9 | 2.1 | 2.1 | 2.0 |
TED | 0.9 | 1.0 | 0.8 | 1.3 | 1.3 | 1.1 | 1.4 | 1.1 | 0.9 |
ML | 6.2 | 5.8 | 6.1 | 7.3 | 7.3 | 7.6 | 7.4 | 7.3 | 7.2 |
FLL | 11.5 | 11.9 | 11.4 | 14.3 | 14.5 | 14.4 | 14.4 | 13.8 | 14.4 |
THL | 12.3 | 11.7 | 12.9 | 14.9 | 16.3 | 15.4 | 15.6 | 14.1 | 14.0 |
TIB | 12.7 | 12.8 | 12.8 | 14.4 | 15.9 | 15.4 | 15.9 | 14.9 | 15.2 |
PL | 11.5 | 11.4 | 11.4 | 13.4 | 14.4 | 14.0 | 14.2 | 13.6 | 13.7 |
FEM | 0.6 | 0.7 | 0.7 | 1.5 | 1.5 | 1.4 | 1.2 | 1.1 | 1.0 |
Leptobrachella wumingensis sp. nov. inhabits evergreen forests in the DMS at elevations of 1000–1200 m. All specimens were discovered over 10 m away from rocky streams (Fig.
To begin with, L. wumingensis sp. nov. is distinguished from Leptobrachella species found south of the Isthmus of Kra, Malay Peninsula by the presence of supra-axillary and ventrolateral glands (vs absent in the latter). Furthermore, Leptobrachella wumingensis sp. nov. and L. damingshanensis are sympatric. Leptobrachella wumingensis sp. nov. can be differentiated from L. damingshanensis by several characters, including smaller body size in males (SVL 26.0–26.7 mm vs 33.6–34.4 mm); indistinct orange glands on the proximal thigh area (vs a pair of distinct tangerine glands on the proximal thigh area); pale brown dorsal skin with brown markings and a shagreened and granular dorsal surface (vs rough dorsal skin with sparse tangerine tubercles and some short longitudinal ridges); the absence of toe webbing and lateral fringes on toes (vs rudimentary toes webbing and narrow lateral fringes on toes); distinct transverse dark brown bars on the dorsal surface of hindlimbs (vs indistinct transverse dark brown bars on the dorsal surface of hindlimbs); and different bioacoustics, including two types of calling models and dominant frequency at 6.0–7.5 kHz (vs a single calling model and dominant frequency at 4.6–5.2 kHz) (Fig.
According to the phylogenetic analysis, L. wumingensis sp. nov., L. damingshanensis, L. nahangensis, and L. nyx constitute a monophyletic group (Fig.
In comparison to other recognized Leptobrachella species from north of the Isthmus of Kra, L. wumingensis sp. nov. is distinguishable from smaller species such as L. applebyi (19.6–22.3 mm in males), L. ardens (21.3–24.7 mm in males), L. aspera (22.4 mm in male), L. feii (21.5–22.8 mm in males), L. korifi (22.7 mm in female), L. melica (19.5–22.7 mm in males), L. murphyi (23.2–24.9 mm in males), L. niveimontis (22.5–23.6 mm in males) and L. pluvialis (21.3–22.3 mm in males) due to its larger size (26.0–26.7 mm in males). Additionally, L. wumingensis sp. nov. is distinct from the notably larger L. sungi (48.3–52.7 mm in males) and L. zhangyapingi (45.8–52.5 mm in males).
Leptobrachella wumingensis sp. nov. can be distinguished from L. aerea, L. alpina, L. dong, L. eos, L. firthi, L. graminicola, L. isos, L. khasiorum, L. laui, L. liui, L. murphyi, L. purpurus, L. shimentaina, L. tamdil, L. yingjiangensis, L. yunkaiensis and L. zhangyapingi by the absence of lateral fringes on its toes (vs. wide lateral fringes); from L. aspera, L. bashaensis, L. bidoupensis, L. bijie, L. botsfordi, L. bourreti, L. chishuiensis, L. damingshanensis, L. dorsospina, L. feii, L. flaviglandulosa, L. fuliginosa, L. jinshaensis, L. jinyunensis, L. korifi, L. mangshanensis, L. maoershanensis, L. niveimontis, L. pelodytoides, L. petrops, L. puhoatensis, L. purpuraventra, L. shangsiensis, L. sinorensis, L. suiyangensis, L. sungi, L. tengchongensis, L. ventripunctata, L. verrucosa, L. wuhuangmontis, L. wulingensis, L. yeae, and L. yunyangensis by the absence of lateral fringes on its toes (vs. narrow lateral fringes). Leptobrachella wumingensis sp. nov. is also differentiated from L. aerea, L. botsfordi, L. crocea, L. graminicola, L. eos, L. firthi, L. isos, L. pallida, L. petrops, and L. tuberosa by the presence of black spots on its flanks (vs. absent). Leptobrachella wumingensis sp. nov. differs from L. applebyi, L. bidoupensis, L. kalonensis, L. melica, L. minima, L. nahangensis, and L. tadungensis by the presence of shagreened and granular dorsal surface (vs smooth).
Lastly, L. wumingensis sp. nov. differs from other Leptobrachella species in terms of acoustic features such as relatively high dominant frequencies and two distinct types of callings (Fig.
Based on morphological characters, molecular data and bioacoustics, we have identified the DMS specimens as a new species. The validation of this assignment is supported by significant genetic divergence (>7.1%), high dominant frequencies, complicated calling styles and substantial morphological characters. The DMS reserve is located in the central region of Guangxi, and previous fieldwork by many investigation teams did not discover any Leptobrachella species until we reported the first one in 2021 (
According to previous research and our new data (
The authors are grateful to the Damingshan National Nature Reserve staff (Li Lin, Chang-Gui Cen, and De-Qi Zhang) for assistance during fieldwork.
The authors have declared that no competing interests exist.
No ethical statement was reported.
This work was supported by the National Natural Science Foundation of China (32060116) and Guangxi Natural Science Foundation, China (2020GXNSFDA 238022).
CWC designed the study and wrote the manuscript. PWX, LP and YGD took part in fieldwork. CWC and LP performed the molecular experiments and analyzed the data. PWX drew the figures.
Wei-Cai Chen https://orcid.org/0000-0002-2398-4079
Wan-Xiao Peng https://orcid.org/0000-0001-5635-9061
Peng Li https://orcid.org/0000-0001-8311-0544
Gui-Dong Yu https://orcid.org/0009-0008-1928-6594
All of the data that support the findings of this study are available in the main text or Supplementary Information.
Supplementary information
Data type: morphological, phylogenetic
Explanation note: table S1: Literature for morphological characters for species of the genus Leptobrachella; table S2: Comparative material examined; table S3: Dominant frequency of advertisement calls of species available in the genus Leptobrachella; table S4: Uncorrected p-distance in Leptobrachella species based on 16S mtDNA fragments.