DSpace at VNU: First Record of the Genus Oreolalax (Anura: Megophryidae) from Vietnam with Description of a New Species

DSpace at VNU: First Record of the Genus Oreolalax (Anura: Megophryidae) from Vietnam with Description of a New Species...

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Description of a New Species

Author(s): Truong Q Nguyen , Trung M Phung , Minh D Le , Thomas Ziegler , and Wolfgang Böhme Source: Copeia, 2013(2):213-222 2013.

Published By: The American Society of Ichthyologists and Herpetologists

DOI: http://dx.doi.org/10.1643/CH-12-021

URL: http://www.bioone.org/doi/full/10.1643/CH-12-021

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First Record of the Genus Oreolalax (Anura: Megophryidae) from Vietnam with Description of a New Species

Truong Q Nguyen1, Trung M Phung2, Minh D Le3,4,5, Thomas Ziegler6, and Wolfgang Bo ¨hme7

The genus Oreolalax is reported from Vietnam for the first time and a new species is described based on morphological differences, molecular divergence, and phylogenetic placement Morphologically, the new species is distinguishable from its congeners on the basis of a combination of the following diagnostic characters: size small; tympanum hidden; toes with webbing at base; dorsum with distinct, round, spiny warts; flanks with white, spiny spots; belly and lower surface of limbs smooth, with dark marbling; interorbital region without dark triangular pattern; upper surface of thigh with dark bars; male with black spines present on margin of lower lip, spinal patches on chest small with fine spines, nuptial spines on fingers small, and without vocal sacs In phylogenetic analyses, the new species is unambiguously nested within the genus Oreolalax.

T HE genus Oreolalax was created by Myers and Leviton

(1962) for four species of Scutiger described by Liu

(1943, 1947) from western China Dubois (1980,

1987) treated Oreolalax as a subgenus of Scutiger Ohler and

Dubois (1992) also assigned Megalophrys weigoldi Vogt, 1924

to the genus Scutiger (subgenus Aelurolalax) However, the

placement of this species in the genus Oreolalax was

mentioned by Fei (1999) and subsequently followed by

Delorme et al (2006), Wei et al (2007, 2009), and Fei et al

(2009, 2010) On the other hand, Oreolalax was recently

considered as a distinct genus based on both morphological

and phylogenetic data (e.g., Wu et al., 1993; Zhao and Adler,

1993; Fu and Murphy, 1997; Zheng et al., 2004; Delorme et

al., 2006; Fu et al., 2007; Wei et al., 2007) Members of the

genus Oreolalax are characterized by having maxillary teeth

short; pupils vertically elliptical; tongue deeply notched

posteriorly; eustachian tubes with large, rounded openings;

tympana hidden beneath skin; fingers free of webbing; toes

with some webbing; outer metatarsals united; dorsum

glandular with numerous warts; males with large, flat, but

prominent pectoral and axillary glands; and palmar

tuber-cles large, flat, and not extending onto first metatarsal

(Myers and Leviton, 1962)

The genus Oreolalax currently consists of 17 species and so

far is known only from southwestern China (Frost, 2011)

Frost (2011) assumed that it might occur in Laos and

Vietnam; however, no voucher specimen has been reported

previously During recent field work in the Hoang Lien

Mountain Range, we discovered a species of frog which

resembled Oreolalax, but it was not assignable to any of the

named species so far Herein, we describe this species as new,

assign it to Oreolalax based on morphological characters and

phylogenetic analysis of nucleotide sequence data, and

report the genus from Vietnam for the first time

MATERIALS AND METHODS Sampling.—A field survey was conducted in April 2010 in the Hoang Lien Mountain Range, Sa Pa District, Lao Cai Province, northern Vietnam Specimens were collected by

T M Phung Samples of muscle tissue were preserved separately in 95% ethanol and voucher specimens were fixed in approximately 80% ethanol, and then later transferred to 70% ethanol for permanent storage DNA sequence data from our new samples were combined with a previously collected mitochondrial sequence dataset for species of Oreolalax and relevant out-groups, including Leptobrachium boringiae, L xanthospilum, Megophrys omei-montis, and Scutiger chintingensis (Zheng et al., 2004; Fu et al., 2007; Table 1) Sequences of Oreolalax granulosus, O pux-iongensis, O weigoldi, and O xiangchengensis are unobtain-able from GenBank; therefore, these species were excluded from our phylogenetic analysis in this study Specimens referred to in this paper are deposited in the collections of the Chengdu Institute of Biology (CIB), Chinese Academy of Sciences, Sichuan, China; the Institute of Ecology and Biological Resources (IEBR), Hanoi, Vietnam; the Institute

of Zoology (IOZ), Chinese Academy of Sciences, Beijing, China; the Royal Ontario Museum (ROM), Toronto, Canada; the Kunming Institute of Zoology (KIZ), Chinese Academy of Sciences, Yunnan, China; and the Zoolo-gisches Forschungsmuseum Alexander Koenig (ZFMK), Bonn, Germany

Nucleotide sequencing and phylogenetic analyses.—We used Le

et al.’s (2006) protocols for extraction, amplification, and DNA sequencing Two fragments of the mitochondrial genes 12S and 16S were amplified using two pairs of primers, L1991 + H1478 for 12S (Kocher et al., 1989) and 16Sar + 16Sbr for 16S (Palumbi et al., 1991) After sequences were

Strasse 47b, D-50674 Cologne, Germany; E-mail: nqt2@yahoo.com Send reprint requests to this address.

2

Dong Khoi 9A, Tam Hiep, Bien Hoa, Dong Nai Province, Vietnam; E-mail: pmytrung@yahoo.com.

minh.le.cres@gmail.com.

6

w.boehme@zfmk.de.

Submitted: 15 February 2012 Accepted: 15 November 2012 Associate Editor: B Stuart.

Copeia 2013, No 2, 213–222

aligned by Clustal X v2 (Thompson et al., 1997), data were

analyzed using maximum parsimony (MP) and Bayesian

analysis (BA), as implemented in PAUP*4.0b10 (Swofford,

2001) and MrBayes v3.2 (Huelsenbeck and Ronquist, 2001),

respectively Settings for these analyses followed Le et al

(2006), except that the number of generations in the

Bayesian analysis was increased to 13107 The optimal

model for nucleotide evolution was set to GTR+I+C as

selected by Modeltest v3.7 (Posada and Crandall, 1998)

Nodal support was evaluated using Bootstrap replication

(BP) as calculated in PAUP and posterior probability (PP) in

MrBayes v3.2 Uncorrected pairwise divergences were

calcu-lated in PAUP*4.0b10

Morphological analysis.—We compared the new species with

other members of the genus (see Material Examined) and

data obtained from the literature (Liu and Hu, 1960; Liu et

al., 1979; Tian, 1983; Fei and Huang, 1983; Yang et al., 1983;

Fei et al., 1992, 1999, 2009, 2010; Ohler and Dubois, 1992;

Fu and Murphy, 1997; Wei et al., 2009) Measurements were

taken with digital callipers to the nearest 0.1 mm The

following abbreviations were used: SVL: snout–vent length,

HL: head length (from posterior margin of lower jaw to the

tip of snout), HW: maximum head width (across angle of

jaws), SNL: snout length (from anterior corner of eye to the

tip of snout), NS: distance from nostril to the tip of snout,

EN: distance from anterior corner of eye to the nostril, IN:

internarial distance, IOD: interorbital distance, ED: eye

diameter, UEW: maximum width of upper eyelid, DAE:

distance between anterior corners of eyes, DPE: distance

between posterior corners of eyes, MAE: distance between

posterior margin of lower jaw and anterior corner of eye,

MPE: distance between posterior margin of lower jaw and

posterior corner of eye, MN: distance from posterior margin

of lower jaw to the nostril, FLL: forelimb length (from axilla

to elbow), HAL: hand length (from elbow to the tip of third finger), FFL: first finger length, TFL: third finger length, IPT: inner palmar tubercle length, OPT: outer palmar tubercle length, FeL: femur length (from vent to knee), TbL: tibia length (from knee to tarsus), FoL: foot length (from tarsus to the tip of fourth toe), FTL: fourth toe length, IMT: inner metatarsal tubercle length, OMT: outer metatarsal tubercle length, ParL: parotoid length (from posterior corner of eye to the end of parotoid gland), SPW: maximum width of spinal patch on chest, CW: chest width (distance between axillae)

We followed Wei et al (2009) to define the size of spinal patches on chest: small if the ratio of SPW/CW ,0.3 and large

if this ratio 0.4 Photos of the preserved specimen were taken with a digital microscope Keyence VHX-500F

RESULTS Phylogenetic analyses.—The combined matrix contains 867 aligned characters, 375 of 12S and 492 of 16S MP analysis of the dataset recovered nine most parsimonious trees with 609 steps (CI 5 0.69, RI 5 0.63) One of the nine trees is shown

in Figure 1 Fifty percent of the in-group nodes in our preferred tree received strong support (Bootstrap value $ 70%; Hillis and Bull, 1993), and the species from Sa Pa is unambigously nested in the genus Oreolalax (Bootstrap value 5 97) In the Bayesian analysis, –lnL scores reached stationarity after 11,000 generations in both runs The Bayesian topology is generally in agreement with the MP topology, but we note a few significant discrepancies In the Bayesian topology, the species of Oreolalax from Sa Pa along with other clades, including O multipunctatus and other species, O jingdongensis and others, and O lichuanensis and others, are unresolved In addition, the positions of O

Species information GenBank no (12S) GenBank no (16S) Locality Voucher Oreolalax chuanbeiensis EF397266 EF397266 Mao Xian, Sichuan, China CIB-XM 074

Oreolalax liangbeiensis EF397253 EF397253 Puxiong, Sichuan, China IOZCAS 3796

Oreolalax major EF397252 EF397252 Hongya, Sichuan, China ROM 40452

Oreolalax multipunctatus EF397268 EF397268 Omei, Sichuan, China ROM 40463

Oreolalax nanjiangensis EF397265 EF397265 Nan Jiang, Sichuan, China CIB-XM 804

Oreolalax omeimontis EF397264 EF397264 Omei, Sichuan, China CIB-XM 439

Oreolalax pingii EF397259 EF397259 Xi Chang, Sichuan, China CIB-XM 980

Oreolalax rhodostigmatus EF397248 EF397248 Da Fang, Guizhou, China CIB-ZYCA 746 Oreolalax rugosus EF397254 EF397254 Shi Mian, Sichuan, China CIB-XM 340

Oreolalax schmidti EF397258 EF397258 Da Yi, Sichuan, China CIB-XM 417

Oreolalax sp (China) EF397256 EF397256 Pengxian, Sichuan, China CIB-XM 092

Oreolalax sterlingae, new

species (Vietnam)

KC569979 KC569981 Sa Pa, Lao Cai, Vietnam IEBR A.2012.1 Oreolalax sterlingae, new

species (Vietnam)

KC569980 KC569982 Sa Pa, Lao Cai, Vietnam ZFMK 92830 Oreolalax xiangchengensis EF397250 EF397250 Li Jiang, Yunnan, China CIB-3LW 008 Leptobrachium boringiae EF397247 EF397247 Leishan, Guizhou, China CIB-XM 594

Leptobrachium xanthospilum EF397245 EF397245 Gia Lai, Vietnam ROM 32184

Megophrys omeimontis EF397242 EF397242 Omei, Sichuan, China ROM 40462

Scutiger chintingensis EF397269 AY526209 Hongya, Sichuan, China ROM 39065 (12S)

/XM 1045 (16S)

chuanbeiensis and O nanjiangensis are interchanged, and O.

liangbeiensis and O major become sister with O rugosus

replacing the position of O liangbeiensis as shown in

Figure 1 Both MP and BA analyses strongly support the

monophyly of the genus Oreolalax including the new species

(BP 5 97, PP 5 98), and demonstrate that the Vietnamese

species is not closely related to any other members of the

genus (Fig 1) This species is largely divergent from others within Oreolalax in terms of genetic distance, with the minimum pairwise divergence of approximately 5% in 16S, the fastest evolving marker in this study (Table 2)

Morphological comparisons.—The new Vietnamese species of Oreolalax differs from previously named species as follows:

0.63) The dataset includes 867 aligned characters of which 156 are potentially parsimony informative Numbers above and below branches are bootstrap (.50%) values and Bayesian posterior probabilities (.95%), respectively.

(except O granulosus, O puxiongensis, O weigoldi, and O xiangchengensis).

Species name

Oreolalax sterlingae, new

species

Oreolalax sterlingae, new

species

Oreolalax sterlingae, new

species

from O chuanbeiensis by having a smaller size (SVL 36.8 mm

[n 5 1] vs 47.5–56.3 mm [n 5 20] in males and 44.6 mm [n

51] vs 55.5–59.0 mm [n 5 3] in females), white spots on

flanks present (vs absent in O chuanbeiensis), and fringes

along the side of toes absent (vs present in O chuanbeiensis);

from O granulosus by having a smaller size (SVL 36.8 mm [n

51] vs 48.6–61.5 mm [n 5 16] in males and 44.6 mm [n 5

1] vs 57.4–60.0 mm [n 5 3] in females), spinal patches on

chest smaller (vs very large in O granulosus), toes with

webbing at base only (vs webbing well developed in O

granulosus), and white spots on flanks present (vs absent in

O granulosus); from O jingdongensis by having a smaller size

(SVL 36.8 mm [n 5 1] vs 48.1–60.3 mm [n 5 16] in males),

spines on chest small (vs large in O jingdongensis), nuptial

spines on fingers small (vs large in O jingdongensis),

webbing on toes basal (vs 1/3 webbed in O jingdongensis),

and triangular pattern between the eyes absent (vs present

in O jingdongensis); from O liangbeiensis by having a smaller

size (SVL 36.8 mm [n 5 1] vs 45.1–56.3 mm [n 5 20] in

males and 44.6 mm [n 5 1] vs 56.0–65.7 mm [n 5 8] in

females), spinal patches on chest small (vs very large in O

liangbeiensis), and belly with dark marbling (vs immaculate

in O liangbeiensis); from O lichuanensis by having a smaller

size (SVL 36.8 mm [n 5 1] vs 50.0–64.8 mm [n 5 2] in males

and 44.6 mm [n 5 20] vs 57.3–62.2 mm [n 5 4] in females),

spines on chest small (vs large in O lichuanensis), nuptial

spines on fingers small (vs large in O lichuanensis), and

white spots on flanks present (vs absent in O lichuanensis);

from O major by having a smaller size (SVL 36.8 mm [n 5 1]

vs 59.2–68.7 mm [n 5 6] in males and 44.6 mm [n 5 1] vs

65.0–70.0 mm [n 5 2] in females), spinal patches on chest

small (vs very large in O major), webbing on toes less

developed (at base vs 1/3–2/3 webbed), fringes along the

side of toes absent (vs present in O major), and white spots

on flanks present (vs absent in O major); from O

multi-punctatus by having a smaller size (SVL 36.8 mm [n 5 1] vs

47.4–49.8 mm [n 5 4] in males), spines on chest small (vs

large in O multipunctatus), nuptial spines on fingers small

(vs large in O multipunctatus), white spots on flanks present

(vs absent in O multipunctatus), and round black spots on

dorsum absent (vs present in O multipunctatus); from O

nanjiangensis by having a smaller size (SVL 36.8 mm [n 5 1]

vs 52.6–60.0 mm [n 5 10] in males and 44.6 mm [n 5 1] vs

53.3–58.2 mm [n 5 8] in females), spines on margin of lower

jaw present (vs absent in O nanjiangensis), spines on chest

small (vs large in O nanjiangensis), nuptial spines on fingers

small (vs large in O nanjiangensis), belly with dark marbling

(vs immaculate in O nanjiangensis), and white spots on

flanks present (vs absent in O nanjiangensis); from O

omeimontis by having a smaller size (SVL 36.8 mm [n 5 1] vs

49.5–58.4 mm [n 5 15] in males and 44.6 mm [n 5 1] vs

51.2–56.1 mm [n 5 3] in females), spines on margin of lower

jaw present (vs absent in O omeimontis), vocal sac absent in

males (vs present in O omeimontis), triangular pattern

between eyes absent (vs present in O omeimontis), and

white spots on flanks present (vs absent in O omeimontis);

from O pingii by having a dorsum with large round warts

(vs small warts in O pingii), belly with dark marbling (vs

immaculate in O pingii), and dark bars on upper side of

limbs present (vs absent in O pingii); from O popei by

having a smaller size (SVL 36.8 mm [n 5 1] vs 60.0–69.0 mm

[n 5 20] in males and 44.6 mm [n 5 1] vs 51.5–67.0 mm [n

5 10] in females), spines on chest small (vs large in O

popei), nuptial spines on fingers small (vs large in O popei),

dark spots with light centers on dorsum absent (vs present

in O popei), and white spots on flanks present (vs absent in

O popei); from O puxiongensis by having a dorsum with round warts (vs back with strongly developed spiny ridges

in O puxiongensis), belly with marbling (vs immaculate in

O puxiongensis), nuptial spines on fingers small (vs large in

O puxiongensis), and triangular pattern between eyes absent (vs present in O puxiongensis); from O rhodostigmatus by having a smaller size (SVL 36.8 mm [n 5 1] vs 57.5–73.5 mm [n 5 7] in males and 44.6 mm [n 5 1] vs 62.4–70.6 mm [n 5 2] in females), tympanum hidden (vs distinct in O rhodostigmatus), and large orange round warts on axilla and at the back of thigh absent (vs present in O rhodostigmatus); from O rugosus by having toes with less developed webbing (at base vs 1/4 webbed in O rugosus), toes with lateral fringes absent (vs present in O rugosus), and light spots on flanks present (vs absent in O rugosus); from O schmidti by having a belly with marbling (vs immaculate in O schmidti), nuptial spines on fingers small (vs large in O schmidti), and dark triangular pattern between the eyes absent (vs present in O schmidti); from

O weigoldi by having a smaller size (SVL 36.8 mm [n 5 1] vs 58.2 mm [n 5 1] in males), two clearly separated spinal patches on chest (vs one large spinalpatch in O weigoldi), toes with webbing at base (vs well developed in O weigoldi), and belly smooth (vs granular in O weigoldi); from O xiangchengensis by having a smaller size (SVL 36.8 mm [n 5 1] vs 45.4–50.6 mm [n 5 10] in males and 44.6 mm [n 5 1]

vs 54.3–61.4 mm [n 5 10] in females), spinal patches on chest small (vs very large in O xiangchengensis), and toes with webbing at base (vs almost completely webbed in O xiangchengensis; Table 3)

Based on the results of the molecular analyses and morphological comparisons, we argue that the Vietnamese specimens represent a new species, which is assigned to the genus Oreolalax and is described below

Oreolalax sterlingae, new species Holotype.—IEBR A.2012.1, adult male, northern Vietnam, Lao Cai Province, Sa Pa District, Hoang Lien Mountain,

2900 m, Phung My Trung, 30 April 2010

Paratype.—ZFMK 92830, adult female, same collection data

as the holotype

Diagnosis.—The new species differs from its congeners by a combination of the following morphological characters: size small (SVL 36.8 mm in male, 44.6 mm in female); tympanum hidden; toes with webbing at base; dorsum with distinct round spiny warts; flanks with white spiny spots; belly and lower surface of limbs smooth, with dark marbling; interorbital region without dark triangular pat-tern; upper surface of thigh with three wide dark bars; male with black spines present on margin of lower lip, spinal patches on chest small with fine spines, nuptial spines on fingers small, and without vocal sacs

Description of holotype.—Size small (SVL 36.8 mm) Head wider than long (HL 13.3 mm, HW 13.7 mm); snout protruding, rounded anteriorly, snout length (SNL 6.1 mm) greater than horizontal diameter of eye (ED 5.1 mm); interorbital region flat, wider (IOD 4.3 mm) than upper eyelid (UEW 3.3 mm); canthus rostralis distinct, loreal region concave; nostril oval, on lateral side, lying midway

Table

Fig 2 Holotype (A: IEBR A.2012.1, male) and paratype (B: ZFMK 92830, female) of Oreolalax sterlingae, new species, in life.

between tip of snout and eye (NS 3.2 mm, EN 3.2 mm); pupil

vertical; tympanum hidden; parotoid gland elongate (ParL

8.3/8.5 mm), about three times longer than wide; canthal,

preorbital, supraorbital, postorbital, supratympanic, and

parietal ridges absent; maxillary teeth present; vomerine

teeth absent; tongue longer than wide, free at the back,

notched posteriorly

Forelimbs: Arm relatively short (FLL 7.4 mm), shorter than

half of hand (HAL 18.4 mm), distinctly enlarged; relative

length of fingers: I,II,IV,III, tip of finger rounded, not

enlarged, without groove; fingers without dermal fringe and

webbing; subarticular tubercles absent on fingers I and II,

indistinct on fingers III and IV; palmar tubercles flat,

rounded; inner and upper sides of fingers I and II with

small black nuptial spines

Hindlimbs: Femur slightly longer than tibia (FeL 17.8 mm, TbL 17.3 mm, TbL/SVL 47%); heels overlapping when held

at right angles to the body; foot long (FoL 28.3 mm); toes long and thin, toe IV (FTL 14.5 mm) shorter than femur; relative length of toes: I,II,V#III,IV; tip of toes rounded, not enlarged, without discs, webbing at base only; dermal fringe along toe V absent; subarticular tubercles indistinct; inner metatarsal tubercle present but not developed, rounded (IMT 2.8 mm); outer metatarsal tubercle absent; tarsal fold absent

Skin: Snout and dorsal head with flattened warts; dorsum and upper flanks with distinct large warts, bearing small black spines; dorsolateral folds absent; upper surface of forelimbs, tarsus, and legs with flattened warts, bearing small black spines; parotoid glands well developed with small black spines; black spines also present in loreal region, upper and lower jaws, tympanic region, and more distinct

on margin of lower lip; flank with white spiny spots; chest with two distinctly separated, oval patches of small black spines (CW 14.2 mm, SPW 4.1 mm, ratio SPW/CW 0.29, narrowest distance between patches 3.3 mm); throat, venter, and lower surface of limbs smooth

Coloration in preservative.—Upper head grayish brown; dorsum and upper flanks grayish brown with small black spines; upper lip grayish brown with some indistinct whitish spots; upper surface of arm and thigh with whitish spots or bars; chest with two patches of black spines; lower zone of flanks grayish brown with whitish spots; chin, throat, and belly cream with brown mottlings; lower surface of limbs grayish brown with whitish spots

Coloration in life.—Dorsum and upper flanks brown with small black spines; upper lip brown with some indistinct whitish spots; upper surface of arm and thigh with whitish spots or bars; lower zone of flanks brown with yellowish white spots; chin, throat, and belly cream with brown mottling; lower surface of limbs dark brown with whitish spots (Figs 2, 3, 4)

Sexual dimorphism.—Male is smaller than the female (Table 4) and has lower lip with distinct black spines, dorsal

A.2012.1) of Oreolalax sterlingae, new species.

A.2012.1) of Oreolalax sterlingae, new species.

warts more prominent, chest with two patches of small

spines, inner and upper sides of fingers I and II with black

nuptial spines Vocal sacs are absent in both sexes

Natural history.—Specimens of Oreolalax sterlingae were

found at night in a rocky rivulet in bamboo forest at an

elevation of 2900 m (Fig 5) Other species of Oreolalax occur

at 700–3550 m (Wei et al., 2007; Fei et al., 2010; Frost, 2011)

The female (ZFMK 92830) contained yellow eggs (3.6 mm in

diameter)

Distribution.—Oreolalax sterlingae is known only from Hoang

Lien Mountain Range, Lao Cai Province, northern Vietnam

(Fig 6)

Etymology.—The specific epithet is dedicated to Dr Eleanor

J Sterling from the Center for Biodiversity and

Conserva-tion, American Museum of Natural History in New York, to

acknowledge her outstanding contribution to biodiversity

research and conservation in Vietnam As common names

we suggest Sterling’s Toothed Toad (English), Co´c nu´ i

s-tec-ling (Vietnamese)

DISCUSSION

Although our morphological and molecular results clearly

show that the new species belongs to the genus Oreolalax, its

position within the genus is still largely unresolved The

sister relationship between O sterlingae and the clade

containing O multipunctatus and other species (see Fig 1)

is only weakly supported in the MP analysis (BP , 50%), and the two groups become unresolved in the Bayesian analysis Moreover, many nodes in the genus receive low support values in both analyses Additional data from both mito-chondrial and nuclear genes might help provide a more robust phylogenetic hypothesis of the genus, as shown in previous study (e.g., Fu et al., 2007)

The first discovery of a member of the genus Oreolalax outside China once again confirms the close affinity between northern Vietnam and southern China as shown

in other taxonomic groups (see Sterling et al., 2006) The new record also highlights the unusual amphibian diversity

of the Hoang Lien Mountain Range, which is known as one

of the most famous regions in terms of herpetofaunal diversity in Vietnam, and situated in one of the highest phylogenetic diversity zones of amphibians in the world (Fritz and Rahbek, 2012) Ohler et al (2000) reported a total

of 42 species of amphibians from Hoang Lien Range with descriptions of two new species, namely Leptolalax pluvialis and Rhacophorus duboisi Orlov et al (2001) listed 20 species

of rhacophorids and discovered another new species of tree frog, viz Rhacophorus hoanglienensis In addition, a number

of new species of amphibians have been discovered from this region during the last decade: Leptobrachium echinatum (Dubois and Ohler, 1998), Amolops cucae (Bain et al., 2006), Amolops minutus and A splendissimus (Orlov and Ho, 2007), and Theloderma lateriticum (Bain et al., 2009)

It is important to note the rarity of this species in Vietnam Although numerous surveys have been conducted in Lai Chau, Lao Cai, and Yen Bai provinces since 1998, this is the first record

of Oreolalax from the Hoang Lien Mountain Range Because the actual distribution of the new species is unknown, we suggest the species should be considered Data Deficient following IUCN’s Red List categories (IUCN, 2001) Among 17 species of Oreolalax known from China, 13 are listed in the IUCN Red List (2012) as near threatened to crictically endangered due to anthropogenic threats and small distribution ranges

MATERIAL EXAMINED

O jingdongensis: (6) China: KIZ 91006–91007, 91011, 91155–

91157, Yunnan, Jingdong Huashan

Oreolalax sterlingae, New Species Abbreviations defined in the text.

IEBR A.2012.1 Holotype (Male)

ZFMK 92830 Paratype (Female)

ParL (left/right) 8.3/8.5 5.5/5.5

Mountain Range, Lao Cai Province, Vietnam.

O liangbeiensis: (1) China: KIZ 65II0684, Sichuan, Puxiong,

Liangbei

O puxiongensis: (1) China: KIZ 19810703001, Sichuan,

Puxiong

O rugosus: (2) China: KIZ 820857, 821129, Yunnan, Yangbi

ACKNOWLEDGMENTS

We thank the directorate of Hoang Lien National Park for

issuing relevant permits For the loan of specimens, we are

grateful to C Le (Hanoi), T Nguyen (Hanoi), J Che and J

Wang (Kunming) Special thanks to D Kizirian for

com-menting on the manuscript, E Sterling and K Koy for

providing the map T Phung thanks T Nguyen, H Vu, and

L Do for assistance in the field T Nguyen would like to

thank M Bonkowski for support of work in Germany and J

Jiang for providing literature M Le was supported by the

National Foundation for Science and Technology

Develop-ment of Vietnam (NAFOSTED: Grant No 106.15-2010.30)

Research of T Nguyen in Germany is funded by the

Alexander von Humboldt Foundation (VIE 1143441)

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new species, from Hoang Lien Mountain Range, Sa Pa District, Lao Cai

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