Corresponding author: Daode Yang (
Academic editor: A. Crottini
A new species,
Zhang B, Li Y, Hu K, Li P, Gu Z, Xiao N, Yang D (2021) A new species of
The odorous frogs of the genus
The Wuling Mountains are a priority area for biodiversity conservation in China (
We conducted a herpetological survey in Zhangjiajie City, Hunan Province, from 23 July to 6 August 2019. A total of five specimens (two adult males and three adult females) were collected in mountain streams on 30 July 2019, in Sangzhi County, Zhangjiajie City, Hunan Province, China (Figure
Collection location of
Genomic DNA was extracted from 95% ethanol-preserved liver tissues using a TSINGKE (
Measurements were taken by a digital caliper (0.01 mm, rounded to the nearest 0.1 mm). Character measurements followed those of
Sexual size dimorphism was represented by the size dimorphism index (
Comparative morphological data of four species (
To reduce the impact of allometry, a corrected value from the ratio of each morphological measurement to
We compared the morphological characters between the new species and its congeners, based on literature values from the following species:
The
Bayesian Inference tree based on the partial DNA sequences of the mitochondrial 12S and 16S genes. Numbers before slashes indicate Bayesian posterior probabilities and numbers after slashes are bootstrap support for Maximum Likelihood (1000 replicates) analyses. The symbol “-” indicates a value below 50%.
Three adult females,
Morphology and colour pattern of living
Adult male,
Forelimbs sturdy (
Hind limbs relatively long, the heels overlapping obviously, tibiotarsal articulation beyond the tip of the snout; tibia length (
Skin slightly rough with dense granules, several large warts on dorsum and flanks; dorsolateral folds absent; two glands behind the back edge of the lips; a small beige dot between anterior corners of the eyes; pale spinules present on throat and chest.
In life, the yellow-green dorsum of head and body with scattered irregular dark brown spots; no light edges around spots; small area of dark brown spots on the dorsum of head; large dark black spots in the centre on dorsum, continuing on to light yellow flank with several brown spots; supratympanic fold slightly distinct, brown; upper and lower lip with vertical brown bars; arms green-yellow with brown transverse bands, thighs and tibias with four brown bands; different widths of brown striations on green-yellow limbs, different distances between striations (Figure
On dorsum, colour fades to grey-blue with dark grey spots, brown bars on upper and lower lips change to dark grey; ventre variable from yellowish to creamy-white; beige dot between anterior corners of the eyes fades to white; underside of limbs varies from brown to beige (Figure
The holotype
Measurements of the five specimens are shown in Table
Measurements (mm) of the morphometric characters for the holotype and paratypes of
Specimen ID | ||||||||
---|---|---|---|---|---|---|---|---|
Sex | Male | Male | Mean (n=2) | Female | Female | Female | Min–Max (n=3) | Mean±SD (n=3) |
Type status | H | P | P | P | P | |||
|
42.1 | 45.1 | 43.6 | 89.7 | 92.7 | 83.3 | 83.3–92.7 | 88.6 ± 4.8 |
|
16.5 | 15.2 | 15.9 | 25.8 | 25.8 | 23.6 | 23.6–25.8 | 25.1 ± 1.3 |
|
12.1 | 12.6 | 12.4 | 21.4 | 22.2 | 21.9 | 21.4–22.2 | 21.8 ±0.4 |
|
6.5 | 5.8 | 6.2 | 12.3 | 11.0 | 11.8 | 11.0–12.3 | 11.7 ±0.7 |
|
5.2 | 5.6 | 5.4 | 8.5 | 8.4 | 8.1 | 8.1–8.5 | 8.3 ± 0.2 |
|
3.9 | 4.6 | 4.3 | 6.5 | 7.4 | 6.5 | 6.5–7.4 | 6.8 ± 0.5 |
|
5.3 | 4.8 | 5.1 | 8.7 | 8.7 | 8.9 | 8.7–8.9 | 8.8 ± 0.1 |
|
3.4 | 3.5 | 3.5 | 5.8 | 6.5 | 6.8 | 5.8–6.8 | 6.4 ± 0.5 |
|
3.4 | 3.3 | 3.4 | 4.6 | 4.9 | 4.3 | 4.3–4.9 | 4.6 ± 0.3 |
|
21.1 | 22.8 | 22.0 | 42.3 | 41.0 | 41.4 | 41.0–42.3 | 41.6 ± 0.7 |
|
4.5 | 4.7 | 4.6 | 8.4 | 8.9 | 7.1 | 7.1–8.9 | 8.1 ± 0.9 |
|
21.6 | 25.9 | 23.8 | 46.9 | 50.2 | 47.6 | 46.9–50.2 | 48.2 ± 1.7 |
|
22.8 | 23.6 | 23.2 | 46.3 | 47.6 | 51.0 | 46.3–51.0 | 48.3 ± 2.4 |
|
5.0 | 5.6 | 5.3 | 12.1 | 12.4 | 12.4 | 12.1–12.4 | 12.3 ± 0.2 |
|
31.5 | 31.1 | 31.3 | 69.0 | 61.4 | 53.2 | 53.2–69.0 | 61.2 ± 7.9 |
|
23.4 | 24.0 | 23.7 | 50.8 | 47.3 | 46.8 | 46.8–50.8 | 48.3 ± 2.2 |
|
1.5 | 1.4 | 1.5 | 3.0 | 3.3 | 2.7 | 2.7–3.3 | 3.0 ± 0.3 |
Within the
Comparisons of morphological characteristics with congeners
Diagnostic characters separating the new species described in this study and its relatives.
Species |
|
Head length1 | Metacarpal tubercles2 | Circum-marginal grooves3 | Vocal sacs4 | Tibiotarsal articulation5 | Relative finger lengths6 | Relative toe lengths7 | |
---|---|---|---|---|---|---|---|---|---|
males (n) | females (n) | ||||||||
42.1–45.1 (2) | 83.3–92.7 (3) | ++ | + | + | + | a | a | a | |
|
50.7–57.8 (8) | 73.3–84.9 (5) | + | ++ | – | + | b | b | a |
|
38.0–46.7 (7) | 76.9–88.6 (6) | – | ++ | + | + | c | a | b |
|
36.2–43.3 (7) | 67.9–78.7 (6) | ++ | + | + | + | b | b | a |
|
37.9–43.4 (7) | 72.1–80.4 (4) | – | – | + | + | b | c | c |
|
39.4–45.9 (23) | 68.1–81.9 (8) | ++ | ++ | + | + | b | a | b |
|
37.4–51.8 (11) | 77.3–111.9 (31) | ++ | ++ | + | / | a | b | a |
|
49.1–61.9 (5) | 75.4–122.5 (8) | ++ | ++ | + | – | d | d | a |
|
52.5–53.5 (2) | 90.5–107.6 (4) | ++ | / | + | + | e | b | / |
1 head longer than wider (++), head longer or almost equal wide (+), head longer almost equal wide (–); 2 three metacarpal tubercles (++), two metacarpal tubercles (+), outer metacarpal tubercle being indistinct (–); 3 present (+), without circum-marginal grooves on finger I (–), outer metacarpal tubercle being indistinct (–); 4 paired external vocal sacs (+), paired internal vocal sacs (–) in males; 5 beyond the tip of the snout (a), reaching to the level between eye to nostril (b), reaching the nostril (c), reaching the anterior corner of the eye or the tip of snout (d), far beyond the snout (e); 6 I ≤ II < IV < III (a), II < I < IV < III (b), II < I ≈ IV < III (c), and II < IV ≈ I < III (d); 7 I < II < III < V < IV (a), I < II < III ≤ V < IV (b), and I < II < III ≈ V < IV (c).
Morphometric comparisons between
Characters |
|
|
|
|
|
---|---|---|---|---|---|
Males (n = 2) | Males (n = 8) | Males (n = 7) | Males (n = 7) | Males (n = 7) | |
|
42.1–45.1 | 50.7–57.8 (54.3 ± 2.4) | 38.0–46.7 (42.2 ± 2.8) | 36.2–43.3 (40.9 ± 2.6) | 37.9–43.4 (41.6 ± 2.0) |
|
15.2–16.5 | 19.2–21.3 (20.3 ± 0.8) | 14.2–18.0 (16.0 ± 1.2) | 14.1–17.3 (15.9 ± 1.3) | 15.4–18.0 (17.2 ± 0.9) |
|
12.1–12.6 | 17.9–21.3 (19.1 ± 1.1) | 12.7–16.0 (14.9 ± 1.1) | 12.5–15.7 (14.4 ± 1.1) | 13.9–15.8 (15.0 ± 0.7) |
|
5.8–6.5 | 8.4–10.1 (9.0 ± 0.6) | 5.9–7.4 (6.7 ± 0.6) | 5.5–7.0 (6.5 ± 0.5) | 6.1–7.3 (6.9 ± 0.4) |
|
5.2–5.6 | 6.0–7.3 (6.6 ± 0.4) | 4.4–6.5 (5.7 ± 0.7) | 4.8–5.9 (5.5 ± 0.5) | 3.6–6.9 (5.8 ± 1.1) |
|
3.9–4.6 | 4.2–5.3 (4.6 ± 0.3) | 3.0–3.9 (3.3 ± 0.4) | 2.6–3.5 (3.1 ± 0.3) | 3.2–4.1 (3.6 ± 0.3) |
|
4.8–5.3 | 5.0–6.7 (5.7 ± 0.6) | 4.1–5.3 (4.6 ± 0.4) | 4.5–5.4 (4.9 ± 0.3) | 4.5–5.1 (4.8 ± 0.2) |
|
3.4–3.5 | 4.2–6.0 (4.9 ± 0.6) | 3.0–4.2 (3.6 ± 0.4) | 3.1–4.0 (3.6 ± 0.4) | 3.3–4.2 (3.8 ± 0.3) |
|
3.3–3.4 | 4.3–5.5 (4.6 ± 0.4) | 3.3–4.4 (4.0 ± 0.5) | 2.9–4.2 (3.8 ± 0.4) | 3.3–4.6 (4.1 ± 0.4) |
|
21.1–22.8 | 24.8–26.8 (26.0 ± 0.7) | 18.5–21.9 (20.3 ± 1.4) | 20.2–24.3 (21.4 ± 1.4) | 19.6–22.2 (20.4 ± 0.9) |
|
4.5–4.7 | 5.0–6.3 (5.6 ± 0.5) | 3.1–5.0 (4.2 ± 0.8) | 3.7–4.4 (4.0 ± 0.2) | 3.8–5.0 (4.3 ± 0.4) |
|
21.6–25.9 | 25.6–30.1 (27.8 ± 1.6) | 19.6–24.0 (22.2 ± 1.4) | 19.6–25.2 (23.0 ± 2.2) | 20.8–24.6 (22.4 ± 1.1) |
|
22.8–23.6 | 30.4–33.9 (31.8 ± 1.0) | 22.6–27.0 (24.7 ± 1.6) | 22.7–28.8 (25.2 ± 2.0) | 23.4–25.0 (24.4 ± 0.6) |
|
5.0–5.6 | 6.4–7.4 (6.8 ± 0.4) | 4.7–5.9 (5.3 ± 0.4) | 4.4–5.6 (5.1 ± 0.5) | 5.0–6.0 (5.6 ± 0.3) |
|
31.1–31.5 | 41.2–43.2 (42.5 ± 0.7) | 29.3–35.9 (32.3 ± 2.3) | 30.1–38.7 (34.7 ± 2.6) | 30.6–36.4 (33.2 ± 1.8) |
|
23.4–24.0 | 28.0–30.8 (29.3 ± 1.0) | 20.4–25.9 (23.3 ± 2.0) | 21.1–26.8 (24.1 ± 1.8) | 22.7–24.6 (23.6 ± 0.6) |
|
1.4–1.5 | 1.4–2.1 (1.7 ± 0.2) | 1.2–2.1 (1.6 ± 0.3) | 1.2–1.9 (1.5 ± 0.2) | 1.4–2.0 (1.6 ± 0.2) |
Plots of principal component analyses of
Morphometric comparisons between
Characters |
|
|
|
|
||||||
---|---|---|---|---|---|---|---|---|---|---|
Females (n = 3) | Females (n = 5) | Females (n = 10) | Females (n = 12) | Females (n = 7) | ||||||
|
83.3–92.7 |
73.3–84.9 |
/ | 75.0–88.6 |
/ | 64.7–81.1 |
/ | 72.1–80.7 |
/ | <0.001 |
|
23.6–25.8 |
26.1–29.1 |
<0.001 | 27.5–32.8 |
<0.001 | 25.6–30.9 |
<0.001 | 27.1–32.3 |
<0.001 | 0.002 |
|
21.3–22.2 |
27.4–28.6 |
<0.001 | 26.9–31.9 |
<0.001 | 23.3–29.5 |
<0.001 | 25.7–28.0 |
<0.001 | <0.001 |
|
11.0–12.3 |
12.5–13.9 |
0.009 | 11.8–14.4 |
<0.001 | 10.6–12.6 |
0.001 | 11.2–13.5 |
0.005 | <0.001 |
|
8.1–8.5 |
8.7–9.9 |
0.052 | 7.8–10.4 |
0.044 | 7.9–9.9 |
<0.001 | 7.5–9.7 |
0.069 | 0.376 |
|
6.5–7.4 |
6.3–8.2 |
0.086 | 4.7–7.4 |
0.985 | 5.2–6.6 |
0.993 | 4.8–7.0 |
0.557 | 0.005 |
|
8.7–8.9 |
8.0–9.3 |
0.243 | 7.7–9.8 |
0.142 | 7.7–9.8 |
0.002 | 7.1–8.7 |
0.417 | 0.231 |
|
5.8–6.8 |
5.8–7.3 |
0.236 | 5.8–7.1 |
0.100 | 5.2–7.2 |
0.041 | 5.3–7.0 |
0.204 | 0.534 |
|
4.3–4.9 |
5.4–6.3 |
0.003 | 3.4–5.7 |
0.233 | 4.3–5.2 |
0.003 | 4.5–5.7 |
0.007 | 0.002 |
|
41.0–42.2 |
34.9–40.0 |
0.773 | 36.0–41.6 |
0.732 | 33.4–41.4 |
0.026 | 35.0–37.4 |
0.808 | 0.003 |
|
7.1–8.9 |
6.0–7.6 |
0.279 | 6.1–8.5 |
0.337 | 5.2–7.8 |
0.155 | 5.8–7.2 |
0.138 | 0.003 |
|
46.9–50.1 |
40.4–47.5 |
0.570 | 41.3–46.2 |
0.241 | 35.8–44.9 |
0.324 | 37.7–42.8 |
0.133 | 0.001 |
|
46.2–51.0 |
46.6–50.7 |
0.128 | 41.8–53.1 |
0.054 | 41.0–49.8 |
0.001 | 43.0–46.5 |
0.122 | 0.054 |
|
12.1–12.4 |
9.6–11.0 |
0.105 | 10.0–13.0 |
0.983 | 8.3–11.2 |
0.256 | 9.6–11.2 |
0.237 | 0.001 |
|
53.2–69.0 |
58.8–68.8 |
0.048 | 61.3–71.4 |
0.001 | 55.9–70.0 |
<0.001 | 60.3–63.5 |
0.009 | 0.103 |
|
46.8–50.8 |
40.8–48.5 |
0.537 | 42.6–48.3 |
0.498 | 37.9–48.1 |
0.014 | 40.2–43.2 |
0.875 | 0.008 |
|
2.7–3.3 |
2.4–3.3 |
0.841 | 2.7–3.5 |
0.014 | 2.4–3.5 |
0.009 | 2.1–3.2 |
0.927 | 0.039 |
The scientific name “
The new species is currently known only from its type locality, Sangzhi County, Zhangjiajie, Hunan Province, China (Figure
Habitat of
Odorous frog species are widely distributed and have many morphological characteristics of derivation or specialisation. Different species of
The Yunnan-Guizhou Plateau is thought to be the centre of
We are much indebted to Thomas Ziegler and one anonymous reviewer for their valuable comments. We thank Qi Gu, Jianjun Xiang, Shihua Li, Tianyu Qian, Xiangyun Ding, Dejia Hou, Yue Cao, Xin Xia and Zhiwei Gao for assistance during the field survey. We thank Shuo Qi, Sikan Chen from the Guizhou Institute of Forest Inventory and Planning, Simin Wu, Chaohui Zeng and Jinyu Ye for providing help with manuscript preparation. We thank Bo Cai from the Chengdu Institute of Biology, Chinese Academy of Sciences; Liang Qiao from the Xinxiang Medical University; Shengchao Shi from the Chengdu Institute of Biology, Chinese Academy of Sciences; Shize Li from the Guizhou University; and Yufan Wang from Zhejiang Forest Resources Monitoring Center for taking and providing photos of the
Table S1. Samples and GenBank accession numbers for molecular analyses used in this study
phylogenetic data
Table S2. The morphometric data of the
morphological data
See abbreviations for characters in the Materials and methods section.
Table S3. Uncorrected
phylogenetic data