DNA barcoding of Vietnamese bent-toed geckos (Squamata: Gekkonidae: Cyrtodactylus) and the description of a new species Author Nguyen, Sang Ngoc Author Yang, Jun-Xiao Author Le, Thanh-Ngan Thi Author Nguyen, Luan Thanh Author Orlov, Nikolai L. Author Hoang, Chung Van Author Nguyen, Truong Quang Author Jin, Jie-Qiong Author Rao, Ding-Qi Author Hoang, Thao Ngoc Author Che, Jing Author Murphy, Robert W. Author Zhang, Ya-Ping text Zootaxa 2014 3784 1 journal volume 46129 10.11646/zootaxa.3784.1.2 e2720913-7963-4303-bff2-e11872684c54 1175-5326 230341 1DF6FF6B-D761-46C4-B3CE-F94DA3DE156E Cyrtodactylus puhuensis sp. nov. Pù Hu bent-toed gecko ( Figs. 3–6 ) Holotype . KIZ 11665, an adult male from Pù Hu Nature Reserve, Thanh Hoa Province, Vietnam , 20° 33’ N , 104° 53’ E , 638 m , collected on 21 August 2011 by Hoang Van Chung, Yang Jun Xiao and Nguyen Ngoc Sang. Diagnosis. Medium sized, snout-vent length 79.24 mm ; body moderately robust; tail 82.59 mm , longer than snout-vent length, with a series of medium enlarged transversal subcaudals; a series of medium enlarged femoral scales continue from precloacal scales; femoral scales without pores; males with five precloacal pores; supralabials 8; infralabials 10; narrow subdigital lamellae on fourth toe 23; ventral scales 36; and dorsal tubercles smooth or slightly keeled. Description of holotype . In addition to diagnosis. Head relatively depressed, height 9.53 mm , width 15.00 mm (height/width ratio 0.64), distinct from neck; supralabials 8, continuous with a row of small scales extending to the corner of the mouth, first contacting nostril; infralabials 10; supra- and infralabials nearest to rostral and mental largest in size; rostral large, width 3.27 mm , height 2.40 mm , vertical sunken groove in middle, contacting nostril; two internasals contact each other; mental nearly triangular, posteriorly contacting two large postmentals that contact one another and first infralabials ( Fig. 5 C–E); gular scales much smaller than ventrals; head scales small, granular, with scattered tubercles in occipital and temporal areas; eye relatively large, 5.85 mm (eye/head width ratio 0.61), pupil vertical; supraciliary scales large, cover the eye anteriorly, superiorly, and posteriorly; ear opening oval, 1.90 mm , obliquely oriented, much smaller than eyes (ear/eye ratio 0.32); eye-nostril distance 7.00 mm, eyesnout distance 8.83 mm , eye-ear distance 6.27 mm , distance between eyes 8.06 mm , distance between nostrils 2.40 mm . Body elongate, trunk length 35.73 mm , with weak ventral-lateral folds; dorsal scales small, granular, not homogeneous; dorsal and lateral tubercles round, smooth or slightly keeled, not in rows; dorsal tubercles between thighs and base of tail conical and keeled; ventral scales fairly large, smooth, and flattened, about 36 midbody transversal ventrals; dorsal surface of limbs with granular scales and slightly keeled conical tubercles; basal subdigital lamellae narrow and irregular, largest in joints of digits; subdigital lamellae well developed at distal phalanx, as broad as the digit. Subdigital lamellae under fourth finger 18 and toe 23; digits unwebbed; forearm length 13.40 mm , crus length 16.00 mm. Precloacal scales large, flattened, and imbricate; about 8 large imbricate scales under a series of 8 precloacal scales arranged in Λ-shape that bears five precloacal scales with pores; a series of moderately enlarged femoral scales on each side of thigh extend from the eight precloacal scales ( Fig. 5 A); precloacal groove and femoral pores absent; tail round in cross-section, 82.59 mm in length, longer than snout-vent length; transversal subcaudals medium size, about a half size of tail width ( Fig. 5 B); dorsum of proximal tail with conical tubercles extending down to the base tail. Color in life ( Fig. 4 ). Dorsal and lateral background of body chocolate-black; dorsum with faint and irregular pale-yellowish transverse bands; dorsal tubercles at the transverse bands yellow to white, others colored as background; flanks with yellow to white tubercles; head dark brown with yellowish pattern; eyelids yellow; eye brown, with black vertical pupil; nuchal band chocolate-black, interrupted at nape; upper surface of legs and digits dark brown with yellow to white bands or blotches; unregenerated tail with seven black and white to yellowish alternating rings; white rings much more narrow than black; lower surfaces of flanks and legs white. Color in preservative ( Fig. 6 A, B). In preservative, color slightly faded and pattern unchanged; body background black or dark brown, dorsal transverse bands white; dorsum and flanks with white tubercles. DNA barcode : GenBank Accession No. KF929529 . Comparisons. The new species was compared to other congeners in Indochina by using the following morphological characters: precloacal pores, subcaudal scales, ventral scales, continuous medium enlarged femoral scales, dorsal tubercles, dorsal pattern, nuchal band, and number of digital lamellae under the 4th toe. Cyrtodactylus puhuensis sp. nov. differs from Vietnamese congeners, from C. bidoupimontis Nazarov, Poyarkov, Orlov, Phung, Nguyen, Hoang & Ziegler 2012 , C. bugiamapensis Nazarov, Poyarkov, Orlov, Phung, Nguyen, Hoang & Ziegler 2012 , C. cattienensis Geissler, Nazarov, Orlov, Böhme, Phung, Nguyen & Ziegler 2009 , C. cryptus Heidrich, Rösler, Vu, Böhme & Ziegler 2007 , C. dati Ngo 2013 , C. huynhi Ngo & Bauer 2008 , C. irregularis ( Smith 1921b , 1935 ), C. pseudoquadrivirgatus Rösler, Nguyen, Vu, Ngo & Ziegler 2008 , and C. ziegleri Nazarov, Orlov, Nguyen & Ho 2008 , in having a series of moderately enlarged subcaudal scales and continuous precloacal-femoral scales. The new species differs from C. martini Ngo 2011 in having a series of transversal and enlarged subcaudal scales ( versus small or medium ones, not transversely enlarged) and a higher number of precloacal pores (5 versus 4 in all four males from the type series). By having series of moderately enlarged femoral scales continuous to the precloacal scales, C. puhuensis sp. nov. differs from C. badenensis Nguyen, Orlov & Darevsky 2006 , C. caovansungi Orlov, Nguyen, Nazarov, Ananjeva & Nguyen 2007 , C. cucphuongensis Ngo & Onn 2011 , C. eisenmanae Ngo 2008 , C. grismeri Ngo 2008 , C. huongsonensis Luu, Nguyen, Do & Ziegler 2011 , C. kingsadai Ziegler, Phung, Le & Nguyen 2013 , C. nigriocularis Nguyen, Orlov & Darevsky 2006 , and C. takouensis Ngo & Bauer 2008 . In addition, the new species differs from C. badenensis , C. cucphuongensis , C. eisenmanae , C. grismeri , and C. nigriocularis in having 5 precloacal pores ( versus 0, or 0–2 in the last species); from C. huongsonensis and C. kingsadai by absence of femoral pores ( versus 17 or 3–7 femoral pores on both thighs, respectively); and from C. caovansungi and C. bichnganae Ngo & Grismer 2010 by lacking femoral pores ( versus 3, or 9 on each thigh, respectively) and having a less precloacal pores (5 versus 9 or 10, respectively). Our examination of the type , paratype and additional specimens collected from the type locality (Appendix 1) showed C. chauquangensis Hoang, Orlov, Ananjeva, Johns, Hoang & Dau 2007 differed from the new species. First, C. chauquangensis has a uniform transversal butterfly-shaped pattern on dorsum ( Fig. 7 ) while the new species shows faint, thin, and irregular dorsal bands. Second, femoral scale rows in C. chauquangensis are undeveloped; they are the same size as other femoral scales. The new species has series of moderately enlarged femoral scales ( Fig. 5 A). Third, C. chauquangensis has 6–7 precloacal pores, except for topotype CQ526, which has 8 pores. In contrast, the new species has 5 pores only. The new gecko differs from C. condorensis ( Smith 1921a ) by the distribution of precloacal pores (in a chevron versus 4–7 in two juxtaposed rows) and dorsal tubercles (smooth or slightly keeled versus subtrihedral) ( Smith 1935 ; Ngo & Grismer 2012 ); from C. paradoxus ( Darevsky & Szczerbak 1997 ) by having more and differently arranged precloacal pores ( 5 in chevron versus 0–4 in two juxtaposed rows) (Orlov et al. 2007); and from C. thochuensis Ngo & Grismer 2012 by the chevron arrangement of its precloacal pores ( versus in two juxtaposed rows), smooth or slightly keeled dorsal tubercles ( versus strongly keeled), normal sized scales on heel ( versus enlarged), and having the dorsal caudal tubercles extending to the base of the tail only ( versus to about 1/3 the length of the tail). Cyrtodactylus puhuensis sp. nov. differs from C. hontreensis Ngo, Grismer & Grismer 2008 , C. intermedius ( Smith 1917 , 1935 ), and C. phuquocensis Ngo, Grismer & Grismer 2010 in having faint, irregular pale yellowish dorsal bands. In contrast, C. hontreensis has 3 broad bands between limbs and C. intermedius and C. phuquocensis have 4. The new species also differs in its absence of nuchal band, which is always present in the other species, and in having 5 precloacal pores versus 7–8 in C. hontreensis , 8–10 in C. intermedius and 6–8 in C. phuquocensis . The new species differs from C. phongnhakebangensis Ziegler, Rösler, Herrmann & Vu 2003 and C. roesleri Ziegler, Nazarov, Orlov, Nguyen, Vu, Dang, Dinh & Schmitz 2010 in lacking femoral pores ( versus 27–42 and 20– 28, respectively, precloaacal-femoral pores situated as one continuous row). The new gecko differs from C. yangbayensis Ngo & Onn 2010 in possessing fewer precloacal pores (5 versus 6–8), more subdigital lamellae under the fourth toe (23 versus 15–17), medium sized scales on heel ( versus 9–10 enlarged scales), and seven pale rings on the tail ( versus 9–11 irregular bands). FIGURE 4. Holotype of Cyrtodactylus puhuensis sp. nov. in life. FIGURE 5. Holotype of Cyrtodactylus puhuensis sp. nov. Precloacal-femoral area (A), ventral surface of tail (B), and lateral (C), dorsal (D), and ventral (E) views of head. FIGURE 6. Holotype of Cyrtodactylus puhuensis sp. nov. dorsal (A) and ventral views (B) in preservative and ventral surface of right hand (C) and foot (D). FIGURE 7. Cyrtodactylus chauquangensis showing butterfly-shaped pattern on dorsum. For Laotian congeners, Cyrtodactylus puhuensis sp. nov. differs from C. buchardi David, Teynié & Ohler 2004 , C. pageli Schneider, Nguyen, Schmitz, Kingsada, Auer & Ziegler 2011 , and C. wayakonei Nguyen, Kingsada, Rösler, Auer & Ziegler 2010 in having a series of enlarged femoral scales ( versus without enlarged femoral scales). The new species also differs from C. buchardi and C. wayakonei in having enlarged transversal subcaudal scales ( versus small or medium scales, not transversally enlarged); and from C. pageli in lacking a nuchal band ( versus dark nuchal band). Cyrtodactylus puhuensis sp. nov. differs from C. jarujini Ulber 1993 and C. lomyenensis Ngo & Pauwels 2010 in lacking femoral pores (52–54 and 39–40, respectively, continuous precloacal-femoral pores). The new gecko differs from C. interdigitalis Ulber 1993 and C. teyniei David, Nguyen, Schneider & Ziegler 2011 in having fewer precloacal pores (5 versus 14) and also differs from the former by lacking femoral pores ( versus 8–9). From Myanmar congeners, Cyrtodactylus puhuensis sp. nov. differs from C. ayeyarwadyensis Bauer 2003 , C. brevidactylus Bauer 2003 , C. gansi Bauer 2003 , C. mandalayensis Mahony 2009 , C. tamaiensis Mahony 2009 , and C. wakeorum Bauer 2003 , in having moderately enlarged transversal subcaudal scales. The new species also differs from C. ayeyarwadyensis , C, brevidactylus , and C. gansi in having few precloacal pores ( versus 10–28, 8, and 16– 29, respectively); from C. gansi in lacking a shallow precloacal groove in males; and from C. tamaiensis in lacking femoral pores ( versus 40 precloacal-femoral pores in an almost continuous series, Mahony 2009 ). From C. pulchellus Gray and C. rubidus ( Blyth 1861 ) the new species differs in lacking a precloacal groove ( Smith 1935 ; Taylor 1963 ). Cyrtodactylus puhuensis sp. nov. differs from C. aequalis Bauer 2003 , in lacking femoral pores ( versus 3–4 minute femoral pores) and possesing more ventral scales (36 versus 24); from C. annandalei Bauer 2003 , in having less precloacal pores (5 versus 11–12), lacking femoral pores ( versus 10–11), and having a dorsal pattern of faint and irregular pale yellowish bands ( versus six dark bands); from C. chrysopylos Bauer 2003 , in having less precloacal pores (5 versus 10 and a single, pored scale posterior to precloacal series); from C. consobrinoides (Annandale) , in having more ventral scales (36 versus 24–30), lacking a nuchal band, and possessing a dorsal pattern of faint and irregular pale yellowish bands ( versus 6–7 narrow dark cross-bars or transverse markings, Smith 1935 ); from C. feae ( Boulenger 1893 ) , in lacking femoral pores (Bauer 2003), and having a dorsal pattern of faint and irregular pale yellowish bands ( versus four dark cross-bars, Smith 1935 ); from C. oldhami (Theoblad 1876) , in having more precloacal pores (5 versus 0–4), absence of a nuchal band, and a having a dorsal pattern of faint and irregular pale yellowish bands versus white spots) ( Smith 1935 ); from C. peguensis ( Boulenger 1893 ) , in having smaller number of precloacal pores (5 versus 7–9) and continuous series of precloacal-femoral scales ( Smith 1935 ; Taylor 1963 ); from C. russelli Bauer 2003 and C. slowinskii Bauer 2003 , in having less precloacal pores (5 versus 15 and 9, respectively) and lacking femoral pores ( versus 16–19 and 11 pores on each thigh, respectively); and from C. variegatus ( Blyth 1859 ) , in having more ventral scales (36 versus 22) and lacking femoral pores ( versus 32 continuous precloacal-femoral pores ( Smith 1935 ). For Thai congeners, Cyrtodactylus puhuensis sp. nov. differs from C. quadrivirgatus Taylor 1962 and C. angularis ( Smith 1921b ) , in having moderately enlarged subcaudal scales ( versus small; Smith 1935 ; Taylor 1963 ). The new gecko differs from C. phuketensis Sumontha, Pauwels, Kunya, Nitikul, Samphanthamit & Grismer 2012 , in lacking a precloacal groove and femoral pores ( versus 33–36 continuous precloacal-femoral pores). The new species differs from the following species in lacking femoral pores: C. astrum Grismer, Wood, Quah, Anuar, Muin, Sumontha, Ahmad, Bauer, Wangkulangkul, Grismer & Pauwels 2012 , C. auribalteatus Sumontha, Panitvong & Deein 2010 , C. brevipalmatus ( Smith 1923 ) , C. chanhomeae Bauer, Sumontha & Pauwels 2003 , C. dumnuii Bauer, Kunya, Sumontha, Niyomwan, Pauwels, Chanhome & Kunya 2010 , C. erythrops Bauer, Kunya, Sumontha, Niyomwan, Panitvong, Pauwels, Chanhome & Kunya 2009 , C. lekaguli Grismer, Wood, Quah, Anuar, Muin, Sumontha, Ahmad, Bauer, Wangkulangkul, Grismer & Pauwels 2012 , C. marmoratus ( Gray 1831 ) , and C. trigroides Bauer, Sumontha & Pauwels 2003 ( Smith 1935 ; Taylor 1963 ). Cyrtodactylus puhuensis sp. nov. can be distinguished from C. papilionoides Ulber & Grossmann 1991 , in having a dorsal pattern of faint and irregular pale yellowish bands ( versus serrated, broad, dark bands with pale margins or transversal rows of blotches), and having a continuous series of 36 digital lamellae under the 4th toes ( versus an interrupted series of 12–16 lamellae; Heidrick et al. 2007; Nguyen et al. 2010); from C. consobrinus ( Peters 1871 ) , in having less precloacal pores (5 versus 9–11) and a dorsal pattern of faint and irregular pale yellowish bands ( versus 8 dark, transverse cross-bands, Boulenger 1885 ); from C. sanook Pauwels, Sumontha, Latinne & Grismer 2013 , in having medium enlarged femoral scale rows ( versus well-developed); from C. sumonthai Bauer, Pauwels & Chanhome 2002 , in having more precloacal pores (5 versus 2 minute pores), and a series of moderately enlarged femoral scales ( versus the absence of enlarged femoral scales); from C. surin Chan-ard & Makchai 2011 , in having more ventral scales (36 versus 25) and smooth or slightly keeled dorsal tubercles ( versus very large, trihedral, and keeled); and from C. thirakhupti Pauwels, Bauer, Sumontha & Chanhome 2004 , in having fewer precloacal pores (5 versus 8–9) and lacking femoral pores ( versus 5–7). Other Thai species, including C. interdigitalis , C. intermedius , C. jarujini , C. oldhami , C. peguensis , C. pulchellus , and C. variegatus , were compared to the new species as above. Two described congeners occur in Cambodia : C. intermedius (Grismer et al. 2008; Smith 1935 ) and C. pseudoquadrivirgatus ( Stuart et al . 2010 ) . Cyrtodactylus bugiamapensis should occur in Cambodia because its habitat extends continuously from the type locality to the border between Vietnam and Cambodia (see also Nazarov et al. 2012 ). These three species were compared above to the new species. Etymology . The specific epithet puhuensis refers to the type locality, Pù Hu Nature Reserve, where the new species was discovered. Distribution and Habitat . Cyrtodactylus puhuensis sp. nov. is known only from the Pù Hu Nature Reserve, in northwestern Thanh Hóa Province, northern Vietnam ( Fig. 2 ). The single known specimen of this new gecko was found at night on a large tree (diameter about 1.5 m ), at about 2.5 m above the forest floor. The habitat was evergreen forest intermixed with bamboo. Limestone was not observed in the area and the recorded elevation was 638 m .