Research Article |
Corresponding author: Ying-Yong Wang ( wangyy@mail.sysu.edu.cn ) Academic editor: Angelica Crottini
© 2019 Zhi-Tong Lyu, Lin-Sheng Huang, Jian Wang, Yuan-Qiu Li, Hong-Hui Chen, Shuo Qi, Ying-Yong 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:
Lyu Z-T, Huang L-S, Wang J, Li Y-Q, Chen H-H, Qi S, Wang Y-Y (2019) Description of two cryptic species of the Amolops ricketti group (Anura, Ranidae) from southeastern China. ZooKeys 812: 133-156. https://doi.org/10.3897/zookeys.812.29956
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Two cryptic species, which were previously reported as Amolops ricketti, are revealed on the basis of significant morphological and genetic divergences. Amolops sinensis sp. n. from central Guangdong, northeastern Guangxi and southwestern Hunan can be distinguished by the longitudinal glandular folds on the skin of the shoulders and other character combinations. Amolops yatseni sp. n. from the coastal hills of west Guangdong can be distinguished by the dense tiny round translucent, or white, spines on the dorsal skin of the body, dorsal and dorsolateral skin of the limbs, and other character combinations. The phylogenetic interrelationships of the A. ricketti group have been inferred as (A. wuyiensis + A. ricketti) + (A. yunkaiensis + (A. albispinus + (A. sinensis sp. n. + A. yatseni sp. n.))). This work indicates that the current records of A. ricketti might be a species complex composed of multiple species, and further work is needed to figure out this puzzle.
Amolops sinensis sp. n., Amolops yatseni sp. n., mitochondrial DNA, morphology, phylogeny, species complex, torrent frog
The torrent frog genus Amolops Cope, 1865, which is comprised of 55 species, is widespread from the southern and eastern Himalayas, eastward to southeastern mainland China, and southward to Malay Peninsula (
During our herpetological surveys in Guangdong, Guangxi and Hunan provinces in southeastern China, we have collected a series of Amolops specimens which were recorded as A. ricketti (
A total of 28 muscle samples of the two new species were collected for molecular analyses, encompassing six from Zhongshan City, Guangdong, two from Shangchuan Island, Guangdong, two from Mt. Gudou, Guangdong, three from Ehuangzhang Nature Reserve, Guangdong, two from Yunkaishan Nature Reserve, Guangdong, seven from Shimentai Nature Reserve, Guangdong, two from Mt. Nankun, Guangdong, two from Mt. Dupangling, Guangxi, one from Mt. Yangming, Hunan, and one from Mt. Hengshan, Hunan. In addition, 36 samples from nine known species of the genus Amolops, namely A. albispinus, A. ricketti, A. wuyiensis, A. yunkaiensis, A. daiyunensis (Liu & Hu, 1975), A. hongkongensis (Pope & Romer, 1951), A. hainanensis(Boulenger, 1900), A. torrentis (Smith, 1923) and A. chunganensis (Pope, 1929), were also collected and incorporated into our dataset. All muscle samples were attained from euthanasia specimens and then preserved in 95% ethanol and stored at -40 °C. Detail information for these materials is shown in Table
Localities, voucher number and GenBank numbers for all samples used in this study.
ID | Species | Localities (* type localities) | Voucher | 16S | CO1 |
1 | Amolops sinensis sp. n. | * China: Shimentai Nature Reserve, Guangdong | SYS a004165 | MK263262 | MK263314 |
2 | * China: Shimentai Nature Reserve, Guangdong | SYS a004722 | MK263278 | MK263318 | |
3 | * China: Shimentai Nature Reserve, Guangdong | SYS a007105 | MK263297 | MK263329 | |
4 | * China: Shimentai Nature Reserve, Guangdong | SYS a007106 | MK263298 | MK263330 | |
5 | * China: Shimentai Nature Reserve, Guangdong | SYS a007107 | MK263299 | MK263331 | |
6 | * China: Shimentai Nature Reserve, Guangdong | SYS a007108 | MK263300 | MK263332 | |
7 | * China: Shimentai Nature Reserve, Guangdong | SYS a007109 | MK263301 | MK263333 | |
8 | China: Mt. Nankun, Guangdong | SYS a005710 | MK263287 | MK263321 | |
9 | China: Mt. Nankun, Guangdong | SYS a005712 | MK263288 | MK263322 | |
10 | China: Mt. Dupangling, Guangxi | SYS a005089 | MK263279 | MK263319 | |
11 | China: Mt. Dupangling, Guangxi | SYS a005111 | MK263280 | MK263320 | |
12 | China: Mt. Yangming, Hunan | SYS a007268 | MK263302 | MK263334 | |
13 | China: Mt. Hengshan, Hunan | SYS a004257 | MK263265 | MK263315 | |
14 | Amolops yatseni sp. n. | * China: Zhongshan City, Guangdong | SYS a006806 | MK263289 | MK263323 |
15 | * China: Zhongshan City, Guangdong | SYS a006807 | MK263290 | MK263324 | |
16 | * China: Zhongshan City, Guangdong | SYS a006808 | MK263291 | MK263325 | |
17 | * China: Zhongshan City, Guangdong | SYS a006810 | MK263292 | MK263326 | |
18 | * China: Zhongshan City, Guangdong | SYS a006811 | MK263293 | MK263327 | |
19 | * China: Zhongshan City, Guangdong | SYS a006857 | MK263296 | MK263328 | |
20 | China: Shangchuan Island, Guangdong | SYS a003633 | MK263250 | MK263304 | |
21 | China: Shangchuan Island, Guangdong | SYS a003634 | MK263251 | MK263305 | |
22 | China: Mt. Gudou, Guangdong | SYS a006818 | MK263294 | MK263306 | |
23 | China: Mt. Gudou, Guangdong | SYS a006819 | MK263295 | MK263307 | |
24 | China: Ehuangzhang Nature Reserve, Guangdong | SYS a003978 | MK263252 | MK263308 | |
25 | China: Ehuangzhang Nature Reserve, Guangdong | SYS a003980 | MK263254 | MK263309 | |
26 | China: Ehuangzhang Nature Reserve, Guangdong | SYS a003981 | MK263255 | MK263310 | |
27 | China: Yunkaishan Nature Reserve, Guangdong | SYS a004642 | MK263269 | MK263316 | |
28 | China: Yunkaishan Nature Reserve, Guangdong | SYS a004643 | MK263270 | MK263317 | |
29 | Amolops albispinus | * China: Mt. Wutong, Guangdong | SYS a003452 | MK263247 | KX507332 |
30 | * China: Mt. Wutong, Guangdong | SYS a003453 | MK263248 | KX507333 | |
31 | * China: Mt. Wutong, Guangdong | SYS a003454 | MK263249 | KX507334 | |
32 | Amolops ricketti | * China: Mt. Wuyi, Fujian | SYS a004141 | MK263259 | MG991927 |
33 | * China: Mt. Wuyi, Fujian | SYS a004142 | MK263260 | MG991928 | |
34 | * China: Mt. Wuyi, Fujian | SYS a004143 | MK263261 | MG991929 | |
35 | China: Mt. Emeifeng, Fujian | SYS a002492 | MK263244 | KX507329 | |
36 | China: Shanghang County, Fujian | SYS a003342 | MK263246 | KX507331 | |
37 | China: Shanghang County, Fujian | SYS a004106 | MK263256 | MK263311 | |
38 | Amolops wuyiensis | * China: Mt. Wuyi, Fujian | SYS a001716 | MK263239 | KX507324 |
39 | * China: Mt. Wuyi, Fujian | SYS a001717 | MK263240 | KX507325 | |
40 | * China: Mt. Wuyi, Fujian | SYS a004139 | MK263257 | MK263312 | |
41 | * China: Mt. Wuyi, Fujian | SYS a004140 | MK263258 | MK263313 | |
42 | China: Jingning County, Zhejiang | SYS a002723 | MK263245 | MK263303 | |
43 | Amolops yunkaiensis | * China: Ehuangzhang Nature Reserve, Guangdong | SYS a003979 | MK263253 | MG991907 |
44 | * China: Ehuangzhang Nature Reserve, Guangdong | SYS a004705 | MK263275 | MG991906 | |
45 | * China: Ehuangzhang Nature Reserve, Guangdong | SYS a004706 | MK263276 | MG991908 | |
46 | * China: Ehuangzhang Nature Reserve, Guangdong | SYS a004707 | MK263277 | MG991909 | |
47 | China: Yunkaishan Nature Reserve, Guangdong | SYS a004681 | MK263271 | MG991910 | |
48 | Amolops yunkaiensis | China: Yunkaishan Nature Reserve, Guangdong | SYS a004682 | MK263272 | MG991911 |
49 | China: Yunkaishan Nature Reserve, Guangdong | SYS a004683 | MK263273 | MG991912 | |
50 | China: Yunkaishan Nature Reserve, Guangdong | SYS a004684 | MK263274 | MG991913 | |
51 | Amolops daiyunensis | * China: Mt. Daiyun, Fujian | SYS a001737 | MK263241 | KX507326 |
52 | * China: Mt. Daiyun, Fujian | SYS a001738 | MK263242 | KX507327 | |
53 | * China: Mt. Daiyun, Fujian | SYS a001739 | MK263243 | KX507328 | |
54 | Amolops hongkongensis | * China: Hong Kong | SYS a004577 | MK263266 | MG991919 |
55 | * China: Hong Kong | SYS a004578 | MK263267 | MG991920 | |
56 | * China: Hong Kong | SYS a004579 | MK263268 | MG991921 | |
57 | Amolops hainanensis | * China: Mt. Wuzhi, Hainan | SYS a005281 | MK263281 | MG991916 |
58 | * China: Mt. Wuzhi, Hainan | SYS a005282 | MK263282 | MG991917 | |
59 | * China: Mt. Wuzhi, Hainan | SYS a005283 | MK263283 | MG991918 | |
60 | Amolops torrentis | * China: Mt. Wuzhi, Hainan | SYS a005289 | MK263284 | MG991930 |
61 | * China: Mt. Wuzhi, Hainan | SYS a005290 | MK263285 | MG991931 | |
62 | * China: Mt. Wuzhi, Hainan | SYS a005291 | MK263286 | MG991932 | |
63 | Amolops chunganensis | China: Mt. Jinggang, Jiangxi | SYS a004212 | MK263263 | MG991914 |
64 | China: Mt. Jinggang, Jiangxi | SYS a004213 | MK263264 | MG991915 |
Genomic DNA was extracted from muscular tissue by using a DNA extraction kit from Tiangen Biotech (Beijing) Co., Ltd. Partial 16S ribosomal RNA gene (16S) and partial cytochrome C oxidase 1 gene (CO1) were amplified. Primers used for 16S were L3975 (5’-CGCCTGTTTACCAAAAACAT-3’) and H4551 (5’-CCGGTCTGAACTCAGATCACGT-3’) following
DNA sequences were aligned by the Clustal W algorithm with default parameters (
We obtained diagnostic characters of known species of the genus Amolops from the literature for comparison. In addition, a total of 67 museum specimens of A. ricketti group were examined for comparison, which are listed in Appendix
Measurements follow
SVL snout-vent length (from tip of snout to posterior margin of vent);
HDL head length (from tip of snout to the articulation of the jaw);
HDW head width (head width at the commissure of the jaws);
SNT snout length (from tip of snout to the anterior corner of the eye);
IND internasal distance (distance between nares);
IOD interorbital distance (minimum distance between upper eyelids);
ED eye diameter (from the anterior corner of the eye to posterior corner of the eye);
TD tympanum diameter (horizontal diameter of tympanum);
TED tympanum-eye distance (from anterior edge of tympanum to posterior corner of the eye);
HND hand length (from distal end of radioulna to tip of distal phalanx III);
RAD radioulna length (from the flexed elbow to the base of the outer palmar tubercle);
FTL foot length (from distal end of tibia to tip of distal phalanx IV);
TIB tibial length (from the outer surface of the flexed knee to the heel);
F3W width of digital disc on finger III;
T4W width of digital disc on toe IV.
Sex was determined by observation of secondary sexual characters, i.e. the presence of nuptial spines in males, following
All specimens were fixed in 10% buffered formalin and later transferred to 70% ethanol, and deposited in The Museum of Biology, Sun Yat-sen University (SYS) and Chengdu Institute of Biology, the Chinese Academy of Sciences (CIB), P.R. China.
The phylogenetic trees strongly supported that the Amolops ricketti group was a monophyletic species group, which can be further divided into six well-supported clades with marked divergences (Fig.
ID | Species | I | II | III | IV | V | VI | VII | IIX | IX | X | XI |
---|---|---|---|---|---|---|---|---|---|---|---|---|
I | Amolops sinensis sp. n. | 0.0–0.6 | – | – | – | – | – | – | – | – | – | – |
II | Amolops yatseni sp. n. | 3.5–4.2 | 0.0–0.8 | – | – | – | – | – | – | – | – | – |
III | Amolops albispinus | 3.8–4.1 | 4.1–4.4 | 0.0–0.0 | – | – | – | – | – | – | – | – |
IV | Amolops ricketti | 6.7–7.2 | 6.4–7.0 | 6.4–6.7 | 0.0–0.7 | – | – | – | – | – | – | – |
V | Amolops wuyiensis | 6.2–6.8 | 6.2–6.8 | 6.8–7.0 | 5.1–5.5 | 0.1–0.5 | – | – | – | – | – | – |
VI | Amolops yunkaiensis | 6.4–6.9 | 6.9–7.7 | 6.7–7.0 | 7.9–8.2 | 8.0–8.3 | 0.0–0.2 | – | – | – | – | – |
VII | Amolops daiyunensis | 12.5–12.9 | 12.6–12.7 | 12.4–12.4 | 12.3–12.8 | 12.6–12.8 | 12.4–12.6 | 0.0–0.0 | – | – | – | – |
IIX | Amolops hongkongensis | 12.5–12.9 | 12.2–12.4 | 12.2–12.3 | 11.8–12.3 | 12.1–12.4 | 12.4–12.7 | 5.5–5.5 | 0.0–0.1 | – | – | – |
IX | Amolops hainanensis | 15.1–15.5 | 15.4–15.8 | 14.6–14.7 | 14.9–15.2 | 14.5–14.8 | 15.6–16.0 | 14.8–14.9 | 15.7–15.8 | 0.0–0.8 | – | – |
X | Amolops torrentis | 15.3–15.7 | 15.5–15.8 | 15.5–15.5 | 15.1–15.5 | 15.3–15.5 | 16.3–16.7 | 14.8–15.0 | 14.6–14.7 | 10.9–11.0 | 0.1–0.4 | – |
XI | Amolops chunganensis | 15.8–16.2 | 15.6–16.0 | 15.8–15.8 | 14.8–15.2 | 14.8–15.0 | 15.3–15.5 | 15.3–15.3 | 15.8–15.9 | 17.7–17.9 | 17.3–17.5 | 0.0 |
Morphologically, the specimens clustered into clade V from central Guangdong, northeastern Guangxi and southwestern Hunan can be distinguished from all known Amolops species by having unique longitudinal glandular folds on the skin of the shoulders and other characters (see diagnosis below). Therefore this clade represents a distinct evolutionary lineage, and is described below as a new species, Amolops sinensis sp. n. The specimens grouped in clade VI from the coastal hills of west Guangdong differ from all known Amolops species by having unique dense tiny round translucent, or white, spines on the dorsal skin of the body, dorsal and dorsolateral skin of the limbs and other characters (see diagnosis below). Therefore, this clade represents a distinct evolutionary lineage, and is described below as a new species Amolops yatseni sp. n.
Amolops ricketti (Boulenger, 1899):
SYS a007107 (Fig.
Ten adult specimens. Male SYS a007105, 7106 and 7108, and female SYS a007109, collected at the same time from the same stream as holotype. Female SYS a004165/CIB 110004, collected by Zhi-Tong Lyu (ZTL) and YQL on 26 July 2015 from Hengshitang, Shimentai Nature Reserve, Guangdong. Male SYS a005710 and female SYS a005712, collected by ZTL and Jian Wang (JW) on 8 April 2017 from Mt. Nankun, Guangdong. Male SYS a005089, collected by JW on 18 July 2016 from Mt. Dupangling, Guangxi. Female SYS a007268, collected by ZTL and Yu-Long Li on 21 June 2018 from Mt. Yangming, Hunan. Female SYS a004257, collected by ZTL and JW on 19 August 2015 from Mt. Hengshan, Hunan.
The new species was assigned to genus Amolops and further to the A. ricketti group morphologically based on the absence of dorsolateral folds, the presence of a circummarginal groove on the disk of the first finger, the absence of tarsal glands, and the presence of nuptial pads with conical nuptial spines on the first finger in males.
Amolops sinensis sp. n. is distinguished from its congeners by a combination of the following morphological characteristics: (1) body stout and robust, SVL 40.2–46.5 (43.1±2.2, n=6) mm in adult males, 47.7–52.7 (50.5±2.0, n=5) mm in adult females; (2) dorsal body olive-brown to dark brown, with irregular light strip-shaped patches or not; (3) ventral surface creamy white or beige, with dark gray patches; (4) dorsal skin of body very rough, granular and scattered with conical tubercles and raised large warts in males; (5) vomerine teeth strong, tongue cordiform, deeply notched posteriorly; (6) absence of dorsolateral folds; (7) a longitudinal glandular fold on skin of shoulder on each side; (8) supernumerary tubercles below base of fingers III and IV indistinct; (9) heels overlapping; (10) absence of outer metatarsal tubercles and tarsal glands; (11) absence of vocal sacs; (12) nuptial pad on first finger prominent with beige spines in breeding males; and (13) white conical spines present on skin of temporal region (including tympanum in several individuals) and loreal region in breeding males.
Body stout, SVL 43.3 mm. Head width slightly smaller than head length (HDW/HDL = 1.04); snout short (SNT/HDL = 0.45) and rounded in profile, projecting beyond lower jaw; nostril closer to tip of snout than eye; loreal region concave; top of head flat; eye large (ED/HDL = 0.32) and convex; canthus rostralis distinct; pineal body distinct; tympanum small, edge distinct; tympanum-eye distance smaller than tympanum, TED/TD 0.90; supratympanic fold distinct, start from back of eye and extending to shoulder, a well-developed gland on end of supratympanic fold; choanae moderate; vomerine teeth present; tongue cordiform, deeply notched posteriorly.
Forelimbs moderately robust; hands moderately long (HND/SVL = 0.33); relative finger lengths I = II < IV < III; finger tips dilated to wide oval disks with circummarginal grooves, relative width of finger disks I < II < III = IV; subarticular tubercles prominent, rounded; supernumerary tubercles below base of fingers III and IV indistinct, below base of fingers I and II absent; inner metacarpal tubercle small, outer metacarpal tubercle prominent and slightly separated; absence of webbing and presence of weak lateral fringes on fingers.
Hindlimbs long and robust (TIB/SVL = 0.60); tibio-tarsal articulation reaching tip of snout when hindlimb stretched alongside of body; relative toe lengths I < II < III = V < IV; tips of all toes expanded to well-developed oval discs with circummarginal grooves; subarticular tubercles oval and distinct; inner metatarsal tubercle prominent, elongated; outer metatarsal tubercles absent; toes fully webbed; lateral fringes of toes I and V developed; tarsal glands absent; heels overlapping when hindlimbs flexed at right angles to axis of body.
Dorsal skin of body rough, granular and scattered with raised tubercles and warts; underdeveloped conical spines on skin of loreal region and temporal region except tympanum; flanks very rough and granular with raised warts; dorsal limbs rough with numerous tubercles; several longitudinal dermal ridges on dorsal surfaces of thigh, tibia and tarsus; dorsolateral fold absent; a longitudinal glandular fold on skin of shoulder; posterior part of upper lip swollen; rictal gland prominent and ellipsoidal, posterior to corner of mouth.
Ventral surface slightly wrinkled with granules; ventral surface of hand smooth, ventral surface of foot granular; large warts surrounding cloaca.
(in mm).SVL 43.3; HDL 15.4, HDW 16.0; SNT 6.9; IND 6.2; IOD 4.3; ED 4.9; TD 1.9; TED 1.7; HND 14.3; RAD 9.4; FTL 36.9; TIB 26.0; F3W 2.9; T4W 2.9.
Dorsal body olive-brown with irregular light yellow patches; longitudinal glands on occipital region light yellow; warts on flanks dark or grayish white; irregular dark patches on dorsal surface of forearms, distinct dark transverse bars on dorsal surface of lower arms and hindlimbs; dorsal discs of digits brown or white; nuptial pads and nuptial spines beige; posterior edge of upper lip and rictal gland light maize-yellow; throat and chest creamy white; belly beige; several dark gray mottling on surface of throat, chest and anterior part of abdomen; ventral surfaces of limbs gray pink grounding; ventral hands and feet dark grey; warts around cloaca yellowish white tubercles and olive-brown.
Dorsal surface dark brown with irregular gray patches, transverse bars on limbs more distinct; longitudinal glands on occipital region more distinct; webs on toes gray, mottling with olive-brown; ventral surface grayish white, mottling on surface of throat, chest and anterior part of abdomen become more distinct; ventral surface of limbs beige.
Measurements of type series specimens are given in Table
Measurements (in mm; minimum-maximum, mean±1SD) of the type series of Amolops sinensis sp. n. and A. yatseni sp. n.
A. sinensis sp. n. | A. yatseni sp. n. | |||
---|---|---|---|---|
Males (n=6) | Females (n=5) | Males (n=6) | Females (n=11) | |
SVL | 40.2–46.5 (43.1±2.2) | 47.7–52.7 (50.5±2.0) | 39.3–44.7 (42.5±2.1) | 42.1–48.9 (46.4±2.0) |
HDL | 14.6–16.7 (15.5±0.7) | 16.1–18.6 (17.1±1.2) | 14.2–16.8 (16.0±1.0) | 15.4–18.1 (16.7±0.9) |
HDW | 15.2–18.4 (16.1±1.2) | 16.2–19.5 (17.7±1.6) | 16.1–17.8 (16.9±0.7) | 16.1–19.0 (17.5±0.9) |
SNT | 6.0–7.5 (6.6±0.6) | 6.6–7.9 (7.1±0.5) | 5.6–7.4 (6.4±0.6) | 6.2–7.2 (6.7±0.4) |
IND | 5.3–6.9 (5.9±0.5) | 5.6–7.4 (6.6±0.8) | 5.7–6.4 (6.1±0.3) | 5.7–6.6 (6.3±0.3) |
IOD | 4.2–4.7 (4.4±0.2) | 4.5–5.4 (4.8±0.4) | 3.9–4.8 (4.3±0.3) | 4.1–4.5 (4.3±0.1) |
ED | 4.8–5.6 (5.1±0.4) | 5.4–6.4 (6.0±0.4) | 4.7–5.3 (5.1±0.3) | 4.9–6.2 (5.5±0.4) |
TD | 1.8–2.2 (2.1±0.2) | 1.8–2.2 (2.1±0.1) | 1.5–1.9 (1.7±0.1) | 1.8–2.3 (2.0±0.2) |
TED | 1.4–1.8 (1.7±0.1) | 1.5–2.0 (1.7±0.2) | 1.4–2.0 (1.8±0.2) | 1.8–2.9 (2.1±0.3) |
HND | 12.0–14.3 (13.0±0.8) | 12.5–14.7 (13.7±0.9) | 11.6–13.8 (12.7±0.9) | 11.8–14.0 (12.9±0.6) |
RAD | 8.4–10.6 (9.3±0.7) | 9.3–10.9 (10.0±0.7) | 7.8–10.9 (8.9±1.1) | 8.2–9.9 (8.8±0.5) |
FTL | 30.7–37.1 (34.3±2.4) | 34.8–39.5 (36.9±1.9) | 27.1–37.0 (32.4±3.3) | 29.1–35.6 (33.2±2.4) |
TIB | 22.4–26.9 (24.7±1.7) | 24.6–29.2 (26.9±1.8) | 20.6–27.0 (24.5±2.5) | 21.5–26.3 (24.6±1.5) |
F3W | 2.4–2.9 (2.7±0.2) | 2.5–3.2 (2.9±0.3) | 2.6–3.4 (2.8±0.3) | 2.2–3.1 (2.8±0.3) |
T4W | 2.3–2.9 (2.5±0.2) | 2.5–3.1 (2.8±0.3) | 2.1–3.2 (2.4±0.4) | 2.0–2.8 (2.4±0.3) |
HDL/SVL | 0.35–0.37 (0.36±0.01) | 0.33–0.36 (0.34±0.01) | 0.36–0.38 (0.38±0.01) | 0.35–0.37 (0.36±0.01) |
HDW/SVL | 0.35–0.40 (0.37±0.02) | 0.33–0.38 (0.35±0.02) | 0.39–0.42 (0.40±0.01) | 0.36–0.39 (0.38±0.01) |
HDW/HDL | 1.00–1.11 (1.04±0.04) | 1.01–1.05 (1.03±0.02) | 1.02–1.13 (1.06±0.04) | 1.02–1.09 (1.05±0.02) |
SNT/HDL | 0.39–0.45 (0.43±0.02) | 0.40–0.43 (0.42±0.01) | 0.38–0.44 (0.40±0.02) | 0.36–0.44 (0.40±0.02) |
SNT/SVL | 0.14–0.16 (0.15±0.01) | 0.13–0.15 (0.14±0.01) | 0.14–0.17 (0.15±0.01) | 0.13–0.15 (0.15±0.01) |
IND/HDW | 0.34–0.39 (0.37±0.02) | 0.35–0.39 (0.38±0.02) | 0.33–0.38 (0.36±0.02) | 0.34–0.39 (0.36±0.01) |
IOD/HDW | 0.26–0.30 (0.28±0.02) | 0.26–0.28 (0.27±0.01) | 0.23–0.28 (0.25±0.02) | 0.24–0.26 (0.25±0.01) |
ED/HDL | 0.31–0.37 (0.33±0.02) | 0.33–0.38 (0.35±0.02) | 0.31–0.33 (0.32±0.01) | 0.30–0.36 (0.33±0.02) |
ED/SVL | 0.11–0.13 (0.12±0.01) | 0.11–0.12 (0.12±0.00) | 0.12–0.12 (0.12±0.00) | 0.11–0.13 (0.12±0.01) |
TD/ED | 0.36–0.46 (0.40±0.04) | 0.29–0.39 (0.35±0.04) | 0.28–0.37 (0.34±0.03) | 0.32–0.40 (0.36±0.03) |
TED/TD | 0.78–0.90 (0.82±0.04) | 0.74–0.97 (0.83±0.08) | 0.81–1.18 (1.05±0.13) | 0.86–1.61 (1.08±0.21) |
HND/SVL | 0.28–0.33 (0.30±0.02) | 0.24–0.28 (0.27±0.02) | 0.28–0.32 (0.30±0.01) | 0.26–0.29 (0.28±0.01) |
RAD/SVL | 0.21–0.23 (0.22±0.01) | 0.19–0.21 (0.20±0.01) | 0.18–0.24 (0.21±0.02) | 0.18–0.21 (0.19±0.01) |
FTL/SVL | 0.76–0.85 (0.80±0.03) | 0.71–0.75 (0.73±0.02) | 0.69–0.83 (0.76±0.04) | 0.66–0.77 (0.72±0.03) |
TIB/SVL | 0.56–0.60 (0.57±0.01) | 0.50–0.55 (0.53±0.02) | 0.52–0.61 (0.57±0.03) | 0.48–0.56 (0.53±0.02) |
Amolops sinensis sp. n. possesses distinct sexual dimorphism: (1) larger body size in females with SVL 47.7–52.7 mm (vs. SVL 40.2–46.5 mm in males); (2) beige nuptial spines on beige nuptial pads in breeding males; (3) dense white conical spines present on skin of temporal region and loreal region in males during breeding season (vs. absent in females); and (4) females bearing light yellow oocytes.
The character of longitudinal glandular folds on skin of shoulders makes Amolops sinensis sp. n. unique when compared with all known congeners within the genus. The new species is further compared with the four recognized species of the A. ricketti species group below (Fig.
Comparisons of morphological characteristics among species in the Amolops ricketti group. A A. sinensis sp. n. B A. yatseni sp. n. C A. albispinus D A. ricketti E A. wuyiensis F A. yunkaiensis 1 dorsolateral view 2 close-up of the head 3 ventral view of the hand 4 ventral view of the foot.
Amolops sinensis sp. n. was previously reported as A. ricketti, but significantly differs from the topotype of A. ricketti by the presence of longitudinal glandular folds on the skin of the shoulders (vs. absent), large raised warts on the dorsal surface of body (vs. relatively smooth), the presence of white conical spines on skin of temporal region and loreal region in breeding males (vs. absent), and nuptial pad and nuptial spines beige (vs. white).
Amolops sinensis sp. n. is phylogenetically close to A. albispinus, but can be distinguished from the later by the presence of longitudinal glandular folds on the skin of the shoulders (vs. absent), the presence of supernumerary tubercles below the base of fingers III and IV (vs. absent), outer metacarpal tubercle slightly separated (vs. completely divided into two tubercles), pineal body distinct (vs. indistinct), ventral surface smooth (vs. presence of tiny, transparent and dispersive conical spines on surface of chest in males), and nuptial spines beige (vs. white).
Amolops sinensis sp. n. can be easily distinguished from A. wuyiensis by the presence of longitudinal glandular folds on skin of shoulders (vs. absent), vomerine teeth present (vs. absent), lacking vocal sacs (vs. present), and nuptial spines beige (vs. black).
Amolops sinensis sp. n. further differs from A. yunkaiensis by the presence of longitudinal glandular folds on the skin of the shoulders (vs. absent), larger body size, SVL 40.2–46.5 mm in adult males and 47.7–52.7 mm in adult females (vs. SVL 31.8–34.1 mm in males and 35.2–39.0 mm in females), vomerine teeth present (vs. absent), lacking vocal sacs (vs. present), and ventral surface smooth (vs. presence of tiny transparent spines on surface of chest).
The specific name “sinensis” refers to “Chinese”, for this new species takes a wide distribution in southern China. We suggest its English common name “Chinese Torrent Frog” and Chinese name “Zhong Hua Tuan Wa (中华湍蛙)”.
Currently, the Chinese Torrent Frog is recognized from the Shimentai Nature Reserve and Mt. Nankun in Guangdong, Mt. Dupangling in Guangxi, and Mt. Yangming and Mt. Hengshan in Hunan, which indicates the potential distribution area of Amolops sinensis sp. n. is from central Guangdong, to northeastern Guangxi and southwestern Hunan.
Amolops sinensis sp. n. inhabits rocky, fast-flowing streams (ca 500–1300 m a.s.l.) surrounded by moist subtropical secondary evergreen broadleaved forests. All individuals were observed from April to August. Males bear nuptial spines from April to July; females bear mature light yellow oocytes from April to August. Nevertheless, much of the ecology and behavior of this species remains unknown.
Amolops ricketti (Boulenger, 1899):
SYS a006807 (Fig.
Sixteen adult specimens. Male SYS a006806, and female SYS a006811/CIB 110005 and SYS a006808–6810, collected at the same time from the same stream as holotype. Male SYS a003634 and 3638, and female SYS a003633, collected by ZTL and JW on 22 April 2015 from Shangchuan Island, Guangdong. Female SYS a006819, collected by JW and HHC on 28 March 2018 from Mt. Gudou, Guangdong. Female SYS a003978 and 3981, collected by ZTL and Chao Huang on 14 May 2015 from Ehuangzhang Nature Reserve, Guangdong. Male SYS a004643 and 4676, and female SYS a004640, 4642 and 4994, collected by ZTL and JW on 14–15 April 2016 from Yunkaishan Nature Reserve, Guangdong.
Juvenile SYS a006857, collected by Yuan-Peng Cen on 26 March 2018 from Daliao, Zhongshan City, Guangdong.
The new species was assigned to genus Amolops and further to the A. ricketti group morphologically based on the absence of dorsolateral folds, the presence of a circummarginal groove on the disk of the first finger, the absence of tarsal glands, and the presence of nuptial pads with conical nuptial spines on the first finger in males.
Amolops yatseni sp. n. is distinguished from its congeners by a combination of the following morphological characteristics: (1) body stout and robust, SVL 39.3–44.7 (42.5±2.1, n=6) mm in adult males, 42.1–48.9 (46.4±2.0, n=11) mm in adult females; (2) dorsal body olive-brown or light brown, with irregular light strip-shaped patches or not; (3) ventral surface creamy white, with nebulous dark gray patches or not; (4) dorsal skin of body very rough, granular and scattered with tubercles and raised large warts, lacking warts on central back of trunk in females; (5) dense tiny round translucent, or white, spines present on dorsal skin of body, dorsal and dorsolateral skin of limbs in males, denser in females; (6) vomerine teeth strong, tongue cordiform, deeply notched posteriorly; (7) absence of the dorsolateral folds; (8) supernumerary tubercles below the base of fingers II, III and IV distinct and prominent; (9) heels just meeting; (10) absence of outer metatarsal tubercles and tarsal glands; (11) absence of vocal sacs; (12) nuptial pad on the first finger prominent with developed white conical spines in breeding males, tip of nuptial spines brown; and (13) dense white conical spines present on the skin of the temporal region (including the tympanum in several individuals), loreal region, snout, lips and chin in males during breeding season, and such spines less developed and rounded only on skin of temporal region except the tympanum and lower lips in females.
Body stout, SVL 41.0 mm. Head width slightly smaller than head length (HDW/HDL = 1.05); snout short (SNT/HDL = 0.38) and rounded in profile, projecting beyond lower jaw; nostril closer to tip of snout than eye; loreal region concave; top of head flat; eye large (ED/HDL = 0.31) and convex; canthus rostralis distinct; pineal body distinct; tympanum small, edge faintly distinct, upper margin of tympanum in contact with supratympanic fold; tympanum-eye distance larger than tympanum, TED/TD 1.08; supratympanic fold distinct, start from back of eye and extending to shoulder; choanae moderate; vomerine teeth present; tongue cordiform, deeply notched posteriorly.
Forelimbs moderately robust; hands moderately long (HND/SVL = 0.31); relative finger lengths I < II < IV < III; finger tips dilated to wide oval disks with circummarginal grooves, relative width of finger disks I < II < III = IV; subarticular tubercles prominent, rounded; supernumerary tubercles below the base of fingers II, III and IV distinct and prominent, below base of fingers I absent; inner metacarpal tubercle elongated and prominent, outer metacarpal tubercle prominent and slightly separated; absence of webbing and presence of weak lateral fringes on fingers.
Hindlimbs long and robust (TIB/SVL = 0.55) ; tibio-tarsal articulation reaching tip of snout when hindlimb stretched alongside of body; relative toe lengths I < II < III = V < IV; tips of all toes expanded to well-developed oval discs with circummarginal grooves; subarticular tubercles oval and distinct; inner metatarsal tubercle prominent, elongated; outer metatarsal tubercles absent; toes fully webbed; lateral fringes of toes I and V developed; tarsal glands absent; heels just meeting when hindlimbs flexed at right angles to axis of body.
Dorsal skin of body very rough, granular and scattered with raised large warts; dense rounded spines present on dorsal body, dorsal limbs, and many well developed and denser ones on sacral region; dense conical spines present on skin of temporal region except tympanum, loreal region, snout, lips and chin, conical spines on skin of lower lips much smaller; flanks very rough and granular with raised warts; dorsal limbs rough with numerous tubercles; several longitudinal dermal ridges on dorsal surfaces of thigh, tibia and tarsus; dorsolateral fold absent; posterior part of upper lip swollen; rictal gland prominent and ellipsoidal, posterior to corner of mouth.
Ventral surface slightly wrinkled with round spines on chest; ventral surface of hand and foot granular; large warts surrounding the vent.
(in mm).SVL 41.0; HDL 15.3, HDW 16.1; SNT 5.8; IND 5.7; IOD 4.0; ED 4.8; TD 1.7; TED 1.8; HND 12.5; RAD 9.0; FTL 31.2; TIB 22.5; F3W 2.7; T4W 2.1.
Dorsal body dark green; faint dark transverse bars on dorsal surface of limbs; dorsal discs of digits yellowish brown; posterior edge of upper lip and rictal gland light maize-yellow; all round spines and conical spines on skin grayish white; throat, chest, and belly creamy white; several dark gray nebulous mottling on surface of throat, chest and anterior part of abdomen; ventral surfaces of limbs gray pink grounding; creamy white blotches on ventral thighs; rear of thighs mottled with dark brown; ventral hands and feet dark grey; yellowish white tubercles and olive-brown warts around cloaca.
Dorsal surface dark brown, irregular light strip-shaped patches present, transverse bars indistinct; ventral surface grayish white, mottling on surface of throat, chest and anterior part of abdomen become more distinct; ventral surface of limbs light brown.
Measurements of type series specimens are given in Table
Amolops yatseni sp. n. possesses significantly-distinct sexual dimorphism: (1) larger body size in females with SVL 42.1–48.9 mm (vs. SVL 39.3–44.7 in males); (2) white nuptial spines with brown tips on white nuptial pads in breeding males; (3) rounded spines on dorsal skin denser and more distinct in females (Fig.
The dense tiny round translucent, or white, spines on dorsal skin of body, dorsal and dorsolateral skin of the limbs makes Amolops yatseni sp. n. unique when compared with all known congeners within the genus. Amolops yatseni sp. n. is further compared with Amolops sinensis sp. n. and other four recognized species within the A. ricketti species group below (Fig.
Amolops yatseni sp. n. is a sister taxon to A. sinensis sp. n. in our phylogenetic trees (Fig.
Amolops yatseni sp. n. was previously reported as A. ricketti, but significantly differs from the topotype A. ricketti by the presence of dense tiny round translucent or white spines on the dorsal skin of the body, dorsal and dorsolateral skin of the limbs (vs. absent), large raised warts on dorsal surface of body (vs. relatively smooth), supernumerary tubercles below the base of fingers II, III and IV distinct and prominent (vs. indistinct below the base of fingers III and IV, absent below the base of finger II), and the presence of white conical spines on skin of temporal region and loreal region in breeding males (vs. absent).
Amolops yatseni sp. n. differs from A. albispinus by the presence of dense tiny round translucent, or white, spines on the dorsal skin of the body, dorsal and dorsolateral skin of the limbs (vs. absent), pineal body distinct (vs. indistinct), the presence of conical spines on skin of the tympanum (vs. absent), the presence of rounded spines on skin of temporal region and lower-lips in females (vs. absent), the presence of supernumerary tubercles below the base of fingers II, III and IV (vs. absent), and ventral surface smooth (vs. presence of tiny, transparent and dispersive conical spines on surface of chest in males).
Amolops yatseni sp. n. can be easily distinguished from A. wuyiensis by the presence of dense tiny round translucent or white spines on dorsal skin of body, dorsal and dorsolateral skin of limbs (vs. absent), vomerine teeth present (vs. absent), lacking vocal sacs (vs. present), and conical nuptial spines white (vs. black).
Amolops yatseni sp. n. further differs from A. yunkaiensis by the presence of dense tiny round translucent or white spines on dorsal skin of body, dorsal and dorsolateral skin of limbs (vs. absent), a significantly larger body size, SVL 39.3–44.7 mm in adult males and 42.1–48.9 mm in adult females (vs. SVL 31.8–34.1 mm in males and 35.2–39.0 mm in females), vomerine teeth present (vs. absent), lacking vocal sacs (vs. present), and ventral surface smooth (vs. presence of tiny transparent spines on surface of chest).
The specific name “yatseni” refers to the founder of Sun Yat-sen University, Dr. Sun Yat-sen, who was born in Cuiheng Village, Zhongshan City, about five kilometers from the type locality, Mt. Wugui. We suggest its English common name “Yat-sen’s Torrent Frog” and Chinese name “Yi Xian Tuan Wa (逸仙湍蛙)”.
Currently, the Yat-sen’s Torrent Frog is known from the Zhongshan City, as well as from Mt. Gudou, Shangchuan Island, Ehuangzhang Nature Reserve, and Yunkaishan Nature Reserve. All these localities are situated in the coastal hills of west Guangdong, indicating the potential distribution area of Amolops yatseni sp. n. is from the west border of Pearl River Delta to the Yunkai Mountains. However, the five known localities of the new species are being threatened by hydropower station construction and tourism development respectively, and surveys are needed in western Guangdong to investigate the accurate population status and the distribution of this species.
Amolops yatseni sp. n. inhabits rocky, fast-flowing streams (ca 250–1000 m a.s.l.) surrounded by moist subtropical secondary evergreen broadleaved forests (Fig.
The species Amolops ricketti was originally described based on two specimens from Mt. Wuyi, Fujian (
Southwestern China has been considered as hotspot area with highest species diversity over time, while southeastern China, which suffers from more human activities, is considered as much less diverse, which may reflect the lack of biodiversity surveys over time. Recently, a number of new amphibian species were described from southeastern China (
We would like to thank Yong-You Zhao, Yao Li, Zhao-Chi Zeng, Hai-Long He, Yu-Long Li, Chao Huang, Si-Ying Mao, Kai-Chi Huang, Yuan-Peng Cen for their help in the field work. This work was supported by the Project of Comprehensive Scientific Survey of Luoxiao Mountains Region of Ministry of Science and Technology, P.R.China (No. 2013FY111500), the Biodiversity Conservation Program of Ministry of Ecology and Environment, P.R.China, the Specimen Platform of Ministry of Science and Technology, P.R.China, teaching specimens sub-platform (No.2005DKA21403-JK), and the Project of Animal Diversity Survey and Monitoring System Construction of Guangdong Shimentai National Nature Reserve.
Specimens examined
Amolops albispinus (n=10): China: Guangdong Province: Shenzhen City: Mt. Wutong (type locality): SYS a003270–3271, 3452–3454, 4511, 5643; Mt. Paiya: SYS a002436, 6898–6899.
Amolops ricketti (n=16): China: Fujian Province: Wuyishan City: Mt. Wuyi (type locality): SYS a004141–4143, 5922–5923; Taining County: Mt. Emeifeng: SYS a002492; Shanghang County: Gutian Town: SYS a003342, 4106; Jiangxi Province: Yanshan County: Wuyishan Nature Reserve: SYS a001605, 1342–1343; Guixi City: Yangjifeng Nature Reserve: SYS a000214, 0240, 0314, 0354–0355.
Amolops wuyiensis (n=20): China: Fujian Province: Wuyishan City: Mt. Wuyi (type locality): SYS a001716–1717, 4139–4140, 5936–5938; Shaowu City: Longhu Forest Station: SYS a004129, 4131; Ninghua County: Mt. Yashu: 5897–5900; Jiangxi Province: Yanshan County: Wuyishan Nature Reserve: SYS a001606; Guixi City: Yangjifeng Nature Reserve: SYS a000324, 0358–0360; Guangfeng County: Tongboshan Nature Reserve: SYS a001668; Zhejiang Province: Jingning County: Makeng Village: SYS a002723.
Amolops yunkaiensis (n=21): China: Guangdong Province: Yangchun City: Ehuangzhang Nature Reserve (type locality): SYS a000773–0774, 3979, 3982, 4696–4701, 4703–4707; Xinyi City: Yunkaishan Nature Reserve: SYS a004674, 4681–4684, 4992.