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Resurrection of genus Nidirana (Anura: Ranidae) and synonymizing N. caldwelli with N. adenopleura, with description of a new species from China

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The taxonomy of Babina sensu lato was controversial in the past decades. In this study, the phylogeny of genus Babina sensu lato was reconstructed based on genetic analysis, morphological comparison and advertisement call analysis. We found that Babina sensu stricto and previous subgenus Nidirana should be two distinct genera in the family Ranidae. N. caldwelli is confirmed to be a synonym of N. adenopleura because of the small genetic divergence and the lack of distinct morphological differences. A new species, Nidirana nankunensis sp. nov. is described based on a series of specimens collected from Mt. Nankun, Guangdong Province, China, which can be distinguished from other known congeners by having a behavior of nest construction, distinctive advertisement calls, significant divergence in the mitochondrial genes, and a combination of morphological characters. Currently, the genus Babina contains two species and the genus Nidirana contains eight species.
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Amphibia-Reptilia 38 (2017): 483-502
Resurrection of genus Nidirana (Anura: Ranidae) and synonymizing
N. caldwelli with N. adenopleura, with description of a new
species from China
Zhi-Tong Lyu, Zhao-Chi Zeng, Jian Wang, Chao-Yu Lin, Zu-Yao Liu, Ying-Yong Wang
Abstract. The taxonomy of Babina sensu lato was controversial in the past decades. In this study, the phylogeny of genus
Babina sensu lato was re-constructed based on genetic analysis, morphological comparison and advertisement call analysis.
We found that Babina sensu stricto and previous subgenus Nidirana should be two distinct genera in the family Ranidae. N.
caldwelli is confirmed to be a synonym of N. adenopleura because of the small genetic divergence and the lack of distinct
morphological differences. A new species, Nidirana nankunensis sp. nov. is described based on a series of specimens collected
from Mt. Nankun, Guangdong Province, China, which can be distinguished from other known congeners by having a behavior
of nest construction, distinctive advertisement calls, significant divergence in the mitochondrial genes, and a combination of
morphological characters. Currently, the genus Babina contains two species and the genus Nidirana contains eight species.
Keywords:Babina, bioacoustic, mitochondrial DNA, morphology, Nidirana nankunensis sp. nov., phylogeny.
Introduction
The ranid genus Babina was established and de-
scribed on the basis of Rana holsti Boulenger,
1892 (type species) and Rana subaspera Bar-
bour, 1908 by Thompson (1912). Dubois (1992)
established subgenus Nidirana under the genus
Rana Linnaeus, 1758, based on morpholo-
gical characters including presence of large
suprabrachial gland in males, absence of a
thumb-like structure on finger I and pres-
ence of well-developed dorsolateral folds. Rana
psaltes Kuramoto, 1985 (type species), Rana
adenopleura Boulenger, 1909, Rana caldwelli
Schmidt, 1925, Rana chapaensis Bourret, 1937,
Rana daunchina Chang, 1933 and Rana pleu-
raden Boulenger, 1904 were placed in this sub-
genus. Moreover, Babina was considered as a
subgenus of Rana, with the characters of pres-
ence of large suprabrachial gland in males,
presence of a thumb-like structure on finger
I and absence of well-developed dorsolateral
State Key Laboratory of Biocontrol/The Museum of Bi-
ology, School of Life Sciences, Sun Yat-sen University,
Guangzhou 510275, P.R. China
Corresponding author;
e-mail: wangyy@mail.sysu.edu.cn
folds (Dubois, 1992). Subsequently, Nidirana
was recognized as a separate genus by Chen
et al. (2005), based on a molecular phyloge-
netic tree of Southeast Asian ranids that only
included one Nidirana species – R. (N.)cha-
paensis. Frost et al. (2006) considered that sub-
genus Babina was a distinct genus, based on
a molecular analysis which included only two
species of subgenus Nidirana R. (N.)adeno-
pleura and R. (N.)chapaensis, and merged Ni-
dirana with genus Babina because Nidirana has
no characters that suggest it as a monophyletic
group from Babina. Chuaynkern et al. (2010)
disagreed with the arrangement of Frost et al.
(2006) and followed Dubois’s (1992) opinions,
considering that these two groups had distinct
morphotypes which indicated different adapta-
tive niches. Fei et al. (2010) provided a phyloge-
netic systematics of Ranidae, in which Babina
and Nidirana were regarded as two genera and
established a new genus Dianrana Fei, Ye, and
Jiang, 2010, but was not widely accepted (Frost,
2017). Kakehashi et al. (2013) provided molec-
ular studies including three species (B. hol-
sti,B. subaspera and B. okinavana), which in-
dicated the genus Babina sensu lato (Babina
sensu stricto and Nidirana) was monophyletic
©Koninklijke Brill NV, Leiden, 2017. DOI:10.1163/15685381-00003130
484 Z.-T. Lyu et al.
and was the sister taxon to genus Odorrana Fei,
Ye, and Huang, 1990.
At present, the monophyletic Babina s. l. is
recognized to contain 10 species occurring in
eastern and southeastern Asia, namely B. holsti
from Okinawa of central Ryukyu, B. subaspera
from Amami of central Ryukyu, B. okinavana
(Boettger, 1895) from Yaeyama of southern
Ryukyu and eastern Taiwan, B. adenopleura
from Taiwan, B. caldwelli from southeastern
mainland China, B. hainanensis (Fei, Ye, and
Jiang, 2007) from Hainan, B. daunchina from
western China, B. chapaensis and B. lini (Chou,
1999) from northeastern Indochina Peninsula,
and B. pleuraden from southwestern China
(Matsui, 2007; Fei et al., 2012; Frost, 2017).
Nevertheless, there is still no consensus on the
phylogenetic placement between Babina s. s.
and Nidirana, for lacking convictive molecular
supports.
In this study, we re-constructed the phy-
logeny of Babina s. l., using mitochondrial
data from all known congeners. Besides, during
our herpetological field surveys in Mt. Nankun
(MNK), south China, we have found a small-
sized frog firstly assigned to Babina s. l. by
possessing large suprabrachial gland in breed-
ing males. Morphological, genetic and acous-
tic analysis indicated that the frog is distinc-
tive from all known species of Babina s. l.
Therefore, we describe it as a new species be-
low.
Material and methods
Taxon sampling
A total of 39 samples from five known species (Babina
caldwelli,B. daunchina,B. hainanensis,B. lini and B.
pleuraden) and an unnamed species of genus Babina s. l.
from China were used for molecular analysis. All samples
were preserved in 95% ethanol and stored at 40°C. In
addition, five sequences from other five known Babina s.
l. species were obtained from GenBank and incorporated
into our dataset. Eleven Ranid and two Dicroglossid species
obtained from GenBank were used as out-groups. Detail
information of these materials is shown in table 1 and
fig. 1.
Extraction, PCR amplification, and sequencing
Genomic DNA were extracted from muscle tissue, us-
ing DNA extraction kit from Tiangen Biotech (Beijing)
Co., Ltd. Three mitochondrion genes namely 16S riboso-
mal RNA gene (16S), 12S ribosomal RNA gene (12S) and
cytochrome C oxidase 1 gene (CO1) were amplified.
Primers used for 16S were L3975 (5-CGCCTGTTTACCA
AAAACAT-3) and H4551 (5-CCGGTCTGAACTCAGAT
CACGT-3), for 12S were L33 (5-CTCAACTTACAMATG
CAAG-3), H56 (5-CGATTATAGAACAGGCTCCT-3),
L1091 (5-CAAACTGGGATTAGATACCCCACTAT-3)
and H1478 (5-TGACTGCAAGGTGACGGGCGGTGTG
T-3 ), and for CO1 were Chmf4 (5-TYTCWACWAAYCAY
AAAGAYATCGG-3)andChmr4(5
-ACYTCRGGRTGR
CCRAARAATCA-3). PCR amplifications were processed
with the cycling conditions that initial denaturing step at
95°C for 4 min, 35 cycles of denaturing at 94°C for 40 s,
annealing at 53°C for 40 s and extending at 72°C for 1 min,
and final extending step at 72°C for 10 min. PCR products
were purified with spin columns and then sequenced with
both forward and reverse primers using BigDye Termina-
tor Cycle Sequencing Kit per the guidelines, on an ABI
Prism 3730 automated DNA sequencer by Shanghai Ma-
jorbio Bio-pharm Technology Co., Ltd. All sequences were
deposited in GenBank (table 1).
Molecular phylogenetic analysis
DNA sequences were aligned in MEGA 6 (Tamura et al.,
2013) by the Clustal W algorithm with default parameters
(Thompson et al., 1997). Three gene segments, which are
1032 base pairs (bp) of 16S, 754 bp of 12S, and 562 bp
of CO1, were concatenated seriatim into a 2348-bp sin-
gle partition. The data was tested in jmodeltest v2.1.2 with
Akaike and Bayesian information criteria, resulting the best-
fitting nucleotide substitution model is GTR+I+G. Se-
quenced data was analyzed using Bayesian inference (BI)
in MrBayes 3.2.4 (Ronquist et al., 2012), and maximum
likelihood (ML) implemented in RaxmlGUI 1.3 (Silvestro
and Michalak, 2012). Three independent runs were con-
ducted in BI analysis, each of which was performed for
2 000 000 generations and sampled every 1000 generations
with the first 25% samples were discarded as burn-in. In ML
analysis, the bootstrap consensus tree inferred from 1000
replicates was used to represent the evolutionary history
of the taxa analyzed. Pairwise distances (p-distance) were
calculated in MEGA 6 using the uncorrected p-distance
model.
Bioacoustic analysis
Bioacoustic data of known Babina s. l. species were ob-
tained from literature (Matsui and Utsunomiya, 1983; Chou,
1999; Chuaynkern et al., 2010; Chuang et al., 2016) for
comparisons. Advertisement calls of the unnamed Babina
s. l. species from MNK and its three congeners (B. cald-
welli,B. daunchina and B. hainanensis) were recorded
in the field by SONY PCM-D50 digital sound recorder,
at the air temperature 18-20°C. The sound files in wave
Resurrection of Nidirana 485
Tab l e 1 . Localities, voucher information and GenBank numbers for all samples used in this study.
ID Species Localities (“*” indicates the type localities) Specimen no. 16S 12S CO1 Cites
1Nidirana nankunensis sp. nov. *China: Mt. Nankun, Guangdong SYS a003618 MF807828 MF807906 MF807867 this study
2Nidirana nankunensis sp. nov. *China: Mt. Nankun, Guangdong SYS a003619 MF807829 MF807907 MF807868 this study
3Nidirana nankunensis sp. nov. *China: Mt. Nankun, Guangdong SYS a004906 MF807835 MF807913 MF807874 this study
4Nidirana nankunensis sp. nov. *China: Mt. Nankun, Guangdong SYS a004907 MF807836 MF807914 MF807875 this study
5Nidirana nankunensis sp. nov. *China: Mt. Nankun, Guangdong SYS a004914 MF807837 MF807915 MF807876 this study
6Nidirana nankunensis sp. nov. *China: Mt. Nankun, Guangdong SYS a005717 MF807838 MF807916 MF807877 this study
7Nidirana nankunensis sp. nov. *China: Mt. Nankun, Guangdong SYS a005718 MF807839 MF807917 MF807878 this study
8Nidirana nankunensis sp. nov. *China: Mt. Nankun, Guangdong SYS a005719 MF807840 MF807918 MF807879 this study
9Nidirana adenopleura China: New Taipei City, Taiwan UMMZ 189963 DQ283117 DQ283117 / Frost et al. (2006)
10 Nidirana caldwelli China: Jingning County, Zhejiang SYS a002725 MF807827 MF807905 MF807866 this study
11 Nidirana caldwelli China: Jinggangshan Nature Reserve, Jiangxi SYS a004025 MF807830 MF807908 MF807869 this study
12 Nidirana caldwelli China: Jinggangshan Nature Reserve, Jiangxi SYS a004026 MF807831 MF807909 MF807870 this study
13 Nidirana caldwelli China: Jinggangshan Nature Reserve, Jiangxi SYS a004027 MF807832 MF807910 MF807871 this study
14 Nidirana caldwelli China: Jiangshi Nature Reserve, Fujian SYS a004112 MF807833 MF807911 MF807872 this study
15 Nidirana caldwelli China: Jiangshi Nature Reserve, Fujian SYS a004132 MF807834 MF807912 MF807873 this study
16 Nidirana caldwelli China: Mt. Yashu, Fujian SYS a005891 MF807841 MF807919 MF807880 this study
17 Nidirana caldwelli China: Mt. Yashu, Fujian SYS a005901 MF807842 MF807920 MF807881 this study
18 Nidirana caldwelli China: Mt. Yashu, Fujian SYS a005902 MF807843 MF807921 MF807882 this study
19 Nidirana caldwelli *China: Yanping District, Nanping City, Fujian SYS a005911 MF807844 MF807922 MF807883 this study
20 Nidirana caldwelli *China: Yanping District, Nanping City, Fujian SYS a005912 MF807845 MF807923 MF807884 this study
21 Nidirana caldwelli *China: Yanping District, Nanping City, Fujian SYS a005913 MF807846 MF807924 MF807885 this study
22 Nidirana caldwelli *China: Yanping District, Nanping City, Fujian SYS a005914 MF807847 MF807925 MF807886 this study
23 Nidirana caldwelli *China: Yanping District, Nanping City, Fujian SYS a005915 MF807848 MF807926 MF807887 this study
24 Nidirana caldwelli *China: Yanping District, Nanping City, Fujian SYS a005916 MF807849 MF807927 MF807888 this study
25 Nidirana caldwelli China: Mt. Wuyi, Fujian SYS a005939 MF807850 MF807928 MF807889 this study
26 Nidirana caldwelli China: Mt. Wuyi, Fujian SYS a005940 MF807851 MF807929 MF807890 this study
27 Nidirana caldwelli China: Mt. Wuyi, Fujian SYS a005941 MF807852 MF807930 MF807891 this study
28 Nidirana caldwelli China: Mt. Wuyi, Fujian SYS a005942 MF807853 MF807931 MF807892 this study
29 Nidirana caldwelli China: Mt. Wuyi, Fujian SYS a005943 MF807854 MF807932 MF807893 this study
486 Z.-T. Lyu et al.
Tab l e 1 . (Continued.)
ID Species Localities (“*” indicates the type localities) Specimen no. 16S 12S CO1 Cites
30 Nidirana chapaensis *Vietnam: Sapa, Lao Cai T2483/2000.4850 KR827711 / KR087625 Grosjean et al. (2015)
31 Nidirana daunchina *China: Mt. Emei, Sichuan SYS a004594 MF807822 MF807900 MF807861 this study
32 Nidirana daunchina *China: Mt. Emei, Sichuan SYS a004595 MF807823 MF807901 MF807862 this study
33 Nidirana daunchina China: Hejiang County, Sichuan SYS a004930 MF807824 MF807902 MF807863 this study
34 Nidirana daunchina China: Hejiang County, Sichuan SYS a004931 MF807825 MF807903 MF807864 this study
35 Nidirana daunchina China: Hejiang County, Sichuan SYS a004932 MF807826 MF807904 MF807865 this study
36 Nidirana hainanensis *China: Mt. Diaoluo, Hainan SYS a003741 MF807821 MF807899 MF807860 this study
37 Nidirana lini *China: Jiangcheng County, Yunnan SYS a003967 MF807818 MF807896 MF807857 this study
38 Nidirana lini *China: Jiangcheng County, Yunnan SYS a003968 MF807819 MF807897 MF807858 this study
39 Nidirana lini *China: Jiangcheng County, Yunnan SYS a003969 MF807820 MF807898 MF807859 this study
40 Nidirana okinavana *Japan: Iriomote Island, Okinawa not given NC022872 NC022872 NC022872 Kakehashi et al. (2013)
41 Nidirana pleuraden China: Mt. Gaoligong, Yunnan SYS a003775 MF807816 MF807894 MF807855 this study
42 Nidirana pleuraden China: Mt. Gaoligong, Yunnan SYS a003776 MF807817 MF807895 MF807856 this study
43 Babina holsti *Japan: Okinawa not given NC022870 NC022870 NC022870 Kakehashi et al. (2013)
44 Babina subaspera *Japan: Amami Island, Kagoshima not given NC022871 NC022871 NC022871 Kakehashi et al. (2013)
45 Amolops mantzorum China: Mt. Xiling, Sichuan not given NC024180 NC024180 NC024180 Shan et al. (2016)
46 Amolops wuyiensis not given not given NC025591 NC025591 NC025591 Huang et al. (2016)
47 Hylarana taipehensis not given not given KX022002 KX022002 KX022002 Chen et al. (2016)
48 Odorrana margaretae China HNNU1207003 NC024603 NC024603 NC024603 Chen et al. (2015)
49 Odorrana nasica Vietnam: Mt. Pomu, Ha Tinh AMNH A161169 DQ283345 DQ283345 / Frost et al. (2006)
50 Odorrana schmackeri not given not given KP732086 KP732086 KP732086 Bu et al. (unpub.)
51 Pelophylax nigromaculatus not given not given KT878718 KT878718 KT878718 Jiang et al. (2017)
52 Pelophylax plancyi not given not given NC009264 NC009264 NC009264 Nie et al. (unpub.)
53 Lithobates catesbeiana Japan: IABHU Lab. strain living individual AB761267 AB761267 AB761267 Kakehashi et al. (2013)
54 Rana japonica Japan: Hiroshima IABHU 17624 AB511305 AB511305 AB511305 Kurabayashi et al. (2010)
55 Sylvirana hekouensis not given not given KX021954 KX021954 KX021954 Chen et al. (2016)
56 Limnonectes fujianensis not given not given NC007440 NC007440 NC007440 Nie et al. (unpub.)
57 Quasipaa boulengeri China: Sichuan XM3632 KX645665 KX645665 KX645665 Yuan et al. (2016)
Resurrection of Nidirana 487
Figure 1. Collecting localities of the samples used in this study. Circles represent type localities. (1) Yanping District,
Nanping City, Fujian; (2) Jingning County, Zhejiang; (3) Mt. Wuyi, Fujian; (4) Jiangshi Nature Reserve, Fujian; (5) Mt.
Yashu, Fujian; (6) Jinggangshan Nature Reserve, Jiangxi; (7) Mt. Nankun, Guangdong; (8) Mt. Diaoluo, Hainan; (9) Hejiang
County, Sichuan; (10) Mt. Emei, Sichuan; (11) Jiangcheng County, Yunnan; (12) Mt. Gaoligong, Yunnan.
format were sampled at 48 kHz with 24 bits in depth.
Raven pro 1.5 (Cornell Lab of Ornithology, 2003-2014) was
used to output the spectrograms and to measure interre-
lated parameters with Fast Fourier transform of 512 points
and a 50% overlap (the calls of B. hainanensis were of
poor quality to measure so that only spectrogram was out-
putted).
The following measurements were taken for each call:
notes number, duration (the difference between begin time
and end time for a selected call/note), peak frequency (PF;
the frequency at which peak power occurs within the se-
lected call/note), inter-quartile range bandwidth (IQR-BW;
the difference between the first and third quartile frequen-
cies within the selected call/note), bandwidth 90% (BW-
90%; the difference between the 5% and 95% frequen-
cies of a selected call/note). The first notes of the calls of
Babina daunchina and the unnamed species from MNK
were unique, so we measured them in the same parameters
as the call; for other notes and the intervals between notes,
we only measured the duration parameter. As for the call
of B. caldwelli whose notes were without significant dif-
ferences, we only measured the duration for each note and
interval.
Morphometrics
Diagnostic characters of all known Babina s. l. species
were obtained from literature for comparisons (Fei et al.,
2009; Chuaynkern et al., 2010). Totally 43 specimens of
five known species (B. caldwelli,B. daunchina,B. haina-
nensis,B. lini and B. pleuraden) were examined (Ap-
pendix).
External measurements were made with digital calipers
(Neiko 01407A Stainless Steel 6-Inch Digital Caliper, USA)
to the nearest 0.1 mm, including snout-vent length (SVL)
from tip of snout to posterior margin of vent in adult frog;
head length (HDL) from tip of snout to the articulation of
the jaw; head width (HDW) at the commissure of the jaws;
snout length (SNT) from tip of snout to the anterior cor-
ner of the eye; internasal distance (IND); interorbital dis-
tance (IOD); eye diameter (ED) from the anterior corner of
the eye to posterior corner of the eye; tympanum horizontal
diameter (TD); tympanum-eye distance (TED) from ante-
rior edge of tympanum to posterior corner of the eye; hand
length (HND) from distal end of radioulna to tip of distal
phalanx III; radioulna length (RAD) from the flexed elbow
to the base of the outer palmar tubercle; foot length (FTL)
from distal end of tibia to tip of distal phalanx IV; and tibial
length (TIB) from the outer surface of the flexed knee to the
heel; body length (BL) from the tip of snout to the origin of
tail, and tail length (TL) from the origin to the tip of tail in
tadpole.
We determined sex by secondary sexual characters, i.e.
the presence of nuptial pads, vocal sacs or suprabrachial
glands in males. Webbing formula was written according
to Savage (1975) and tadpole stage was identified followed
Gosner (1960).
All specimens were fixed in 10% buffered formalin and
later transferred to 70% ethanol. All studied specimens and
all muscle tissues samples are 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.
488 Z.-T. Lyu et al.
Figure 2. Bayesian inference and maximum-likelihood phylogenies. Numbers above branches indicate Bayesian posterior
probabilities and numbers below branches are bootstrap support for maximum likelihood (1000 replicates) analysis.
Results
Phylogenetic analysis
The ML and BI analyses resulted in essentially
identical topologies which were integrated in
fig. 2. All major nodes were sufficiently sup-
ported with the Bayesian posterior probabilities
(BPP) >0.95 and the bootstrap supports for
maximum likelihood analysis (BS) >70. The
p-distances among all samples are given in the
online supplementary table S1.
All 10-known species of Babina s. l. and
the population from MNK grouped in a single
clade, which was the sister taxon of the genus
Odorrana within Ranidae. The clade was fur-
ther divided into two major, deeply divergent
(p-distance 7.0%-12.4%), and strongly sup-
ported monophyletic groups, which completely
correspond to two former subgenera Babina and
Nidirana (designated here as Babina group and
Nidirana group). The Babina group was com-
posed of two original members, B. holsti and B.
subaspera from central Ryukyu. The Nidirana
group consisted of the remaining eight known
species and an undescribed species from MNK,
which were further divided into four subclades
(designated here as A, B, C and D). The phylo-
genetic interrelationships of Babina s. l. inferred
as following: Babina group +Nidirana group
(subclade A +(subclade B +(subclade C +
subclade D))).
Within subclade D, Babina adenopleura from
Taiwan and B. caldwelli from east mainland
Resurrection of Nidirana 489
Figure 3. Advertisement call spectrograms. (A) Nidirana nankunensis sp. nov. from Mt. Nankun; (B) N. hainanensis from
Mt. Diaoluo; (C) N. daunchina from Hejiang County; (D) N. caldwelli from Yanping District.
China significantly clustered into a mono-
phyletic branch with strong supports and small
divergence (p-distance 0.3%-1.0%). Further-
more, the individuals from MNK grouped into
a monophyletic branch with strong supports
and small divergence (p-distance 0.0%-0.3%),
forming the sister taxon to ((B. caldwelli +
B. adenopleura)+B. okinavana). The signif-
icantly deep divergences between this branch
and all other species of Nidirana group (the
smallest interspecific p-distance is 4.8%-5.1%
between branch of MNK and B. okinavana)in-
dicated that this population represented a sepa-
rately evolving lineage.
Bioacoustic analysis
The call spectrograms were shown in fig. 3.
The literature and bioacoustic analysis in-
ferred the following call pattern differences
among species of Babina group and Nidirana
group.
Within Babina group, breeding males emitted
short advertisement calls (0.27 ±0.01 s for both
species) composed of a single note (Matsui and
Utsunomiya, 1983); whereas within Nidirana
group, all mating calls composed of several
notes (Chuaynkern et al., 2010).
490 Z.-T. Lyu et al.
Among species of Nidirana group, there are
three advertisement call patterns differing re-
markably from each other. The first pattern con-
tains a first note which is significantly different
in timbre characteristics, such as the calls from
MNK (MNK call; fig. 3A) and that of Babina
daunchina (fig. 3C): MNK call lasts about 2.30-
2.74 s (2.52 ±0.17 s, n=11) and contains 13-
15 (14.0 ±0.94, n=11) fast-repeated notes,
of which the first note continues for 108-135 ms
(122.9 ±8.4 ms, n=11) while others continue
for 38-56 ms (46.1 ±3.7 ms, n=50) and the
intervals between notes last 12-166 ms (138.7 ±
10.5 ms, n=50); the call of B. daunchina lasts
0.51-1.57 s (1.01 ±0.42 s, n=11) and con-
tains 2-5 (3.5 ±1.21, n=11) notes, of which
the first note continues for 162-197 ms (182.7 ±
12.4 ms, n=11) while others continue for 131-
150 ms (140.6 ±5.6 ms, n=23) and the inter-
vals between notes last 159-259 ms (193.6 ±
26.3 ms, n=23). B. hainanensis (fig. 3B)
has a unique call pattern that contains 2-4 fast-
repeated double-notes. B. caldwelli (fig. 3D) has
the third call pattern which contains several al-
most identical regular notes: 2-3 (2.27 ±0.45,
n=30) notes with the call duration of 0.53-
0.90 s (0.63 ±0.14 s, n=30), the notes
continuing for 159-252 ms (209.9 ±22.5 ms,
n=68) and the intervals between notes lasting
98-213 ms (122.4 ±23.1 ms, n=38). From the
literatures (Chou, 1999; Chuang et al., 2016),
the remaining congeners are recognized to have
similar call pattern to B. caldwelli: 3 notes in
B. chapaensis; 5-7 notes in B. lini; 17-25 fast-
repeated notes in B. okinavana; 4-7 notes in B.
pleuraden; 3-5 notes in B. adenopleura with the
call duration of 0.65-1.18 s and the notes con-
tinuing for 115-182 ms.
Morphological comparison
Measurements and body proportions of ex-
amined specimens of Babina caldwelli,B.
daunchina,B. hainanensis,B. lini,B. pleuraden
and that from MNK were given in table 2 and
the detail comparative data of species of Ni-
dirana group were shown in table 3. The sis-
ters Babina and Nidirana group shared synapo-
morphy of suprabrachial gland, and showed
distinct morphological differences from each
other in following characters: body size large
in Babina group (SVL 90-140 mm in adult B.
subaspera and 100-125 mm in adult B. holsti)
vs. body size small or moderate in Nidirana
group (maximum SVL 68.6 mm in adult female
B. lini); presence of a thumb-like structure on
finger I in Babina group vs. absent in Nidirana
group; dorsolateral folds not well developed in
Babina group vs. well developed in Nidirana
group.
Discussion
Babina caldwelli is synonymous with
B. adenopleura
The Babina caldwelli was originally described
as Rana caldwelli based on one holotype from
the locality probably near Yenping (=Yanping
District, Nanping City), Fujian, China, differing
from R. adenopleura by more projecting snouts,
rougher skin, and posteriorly broken up dorso-
lateral glandular folds (Schmidt, 1925). Subse-
quently, Pope (1931) placed it in the synonymy
of R. adenopleura and was followed by sev-
eral authors (Liu, 1950; Kuramoto, 1985; Chou,
1999; Fei et al., 2009; Fei et al., 2012). Dubois
(1992) and Chuaynkern et al. (2010) resurrected
population from Fujian as a valid species R.
caldwelli and considered R. adenopleura to be
restricted in Taiwan, further designating that R.
caldwelli differs by having shorter loreal region,
shorter forelimb, wider shank, narrower disc of
the fifth toe and spinules on the posterior half of
dorsum (vs. spinules scattered from tip of dorsal
snout to vent in R. adenopleura) (Chuaynkern
et al., 2010); besides, labial tooth row formula
in tadpoles of R. caldwelli was 1:0+0/1+1:2
(Pope, 1931) compared to 1:1+1/1+1:2 of R.
adenopleura (Chou and Lin, 1997).
Resurrection of Nidirana 491
Tab l e 2 . Measurements (in mm; minimum-maximum, mean ±1SD) and body proportions of the examined specimens of Babina sensu lato. F: females; M: males.
N. nankunensis sp. nov. N. caldwelli N. daunchian N. hainanensis N. lini N. pleuraden
M(n=10) F (n=2) M (n=18) F (n=3) M (n=4) F (n=1) M (n=1) M (n=2) F (n=1) M (n=3) F (n=1)
SVL 33.3-37.1
(35.6 ±1.3)
37.8-39.5
(38.6 ±1.2)
44.5-57.6
(51.4 ±3.8)
57.6-60.7
(58.9 ±1.6)
42.9-47.9
(45.8 ±2.5)
49.644.4 60.1-63.1
(61.6 ±2.2)
68.6 47.5-53.0
(50.2 ±2.8)
59.4
HDL 12.4-14.2
(13.2 ±0.7)
13.8-14.8
(14.3 ±0.7)
17.0-21.4
(19.1 ±1.3)
20.3-22.5
(21.4 ±1.1)
15.5-18.4
(16.8 ±1.6)
18.417.5 21.5-22.5
(22.0 ±0.8)
25.0 18.6-19.0
(18.9 ±0.2)
21.1
HDW 11.0-13.0
(11.9 ±0.7)
12.0-13.3
(12.6 ±0.9)
15.1-19.5
(17.6 ±1.2)
19.0-20.9
(19.9 ±1.0)
13.2-16.8
(14.8 ±1.8)
16.416.1 18.1-19.6
(18.8 ±1.1)
21.0 16.3-16.6
(16.5 ±0.1)
19.1
SNT 5.2-6.0
(5.6 ±0.3)
5.8-6.4
(6.1 ±0.4)
6.7-8.5
(7.8 ±0.6)
7.5-8.8
(8.3 ±0.7)
6.3-7.7
(7.0 ±0.6)
7.87.3 9.2-9.3
(9.2 ±0.0)
10.3 7.3-7.6
(7.5 ±0.2)
8.1
IND 4.2-4.8
(4.5 ±0.2)
4.6-4.6
(4.6 ±0.0)
4.8-6.7
(5.6 ±0.5)
5.9-6.4
(6.1 ±0.2)
4.8-6.1
(5.5 ±0.6)
6.15.6 6.2-6.4
(6.3 ±0.1)
7.2 4.9-5.3
(5.1 ±0.2)
6.6
IOD 3.3-4.1
(3.7 ±0.3)
3.9-4.0
(3.9 ±0.1)
4.0-5.4
(4.6 ±0.4)
4.8-5.1
(5.0 ±0.2)
4.1-4.2
(4.2 ±0.1)
4.44.8 5.4-5.6
(5.5 ±0.1)
5.6 3.7-4.4
(4.0 ±0.3)
4.6
ED 3.8-4.2
(4.0 ±0.1)
4.1-4.1
(4.1 ±0.0)
4.7-6.0
(5.2 ±0.3)
5.4-5.9
(5.7 ±0.3)
4.7-5.1
(4.9 ±0.2)
5.05.3 5.6-5.8
(5.7 ±0.1)
6.4 5.2-5.3
(5.3 ±0.0)
5.4
TD 3.2-3.4
(3.3 ±0.1)
3.3-3.4
(3.3 ±0.1)
3.4-5.2
(4.4 ±0.5)
4.5-5.2
(4.9 ±0.4)
3.8-4.4
(4.1 ±0.3)
4.44.5 4.9-5.9
(5.4 ±0.7)
4.6 4.1-4.3
(4.2 ±0.1)
4.9
TED 1.0-1.2
(1.2 ±0.1)
1.2-1.2
(1.2 ±0.0)
1.3-1.5
(1.4 ±0.1)
1.5-1.7
(1.6 ±0.1)
1.5-1.5
(1.5 ±0.0)
1.61.2 1.9-2.0
(2.0 ±0.1)
2.4 1.8-1.9
(1.8 ±0.0)
1.8
HND 8.9-10.2
(9.5 ±0.4)
9.6-10.2
(9.9 ±0.4)
11.6-15.0
(13.6 ±1.0)
14.0-17.2
(15.2 ±1.8)
10.6-13.2
(12.0 ±1.1)
12.411.9 14.1-16.1
(15.1 ±1.4)
17.3 12.0-13.9
(12.9 ±1.0)
14.6
RAD 6.0-6.4
(6.2 ±0.1)
6.4-6.6
(6.5 ±0.1)
18.0-25.0
(21.9 ±1.7)
20.0-26.2
(23.6 ±3.2)
17.1-21.9
(19.7 ±2.0)
21.320.1 25.3-25.6
(25.5 ±0.2)
29.7 19.3-21.8
(20.2 ±1.4)
22.9
FTL 25.6-28.3
(27.3 ±0.8)
27.7-29.8
(28.8 ±1.5)
23.0-29.9
(27.3 ±1.8)
28.8-33.5
(30.8 ±2.4)
23.5-27.2
(24.8 ±1.7)
24.625.3 29.8-32.6
(31.2 ±1.9)
36.8 22.5-25.0
(23.8 ±1.3)
29.3
TIB 17.7-18.7
(18.4 ±0.3)
18.8-20.8
(19.8 ±1.4)
34.3-43.7
(39.6 ±2.6)
42.5-48.5
(44.7 ±3.3)
33.3-37.9
(35.1 ±2.1)
36.135.6 43.0-50.6
(46.8 ±5.4)
53.5 34.7-37.6
(36.0 ±1.5)
42.2
HDL/SVL 0.35-0.39
(0.37 ±0.01)
0.36-0.37
(0.37 ±0.01)
0.35-0.39
(0.37 ±0.01)
0.35-0.37
(0.36 ±0.01)
0.35-0.38
(0.37 ±0.02)
0.37 0.39 0.36-0.36
(0.36 ±0.00)
0.36 0.36-0.39
(0.38 ±0.02)
0.36
492 Z.-T. Lyu et al.
Tab l e 2 . (Continued.)
N. nankunensis sp. nov. N. caldwelli N. daunchian N. hainanensis N. lini N. pleuraden
M(n=10) F (n=2) M (n=18) F (n=3) M (n=4) F (n=1) M (n=1) M (n=2) F (n=1) M (n=3) F (n=1)
HDW/SVL 0.32-0.35
(0.33 ±0.01)
0.32-0.34
(0.33 ±0.01)
0.32-0.37
(0.34 ±0.01)
0.33-0.34
(0.34 ±0.01)
0.30-0.35
(0.32 ±0.02)
0.33 0.36 0.30-0.31
(0.31 ±0.01)
0.31 0.31-0.35
(0.33 ±0.02)
0.32
HDW/HDL 0.87-0.92
(0.90 ±0.02)
0.87-0.90
(0.89 ±0.02)
0.87-0.95
(0.92 ±0.02)
0.92-0.93
(0.93 ±0.01)
0.85-0.92
(0.88 ±0.03)
0.89 0.92 0.84-0.87
(0.86 ±0.02)
0.84 0.86-0.89
(0.87 ±0.02)
0.91
SNT/HDL 0.41-0.45
(0.43 ±0.01)
0.42-0.43
(0.43 ±0.01)
0.36-0.43
(0.41 ±0.02)
0.37-0.40
(0.39 ±0.02)
0.40-0.43
(0.42 ±0.02)
0.42 0.42 0.41-0.43
(0.42 ±0.01)
0.41 0.38-0.41
(0.40 ±0.02)
0.38
SNT/SVL 0.15-0.16
(0.16 ±0.00)
0.15-0.16
(0.16 ±0.01)
0.14-0.16
(0.15 ±0.01)
0.13-0.15
(0.14 ±0.01)
0.14-0.16
(0.16 ±0.01)
0.16 0.16 0.15-0.15
(0.15 ±0.00)
0.15 0.14-0.16
(0.15 ±0.01)
0.14
IND/HDW 0.37-0.40
(0.38 ±0.01)
0.35-0.38
(0.37 ±0.02)
0.28-0.37
(0.32 ±0.02)
0.30-0.31
(0.30 ±0.01)
0.35-0.38
(0.37 ±0.02)
0.37 0.35 0.33-0.34
(0.34 ±0.01)
0.34 0.29-0.33
(0.31 ±0.02)
0.35
IOD/HDW 0.29-0.33
(0.31 ±0.01)
0.30-0.32
(0.31 ±0.01)
0.24-0.30
(0.26 ±0.02)
0.24-0.27
(0.25 ±0.02)
0.25-0.32
(0.28 ±0.03)
0.27 0.30 0.27-0.31
(0.29 ±0.03)
0.27 0.22-0.27
(0.24 ±0.03)
0.24
ED/HDL 0.27-0.32
(0.30 ±0.02)
0.28-0.29
(0.29 ±0.01)
0.24-0.31
(0.27 ±0.02)
0.26-0.27
(0.27 ±0.01)
0.28-0.32
(0.30 ±0.02)
0.27 0.30 0.26-0.26
(0.26 ±0.00)
0.26 0.27-0.28
(0.28 ±0.01)
0.26
ED/SVL 0.11-0.12
(0.11 ±0.00)
0.10-0.11
(0.11 ±0.01)
0.09-0.11
(0.10 ±0.01)
0.09-0.10
(0.10 ±0.01)
0.10-0.11
(0.11 ±0.01)
0.10 0.12 0.09-0.09
(0.09 ±0.00)
0.09 0.10-0.11
(0.10 ±0.01)
0.09
TD/ED 0.80-0.85
(0.83 ±0.02)
0.81-0.83
(0.82 ±0.01)
0.67-0.99
(0.85 ±0.09)
0.76-0.96
(0.86 ±0.10)
0.78-0.86
(0.84 ±0.04)
0.88 0.85 0.86-1.02
(0.94 ±0.11)
0.72 0.78-0.81
(0.80 ±0.02)
0.91
TED/TD 0.32-0.38
(0.35 ±0.02)
0.35-0.37
(0.36 ±0.01)
0.28-0.38
(0.33 ±0.03)
0.29-0.34
(0.32 ±0.03)
0.34-0.40
(0.37 ±0.03)
0.36 0.27 0.34-0.39
(0.37 ±0.04)
0.52 0.42-0.46
(0.44 ±0.02)
0.37
HND/SVL 0.24-0.28
(0.26 ±0.01)
0.25-0.26
(0.26 ±0.01)
0.24-0.29
(0.26 ±0.01)
0.24-0.28
(0.25 ±0.02)
0.25-0.28
(0.27 ±0.01)
0.25 0.27 0.23-0.25
(0.24 ±0.01)
0.25 0.25-0.26
(0.26 ±0.01)
0.25
RAD/SVL 0.17-0.18
(0.18 ±0.01)
0.17-0.17
(0.17 ±0.00)
0.39-0.46
(0.43 ±0.02)
0.35-0.43
(0.40 ±0.04)
0.40-0.46
(0.43 ±0.03)
0.43 0.45 0.41-0.42
(0.42 ±0.01)
0.43 0.39-0.41
(0.40 ±0.01)
0.39
FTL/SVL 0.72-0.80
(0.77 ±0.02)
0.73-0.75
(0.74 ±0.01)
0.47-0.58
(0.53 ±0.03)
0.50-0.55
(0.52 ±0.03)
0.52-0.57
(0.54 ±0.02)
0.50 0.57 0.50-0.52
(0.51 ±0.01)
0.54 0.45-0.50
(0.47 ±0.03)
0.49
TIB/SVL 0.50-0.54
(0.52 ±0.01)
0.50-0.53
(0.52 ±0.02)
0.70-0.87
(0.77 ±0.05)
0.74-0.80
(0.76 ±0.03)
0.74-0.79
(0.77 ±0.02)
0.73 0.80 0.72-0.80
(0.76 ±0.06)
0.78 0.71-0.73
(0.72 ±0.01)
0.71
Resurrection of Nidirana 493
Tab l e 3 . Diagnostic characters separating Nidirana nankunensis sp. nov. from its congeners.
Characteristics N. nankunensis N. adenopleura N. chapaensis N. daunchina N. hainanensis N. lini N. okinavana N. pleuraden
SVL of male 33.3-37.1 43.1-57.6 35.5-42.5 40.6-51.0 32.8-44.4 44.1-63.1 35.5-42.8 45.4-58.7
SVL of female 37.8-39.5 47.6-60.7 41.0-51.8 44.0-53.0 ? 57.7-68.6 44.6-48.8 45.5-62.5
Body habitus stocky elongated stocky stocky stocky elongated stocky elongated
Fingers tips dilated dilated dilated dilated dilated dilated dilated not dilated
Lateroventral
groove on fingers
present or absent present or absent present or absent absent or rarely
present
present present or absent present or absent absent
Relative length of
fingers
II <I<IV <III II <I<IV <III II <I=IV <III II <I<IV <III II <I<IV <III II <I<IV <III II <I<IV <III II <I<IV <III
Toes tips dilated dilated dilated dilated dilated dilated dilated not dilated
Lateroventral
groove on toes
present present present present present present present absent
Relative length of
toes
I<II <V<III <
IV
I<II <V<III <
IV
I<II <V<III <
IV
I<II <V<III <
IV
I<II <V<III <
IV
I<II <V<III <
IV
I<II <V<III <
IV
I<II <V<III <
IV
Tibio-tarsal
articulation
nostril snout tip or
eye-snout
nostril nostril nostril beyond snout eye center-near
nostril
eye-snout
Subgular vocal sacs present present present present present present absent present
Nuptial pad one one two one absent one poorly one one
Spinules on dorsal
skin
absent or few
above vent
entire or posterior
dorsal skin
absent or few
above vent
absent absent posterior dorsal
skin
absent posterior dorsal
skin
Nest construction present absent present present ? absent present absent
Calling 13-15 fast-repeated
notes
2-4 notes 3 notes 2-5 notes 2-4 fast-repeated
double-notes
5-7 notes 17-25 fast-repeated
notes
4-7 notes
Tadpole labial tooth
row formula
1:1+1/1+1:2 1:1+1/1+1:2 or
1:0+0/1+1:1
1:1+2/1+1:2 1:1+1/1+1:2 or
1:1+1/2+2:1
?1:1+1/1+1:2 1:1+1/1+1:2 1:1+1/1+1:2 or
1:1+1/2+2:1
Cites this study this study; Pope
(1931);
Chuaynkern et al.
(2010)
Chuaynkern et al.
(2010)
this study; Liu
(1950); Fei et al.
(2009)
this study; Fei et
al. (2009)
this study; Chou
(1999); Fei et al.
(2009)
Matsui and
Utsunomiya
(1983);
Chuaynkern et al.
(2010)
this study; Fei et
al. (2009)
494 Z.-T. Lyu et al.
Our examined specimens of Babina caldwelli
from Fujian, Zhejiang and Jiangxi did not show
significant differences from the revision from
Chuaynkern et al. (2010), in which the frogs
from continent and island were found several
differences in statistically morphometric data.
However, we found that the dorsolateral glan-
dular folds were not steadily broken up or con-
tinuous in Taiwanese B. adenopleura (Xiang et
al., 2009; Lin, personal communication), which
was not mentioned by the original description
(Boulenger, 1909). In addition, the labial tooth
row formula of tadpoles from Mt. Wuyi were
1:0+0/1+1:2 (SYS a005942) or 1:1+1/1+1:2
(SYS a005943), which were both identified as
B. caldwelli in the molecular trees. Therefore,
the characteristics separating B. caldwelli and B.
adenopleura were not diagnostic, though there
were statistically morphometric variations be-
tween continent and island populations. Further,
the molecular analysis suggested that the sam-
ples from both Taiwan and mainland gather to-
gether with strong supports (BPP =1.00, BS =
100) and small divergence (p-distance 0.3%-
1.0%). Hence, we regard the populations from
Taiwan and mainland China as the same species
and B. caldwelli is synonymous with B. adeno-
pleura.
Babina and Nidirana are two distinct genera
The Babina group, including B. holsti and B.
subaspera, occurs in central Ryukyu, sharing
the derived characters of large-sized body and
thumb-like structure on finger I which is im-
portant during mating and regarded as the sec-
ondary sexual characteristics (Iwai, 2012). The
Nidirana group is sister taxon to Babina with
strongly supported and deep divergences (p-
distance 7.0%-12.4%) in our molecular phy-
logenetic trees; morphologically, Nidirana sig-
nificantly differs from Babina by the absence
of the thumb-like structure, having relatively
small-sized body and well developed dorso-
lateral folds; bioacoustically, as a symbol for
reproductive isolation, Nidirana has obviously
different call pattern from Babina; geographi-
cally, basal lineages A, B and C of Nidirana
group occur from Sichuan, Yunnan and Hainan
of China and extend to Indochina peninsula,
showing that Nidirana and Babina have been
isolated and undergone different geographical
evolution processes.
The definition of a genus should fulfill the
following three criteria to be descriptively use-
ful: monophyletic, reasonably compact, and
ecologically, morphologically or biogeograph-
ically distinct (Gill et al., 2005). Based on our
data, Nidirana and Babina groups can be easily
and steadily distinguished by the morphological
and bioacoustic differences which are indicating
that these two groups have distinct adaptative
niches. Though the genetic divergence between
them is relatively small when compared with
other ranid genera, but the result exposed differ-
ent geographical evolution processes of the two
groups. Therefore, we consider the two groups
should be treated as two genera within family
Ranidae.
We here resurrect the genus Nidirana Dubois,
1992, and suggest its English common name
“Music Frogs” and Chinese name “Qin Wa
Shu ( )” (Fei et al., 2012), including seven
known species: Nidirana okinavana comb. nov.
(Ryukyu Music Frog), Nidirana adenopleura
(East China Music Frog), Nidirana hainanen-
sis (Hainan Music Frog), Nidirana chapaen-
sis (Chapa Music Frog), Nidirana daunchina
(Emei Music Frog), Nidirana lini (Lin’s Music
Frog) and Nidirana pleuraden (Yunnan Music
Frog).
The population from MNK is a new member of
genus Nidirana
The comprehensive evidences of molecular
analysis, morphological comparison and bioa-
coustics support the population from MNK to
be a distinct species of genus Nidirana and we
describe it as a new species Nidirana nankunen-
sis sp. nov. as below.
Resurrection of Nidirana 495
Taxonomic account
Nidirana nankunensis sp. nov.
Holotype. SYS a005719 (fig. 4), adult male,
collected by Zhi-Tong Lyu (ZTL) and Jian
Wang (JW) on 9 April 2017 from Mt. Nankun
(23°3812N, 113°5115E; 506 m a.s.l.), Long-
men County, Guangdong Province, China.
Paratypes. Eleven adult specimens from the
same locality as the holotype (500-600 m a.s.l.).
SYS a003615, 3617-3620, five adult males, col-
lected on 14-15 April 2015 by Run-Lin Li,
ZTL and JW; SYS a004019/CIB 106879, adult
male, collected on 10 June 2015 by Ying-Yong
Wang (YYW), ZTL and JW; SYS a004905-
4907, three adult males, collected on 5 June
2016 by ZTL and YYW; SYS a005717-5718,
two adult females, collected on 9 April 2017 by
ZTL and JW.
Other examined materials. SYS a004914, two
tadpoles, collected from the same locality as the
holotype by ZTL and YYW on 5 June 2016.
Etymology. The specific name “nankunensis
refers to the type locality of the new species, the
Mt. Nankun. We suggest its English common
name “Mt. Nankun Music Frog” and Chinese
name “Nan Kun Shan Qin Wa ( )”.
Diagnosis. Nidirana nankunensis sp.nov.is
distinguished from its congeners by following
combination of the morphological characteris-
tics: (1) body small and stocky, with SVL 33.3-
37.1 mm in adult males and 37.8-39.5 mm
in adult females; (2) disks of digits dilated,
pointed; (3) lateroventral grooves present on
every digit except finger I; (4) heels meet-
ing; (5) tibio-tarsal articulation reaching for-
ward the nostril; (6) mid-dorsal stripe present;
(7) spinules on dorsal skin absent or few
above vent; (8) a pair of subgular vocal sacs
present; (9) one single distinct prominent nup-
tial pad present on the first finger, nuptial
spinules invisible; (10) suprabrachial gland
present and large; (11) nest construction behav-
ior present; (12) tadpole labial tooth row for-
mula: 1:1+1/1+1:2; (13) calling: 13-15 fast-
repeated notes.
Comparisons. Nidirana nankunensis sp. nov.
differs from Nidirana congeners by following
characteristics: (1) small-sized body, SVL 37.8-
39.5 mm in adult females, 33.3-37.1 mm in
adult males vs. 57.7-68.6 mm in adult females
and 44.1-63.1 mm in adult males in N. lini
(this study; Chuaynkern et al., 2010); 47.6-
60.7 mm in adult females and 43.1-57.6 mm
in adult males in N. adenopleura (this study;
Chuaynkern et al., 2010); 45.5-62.5 mm in adult
females and 45.4-58.7 mm in adult males in
N. pleuraden (this study; Chuaynkern et al.,
2010); 44.0-53.0 mm in adult females and 40.6-
51.0 mm in adult males in N. daunchina (this
study; Chuaynkern et al., 2010); 44.6-48.8 mm
in adult females in N. okinavana (Chuaynkern
et al., 2010); 41.0-51.8 mm in adult females
in N. chapaensis (Chuaynkern et al., 2010);
(2) tibio-tarsal articulation reaching forward the
nostril vs. beyond the snout tip in N. lini;atthe
snout tip or eye-snout in N. adenopleura; eye-
snout in N. pleuraden; eye center-near nostril
in N. okinavana; (3) the presence of a single
nuptial pad well developed, raised vs. poorly
developed in N. okinavana; divided into two
parts in N. chapaensis; absent in N. hainanen-
sis; (4) having a behavior of nest construction
vs. absent in N. adenopleura,N. lini and N.
pleuraden; (5) the tadpole labial tooth row for-
mula 1:1+1/1+1:2 vs. 1:1+2/1+1:2 in N. cha-
paensis;1:1+1/1+1:2 or 1:0+0/1+1:1 in N.
adenopleura;1:1+1/1+1:2 or 1:1+1/2+2:1 in
N. pleuraden. Further, N. nankunensis sp. nov.
is different from N. pleuraden by its dilated dig-
its (vs. not dilated) and lateroventral grooves
present on digits expect finger I (vs. all absent);
from N. chapaensis by its finger IV longer than
finger I (vs. equal); from N. okinavana by hav-
ing a pair of subgular vocal sacs (vs. absent);
from N. daunchina by presence of lateroventral
grooves on fingers expect finger I (vs. absent
496 Z.-T. Lyu et al.
Figure 4. Morphological features of the adult male holotype SYS a005719 of Nidirana nankunensis sp.nov.inlife.
(A) dorsolateral view; (B) ventral view; (C) right hand; (D) nuptial pad; (E) right foot; (F) posterior part of dorsal surface.
or rarely present); from N. hainanensis by ab-
sence of lateroventral grooves on finger I (vs.
present).
Description of holotype. Adult male. Body
stocky, SVL 36.3 mm; head longer than wide
(HDL/HDW 1.10), flat above; snout rounded
Resurrection of Nidirana 497
in dorsal and lateral views, slightly protruding
beyond lower jaw, longer than horizontal dia-
meter of eye (SNT/ED 1.33); canthus rostralis
distinct, loreal region concave; nostril round,
directed laterally; a longitudinal mandibular
ridge extending from tip of snout through lower
edges of nostril, eye and tympanum to axial
region, forming a maxillary gland in poste-
rior corner of mouth; supratympanic fold ab-
sent; interorbital space flat, narrower than in-
ternarial distance (IND/IOD 1.17); pupil el-
liptical, horizontal; tympanum distinct, round,
TD/ED 0.81, and close to eye, TED/TD 0.32;
pineal ocellus present, in middle point between
anterior borders of eyelids; vomerine ridge
present, bearing numerous small teeth; tongue
large, cordiform, longer than wide, notched be-
hind.
Forelimbs moderately robust, lower arm 17%
of SVL and hand 28% of SVL; fingers thin, rel-
ative finger lengths II <I<IV <III; tip of
fingers weakly dilated, forming elongated and
pointed disks; lateroventral grooves on finger
II, III and IV, but absent on finger I; lateroven-
tral grooves not meeting at the tip of disks; fin-
gers free of webbing, with weak lateral fringes;
subarticular tubercles present, prominent and
rounded; palmar tubercles three, large, distinct
and elliptic.
Hindlimbs relatively robust, tibia 51% of
SVL and foot 78% of SVL; heels meeting
when hindlimbs flexed at right angles to axis of
body; tibio-tarsal articulation reaching forward
the nostril when hindlimb is stretched along the
side of the body; toes relatively long and thin,
relative lengths I <II <V<III <IV; tip of
toes weakly dilated, forming significantly elon-
gated and pointed disks; distinct lateroventral
grooves on toes; webbing moderate, webbing
formula: I 2-21
2II 12
3-3 III 21
3-31
2IV 31
2-2 V;
toes with lateral fringes, distinct dermal flap
running along lateral edge of 5th toe; subartic-
ular tubercles oval or rounded, prominent; in-
ner metatarsal tubercle elliptic, three times as
long as wide; outer metatarsal tubercle indis-
tinct, small and rounded; tarsal folds absent,
tarsal tubercle present.
Dorsal skin of head and body smooth with
tiny granules on dorsal head and body, exclud-
ing the snout; posterior part of back with sev-
eral tubercles, not bearing horny spinules; de-
veloped intermittent dorsolateral fold from pos-
terior margin of upper eyelid to above groin;
flanks smooth, a large and smooth suprabrachial
gland behind base of forelimb; two longitudinal
ridges on dorsal side of upper arm and slightly
extending on to lower arm; several longitudinal
dermal ridges with horny spinules on the dorsal
surfaces of thigh, tibia and tarsus. Ventral sur-
face of head, body and limbs smooth, large flat-
tened tubercles densely arranged on the rear of
thigh and around vent.
Measurement of holotype (in mm). SVL 36.3;
HDL 13.2, HDW 12.0; SNT 5.6; IND 4.7; IOD
4.0; ED 4.2; TD 3.4; TED 1.1; HND 10.2; RAD
6.0; FTL 28.3; TIB 18.6.
Color in life of holotype. Dorsal surface of
head and body light brown; light brown mid-
dorsal stripe edged with broad dark brown
stripes on two sides from pineal ocellus to vent;
several black spots on the top of tip of snout,
upper eyelids and posterior dorsum of body;
dorsolateral fold bicolor, red brown upper and
black lower part; upper flank light brown with
large black spots; lower flank yellowish white;
suprabrachial gland pale brown, feebly tinged
with pink. Dorsal forelimbs reddish brown, with
a black crossbar on the lower arm; one black
stripe in front of the base of forelimb; irregular
black marks on lower arm and dorsal hand; dor-
sal hindlimbs non-uniform reddish brown, three
black crossbars edged with light-colored on the
thigh, two on the tibia and three on the tarsus; ir-
regular black marks on dorsal toes. Dark brown
stripe from tip of snout through nostril to ante-
rior border of eye; tympanum and temporal re-
gion dark brown; pupil black, upper 1
3iris bright
brownish white and lower 2
3iris dark red; lips
yellowish white with dense tiny black specks;
maxillary gland white. Ventral surface creamy
498 Z.-T. Lyu et al.
white; faint dark stripes on the throat; rear thigh
pale yellow, tinged with pink; ventral hand pale
white with dense tiny black specks; ventral foot
mottled with grey-brown.
Color in preservative of holotype. Dorsal sur-
face faded lighter, but dark brown stripes on
two sides of the mid-dorsal stripe more distinct;
black spots on dorsum more distinct; limbs
Figure 5. Paratypes and tadpoles of Nidirana nankunensis sp. nov. (A) adult female paratype SYS a005717; (B) adult male
paratype SYS a004905; (C) dorsolateral view of the 36th stage tadpole; (D) dorsolateral view of the 29th stage tadpole;
(E) labial tooth row formula of the 29th stage tadpole.
Resurrection of Nidirana 499
faded light brown and the crossbars becoming
clearer; ventral surface faded pale.
Variation. Measurements of type series are
given in table 2. All specimens were similar
in morphology and color pattern. Body size of
two females (SYS a005717, 5718) are larger
than that of males (minimum 37.8 mm vs.
maximum 37.1 mm in males) and the skin
around vent are more rough in the females.
Mid-dorsal stripe extends to the tip of snout
in SYS a003615, 3617, 3618, 5717 (fig. 5A),
5718. White horny spinules present above the
vent in SYS a003618, 5717, 5718. Longitudinal
dermal ridges on limbs not well developed in
SYS a004905 (fig. 5B), 4906, 5717. Contorted
mid-dorsal stripe and dark brown upper flank in
SYS a004905.
Male secondary sexual characteristics. A pair
of subgular vocal sacs, a pair of slit-like open-
ings at posterior of jaw; a creamy white sin-
gle nuptial pad prominent on the dorsal sur-
face of first finger, nuptial spinules invisible;
suprabrachial gland present.
Tadpole. Body oval, flattened above; snout
rounded in dorsal aspect and profile; eyes lat-
eral; labial tooth row formula: 1:1+1/1+1:2
(fig. 5E); spiracle on left side of body, directed
dorsoposteriorly; vent tube long, dextral, at-
tached to ventral fin; tail depth slightly larger
than body depth; dorsal fin arising just before
Figure 6. (A) Habitat of Nidirana nankunensis sp. nov. in the type locality from Mt. Nankun; (B) The nest opening at the
bottom of the pond; (C) The nest opening on the bank; (D) The holotype SYS a005719 in the wild.
500 Z.-T. Lyu et al.
origin of tail, maximum depth near mid-length,
tapering gradually to narrow pointed tip; BL
16.3 mm and TL 36.1 mm in the 36th stage tad-
pole (fig. 5C); BL 7.1 mm and TL 14.5 mm in
the 29th stage tadpole (fig. 5D).
Vocalization. Advertisement calls of Nidirana
nankunensis sp. nov. were recorded on 9 April
2017 by ZTL from MNK at the air tempera-
ture 18°C. The call of N. nankunensis sp. nov.
has a duration of 2.30-2.74 s (2.52 ±0.17 s,
n=11) and consisted of 13-15 (14.0 ±0.94,
n=11) fast-repeated notes. The PF of calls
is 1406.2 Hz generally (91%). The IQR-BW of
calls is 281.2 Hz and the BW-90% is 1312.5 Hz.
The first note of a call is obviously different
from others. The first note lasts for 108-135 ms
(122.9 ±8.4 ms, n=11), with IQR-BW 375
Hz generally (82%) and BW-90% 1218.8 Hz
generally (91%). The PF of the first note is
equal to that of the call generally (73%). Other
notes of the call each have a duration of 38-
56 ms (46.1 ±3.7 ms, n=50) and the inter-
val between them lasts for 112-166 ms (138.7 ±
10.5 ms, n=50).
Distribution and ecology. Currently, Nidirana
nankunensis sp. nov. is known only from the
type locality, the Mt. Nankun in southern China.
This frog appeared to be very rare which is
found in only four unconnected ponds and un-
der the threats of tourism development and road
construction. The extent of occurrence is esti-
mated to be less than 5000 km2, and the area of
occupancy is estimated to be less than 500 km2.
It was only found in small ponds with sludge
bottom and covered by plants (fig. 6A). From
April to June, males call from the dusk to mid-
night, more active during rainfall. Females bear
pigmented oocytes with white animal pole and
brown vegetative pole brown in the oviduct on
April. The frog was observed to have the be-
havior of nest construction for oviposition. The
nest is a soil burrow, 60 mm of diameter and lo-
cated at the bottom of the pond bank. Half of the
burrow is filled with water. The burrow is con-
nected by two narrow tunnels in opposite direc-
tions to two openings: the small one is 25 mm
of diameter and open on the bank and exposed
(fig. 6C); another one is 40 mm of diameter and
open at the bottom of the pond for water diver-
sion (fig. 6B).
Acknowledgements. We thank Si-Min Lin from National
Taiwan Normal University for his help in providing images
and videos of Nidirana adenopleura from Taiwan. We thank
Yik-Hei Sung from Hong Kong Baptist University for his
help in polishing the article. This work was partially sup-
ported by Project of Comprehensive Scientific Survey of
Luoxiao Mountains Region (No. 2013FY111500) of Min-
istry of Science and Technology of P.R. China to Ying-Yong
Wang.
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Appendix. Specimens examined
Nidirana caldwelli (N. adenopleura) (29): Chi-
na: Fujian Province: Nanping City: Yanping
District (type locality): SYS a005911-5916;
Wuyishan City: Mt. Wuyi: SYS a005939-
5943; Shaowu City: Jiangshi Nature Re-
serve: SYS a004112, 4132; Ninghua County:
Mt. Yashu: SYS a005890-5891, 5901-5902;
Jiangxi Province: Guangfeng County: Tong-
boshan Nature Reserve: SYS a001663-1665,
1667, 1698; Guixi City: Yangjifeng Nature
502 Z.-T. Lyu et al.
Reserve: SYS a0000317, 0334; Jinggang-
shan City: Jinggangshan Nature Reserve:
SYS a004025-4027; Zhejiang Province:
Jingning County: Dongkeng Town: SYS
a002725-2726.
Nidirana daunchina (5): China: Sichuan Pro-
vince: Emeishan City: Mt. Emei (type local-
ity): SYS a004594-4595; Hejiang County:
Zihuai Town: SYS a004930-4932.
Nidirana hainanensis (1): China: Hainan Pro-
vince: Lingshui County: Mt. Diaoluo (type
locality): SYS a003741.
Nidirana lini (4): China: Yunnan Province:
Jiangcheng County: Hongjiang Town (type
locality): SYS a003967-3970.
Nidirana pleuraden (4): China: Yunnan Provin-
ce: Tengchong City: Mt. Gaoligong: SYS
a003775-3778.
... daunchina, N. adenopleura, N. hainanensis, N. okinavana and N. lini) with a wide allopatrically geographic distribution in east Asia 60,61 , although some new species (N. nankunensis, N. yaoica, N. guangdongensis, N. mangveni, N. xiangica and N. leishanensis) have been described in very recent years [62][63][64][65] . The Emei music frog (N. ...
... Among these species, the Emei music frog (N. daunchina) is a relative "recent" species in the genus according to the present results (Fig. 1a) and previously published phylogenies [62][63][64][65][66] . Previous phylogenetic trees have showed somewhat inconsistent topological patterns, likely resulting from the species members and/or models used in the analyses 62,63,65,67 . ...
... Thus, we created our own molecular phylogenetic tree using the first five confirmed species in the genus Nidirana 60,61 and calculated their genetic distances from each other. The structural profile of our strongly supported phylogenetic tree is topologically the same as three previous trees [62][63][64] and only slightly different from the other two 65,67 . Here we take the species relationships between the subject and other species in further analyses based on the following considerations. ...
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Nidirana guangxiensis sp. nov. , a new music frog species, is proposed, based on a series of specimens collected from Mt Daming, Guangxi, southern China. The new species is close to N. yeae , N. daunchina , N. yaoica , and N. chapaensis from southwestern and south-central China and northern Indochina, while the relationships among these species remain unresolved. Nidirana guangxiensis sp. nov. can be distinguished from all known congeners by the genetic divergences in the mitochondrial 16S and COI genes, the behavior of nest construction, the advertisement call containing 6–11 rapidly repeated regular notes, and a combination of morphological characteristics. Furthermore, the Nidirana populations recorded in Guangxi are clarified in this work, providing valuable new information on the knowledge of the genus Nidirana .
... Babina Thompson, 1912 (Fig. 29) This sister-genus of Nidirana Dubois, 1992 Lyu et al., 2017) is endemic to the Ryukyu Archipelago, including Babina subaspera (Barbour, 1908) in the central islands (Amami and Kakeromajima) and Babina holtsi (Boulenger, 1892) in the northern islands (Okinawa, Tokashiki). Monophyletic in mitochondrial phylogenetic analyses (Kakehashi et al., 2013;Lyu et al., 2017Lyu et al., , 2020 taxa diverged ~2-6 Mya depending on the calibration used (Tominaga et al., 2014), while featuring 5%, 3% and 2% of sequence divergence at Cytb, 16S/COI and (partial) 16S, respectively (Kuramoto et al., 2011;Tominaga et al., 2014;Lyu et al., 2020). ...
... Babina Thompson, 1912 (Fig. 29) This sister-genus of Nidirana Dubois, 1992 Lyu et al., 2017) is endemic to the Ryukyu Archipelago, including Babina subaspera (Barbour, 1908) in the central islands (Amami and Kakeromajima) and Babina holtsi (Boulenger, 1892) in the northern islands (Okinawa, Tokashiki). Monophyletic in mitochondrial phylogenetic analyses (Kakehashi et al., 2013;Lyu et al., 2017Lyu et al., , 2020 taxa diverged ~2-6 Mya depending on the calibration used (Tominaga et al., 2014), while featuring 5%, 3% and 2% of sequence divergence at Cytb, 16S/COI and (partial) 16S, respectively (Kuramoto et al., 2011;Tominaga et al., 2014;Lyu et al., 2020). Although borderline with speciation thresholds and probabilities [P(S) ~0.5], we tend to follow current arrangements where they are treated as species. ...
... Nidirana Dubois, 1992 (Fig. 29) A widespread group present in southern China, Taiwan and the Ryukyus, in which several new species were described based on molecular and phenotypic evidence (Lyu et al., 2017(Lyu et al., , 2019Li et al., 2019). In the EP, Nidirana adenopleura (Boulenger 1909) occurs only in Taiwan. ...
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Biodiversity analyses can greatly benefit from coherent species delimitation schemes and up-to-date distribution data. In this article, we have made the daring attempt to delimit and map described and undescribed lineages of anuran amphibians in the Eastern Palaearctic (EP) region in its broad sense. Through a literature review, we have evaluated the species status considering reproductive isolation and genetic divergence, combined with an extensive occurrence dataset (nearly 85k localities). Altogether 274 native species from 46 genera and ten families were retrieved, plus eight additional species introduced from other realms. Independent hotspots of species richness were concentrated in southern Tibet (Medog County), the circum-Sichuan Basin region, Taiwan, the Korean Peninsula and the main Japanese islands. Phylogeographic breaks responsible for recent in situ speciation events were shared around the Sichuan Mountains, across Honshu and between the Ryukyu Island groups, but not across shallow water bodies like the Yellow Sea and the Taiwan Strait. Anuran compositions suggested to restrict the zoogeographical limits of the EP to East Asia. In a rapidly evolving field, our study provides a checkpoint to appreciate patterns of species diversity in the EP under a single, spatially explicit, species delimitation framework that integrates phylogeographic data in taxonomic research.
... After prolonged taxonomic controversy, the Music Frog genus Nidirana Dubois, 1992 was recently resurrected as a distinct genus (previously considered a subgenus of Rana Linnaeus, 1758 or synonym of Babina Thompson, 1912). Species of this genus are characterized by having a relatively small-sized body, absence of the thumb-like structure on finger I, presence of well-developed dorsolateral folds, and the presence of suprabrachial glands in breeding males (Lyu et al. 2017). Currently, the genus contains 14 recognized species, widely distributed in subtropical eastern and southeastern Asia: N. okinavana (Boettger, 1895) occurring in Yaeyama of southern Ryukyu, and eastern Taiwan; N. adenopleura (Boulenger, 1909), N. guangdongensis Lyu, Wan, and Wang, 2020, N. hainanensis (Fei, Ye, and Jiang, 2007), N. mangveni Lyu, Qi, andN. ...
... Fei et al. (2010) further established a monotypic genus Dianrana Fei, Ye, and Jiang, 2010 for N. pleuraden. In the most recent and species-comprehensive analysis based on a molecular phylogeny, morphology, bioacoustics, and biogeography, N. pleuraden was inferred as the basal lineage of genus Nidirana, forming an independent species group, and Dianrana was considered invalid (Lyu et al. 2017(Lyu et al. , 2019. The remaining 13 species of Nidirana were assigned to the N. adenopleura group, characterized by the presence of a lateroventral groove on the toes (Dubois 1992;Lyu et al. 2019Lyu et al. , 2020. ...
... Tissue samples were dissected from euthanized specimens, preserved in 95% ethanol and stored at -40 °C. In addition, 35 sequences from all known Nidirana species and two out-group Babina species (following Lyu et al. 2017) were obtained from GenBank and incorporated into our dataset (Table 1; Fig. 1). ...
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The Music Frog genus Nidirana was recently resurrected as a distinct genus and contains 14 species distributed in subtropical eastern and southeastern Asia. The species diversity of Nidirana is dramatically underestimated, and half of its species was described in the last five years. In this study, Nidirana occidentalis sp. nov., a new species of Music Frog from western Yunnan, China, is proposed based on morphological and molecular evidences. The new species was previously misidentified as N. pleuraden, but can be distinguished from the true N. pleuraden from eastern Yunnan, and all other congeners, by a combination of morphological characteristics, and significant divergence in the mitochondrial genes (≥ 5.1% in 16S and ≥ 8.9% in CO1). Nidirana occidentalis sp. nov. is assigned to the N. pleuraden group on the basis of morphological characters, but its phylogenetic placement remains unresolved due to weak branch support. Geographically, these two species are isolated by the Red River in Yunnan, supporting the hypothesis that the Red River is an important geographical barrier that drives speciation in flora and fauna. Nidirana occidentalis sp. nov. represents the second species of N. pleuraden group and the 15th species of the genus.
... Nidirana pleuraden, the Yunnan pond frog [47] or Yunnan music frog [48], is endemic to Southwestern China and inhabits paddy fields, ditches, and ponds at elevations from 1150 to 2300 m asl. on the Yunan-Guizhou subtropical plateau. Male N. pleuraden possesses a pair of subgular external vocal sacs, and during the breeding season (June to July) [47], it develops a pair of male-specific postaxillary glands. ...
... Male N. pleuraden possesses a pair of subgular external vocal sacs, and during the breeding season (June to July) [47], it develops a pair of male-specific postaxillary glands. The SDSG, postaxillary glands, have been documented in several anuran amphibians as round or irregularshaped cutaneous swellings lying behind base of forelimb, colored pale brown (in Nidirana) or whitish (in Hymenochirus) [7,46,48]. The postaxillary glands in a congeneric species of N. pleuraden, east China music frog (N. ...
... These may extend backwards to the anterior end of the ilium, become thinner and eventually end in the front half of the dorsolateral regions. Surfaces of the postaxillary glands in N. pleuraden are normally rough and granular, which differ from the smooth surfaces described in other Nidirana species [48]. Visually, the postaxillary glands are yellowish brown or pale brown with dark brown or black irregular spots (Fig. 1a, c). ...
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Background: Owing to their incomplete adaptation to the terrestrial environment, amphibians possess complex cutaneous glandular systems. The skin glands not only regulate water loss and respiratory gas and salt exchange, but are also involved in defense against predators and microorganisms, social communication, and reproduction. These glands are distributed throughout the integument, but can accumulate in specific regions, forming visible outgrowths known as macroglands. Some macroglands are sexually dimorphic and mediate intersexual communication and reproductive success. The postaxillary gland is a sexually dimorphic macrogland in Nidirana pleuraden. Its biological function and its morphological and histochemical characteristics are unclear. In the present study, we describe the structure and ultrastructure of the postaxillary gland, and explore its main function. Results: The postaxillary gland has a thinner epidermis than the dorsal region of N. pleuraden. In addition to ordinary serous glands (OSG), type I and II mucous gland (I MG & II MG), a type of specialized mucous gland (SMG) is also found to constitute the postaxillary gland. The SMG is larger than other gland types, and consists of high columnar mucocytes with basal nuclei arranged radially toward a lumen. SMGs are positive to periodic acid-Schiff stain and stained blue in Masson's trichrome stain. A discontinuous myoepithelial sheath lacking innervation encircles SMG mucocytes, and the outlets of such glands are X- or Y-shaped. Transmission electron microscopy reveals abundant secretory granules in SMG, which are biphasic, composed of an electron-opaque outer ring and a less electron-dense core. Lipid droplets, and organelles, such as rough endoplasmic reticulum and Golgi stacks, are located in the supranuclear cytoplasm of the mucocytes in SMG. Female N. pleuraden exhibits chemotaxis toward homogenate of the postaxillary gland, but male does not. On treatment with trypsin, this sexual attraction disappears. Conclusions: The postaxillary gland of N. pleuraden is a male-specific macrogland that consists primarily of SMGs, together with OSGs, I MGs and II MGs. Other than their extremely large size, SMGs structurally and histochemically resemble many reported specialized gland types in amphibian sexually dimorphic skin glands. Secretions of the postaxillary gland are proteinaceous sexual pheromones, which are believed to attract females at male calling intermissions.
... The Music frog genus Nidirana Dubois, 1992 was recently reconsidered as a distinct genus based on comprehensive approaches (Lyu et al. 2017). Ten species are currently recognized from subtropical eastern and southeastern Asia: N. okinavana (Boettger, 1895) from Yaeyama of southern Ryukyu, and eastern Taiwan; N. adenopleura (Boulenger, 1909) from Taiwan and southeastern mainland China; N. nankunensis Lyu, Zeng, Wang, Lin, Liu, & Wang, 2017 from Mt Nankun of Guangdong; N. yaoica Lyu, Mo, Wan, Li, Pang, & Wang, 2019 from Mt Dayao of Guangxi; N. hainanensis (Fei, Ye, & Jiang, 2007) from Mt Diaoluo of Hainan; N. leishanensis Li, Wei, Xu, Cui, Fei, Jiang, Liu, & Wang, 2019 from Mt Leigong of Guizhou; N. daunchina (Chang, 1933) from western China; N. pleuraden (Boulenger, 1904) from southwestern China; and N. chapaensis (Bourret, 1937) and N. lini (Chou, 1999) from the northeastern Indochinese peninsula. ...
... The Music frog genus Nidirana Dubois, 1992 was recently reconsidered as a distinct genus based on comprehensive approaches (Lyu et al. 2017). Ten species are currently recognized from subtropical eastern and southeastern Asia: N. okinavana (Boettger, 1895) from Yaeyama of southern Ryukyu, and eastern Taiwan; N. adenopleura (Boulenger, 1909) from Taiwan and southeastern mainland China; N. nankunensis Lyu, Zeng, Wang, Lin, Liu, & Wang, 2017 from Mt Nankun of Guangdong; N. yaoica Lyu, Mo, Wan, Li, Pang, & Wang, 2019 from Mt Dayao of Guangxi; N. hainanensis (Fei, Ye, & Jiang, 2007) from Mt Diaoluo of Hainan; N. leishanensis Li, Wei, Xu, Cui, Fei, Jiang, Liu, & Wang, 2019 from Mt Leigong of Guizhou; N. daunchina (Chang, 1933) from western China; N. pleuraden (Boulenger, 1904) from southwestern China; and N. chapaensis (Bourret, 1937) and N. lini (Chou, 1999) from the northeastern Indochinese peninsula. ...
... Among the species in genus Nidirana, N. adenopleura has the widest distribution area and has been reported from Taiwan, Fujian, Zhejiang, Anhui, Jiangxi, Guangdong, Guangxi, Hunan and Guizhou (Fei et al. 2009(Fei et al. , 2012. In the previous study (Lyu et al. 2017), the populations from Taiwan, northern Fujian, Jingning County of Zhejiang, and Mt Jinggang of Jiangxi were confirmed as the same species, which also synonymized N. caldwelli Schmidt, 1925 with N. adenopleura. Besides, it is worth noting that the frogs previously considered as N. adenopleura from Mt Dayao of Guangxi and Mt Leigong of Guizhou were respectively revealed as two new species, N. yaoica and N. leishanensis, most recently (Lyu et al. 2019;Li et al. 2019). ...
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Three cryptic species, which were previously reported as Nidirana adenopleura , are revealed on the basis of comprehensive approaches. Nidirana guangdongensis Lyu, Wan, and YY Wang, sp. nov. is distributed in Nanling Mountains and southern Luoxiao Mountains, Nidirana mangveni Lyu, Qi, and YY Wang, sp. nov. is known from northern Zhejiang, and Nidirana xiangica Lyu and YY Wang, sp. nov. occurs in Xiangjiang River Basin, while the true Nidirana adenopleura is designated from Taiwan Island, northern Fujian, southern Zhejiang, and central Jiangxi. These three new species can be distinguished from all congeners by significant divergences in the mitochondrial 16S and CO1 genes, differences in advertisement calls, and the combination of multiple characteristics. This work indicates that the current records of Nidirana adenopleura should be of a species complex composed of multiple species and have clarified the true identity of N. adenopleura .
... nov. Fei & Ye 2016 16 Panophrys insularis (Wang, Liu, Lyu, Zeng & Wang, 2017) (Lyu, Zeng & Wang, 2020) comb. nov. ...
... The identification of Asian horned toad species based solely on traditional morphological characters has been somewhat ambiguous (Li et al. 2014;Mahony et al. 2017;Liu et al. 2018), which led previously to underestimated diversity and the controversial systematics. However, anurans are characterized by generally conservative, plesiomophic phenotypes (Cherty et al. 178), and many other higher frog taxa have also been the subject of similar debates (e.g., Frost 2006;Duellman et al. 2016;Lyu et al. 2017;Arifin et al. 2018). Furthermore, despite extensive stasis in evolution of morphological features, other aspects of these organisms (e.g., the genotype) continue differentiating (Wilson et al. 177;Schopf 184;Avise et al. 14). ...
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The diversity of Panophrys horned toads is considered highly underestimated with a large number of undescribed cryptic species. In this work, we describe four Panophrys species from eastern China which were proposed as cryptic species by molecular data in previous study, additionally provide new information on the biogeography of these four species. Panophrys daiyunensis sp. nov. from southern Fujian, Panophrys daoji sp. nov. from eastern Zhejiang, Panophrys sanmingensis sp. nov. from the hilly area among Fujian, Jiangxi and Guangdong, and Panophrys tongboensis sp. nov. from northeastern Jiangxi, can be distinguished from all recognized congeners by a combination of morphological characteristics. The descriptions of these four new species take the recognized species of Panophrys to 51, which is the largest genus within the Asian horned toads subfamily Megophryinae. Considered as an appropriate arrangement for the Asian horned toads currently and applied in this study to describe the new species, the generic recognition of Panophrys is also discussed.
... Primers used for 16S were L3975 (5'-CGCCTGTTTACCAAAAACAT-3') and H4551 (5'-CCGGTCTGAACTCAGATCACGT-3'), for 12S were L33 (5'-CT-CAACTTACAMATGCAAG-3') and H56 (5'-CGATTATAGAACAGGCTCCT-3'). PCR sequencing methods followed Lyu et al. (2017). ...
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Karstic landscapes play an important role in biodiversity formation and often contain high levels of endemism. However, site-endemic taxa in karstic landscapes are being threatened by exploitation and weak legal protection. In this study, we describe Odorrana concelata Wang, Zeng, & Lin, sp. nov. , a limestone karst-restricted odorous frog from northern Guangdong, China. This new species shows distinctive genetic divergence and morphological differences from its congeners. Phylogenetic results suggest that the new species represents an independent lineage that is grouped with O. lipuensis and O. liboensis based on the mitochondrial 16S and 12S ribosomal RNA genes. We recommend the new species be listed as Vulnerable (VU) in the IUCN categorization as it is only known from the type locality with limited microhabitats and is threatened by habitat degradation.
... ;Nguyen S. V. et al., 2009;Chuaynkern et al., 2010;Luu et al., 2013;Oliver et al., 2015;Lyu et al., 2017;Yuan et al., 2019]. ...
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A new species of the genus Rana (subgenus 'Nidirana'; Ranidae) is described from Yunnan, China. Rana lini is distinguished from the other species of 'Nidirana' by a combination of relatively large size in females, dilated tips of digits, prominent vocal sacs, extensive toe webbing, dermal folds on inner margins of 2nd and 3rd fingers wider than those of outer margins, and a distinct dermal flap along the lateral edge of the 5th toe. The new species is currently known from the subtropical forests of southern Yunnan, China, at an elevation of 1400-1650 m. A key to the species of the subgenus 'Nidirana' is provided.
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