Zootaxa 3974 (3): 377 390 www.mapress.com/zootaxa/ Copyright 2015 Magnolia Press Article http://dx.doi.org/10.11646/zootaxa.3974.3.5 http://zoobank.org/urn:lsid:zoobank.org:pub:715826e0-4a40-44ad-a07d-d12f8185c132 ISSN 1175-5326 (print edition) ZOOTAXA ISSN 1175-5334 (online edition) A new species of Hemiphyllodactylus Bleeker, 1860 (Squamata: Gekkonidae) from western Yunnan, China WEIBO GUO 1, KAIYA ZHOU 1,2, JIE YAN 1 & PENG LI 1 1 Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing 210023, China. E-mail: guoweibo901201@126.com, kyzhounj@126.com, janeiam@163.com and biolipeng@163.com 2 Corresponding author Abstract A new species of the genus Hemiphyllodactylus is described from mountainous area of Changning County, Yunnan Province, China. Hemiphyllodactylus changningensis sp. nov. is distinguished from all other congeners by morphology and a significant genetic divergence of greater than 17% (ND2 gene). The new species from Changning is characterized by the following features: a maximum SVL of 40.1 mm in males and 43.8 mm in females; 11 15 dorsal scale rows; 6 8 ventral scale rows; a forefoot lamellar formula of 3-3/4-3/4-3; a hindfoot lamellar formula of 3-4-4-4 or 3-3-3-3; precloacal and femoral pore series continuous; cloacal spurs present in both sexes; dark dorsal transverse blotches; dark postorbital stripe; a brown postsacral mark bearing anteriorly projecting arms; and unpigmented caecum and gonads. The new species occurs also in Longyang District of Baoshan City, Yunnan Province, China. Key words: Slender Gecko, Hemiphyllodactylus changningensis sp. nov., molecular data, Changning County, Yunnan Province Introduction The first record of the slender gecko, Hemiphyllodactylus in China was based on specimens collected from the Yunnan Province. It was described by Boulenger (1903) as Gehyra yunnanensis and was recognized as Hemiphyllodactylus yunnanensis (Boulenger) in Smith s work on the lizards of British India (Smith 1935). Zhou et al. (1981) examined specimens of H. yunnanensis from Yunnan, Guizhou, and Guangxi Zhuang Autonomous Region, China, and recognized three new subspecies of Hemiphyllodactylus based on digital lamellae patterns: H. y. dushanensis Zhou & Liu, H. y. jinpingensis Zhou & Liu, and H. y. longlingensis Zhou & Liu. These subspecies were elevated to full species status in an integrative taxonomy study of Hemiphyllodactylus (see Grismer et al. 2013 for details). So far, four species of Hemiphyllodactylus have been described in Yunnan, Guizhou, and Guangxi Zhuang Autonomous Region in China. During fieldwork in the mountainous area of Changning County, western Yunnan, we collected specimens of an unnamed species of Hemiphyllodactylus with a slender body and unique U-shaped digital lamellae. The population differs from any known species of Hemiphyllodactylus in molecular and morphological characteristics. Therefore, we describe it here as a new species. Material and methods Sampling. Field surveys were conducted in July 2012 in Changning County of Yunnan Province in China. Geographic coordinates of sampling sites were recorded with a Geographic Positioning System unit (Garmin GPSmap 60CSX, GARMIN, USA) using map datum WGS84. All specimens were collected in the evening and were euthanized with ether. Liver tissues were preserved in 95% ethanol and voucher specimens were fixed and stored in approximately 10% formalin. Specimens were deposited in the collections of the College of Life Sciences, Nanjing Normal University, Nanjing, China. Accepted by A. Bauer: 25 May 2015; published: 23 Jun. 2015 377
Molecular data and phylogenetic analyses. Total DNA was extracted from liver tissues of nine individuals following standard phenol/chloroform extraction methods. A 594 bp fragment of mitochondrial gene ND2, NADH dehydrogenase subunit 2, was amplified and sequenced using a pair of primers designed in our lab, ND2L-142 (5 - CACCACCCACGAGCAACAGA-3 ) and ND2H-859 (5 -GAAAAATAGGCTTGGTAGGC-3 ). PCRs were performed in 50 µl reactions containing 1 µl of each primer, 10 µm; 1.8 µl DNA template, ca. 100 ng; 5 µl MgCl 2, 25 mm; 5 µl 10 PCR buffer; 4 µl dntp mixture; 0.25 µl Takara Taq (Takara Biomedical, Dalian, China). Amplification was run under the following thermal cycle program: 4 min at 94 C followed by 35 cycles of 40 s at 94 C, 50 s at 48 C, 50 s at 72 C, and a subsequent 10 min final extension step at 72 C. PCR products were checked by electrophoresis in 1.5% agarose gel and purified with AxyPrep PCR Cleanup Kit (AxyGen MA, USA). Sequencing was carried out bidirectionally on an ABI 3730 xl DNA Analyzer. Sequences were assembled by DNASTAR. Lasergene. v7.1 (DNASTAR Inc., Madison, WI, USA) after manual editing, all sequences were aligned by MEGA5.04 (Tamura et al. 2011) and there was no insert/miss sites and stop codons in each sequence, unique haplotypes were identified using DnaSP v5.10 (Librado & Rozas 2009), a total of 4 haplotype sequences were deposited in GenBank, accession numbers KP732436 KP732439. Uncorrected pairwise divergences were calculated using MEGA5.04 (Tamura et al. 2011). For the phylogenetic analyses, we using maximum likelihood (ML) and Bayesian inference (BI). The Akakike information criterion (AIC) and Bayesian information criterion (BIC) as implemented in PartitionFinder v1.1.0 (Lanfear et al. 2012), were used to calculate the best-fitting model of evolution for each codon position (Table 2). TABLE 1. Samples used in molecular analyses. Species GenBank number Locality Voucher information Hemiphyllodactylus banaensis KF219783 Ba Na-Nui Chua, Vietnam ITBCZ2450 H. changningensis sp. nov. KP732436 9 Changning, Yunnan, China h00315, h00325 326, h00328, h00331, h00334; h00321; h00323; h00349 H. chiangmaiensis KF219782 Chang Mai, Thailand LSUHC9504 H. dushanensis FJ971016 Dushan, Guizhou, China h00001 H. engganoensis KF219776 Pulau, Engano, Sumatra MVZ239346 H. ganoklonis JN393950 Ngercheu, Palau USNM 563671 H. harterti KF219760 Bukit Larut, Malaysia LSUHC10383 H. jinpingensis FJ971041 Jinping, Yunnan, China h00138 H. kiziriani KJ676800 Luang Prabang, Laos IEBR A.2014.3 H. longlingensis FJ971046 Longyang District, Baoshan, h00104 Yunnan, China H. longlingensis inserta sedis FJ971049 Longyang District, Baoshan, h00103 Yunnan, China H. titiwangsaensis KF219785 Cameron Highlands, Malaysia LSUHC10717 H. typus GQ257745 Suva, Fiji ABTC 32736 H. yunnanensis FJ971021 Kunming,Yunnan, China h00056 H. zugi KF575152 Ha Lang, Cao Bang, Vietnam IEBR A.2013.21 H. sp. nov. 8 JN393949 Mandalay, Pyin Oo Lwin, Myanmar USNM-FS 36836 378 Zootaxa 3974 (3) 2015 Magnolia Press GUO ET AL.
Maximum likelihood (ML) analysis for all datasets was performed using RAxML-HPC2 on XSEDE (Stamatakis 2006a) through the Cipres Science Gateway (Miller et al. 2010). Given the advantages of CAT over Gamma (Stamatakis 2006b) and the proportion of invariable sites estimate not recommended by the developer of RAxML (Mayrose et al. 2005; Stamatakis 2006a), the model GTR + CAT for all codon positions was used. Node reliability was obtained after 1000 bootstrap replicates. The Bayesian inference (BI) analysis was carried out with the software MrBayes v3.12 (Ronquist et al. 2005) by running four Markov chains (one cold and three heated) for 1,000,000 generations, saving trees every 100 generations. The rates of different partitions were allowed to vary under a flat Dirichlet prior by setting ratepr = variable. The analysis was halted after the average standard deviation split frequency was below 0.01 which is considered to converge to the same steady state distribution. The first 25 % of the trees samples from each run were discarded as a conservative measure to avoid the possibility of including random, suboptimal trees. Samples used in molecular analyses are shown in Table 1. Automatic Barcode Gap Discovery (ABGD). ABGD is an automated iterative process to sort sequences into putative species based on pairwise distances (Puillandre et al. 2012). Aligned sequences of ND2 were uploaded to the web interface at http://wwwabi.snv.jussieu.fr/public/abgd/abgdweb.html (last modification date: 28 October 2014). P (prior limit to intraspecific diversity) had a minimum value of 0.03 and a maximum value of 0.1. The minimum relative gap width (X) had a default value of 1.5. All available models [Jukes-Cantor (JC86), Kimura (K80)] were tested. Morphological characters. Mensural data were taken with a digital caliper to the nearest 0.1 mm under a Nikon SMZ1500 dissecting microscope on the right side of the body following the methods of Zug (2010): snoutvent length (SVL, distance from the tip of snout to the vent), tail length (TailL, from the vent to the tip of tail), trunk length (TrunkL, taken from the posterior margin of the forelimb at its insertion point on the body to the anterior margin of the hind limb at its insertion point on the body), head length (HeadL, distance between retroarticular process of jaw and snout-tip), head width (HeadW, maximum width of head), ear opening diameter (EarD, longest dimension of ear); eye diameter (EyeD, the horizontal diameter of the eyeball), nares-eye length (NarEye, measured from the anterior margin of the eye ball to the posterior margin of the external nares), snout-eye length (SnEye, distance between anteriormost point of eye and tip of snout), internarial distance (SnW, shortest distance between left and right nares). Scale counts and color pattern characters observations were made following Zug (2010) and Grismer et al. (2013). TABLE 2. Models of molecular evolution used for maximum-likelihood and Bayesian analyses. Gene and For RAxML For BI codon positions Best-fitting model Model selected Best-fitting model Model selected ND2 1 st pos GTR + I + Gamma GTR + CAT GTR + Gamma GTR + Gamma 2 nd pos GTR + I + Gamma GTR + CAT HKY + Gamma HKY + Gamma 3 rd pos GTR + Gamma GTR + CAT HKY + Gamma HKY + Gamma Results Phylogenetic analyses. ML and BI analyses of the ND2 data produced the identical topological structure tree (Fig. 1). The lineage containing the new species and H. longlingensis inserta sedis (Grismer et al. 2013) is supported as sister taxon to H. longlingensis (Bootstrap value (BP) = 50, Bayesian posterior probability (PP) = 0.53). The genetic divergence between H. longlingensis inserta sedis and Hemiphyllodactylus sp. nov. is 1.2-1.5% which is far less than genetic divergence between Hemiphyllodactylus species (Table 3). The minimal genetic divergence between the new species and other members within the genus Hemiphyllodactylus was ca. 17% (between Hemiphyllodactylus sp. nov. and H. jinpingensis) (Table 3). The molecular evidence suggests that H. longlingensis inserta sedis from Longyang District, Baoshan City (about 100 km from the type locality of Hemiphyllodactylus sp. nov.) is actually a population of the new species. Hemiphyllodactylus sp. nov. and Hemiphyllodactylus longlingensis are the sister group to another lineage, i.e., Hemiphyllodactylus chiangmaiensis + (H. jinpingensis + Hemiphyllodactylus sp. nov. 8) (BP = 98 and PP = 1.00). A NEW HEMIPHYLLODACTYLUS FROM CHINA Zootaxa 3974 (3) 2015 Magnolia Press 379
TABLE 3. Uncorrected ( p ) distance matrix showing pairwise genetic divergence (ND2) between Hemiphyllodactylus species computed in MEGA5.04 (Tamura et al. 2011). Species name 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 1. Hemiphyllodactylus changningensis sp. nov. (KP732436 9) - 2. H. banaensis (KF219783) 26.9 27.3-3. H. chiangmaiensis (KF219782) 18.9 19.2 25.6-4. H. dushanensis (FJ971016) 25.1 25.6 17.4 27.2-5. H. engganoensis (KF219776) 27.8 27.9 31.3 28.3 29.5-6. H. ganoklonis (JN393950) 27.9 28.1 30.7 28.8 30.3 22.7-7. H. harterti (KF219760) 30.8 31.1 32.7 30.2 31.8 31.8 30.1-8. H. jinpingensis (FJ971041) 17.3 17.7 27.5 14.8 25.9 27.8 27.4 30.5-9. H. kiziriani (KJ676800) 25.8 26.1 22.7 27.3 20.5 29.8 30.7 32.4 28.0-10. H. longlingensis (FJ971046) 18.2 18.5 30.3 18.7 29.5 28.3 28.8 32.7 18.5 27.6-11. H. longlingensis inserta sedis (FJ971049) 1.2-1.5 27.3 19.6 26.3 27.9 28.8 30.6 18.0 26.1 18.7-12. H. titiwangsaensis (KF219785) 27.8 28.3 29.3 30.0 30.0 30.3 29.6 25.4 28.1 32.4 28.6 28.5-13. H. typus (GQ257745) 29.1 29.3 31.7 27.7 30.1 19.5 19.0 31.1 28.3 28.6 27.9 24.1 29.8-14. H. yunnanensis (FJ971021) 23.9 24.4 22.0 25.0 21.0 28.3 27.9 29.8 25.6 20.7 26.4 29.3 29.5 27.9-15. H. zugi (KF575152) 25.5 26.0 16.2 26.7 5.9 30.4 31.9 31.4 25.8 20.5 30.4 26.0 30.7 31.0 21.2-16. H. sp. nov. 8 (JN393949) 20.1 20.3 27.3 14.4 27.9 28.2 27.0 31.1 13.2 28.1 18.9 20.3 28.4 25.7 25.7 27.2 380 Zootaxa 3974 (3) 2015 Magnolia Press GUO ET AL.
Automatic Barcode Gap Discovery (ABGD). The primary species delimitation analysis based on 24 sequences of ND2 with ABGD resulted in 14 groups (i.e. species) whether using primary and recursive partition. The results are consistent with accepted species classification except H. dushanensis Zhou & Liu and H. zugi Nguyen, Lehmann, Le, Duong, Bonkowski & Ziegler were placed into the same group because of low genetic distances (see Table 3). The nine sequences from Hemiphyllodactylus sp. nov. and one sequence of H. longlingensis inserta sedis were assigned to a separate group. FIGURE 1. Bayesian phylogenetic tree based on 594 bp of the ND2 gene. Numbers by the nodes indicate (in this order): ML bootstrap support values and posterior probability values of the BI. * Hap-1 contains descriptions for h00315, h00325 326, h00328, h00331, h00334. ** GenBank records of FJ971038, FJ971040 971042, and FJ971045 971048 were updated on 9 December 2014. In these updated GenBank records, Hemiphyllodactylus yunnanensis jinpingensis is the source organism of sequences FJ971038, FJ971040-971042; Hemiphyllodactylus yunnanensis longlingensis is the source organism of sequences FJ971045 971048. Based on morphological differences and molecular divergence, we herein describe this population as a new species. Hemiphyllodactylus changningensis sp. nov. Changning Slender Gecko Holotype. h (Nanjing Normal University herpetology) 00315, adult male, collected on 8 July, 2012 by P. Li and J. Yan in the vicinity of Changning County, Yunnan Province, China (24 49.224 N, 99 36.523 E, at an elevation of 1682 m). Paratypes. Nine specimens from the same locality and with the same collection data as the holotype: three adult males h00328, h00331 and h00334, and six females h00321, h00323, h00325 00326, h00335 and h00349. A NEW HEMIPHYLLODACTYLUS FROM CHINA Zootaxa 3974 (3) 2015 Magnolia Press 381
Diagnosis. The new species is distinguished from the remaining congeners by a combination of the following characters: a bisexual taxon; a maximum SVL of 40.1 mm in males and 43.8 mm in females; 7 or 8 chin scales; enlarged postmental scales; 3 or 4 circumnasal scales; 2 or 3 scales between supranasals; 8 11 supralabials; 8 10 infralabials; 11 15 dorsal scale rows; 6 8 ventral scale rows; a forefoot lamellar formula of 3-3/4-3/4-3; a hindfoot lamellar formula of 3-4-4-4 or 3-3-3-3; 3 or 4 subdigital lamellae on the first finger; 3 or 4 subdigital lamellae on the first toe; precloacal and femoral pore series continuous; 1 or 2 cloacal spurs on each side in both males and females; no enlarged subcaudal scales; dark dorsal transverse blotches; dark postorbital stripe; a brown postsacral mark bearing anteriorly projecting arms; and unpigmented caecum and gonads. FIGURE 2. Subcaudals in Hemiphyllodactylus longlingensis (h00198) (left) and H. changningensis sp. nov. (h00315) (right). FIGURE 3. Holotype (h00315, male) of Hemiphyllodactylus changningensis sp. nov. from Changning County, Yunnan Province, China. Description of holotype. Adult male; body slender; size small SVL 39.8 mm, tail 32.7 mm; trunk length (TrunkL) 18.7 mm; eye diameter (EyeD) 2.1 mm; head longer than wide (HeadL 8.9 mm, HeadW 7.1 mm); nareeye length (NarEye) 2.8 mm; snout-eye length (SnEye) 4.4 mm; internarial distance (SnW) 1.4 mm; ear opening 382 Zootaxa 3974 (3) 2015 Magnolia Press GUO ET AL.
distance (EearD) 0.7 mm. Proportions: TrunkL/SVL 47.0%, HeadL/SVL 22.4%, HeadW/SVL 17.8%, HeadW/ HeadL 79.8%, SnEye/HeadL 49.4%, NarEye/HeadL 31.5%, EyeD/HeadL 23.6%, SnW/HeadL 15.7%, EyeD/ NarEye 75.0%, SnW/HeadW 19.7%. Scalation. Mental triangular, three postmentals, outer ones distinctly larger than the middle scale, in contact with mental and first infralabials anteriorly, behind the middle small postmental, there was a large scale in same size with two other postmentals; eight chin scales; labial scales enlarged from rostral to below eye, supralabials 9 (right) and 10 (left), infralabials 9 (right) and 10 (left); three circumnasal scales and three scales between supranasals; ventral trunk scales larger than dorsal ones, dorsal scale rows 12 at midbody (contained within one eye diameter), and ventral scales rows 6 at midbody (contained within one eye diameter); subcaudal scales slightly larger than dorsal caudal scales but not plate-like; one cloacal spur on each side; precloacal and femoral pore series continuous, 20 in total; subdigital lamellae 3 on the first finger and 3 on the first toe, respectively; digital formula 3-3-3-3 (forefoot) and 3-4-4-4 (hindfoot). Coloration in preservative. Ground color of dorsal surface of head grey, two dark brown stripes extend from the posterior corner of the orbit on each side, passing above the ear opening to the occiput. Dark brown transverse bands present along dorsum. Dorsal surface of the limbs grey, with irregular brown markings. Dorsal surface of the tail is yellowish grey with several transverse brown bands. Ventral surface of the tail is light yellow. Ventral surface is cream grey, with dark dots on the head and body; dark dots occurring in high density on hind limbs and base of the tail. Variation. Variation of measurements data and scalation characters are presented in Table 5 and Table 6. Coloration of this species on body varies from grey (h00315, h00323, h00326, h00349) to tan (h00328, h00331, h00334). Etymology. These slender geckos occur in the mountainous area of Changning County and the specific epithet refers to this region. Natural history. Changning is located in the southern part of Hengduan Mountains. The specimens were collected in the evening on the walls of the village houses located near terraced fields about 1680 m above sea level. Two gravid females (h00321, h00323) each with two eggs. Distribution. The species is currently known from Changning County and Longyang District of Baoshan City in western Yunnan, China. Morphological comparisons. We compared the undescribed gecko species from western Yunnan with selected members of the genus Hemiphyllodactylus based on examination of specimens (see Appendix) and data obtained from the literature (Zhou et al. 1981 with English translation of Ota 1996; Zug 2010; Grismer et al. 2013, 2014; Nguyen et al. 2013, 2014; Ngo et al. 2014). Diagnostic characters separating the species from other nominal taxa of Hemiphyllodactylus are shown in Table 4. The specimen of H. longlingensis inserta sedis is seriously damaged and many characters cannot be determined, but the lack of bright spots on the back is consistent with H. changningensis sp. nov., and different from H. longlingensis which has a series of white spots on the back of the trunk. The undescribed species from western Yunnan differs from its closest sister taxon, Hemiphyllodactylus longlingensis (10 samples) by having the digital lamellar formulae (hindfoot) 3-4-4-4/3-3-3-3 (versus 4-4/5-4/5-4 in H. longlingensis), postsacral mark bearing anteriorly projecting arms (versus lacking anteriorly projecting arms in H. longlingensis), without light longitudinal series of dorsal spots (versus white dorsal spots in H. longlingensis), and smaller subcaudals than in H. longlingensis (Fig. 2). It differs from other mainland China species in having the postsacral mark bearing anteriorly projecting arms (versus lacking anteriorly projecting arms in H. dushanensis (4 samples), H. jinpingensis (4 samples) and H. yunnanensis (4 samples)). It further differs from H. jinpingensis by its maximum SVL 43.8 mm (versus maximum SVL 39.6 mm in H. jinpingensis), hindfoot lamellar formula 3-4-4-4/3-3-3-3 (versus 4-4-4-4 in H. jinpingensis); from H. yunnanensis by having the hindfoot lamellar formula 3-4-4-4/3-3-3-3 (versus 4-5-5-4 or 3-4-5-5 in H. yunnanensis); from H. dushanensis by its maximum SVL 43.8 mm (versus 50.6 mm in H. dushanensis), lamellar formula 3-3/4-3/4-3 (forefoot) and 3-4-4-4 or 3-3-3-3 (hindfoot) (versus 3-4- 5-4 or 4-5-5-4/5 and 4-5-5-5 or 5-6-6-5, respectively, in H. dushanensis), fewer total pores in males (19 22 versus 24 26 in H. dushanensis), presence of dark dorsal transverse blotches (versus absent in H. dushanensis); from H. banaensis Ngo, Grismer, Pham & Wood by its maximum SVL 43.8 mm (versus maximum SVL 51.0 mm in H. banaensis), having fewer scales between supranasals (2 3 versus 4 11 in H. banaensis), fewer dorsal and ventral scale rows (11 15 and 6 8 versus 17 20 and 9 12, respectively, in H. banaensis), fewer subdigital lamellae on first finger and first toe (3 4 and 3 4 versus 5 and 5, respectively, in H. banaensis); from H. chiangmaiensis by having A NEW HEMIPHYLLODACTYLUS FROM CHINA Zootaxa 3974 (3) 2015 Magnolia Press 383
caecum and gonadal peritoneum unpigmented (versus pigmented in H. chiangmaiensis); from H. kiziriani Nguyen, Botov, Le, Nophaseud, Zug, Bonkowski & Ziegler by having fewer dorsal and ventral scale rows (11 15 and 6 8 versus 18 27 and 11 15, respectively, in H. kiziriani), by having hindfoot lamellar formula 3-4-4-4/3-3-3-3 (versus 4-4/5-4/5-4 in H. kiziriani), fewer subdigital lamellae on first finger and first toe (3 4 and 3 4 versus 5 and 5, respectively, in H. kiziriani); from H. zugi by having fewer chin scales (7 8 versus 9 12 in H. zugi), fewer dorsal and ventral scale rows (11 15 and 6 8 versus 20 22 and 15 16, respectively, in H. zugi), and by having the hindfoot lamellar formula 3-4-4-4/3-3-3-3 (versus 4-5-5-5 in H. zugi). FIGURE 4. Map showing the type localities of Hemiphyllodactylus changningensis sp. nov. at Changning, Yunnan Province, China, and congeneric species of Hemiphyllodactylus in China and adjacent countries. H. longlingensis from Longling, Yunnan Province, China; H. dushnanesis from Dushan, Guizhou Province, China; H. jinpingensis from Jinping, Yunnan Province, China; H. yunnanensis from Kunming, Yunnan Province, China; H. banaensis from Ba Na-Nui Chua Nature Reserve, Da Nang Province, Vietnam; H. chiangmaiensis from the vicinity of Chiang Mai, Chiang Mai Province, Thailand; H. kiziriani in Ban Xieng Muak Village, Luang Prabang District, Luang Prabang Province, Laos; H. zugi from Ha Lang District, Cao Bang Province, northern Vietnam; H. sp. nov. 8 from Pyin Oo Lwin, Mandalay Division, Myanmar. Discussion Our molecular analysis shows that the new species is supported as a member of the clade containing H. longlingensis and H. jinpingnensis from China, H. chiangmaiensis from northern Thailand and Hemiphyllodactylus sp. nov. 8 from Myanmar. Hemiphyllodactylus changningensis + H. longlingensis comprise the sister group to H. jinpingnensis + H. changningensis. The molecular evidence suggests that H. longlingensis inserta sedis from the Longyang District of Baoshan City is actually a population of the new species. The Yunnan-Guizhou Plateau in the southwest China has the highest species diversity of Hemiphyllodactylus. There are five species of slender geckos, H. changningensis, H. longlingensis, H. yunnanensis, H. jinpingensis and H. dushanensis, known to occur in southwestern China. Further studies on geographic distribution and cryptic diversity of the slender geckos in southwestern China are needed. 384 Zootaxa 3974 (3) 2015 Magnolia Press GUO ET AL.
TABLE 4. Diagnostic characters (grey highlight) separating the species from other nominal taxa of Hemiphyllodactylus. /=data unavailable. banaensis chiangmaiensis dushanensis jinpingensis kizirianis longlingensis yunnanensis zugi changningensis sp. nov. Max SVL 51.0 41.2 50.6 39.6 40.8 45.7 45.8 46.6 43.8 Chin sacles 6 7 8 12 8 10 7 9 6 9 7 9 7 8 9 12 7 8 Postmentals distinctyly enlarged (1) or not (0) 1 1 1 1 1 1 1 1 1 Circumnasal scales 3 3 4 3 4 3 4 4 4 5 3, 4 2, 3 3, 4 Scales between supranasals 4 11 1 3 2 3 2 3 2, 3 1 3 2 4 3 5 2, 3 Supralabial scales 9 12 9 11 11 13 9 11 10 11 9 10 9 12 10 13 8 11 Infralabial sacles 9 11 9 12 10 11 9 11 9 11 8 10 9 11 10 or 11 8 10 Dorsal scales 17 20 11 21 14 15 11 12 18 27 10 14 8 14 20 22 11 15 Ventral scales 9 12 6 10 8 9 5 7 11 15 6 7 5 7 15 or 16 6 8 Lamellar formula on forefoot 3444, 4554 3333, 3433 Lamellar formula on hindfoot 4555 3333, 3444 3454, 4554 4555, 5665 3444, 4444, 3343 3444 3444(3), 444(5)4 4444 44(5)4(5)4 44(5)4(5)4 4554, 3455 3444, 3444 33(4)3(4)3 3444(3) 4555 3444, 3333 Subdigital lamellae on first finger 5 3 4 4 5 4 5 4 5 4 6 4 5 3 4 Subdigital lamellae on first toe 5 3 4 5 6 4 5 5 4 6 4 6 4 5 3 4 Precloacal and femoral pore series separate (1) or continuous (0) 0 0 0 0 0 0 0 0 0 Precloacal and femoral pores 18 21 17 25 24 26 20 24 10 13 16 27 0 20 18 21 19 22 Cloacal spurs on each side 1 1 1 1 1 2 1 2 1 1 1 2 Subcaudals enlarged, plate-like (1) or not (0) 0 0 0 0 0 0 0 0 0 Dark postorbital stripe present (1) or absent (0) 1 1 0 1 1 1 1 1 1 Light postocular or trunk spots (1) or absent (0) 1 1 1 0 1 1 0 1 0 Dark dorsolateral stripe on trunk (1) or not (0) 1 0 0 1 1 1 0 or 1 0 1 continued on the next page A NEW HEMIPHYLLODACTYLUS FROM CHINA Zootaxa 3974 (3) 2015 Magnolia Press 385
TABLE 4. (Continued) banaensis chiangmaiensis dushanensis jinpingensis kizirianis longlingensis yunnanensis zugi changningensis sp. nov. Dorsal pattern unicolor (1) or not (0) 0 0 1 0 0 0 0 0 0 Dark dorsal transverse blotches (1) or not (0) 1 1 0 1 1 1 1 0 1 Longitudinal series of white (1) or yellow or red (0) dorsal spots 0 0 / / / 1 / 0 / Postsacral mark brown or orange (2), outer edge yellow or red (1), 0 2 2 2 2 2 2 0 2 outer edge red (0) Postsacral mark lacking anterior arms (1) or arms present (0) 1 0 1 1 0 1 1 0 0 Caecum pigmented (1) or not (0) 0 1 0 0 0 0 0 0 0 Gonads pigmented (1) or not (0) 0 1 0 0 0 0 0 0 0 TrunkL/SVL 0.44 0.49 0.46 0.56 0.45 0.49 0.47 0.51 0.45 0.55 0.47 0.52 048 0.53 0.50 0.56 0.46 0.51 HeadL/SVL 0.22 0.24 0.25 0.43 0.22 0.23 0.23 0.24 0.17 0.18 0.22 0.24 0.22 0.23 0.21 0.24 0.22 0.25 HeadW/SVL 0.15 0.16 0.18 0.23 0.17 0.18 0.17 0.17 0.18 0.17 0.19 0.17 0.19 0.17 0.18 0.17 0.18 HeadW/HeadL 0.65 0.7 0.41 0.8 0.73 0.80 0.73 0.76 0.93 1.10 0.75 0.80 0.76 0.81 0.74 0.79 0.72 0.80 SnEye/HeadL 0.34 0.45 0.23 0.49 0.42 0.46 0.41 0.43 0.56 0.62 0.42 0.45 0.41 0.47 0.43 0.49 0.41 0.49 NarEye/HeadL 0.25 0.32 0.17 0.33 0.30 0.35 0.29 0.32 0.45 0.48 0.29 0.34 0.30 0.33 0.29 0.39 0.30 0.37 EyeD/HeadL 0.22 0.25 0.13 0.24 0.24 0.25 0.22 0.26 0.36 0.39 0.22 0.25 0.20 0.24 / 0.21 0.25 SnW/HeadL 0.13 0.17 0.08 0.23 0.14 0.15 0.13 0.15 0.19 0.23 0.12 0.14 0.13 0.15 0.16 0.17 0.12 0.16 EyeD/NarEye 0.71 0.84 0.68 0.81 0.76 0.82 0.71 0.83 0.79 0.83 0.66 0.82 0.64 0.72 / 0.61 0.77 SnW/HeadW 0.2 0.24 0.17 0.32 0.19 0.18 0.21 0.19 0.21 0.15 0.18 0.15 0.19 0.21 0.23 0.16 0.20 386 Zootaxa 3974 (3) 2015 Magnolia Press GUO ET AL.
TABLE 5. Measurements (in mm) and proportions of the type series of Hemiphyllodactylus changningensis sp. nov. from Changning County, Yunnan Province, China (for abbreviations see Materials and Methods). Holotype Paratypes h00315 h00328 h00331 h00334 Sex male male male male female female female female female female h00321 h00323 h00325 h00326 h00335 h00349 SVL 39.8 40.1 40.0 39.1 40.8 42.8 43.8 36.7 38.3 36.3 TaL (*regenerated, **tail tip lost) 32.7 17.5 - - 5.9+20.1* 15.8* - 22.1** 31.6 27.3 TrunKL 18.7 19.5 20.1 19.1 19.9 19.6 22.5 17.9 19.0 17.4 EyeD 2.1 2.2 2.0 2.2 2.3 2.2 2.3 1.9 2.0 1.7 HeadL 8.9 8.9 9.0 9.2 10.0 9.8 10.1 8.7 8.9 8.2 HeadW 7.1 6.9 7.0 6.8 7.5 7.3 7.3 6.6 6.6 6.1 Nare-eye length (NarEye) 2.8 3.1 3.3 3.0 3.0 3.4 3.3 3.0 2.9 2.8 Snout-eye length (SnEye) 4.4 4.1 3.9 4.1 4.1 4.5 4.3 4.1 3.7 3.5 Internarial distance (SnW) 1.4 1.2 1.4 1.3 1.2 1.3 1.3 1.2 1.1 1.1 Ear opening distance (EarD) 0.7 0.4 0.5 0.4 0.6 0.4 0.5 0.6 0.5 0.4 EarD/EyeD 0.33 0.18 0.25 0.18 0.26 0.18 0.22 0.32 0.25 0.24 TrunKL/SVL 0.47 0.49 0.50 0.49 0.49 0.46 0.51 0.49 0.50 0.48 HeadL/SVL 0.22 0.22 0.23 0.24 0.25 0.23 0.23 0.24 0.23 0.23 HeadW/SVL 0.18 0.17 0.18 0.17 0.18 0.17 0.17 0.18 0.17 0.17 HeadW/HeadL 0.80 0.78 0.78 0.74 0.75 0.74 0.72 0.76 0.74 0.74 SnEye/HeadL 0.49 0.46 0.43 0.45 0.41 0.46 0.43 0.47 0.42 0.43 NarEye/HeadL 0.31 0.35 0.37 0.33 0.30 0.35 0.33 0.34 0.33 0.34 EyeD/HeadL 0.24 0.25 0.22 0.24 0.23 0.22 0.23 0.22 0.22 0.21 SnW/HeadL 0.16 0.13 0.16 0.14 0.12 0.13 0.13 0.14 0.12 0.13 EyeD/NarEye 0.75 0.71 0.61 0.73 0.77 0.65 0.70 0.63 0.69 0.61 SnW/HeadW 0.20 0.17 0.20 0.19 0.16 0.18 0.18 0.18 0.17 0.18 A NEW HEMIPHYLLODACTYLUS FROM CHINA Zootaxa 3974 (3) 2015 Magnolia Press 387
TABLE 6. Scalation of the type series of Hemiphyllodactylus changningensis sp. nov. from Changning County, Yunnan Province, China. Holotype Paratypes h00315 h00328 h00331 Sex male male male male female female female female female female Dorsal scales 12 15 12 14 11 13 13 12 11 11 Ventral scales 6 7 6 7 7 6 6 7 8 7 Cloacal spurs on each side 1 2 1 2 2 2 1 2 1 1 Circumnasal scales 3 3 4 4 3 3 3 3 4 3 Scales between supranasals 3 3 2 2 2 2 3 2 2 2 Supralabial scales 9/10 9/8 9/10 9/9 10/10 10/- 11/10 10/9 9/9 10/9 Infralabial sacles 9/10 9/8 9/9 9/8 10/10 8/- 10/10 8/10 9/9 9/9 Chin sacles 8 8 8 8 7 7 7 8 8 8 Lamellar formula on forefoot 3333 3443 3343 3443 3443 3443 3443 3333 3443 3333 Lamellar formula on hindfoot 3444 3444 3444 3444 3444 3444 3444 3333 3444 3444 Subdigital lamellae on first finger 3 4 4 4 4 4 3 3 4 3 Subdigital lamellae on first toe 3 4 4 4 4 4 4 4 4 4 Precloacal and femoral pores (pitted scales in females) 20 22 19 22 10 5 7 0 9 10 h00334 h00321 h00323 h00325 h00326 h00335 h00349 388 Zootaxa 3974 (3) 2015 Magnolia Press GUO ET AL.
Acknowledgments We express special appreciation to Yingying Liu, Yubo Lin, Xiaotong Wu and Yanqing Wu for their assistance in lab work. We are grateful to reviewers for valuable comments and language corrections that greatly improved the manuscript. Support for this research was provided by the Priority Academic Program Development of Jiangsu Higher Education Institutions to KZ. References Boulenger, G.A. (1903) Description of new lizards in the collection of the British Museum. Annals and Magazine of Natural History, Series 7, 12 (70), 429 435. http://dx.doi.org/10.1080/00222930308678877 Grismer, L.L., Wood, P.L. Jr., Anuar, S., Muin., M.A., Quah, E.H., McGuire, J.A., Brown, R.M., Ngo, V.T. & Pham, H.T. (2013) Integrative taxonomy uncovers high levels of cryptic species diversity in Hemiphyllodactylus Bleeker, 1860 (Squamata: Gekkonidae) and the description of a new species from Peninsular Malaysia. Zoological Journal of the Linnean Society, 169, 849 880. http://dx.doi.org/10.1111/zoj.12064 Grismer, L.L., Wood, P.L. Jr. & Cota, M. (2014) A new species of Hemiphyllodactylus Bleeker, 1860 (Squamata: Gekkonidae) from northwestern Thailand. Zootaxa, 3760 (1), 67 78. http://dx.doi.org/10.11646/zootaxa.3760.1.4 Lanfear, R., Calcott, B., Ho, S.Y. & Guindon, S. (2012) PartitionFinder: combined selection of partitioning schemes and substitution models for phylogenetic analyses. Molecular Biology and Evolution, 29 (6), 1695 1701. http://dx.doi.org/10.1093/molbev/mss020 Librado, P. & Rozas, J. (2009) DnaSP 5.10. 00: A software for comprehensive analysis of DNA polymorphism data. Bioinformatics, 25, 1451 1452. http://dx.doi.org/10.1093/bioinformatics/btp187 Mayrose, I., Friedman, N. & Pupko, T. (2005) A Gamma mixture model better accounts for among site rate heterogeneity. Bioinformatics, 21 (2), 151 158. http://dx.doi.org/10.1093/bioinformatics/bti1125 Miller, M., Pfeiffer, W., & Schwartz, T. (2010) Creating the CIPRES Science Gateway for inference of large phylogenetic trees. Gateway Computing Environments Workshop (GCE), IEEE, 2010, 1 8. http://dx.doi.org/10.1016/j.ympev.2005.10.001 Ngo, V.T., Grismer, L.L., Pham, H.T. & Wood, P.L. Jr. (2014) A new species of Hemiphyllodactylus Bleeker, 1860 (Squamata: Gekkonidae) from Ba Na Nui Chua Nature Reserve, Central Vietnam. Zootaxa, 3760 (4), 539 552. http://dx.doi.org/10.11646/zootaxa.3760.4.3 Nguyen, T.Q., Lehmann, T., Le, M.D., Duong, H.T., Bonkowski, M. & Ziegler, T. (2013) A new species of Hemiphyllodactylus (Reptilia: Gekkonidae) from northern Vietnam. Zootaxa, 3736 (1), 89 98. http://dx.doi.org/10.11646/zootaxa.3736.1.5 Nguyen, T.Q., Botov, A., Le, M.D., Nophaseud, L., Zug, G., Bonkowski, M. & Ziegler, T. (2014) A new species of Hemiphyllodactylus (Reptilia: Gekkonidae) from northern Laos. Zootaxa, 3827 (1), 45 56. http://dx.doi.org/10.11646/zootaxa.3827.1.4 Nylander, J.A.A. (2004) MrModeltest 2.2. Evolutionary Biology Centre, Uppsala University: Computer program and documentation distributed by the author. Puillandre, N., Lambert, A., Brouillet, S. & Achaz, G. (2012) ABGD, Automatic Barcode Gap Discovery for primary species delimitation. Molecular Ecology, 21, 1864 1877. http://dx.doi.org/10.1111/j.1365-294x.2011.05239.x Ronquist, F., Huelsenbeck, J.P. & van der Mark, P. (2005) MrBayes 3.1 Manual. Distributed with the program by the authors. Smith, M.A. (1935) The Fauna of British India, including Ceylon and Burma. Reptilia and Amphibia. Vol. II. Sauria. Taylor and Francis, London, 440 pp. Stamatakis, A. (2006a) RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics, 22 (21), 2688 2690. Stamatakis, A. (2006b) Phylogenetic models of rate heterogeneity: a high performance computing perspective. Parallel and Distributed Processing Symposium, 2006. IPDPS 2006. 20th International, IEEE, 2006, 1 8. Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., & Kumar, S. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28, 2731 2739. http://dx.doi.org/10.1093/molbev/msr121 Zhou, K.Y., Liu, Y.Z. & Yang, G.P. (1981) Three new subspecies of Hemiphyllodactylus yunnanensis (Boulenger) from China (Lacertiformes: Gekkonidae). Acta Zootaxonomica Sinica, 6, 202 209. [English translation by H. Ota (1996) in A NEW HEMIPHYLLODACTYLUS FROM CHINA Zootaxa 3974 (3) 2015 Magnolia Press 389
Smithsonian Herpetological Information Services, 110, 1 8, pl. 1.] Zug, G.R. (2010) Speciation and dispersal in a low diversity taxon: the Slender Geckos Hemiphyllodactylus (Reptilia, Gekkonidae). Smithsonian Contributions to Zoology, 631, 1 70. http://dx.doi.org/10.5479/si.00810282.631 APPENDIX. Specimens examined. Hemiphyllodactylus yunnanensis: China: Yunnan: h00567, h00574, h00616 00617. H. longlingensis: China: Yunnan: h00188 00189, h00192 00193, h00201, h00204, h00207, h00198, h00213, h00216. H. jinpingensis: China: Yunnan: h00366, h00371, h00377, h00383. H. dushanensis: China: Guizhou: h00173 00174, h00178 00179. 390 Zootaxa 3974 (3) 2015 Magnolia Press GUO ET AL.