Title First Record of the Tree-Frog Genus Liuixalus from Vietnam With

First Record of the Tree-Frog Genus Liuixalus from Vietnam Title with the Description of a New Species (Amphibia: Rhacophoridae)

Author(s) Nguyen, Tao Thien; Matsui, Masafumi; Yoshikawa, Natsuhiko

Citation Current Herpetology (2014), 33(1): 29-37

Issue Date 2014-02

URL http://hdl.handle.net/2433/216852

Type Journal Article

Textversion publisher

First Record of the Tree-frog Genus Liuixalus from Vietnam with the Description of a New Species (Amphibia: Rhacophoridae)

TƵǄ Tƾƿƻǃ NGUYEN1, MƵǈƵƼǊǑƿ MATSUI2*, Ƶǃƺ NƵǉǈǊƾƿǁǄ YOSHIKAWA2

1Vietnam National Museum of Nature, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet Road, Hanoi, VIETNAM 2Graduate School of Human and Environmental Studies, Kyoto University, Sakyo, Kyoto 606–8501 JAPAN

Abstract: We record a tree frog of the genus LiuixalusCLOQEBÖOPQQFJBCOLJ outside of China and describe it as a new species, Liuixalus catbaensis, on the basis of a single juvenile specimen collected from Cat Ba Island, northern Vietnam. The new species is easily distinguished from all other members of the genus Liuixalus by its uniformly brick-red dorsum lacking dark markings. The ?FLDBLDO>MEF@>I PFDKFÖ@>K@B LC ÖKAFKD LC QEFP PMB@FBP FK 3FBQK>J FP ?OFB×V discussed.

Key words: Liuixalus catbaensis; Mitochondrial phylogeny; Southeast Asia; Taxonomy; Zoogeography

IǃǉǇǄƺǊƹǉƿǄǃ orids (Li et al., 2008), they were moved to a distinct genus, Liuixalus, with L. romeri as the The Old-World tree-frog genus Liuixalus Li, type species. Che, Bain, Zhao, and Zhang, 2008 currently Although Frost (2013) listed Vietnam in the contains three species L. romeri (Smith, 1953), distribution range of the genus Liuixalus, no L. ocellatus (Liu and Hu in Liu, Hu, Fei, and formal report of occurrence of the genus has Huang, 1973), and L. hainanus (Liu and Wu, been given outside Southeast China. However, 2004) occurring in southeastern China (e.g., Fei a juvenile specimen of a small rhacophorid et al., 2009). These species were formerly placed RODBHDR BNKKDBSDC HM NTQ QDBDMS ëDKC RTQUDX HM in various genera, Philautus Gistel, 1848, northern Vietnam (Fig. 1) proved to be a mem- Chirixalus Boulenger, 1893, Chiromantis ber of the genus Liuixalus through subsequent Peters, 1854, and Aquixalus Delorme, Dubois, molecular analysis. The specimen formed a Grosjean, and Ohler, 2005 (Smith, 1953; Liu et clade on mtDNA trees with members of the al., 1973; Bossuyt and Dubois, 2001; Frost et genus, but was placed in a unique position dif- al., 2006; Fei et al., 2010). However, based on ferent from available sequences of its conge- results of recent molecular studies on rhacoph- MDQR !DB@TRDSGDRODBHLDM@KRNCHðDQDCLNQ- phologically from all the known members of * Corresponding author. Tel: +81–75–753–6846; Liuixalus, we describe it as a new species of the Fax: +81–75–753–6846; FDMTR 3GHRHR@KRNSGDëQRSQDBNQCNESGDFDMTR E-mail address: [email protected] outside China. 30 Current Herpetol. 33(1) 2014

a partial sequence of 16S rRNA (ca. 800 bp) with GenBank data of rhacophorid species whose congeners are recorded from Vietnam (Orlov 2012), i.e., Chiromantis hansenae (Cochran, 1927) (AB813161), Feihyla kajau (Dring, 1984) (AB847122), Gracixalus gra- cilipes (Bourret, 1937) (DQ283051), Kurixalus appendiculatus (Günther, 1859) (AB847125), Liuixalus romeri (EU215528), Nyctixalus pictus (Peters, 1871) (DQ283133), Philautus abditus Inger, Orlov, and Darevsky, 1999 (GQ285673), Polypedates leucomystax (Gravenhorst, 1829) (AB728138), Rhacophorus kio Ohler and Delorme, 2006 (AB781695), Rhaorchestes parvu- lus (Boulenger, 1893) (AB871421), Theloderma leprosum Tschudi, 1838 (AB847128), and Buergeria buergeri (Temminck and Schlegel, 1838) (AB127977). Following the result obtained (see results), we then determined the relationships of the specimen from Cat Ba Island, Vietnam, by comparing the partial sequence of 16S rRNA (ca. 500 bp) with those of Liuixalus romeri from Hong Kong (CIB [Chengdu Institute of Biology] 20080048; AB871412) and L. ocellatus from Wuzhishan, Shuiman, Fƿƽ. 1. Map of Vietnam showing Cat Ba Island and Diaoluoshan, Hainan Island, China ëKKDCRS@Q VGDQDSGDQG@BNOGNQHCRO V@RENTMC (CIB 20081062, 63, 65–69; AB871413– 871419), and additional GenBank data (L. MƵǉƻǇƿƵǂǈ Ƶǃƺ MƻǉƾǄƺǈ hainanus [GQ285671, KC465826], L. ocellatus [GQ285672, GU120328, KC465829] and Specimens examined are stored in the collec- L. romeri [EF564535, EF564536]), as well tion of the Vietnam National Museum of as Chiromantis hansenae (AB813161) and Natural History (VNMN) and Chengdu Institute Buergeria buergeri (AB127977) as out group. of Biology (CIB). We obtained DNA sequence For tree construction and calculation of genetic data from tissues preserved in 99% ethanol for distances (uncorrected p-distance), we followed a specimen of rhacophorid species from Vietnam Kuraishi et al. (2013). 5-,- 3' :3RTSNLT 'HJHC@¥R ëDKC We took the following 32 body measure- number] 2155; GenBank accession number ments to the nearest 0.1 mm with a dial caliper AB871420). Methods for DNA extraction, and under a binocular microscope, and to the @LOKHëB@SHNM @MC RDPTDMBHMF NE SGD LS#- nearest 0.01 mm using a micrometer, mainly fragments are the same as those reported by following Matsui (1984, 1994): (1) snout-vent Kuraishi et al. (2013). We deposited the resul- length (SVL); (2) head length (HL) from tip of tant sequence (858 base pairs [bp] of partial snout to hind border of angle of jaw, not sequences of mitochondrial 16S rRNA gene) measured parallel with the median line; (3) in GenBank (Accession numbers as shown snout-nostril length (S-NL); (4) nostril-eyelid above). length (N-EL); (5) snout length (SL); (6) eye In order to generically assign the rhacopho- length (EL); (7) eye diameter (ED), diameter QHCRODBHLDMEQNL5HDSM@L VDëQRSBNLO@QDC of the exposed portion of the eyeball; (8) NGUYEN ET AL.—A TREE-FROG GENUS NEW TO VIETNAM 31

tympanum-eye length (T-EL); (9) tympanum Rhacophorid sp. from Vietnam substantially diameter (TD); (10) head width (HW); (11) CHðDQDC EQNL RODBHDR NE SGD HMFQNTO FDMDQ@ internarial distance (IND); (12) interorbital (16.3% from Gracixalus to 22.6% from distance (IOD); (13) upper eyelid width (UEW); Kurixalus) other than Liuixalus (2.4%). (14) forelimb length (FLL); (15) lower arm and These results clearly suggest placement of the G@MC KDMFSG + + ëQRS SN ENTQSG rhacophorid sp. from Vietnam in the genus ëMFDQCHRJCH@LDSDQR %#6HMMDQ Liuixalus. palmar tubercle length (IPTL); (21) outer After adding sequences both from our own palmar tubercle length (OPTL); (22) hindlimb data and from GenBank, support for the mono- length (HLL); (23) thigh length (THIGH); phyly of Liuixalus including the rhacophorid (24) tibia length (TL); (25) foot length (FL); RO EQNL 5HDSM@L V@R RTîBHDMS !// ëQRSSNDKDMFSG3.$+HMMDQLDS@- BS=99%: Fig. 2), but the relationships within S@QR@KSTADQBKDKDMFSG(,3+ ëQRSSN SGDBK@CDVDQDMNSQDRNKUDC@MCëUDFQNTORL. ëESGSNDCHRJCH@LDSDQR 3#6 6DENKKNVDC hainanus, L. romeri from Jinxiu, Gungxi, the the system of description of toe-webbing states rhacophorid sp. from Vietnam, L. romeri from used by Savage (1975). Hong Kong and Shiwanshan, Guangxi, and L. ocellatus) were recognized only in a maximum SǎǈǉƻǑƵǉƿƹǈ likelihood (ML) tree. The rhacophorid sp. from Vietnam was close to L. romeri from Jinxiu, In the preliminary analyses using all known Guangxi (EF564535, EF564536), but their 11 genera from Vietnam and an outgroup monophyly was not supported (BPP=0.53, Buergeria, the rhacophorid sp. from northern BS=35%). Vietnam formed a clade with Liuixalus romeri 3GDQG@BNOGNQHCRO EQNL5HDSM@LCHðDQDC with full support (Bayesian posterior probabil- from the species of Liuixalus by small genetic ity [BPP]=1.00, bootstrap support [BS]=100%). distances (2.2–3.0%). Because the distances

Fƿƽ. 2. Maximum-likelihood (ML) tree from ca. 500 bp partial sequence of mitochondrial 16S rRNA gene for the rhacophorid sp. from Vietnam and Liuixalus. A rhacophorine Chiromantis hansenae and a buerge- riine Buergeria buergeri were used as outgroup. Numbers above or below branches represent bootstrap supports for ML inferences and Bayesian posterior probabilities (ML-BS/BPP). 32 Current Herpetol. 33(1) 2014 between the three species of Liuixalus were juvenile, collected by An Thi Hang, between also small, ranging from 1.8 to 3.0%, the 9:00–13:00 h on 18 September 2013 from Ao rhacophorid sp. from Vietnam can be geneti- Ech (20q47'56'' N, 106q59'52'' E; ca. 45 m cally regarded as a good species. Furthermore, a.s.l.) of the Cat Ba National Park, Cat Ba it is also clearly separated morphologically Island, Haiphong City, northern Vietnam. from all nominal species of Liuixalus. Thus, we conclude that the rhacophorid sp. from Description of holotype Vietnam is a distinct species in the genus Unsexed juvenile; SVL 12.8 mm; head longer Liuixalus and describe it as follows. (HL 5.3 mm, 41.5%SVL) than wide (HW 5.1 mm, 39.8%SVL), wider than body; snout Liuixalus catbaensis sp. nov. rounded in dorsal and lateral view, length (Figs. 3–5) (SL 2.3 mm, 18.1%SVL) longer than eye length (EL 2.2 mm, 17.2%SVL, ED 1.9 mm, Diagnosis 14.6%SVL), projecting beyond mouth; can- The new species is assigned to the genus thus rostralis clear; loreal region vertical and Liuixalus based on molecular evidence, and concave; nostrils slightly nearer to eye (N-EL has morphological characteristics common to 1.0 mm, 7.8%SVL) than to tip of snout (S-NL its congeners: intercalary cartilage present 1.1 mm; 8.4%SVL); internarial distance (IND between terminal and penultimate phalanges 1.8 mm, 13.9%SVL) greater than interorbital of digits; tips of digits expanded into large discs distance (IOD 1.6 mm, 12.7%SVL) and upper AD@QHMFBHQBTL L@QFHM@KFQNNUDRëMFDQREQDD eyelid (UEW 1.2 mm, 9.2%SVL); eye large, of webbing; toes very poorly webbed; no web protuberant; pupil horizontal; tympanum dis- between outer metatarsals; vomerine teeth tinct, circular, diameter (TD 0.8 mm, 6.4%SVL) absent. Liuixalus catbaensis is distinguishable about one-third of eye length and separated from all its congeners by the brick-red dorsum from eye by one-third of tympanum diameter lacking dark markings. (T-EL 0.2 mm, 1.8%SVL); vomerine teeth absent; choana oval; tongue notched posteri- Etymology orly; vocal sac absent. 3GDRODBHëBM@LDQDEDQRSNSGD"@S!@(RK@MC Forelimb slender (FLL 7.8 mm, 61.2%SVL); of northern Vietnam, where the new species hand and forearm long (LAL 6.2 mm, was found. 25+ëMFDQKDMFSGENQLTK@((((5((( (Fig. 5A); expanded disks each with a circum- Holotype marginal groove and a transverse ventral VNMN 3684 (formerly TH 2155), unsexed FQNNUDCHRJRNMSGHQCëMFDQ%#6 LL 3.0%SVL,) slightly wider than those on second (2FDW 0.35 mm, 2.7%SVL) and fourth (4FDW 0.33 mm, 2.6%SVL), and much wider SG@M ëQRS %#6 LL 25+ @KK much narrower than tympanum; webbing ADSVDDMëMFDQR @MC K@SDQ@K EQHMFDR NM ëMFDQR absent; subarticular tubercles moderately CDUDKNODC HMMDQ O@KL@Q STADQBKD ì@S (/3+ 0.39 mm, 3.0%SVL), outer one much smaller (OPTL 0.23 mm, 1.8%SVL); nuptial pad absent. Hindlimb moderately long (HLL 19.5 mm, 152.3%SVL); tibiotarsal articulation reaching to point between eye and nostril when fully Fƿƽ. 3. Juvenile holotype (VNMN 3684) of Liuixalus catbaensis sp. nov. in life. stretched leg adpressed to body; heels touching NGUYEN ET AL.—A TREE-FROG GENUS NEW TO VIETNAM 33 each other when thigh (THIGH 6.8 mm, NEëQRSSND3.$+ LL 25+MN 53.2%SVL) and tibia (TL 6.9 mm, 53.6%SVL) outer metatarsal tubercle. placed at right angle to body; foot (FL 5.3 mm, Dorsal surface nearly smooth, sparsely scat- 41.7%SVL) much shorter than tibia; toe tered with minute, blunt tubercles; strongly KDMFSG ENQLTK@ (((5((((5 SND SHOR developed supratympanic fold between eye and expanded into enlarged disks each with a cir- @QLHMRDQSHNMì@MJRB@SSDQDCVHSGì@SSTADQ- cummarginal groove and a transverse ventral cles; ventral surface smooth on lower jaw and groove; width of fourth and third toe disks AQD@RS BNLONRDCNEì@SFQ@MTKDRNM@ACNLDM (4TDW 0.41 mm, 3.2%SVL; 3TDW 0.39 mm, no protruding tubercle at heel. 25+ FQD@SDQ SG@M SG@S NE ëMFDQ CHRJR and remaining toe disks (1TDW 0.25 mm, Color 2.0%SVL; 2TDW 0.30 mm, 2.4%SVL; 5TDW In life, dorsal ground color on body brick-red 0.32 mm, 2.5%SVL); webbing formula I with a few faint brown spots (Figs. 3, 4A); (( (((Î (5ñ Ð5 narrow dark stripe between eye and nostril (Fig. 5B); subarticular tubercles rounded; inner below canthus; lips barred with white and metatarsal tubercle small and oval, (IMTL black; side of head from snout and below eye 0.39 mm, 3.0%SVL), less than half the length to temporal region dusted with brown; darker

Fƿƽ. 4. (A) Dorsal, (B) ventral, and (C) lateral views of juvenile holotype (VNMN 3684) of Liuixalus catbaensis sp. nov. in an anesthetized condition. Scale bar=10 mm. 34 Current Herpetol. 33(1) 2014 brown stripe on supratympanic fold between Archipelago in Ha Long Bay, northern Vietnam eye and arm insertion, including posterodorsal (Fig. 1). one-third of tympanum and continuing to dark mark at base of upper arm; dorsal red fading Natural history to ventral gray at side of trunk, where several In the type locality, Cat Ba National Park, irregular dark markings are scattered (Fig. the holotype was found during the daytime, in 4C); forelimb and hindlimb dorsally brick-red a disturbed primary forest surrounded by large with faint dark brown bars on forearm and limestone rocks. There was a small pool formed hindlimb; black mark at groin extending to base in a small stream (ca. 1 m wide) in the forest, of thigh; thigh posteriorly dark brown, scattered near which the frog was found. The air tem- with white posteroventrally and forming black ODQ@STQD@SSGDSHLDNEëMCHMFV@RMNSQDBNQCDC marking below cloaca; distal half of toe tips but was 27C at 19:40 h. At night Hylarana darker; ventral side grayish white sparsely guentheri (Boulenger, 1882), +GAPMFWJ?ØQQGNCQ dotted with melanophores, more densely on Boulenger, 1884, Microhyla heymonsi Vogt, throat than on abdomen (Fig. 4B); iris golden 1911, Polypedates mutus (Smith, 1940), and with black reticulations. In preservative, pattern Chiromantis hansenae (Cochran, 1927) were has not obviously changed, although all dorsal found there, but no additional specimens of surfaces are darkened. the new species were encountered.

Range Comparisons Known from the type locality, Ao Ech (Frog Because only a single juvenile specimen is Pond) within the Cat Ba National Park, Cat available for Liuixalus catbaensis, it is impos- Ba Island, the largest island of the Cat Ba sible to compare its adult body size with those of congeneric species. However, the new species is easily distinguished from L. romeri from Hong Kong and Guangxi, China, L. ocellatus from Hainan Island, and L. hainanus from Hainan Island by: having a dorsally and later- ally rounded snout, whose length is slightly longer than the length of the eye (vs. snout slightly constricted in L. hainanus and tip bluntly pointed in L. romeri, snout shorter than eye in L. ocellatus and L. hainanus), tympanum diameter about one-third of eye length and separated from eye by one-third of tympanum diameter (vs. tympanum diameter about half eye diameter in L. romeri and L. hainanus, distance between tympanum and eye about half tympanum diameter in L. romeri), hand and forearm 49% of body length (vs. about 40% in L. romeri), no VDAAHMFADSVDDM ëMFDQR UR RKHFGS VDAAHMF ADSVDDMNTSDQSVNëMFDQRHML. romeri), tibiotarsal articulation reaching between eye and nostril (vs. articulation reaching to anterior F . 5. Ventral views of left (A) hand and (B) ƿƽ corner of eye in L. ocellatus, to nostril or foot of juvenile holotype (VNMN 3684) of Liuixalus catbaensis sp. nov. after preservation. Scale bar= snout tip in L. romeri, and exceeding snout tip 3 mm. in L. hainanus), heels touching each other NGUYEN ET AL.—A TREE-FROG GENUS NEW TO VIETNAM 35 when thigh and tibia placed at right angle to 2011a, b). In the case of the new species body (vs. heels remarkably overlapping in L. described here, only a single juvenile is avail- hainanus VDAAHMFADSVDDMENTQSG@MCëESG able and biological data except other than toes not reaching to third articulation of fourth LNKDBTK@QC@S@@QDHMRTîBHDMS 'NVDUDQ SGD (vs. reaching to second articulation in L. ocel- molecular, as well as morphological, data latus), outer metatarsal tubercle absent (vs. obtained clearly demonstrate its distinct spe- tubercle small or indistinct in L. romeri, and cies status. small in L. ocellatus), dorsal surface nearly The discovery of Liuixalus from northern smooth (vs. skin with a few tubercles in L. Vietnam is not surprising, since the region is romeri and relatively rough, scattered with close to southern China, where the genus was large and small tubercles in L. hainanus) with restricted previously. In a megophryid frog, only sparsely scattered, minute, blunt tubercles Leptobrachium (Vibrissaphora) chapaense between shoulder and sacral regions (vs. upper (Bourret, 1937), some Vietnamese populations eyelid with tubercles relatively clear in L. ocel- OQNUDC SN AD BNMRODBHëB VHSG L. (V.) haina- latus and numerous in L. hainanus, and a pair nense Ye and Fei in Ye, Fei, and Hu, 1993 of small, black, rounded warts posterior to eye from Hainan Island by Matsui et al. (2010), in L. ocellatus), dorsal ground color brick-red who estimated their separation at 2.6 MYBP, VHSG@EDV U@FTDKXCDëMDCRL@KKAQNVMRONSR conforming to the date of separation of these (vs. ground color mostly olive brown or light regions in the early Pleistocene. Genetic dis- reddish brown in L. romeri, yellowish, reddish, tances on which this dating was based were or dark brown in L. ocellatus, and brown in L. 1.8–3.0% in 12S and 16S rRNA genes (Matsui hainanus, with dark horizontal or triangle et al., 2010). Compared with these values, dis- mark between eyes and a pair of X-shaped tances obtained for 16S rRNA genes between dark marks on shoulder, followed by a reverse- Liuixalus catbaensis and L. romeri from V mark in L. romeri, a pair of reverse triangu- Jinxiu are small (1.8%), suggesting their sepa- lar or V-shaped dark markings between eyes, ration later than in L. (V.) hainanense. posteriorly with a pair of small black marking However, 12S and 16S rRNA genes on which in L. ocellatus, and with irregular or X-shaped dating for Leptobrachium (Vibrissaphora) blackish brown markings, medially with a clear, was based in Matsui et al. (2010) would gener- light brown elliptical mark, followed by a trian- ally give lower values than 16S rRNA gene gular dorsal brown mark in L. hainanus), and alone. Apart from this, it is noted that inter- grayish white ventrum sparsely dotted with RODBHëBFDMDSHBCHRS@MBDRQDONQSDCENQLiuixalus melanophores (vs. ventrally cream white with are very small, even smaller than in other rha- gold-yellow tinge, scattered with several dark cophorine genera (e.g., Li et al., 2008, 2013; spots in L. romeri). this study). The family Rhacophoridae is split into two DƿǈƹǊǈǈƿǄǃ subfamilies, Buergeriinae Channing, 1989 and 1G@BNOGNQHM@D 'NðL@M "G@MMHMF The family Rhacophoridae is the most diverse 1989). Members of Buergeriinae are restricted group of amphibians in Vietnam with a total to the eastern periphery of the Asian conti- of 10 genera and 61 species recognized so far nent. Within the Rhacophorinae, Liuixalus is (Orlov, 2012; Rowley et al., 2010, 2011b; Nguyen now generally considered as the sister taxon of et al., 2013; Nguyen, unpublished data). Indeed, all the remaining rhacophorines (Li et al., the increase in the number of Vietnamese 2008, 2009; Yu et al., 2008, 2009; Pyron and amphibian species is mainly due to the discov- Wiens, 2011; Hertwig et al., 2012). Meanwhile ery of new rhacophorids. The description of the range of the genus is restricted to south- recent new species is usually made based on eastern China from Hong Kong through Guangxi molecular and acoustic data (e.g., Rowley, to Hainan, and now northern Vietnam, all in 36 Current Herpetol. 33(1) 2014 the eastern periphery of the Asian continent. LƿǉƻǇƵǉǊǇƻ Cƿǉƻƺ Although the origin and subsequent evolution- ary history of the Rhacophoridae are in debate BǄǈǈǊǎǉ, F. Ƶǃƺ DǊƸǄƿǈ, A. 2001. A review of the (e.g., Bossuyt and Milinkovitch, 2001; Li et al., frog genus Philautus Gistel, 1848 (Amphibia, 2013), the fact that phylogenetically very old Anura, Ranidae, Rhacophorinae). Zeylanica 6: groups, subfamily Buergeriinae and the genus 1–112. Liuixalus, occur on the eastern periphery of BǄǈǈǊǎǉ, F. Ƶǃƺ MƿǂƿǃǁǄǋƿǉƹƾ, M. C. 2001. the Asian continent is biogeographically inter- Amphibians as indicators of early tertiary “out- esting. of-India” dispersal of vertebrates. Science 292: In spite of its remarkable phylogenetic antiq- 93–95. uity, the degree of genetic divergence within CƾƵǃǃƿǃƽ, A. 1989. A re-evaluation of the phy- the genus Liuixalus is very small as shown logeny of Old World treefrogs. South African above. This low genetic diversity is suspected Journal of Zoology 24: 116–131. to be the result of past population bottlenecks. Fƻƿ, L., HǊ, S.-Q., Yƻ, C.-Y., Ƶǃƺ HǊƵǃƽ, Y.-Z. Probably, after very old separation from the 2009. Fauna Sinica. Amphibia. Volume 2. Anura. ancestral stock leading to the other rhacopho- Chinese Academy of Science. Science Press, rine genera in the Eocene (around 50.0-36.9 Beijing. MYBP: Li et al., 2013), the ancestors of Fƻƿ, L., Yƻ, C.-Y., Ƶǃƺ JƿƵǃƽ, J.-P. 2010. Colored Liuixalus might have experienced very limited Atlas of Chinese Amphibians. Sichuan Publishing range expansion, severe range fragmentation, Group/Sichuan Publishing House of Science and local extinctions before much more recent and Technology, Chengdu. speciation events occurred at 8.3-3.5 MYBP in FǇǄǈǉ, D. R. 2013. Amphibian Species of the the late Miocene (Li et al., 2013). In order to World: an Online Reference. Version 5.6 (9 test this hypothesis, further analyses including January, 2013). American Museum of Natural additional specimens, especially adults, of L. History, New York. http://research.amnh.org/ catbaensis from Vietnam are necessary. herpetology/amphibia/index.html FǇǄǈǉ, D. R., GǇƵǃǉ, T., FƵƿǋǄǋƿƹƾ, J. N., BƵƿǃ, AƹǁǃǄǌǂƻƺƻǑƻǃǉǈ R. H., HƵƵǈ, A., HƵƺƺƵƺ, C. F. B., ƺƻ Sѯ, R. O., CƾƵǃǃƿǃƽ, A., WƿǂǁƿǃǈǄǃ, M., DǄǃǃƻǂǂƵǃ, S. We thank the directorate of the Cat Ba C., RƵǍǌǄǇǉƾǎ, C. J., CƵǑǅƸƻǂǂ, J. A., BǂǄǉǉǄ, National Park for issuing relevant permits and B. L., MǄǂƻǇ, P., DǇƻǌƻǈ, R. C., NǊǈǈƸƵǊǑ, R. RTOONQS NE NTQ ëDKCVNQJ 6D @KRN SG@MJ 3 A., Lǎǃƹƾ, J. D., GǇƻƻǃ, D. M., Ƶǃƺ WƾƻƻǂƻǇ, 'HJHC@@MC' 3 MENQëDKCBNLO@MHNMRGHO W. C. 2006. The amphibian tree of life. Bulletin and Y. Kawahara and K. Eto for laboratory of the American Museum of Natural History assistance. J.-P. Jiang and K. Nishikawa kindly 297: 1–370. allowed us to use Chinese materials, and E. HƻǇǉǌƿƽ, S. T., DƵǈ, I., SƹƾǌƻƿǏƻǇ, M., BǇǄǌǃ, Sterling and K. Koy provided the map. Fieldwork R. M., Ƶǃƺ HƵƵǈ, A. 2012. Phylogenetic rela- by T. T. Nguyen and N. Yoshikawa was sup- tionships of the Rhacophorus everetti-group ported by a grant from the Japan Society for and implications for the evolution of reproduc- the Promotion of Science (JSPS AA Science tive modes in Philautus (Amphibia: Anura: platform Program, FY2011-2013) to M. Rhacophoridae). Zoologica Scripta 41: 29–46. Motokawa, and partly by a Grant-in Aid from KǊǇƵƿǈƾƿ, N., MƵǉǈǊƿ, M., HƵǑƿƺǎ, A., BƻǂƵƸǊǉ, the Monbusho through the JSPS (Field D. M., AƾǑƵƺ, N., PƵǃƾƵ, S., SǊƺƿǃ, A., YǄǃƽ, Research, No. 23405014) to M. Matsui. Research H.-S., JƿƵǃƽ, J.-P., OǉƵ, H., TƾǄǃƽ, H. T., Ƶǃƺ of T. T. Nguyen in Japan is funded by the NƿǈƾƿǁƵǌƵ, K. 2013. Phylogenetic and taxo- JSPS RONPAKU Program (VAST-11224). nomic relationships of the Polypedates leucomystax complex (Amphibia). Zoologica Scripta 42: 54–70 NGUYEN ET AL.—A TREE-FROG GENUS NEW TO VIETNAM 37

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