Zoosyst. Evol. 101 (1) 2025, 245-255 | DOI 10.3897/zse.101.138964 eee BERLIN A new species of Hemiphyllodactylus (Squamata, Gekkonidae) from western Yunnan, China Hongxin Zhou! *", Xiaohan Wang"", Liangwei Cui', Dongru Zhang?, Ziqi Shen*, Xiuyan Li*, Shuo Liu**, Dingqi Rao? 1 Key Laboratory for Forest Resources Conservation and Utilization in the Southwest Mountains of China, Southwest Forestry University, Kunming, Yunnan 650224, China 2 Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan 650201, China 3 School of Biological Science and Technology, Liupanshui Normal University, Liupanshui, Guizhou 553004, China 4 Kunming Natural History Museum of Zoology, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan 650223, China https://zoobank. org/5C83D016-F 105-4D97-83 17-89 16B792A9AS& Corresponding authors: Dingqi Rao (raodq@mail.kiz.ac.cn); Shuo Liu (liushuo@mail.kiz.ac.cn) Academic editor: Justin Bernstein # Received 10 October 2024 # Accepted 29 December 2024 @ Published 6 February 2025 Abstract A new species of gekkonid, Hemiphyllodactylus laowozhenensis sp. nov., is described based on three specimens from Lushui City, Yunnan, China. The new species can be distinguished from its congeners by a significant genetic divergence of greater than 4.8% in the mitochondrial ND2 gene and a combination of the following characters: maximum SVL of 38.5 mm; 10 or 11 chin scales; enlarged postmentals; 6 circumnasal scales; 2 or 3 internasal scales; 8 or 9 supralabial scales; 8 or 9 infralabial scales; 15 or 16 dor- sal scales; 9 ventral scales; a manual lamellar formula of 3-4-4-4 and a pedal lamellar formula of 3(4)-4-4-4; 26-28 precloacal and femoral pore-bearing scales contiguous in males; 1—3 cloacal spur on each side; a dark postorbital stripe extending to at least base of neck; dorsolateral light-colored spots on trunk present; dark ventrolateral stripe on trunk present; dark dorsal transverse blotches present; and a dark and brown postsacral mark bearing anteriorly projecting arms. Key Words Hemiphyllodactylus laowozhenensis sp. nov., integrative taxonomy, Lushui City, molecular phylogeny, slender gecko Introduction The genus Hemiphyllodactylus currently comprises a total of 59 species (Uetz et al. 2024), primarily distrib- uted in South Asia, Southeast Asia, and the Indo-Pacific islands (Zug 2010; Grismer et al. 2013, 2018; Agarwal et al. 2019; Eliades et al. 2019; Agung et al. 2021, 2022; Luu et al. 2023, 2024). It was previously considered a “low-diversity group” (Zug, 2010). However, an integra- tive taxonomic study by Grismer et al. (2013) revealed that the diversity of Hemiphyllodactylus species 1s much * These authors contributed equally to this work. greater than previously thought. Subsequently, the num- ber of Hemiphyllodactylus species entered a period of rapid increase (Uetz et al. 2024). Currently, the majority of newly discovered species of Hemiphyllodactylus from Southeast Asia, while the number of species published within China is significantly lower (Uetz et al. 2024). Considering the similar climates between southern China and Southeast Asia, it 1s plau- sible that a similar pattern may exist in China (Grismer et al. 2018). Therefore, a large-scale survey is needed in southern China. Yunnan Province, as the most diverse Copyright Zhou, H. et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 246 province for Hemiphyllodactylus species in China (8/15), may still harbor numerous cryptic species (Agung et al. 2022; Zhou et al. 2024a, b). During a recent field survey in Laowo Town, Lushui City, Nujiang Prefecture, Yunnan Province, we collected three specimens of Hemiphyllodactylus. The phylogenetic topology resulting from both Maximum Likelihood (ML) and Bayesian Inference (BI) analyses based on ND2 se- quences indicates that we have reassigned the collected specimens to a new lineage within Clade 3 of Agung et al. (2022), with Hemiphyllodactylus longlingensis as their sister group. Additionally, they exhibit significant mor- phological differences from other known species. There- fore, we hereby describe them as a new species. Materials and methods Specimens and morphology Three specimens were collected from Laowo Town (Fig. 1), Lushui City, Yunnan Province, China on August 2024. We took samples of liver tissue from each individual 96° O'0"E 99° Q'0"E 27° 0' 0"N 24° 0'0"N H. changningensis H.. gengmaensis H. jinpingensis H. laowozhenensis sp. nov. H. longlingensis H. simaoensis 21° 0° 0°N H., yanshanensis H. yunnansis & | A & ¢ 8 * 2 * H. zhutangxiangensis 96° 0' 0"E 99° O° 0°E Zhou, H. et al.: Hemiphyllodactylus laowozhenensis sp. nov. from Yunnan specimen and preserved them in 95% ethanol for genetic analysis. The specimens were then stored in 80% ethanol for morphological measurements. All specimens were de- posited in Kunming Institute of Zoology (KIZ), Chinese Academy of Sciences (CAS). The raw lengths of mensural data and meristic charac- ters were taken with a digital calipers to the nearest 0.01 mm under a dissecting microscope (Jiangnan XTB-01) following Zug (2010), Grismer et al. (2013), and Agung et al. (2021, 2022): snout—vent length (SVL), taken from the tip of the snout to the vent; tail length (TL), taken from the vent to the tip of the tail; trunk length (TrunkL), taken from the posterior margin of the forelimb at its in- sertion point on the body to the anterior margin of the hind limb at its insertion point on the body; head length (HL), measured from the posterior margin of the retroar- ticular process of the lower jaw to the tip of the snout; head width (HW), measured at the angle of the jaws; eye diameter (ED), the greatest horizontal diameter of the eyeball; snout—-eye length (SnEye), measured from anterior-most margin of the eyeball to the tip of snout; nares—eye length (NarEye), measured from the anterior margin of the eyeball to the posterior margin of the ex- 102° 0'0°E 105° 0°0°E 0’ O"N 27 24° 0° 0"N 21° 0'0"N |e ee (ee ] 102° 0'0°E 105° 0°0°E Figure 1. Distribution map of the genus Hemiphyllodactylus in Yunnan Province, China. zse.pensoft.net Zoosyst. Evol. 101 (1) 2025, 245-255 ternal nares; and snout width (SnW), measured between the external nares; chin scales (Chin); circumnasal scales (CN); internasal scales (IS); supralabial (SL); infralabial (IL); ventral scales (VS); dorsal scales (DS); subdigital lamellae wider than long on first finger (SLIF) and toe (SLIT); subdigital lamellae formula determined as num- ber of U-shaped digital pads on digits II-V of hands and feet; number of femoroprecloacal pores; and number of cloacal spurs (CloacS) on each side of hemipenial swell- ing; We also noted coloration and pattern on the dorsum: presence or absence of dark postorbital stripe; presence or absence of dorsolateral light-colored spots on trunk; presence or absence of dark dorsolateral and ventrolat- eral stripe on trunk; presence or absence of dark dorsal transverse blotches; presence or absence of dark reticu- late pattern on dorsum; presence or absence of postsacral marking anteriorly projecting arms. For the statistical analysis of morphological character- istics, we conducted a statistical analysis on the morpho- logical characteristics within clade 3 (H. zhutangxiangen- sis, H. changningensis, H. gengmaensis, H. Jinghongensis and H. laowozhenensis sp. nov.). due to the lack of de- tailed traits of H. Jonglingensis and the availability of only one specimen for H. zalonicus, both H. longlingensis and H. zalonicus were excluded from the statistical analysis. 247 First, we utilized the GroupStruct package in R to adjust the measurement characteristics following Agung et al. (2021, 2022), Luu et al. (2023, 2024), Zhou et al. (2024b); then, we employed Principal Component Analysis (PCA) based on the corrected measurement values and meristic features in the ADEGENET package in R (R Core Team 2021); Finally, we utilized ggplot 2 to visualize the results. Phylogenetic sampling and analyses 25 sequences of genus Hemiphyllodactylus were obtained from GenBank for phylogenetic analyses. GenBank ac- cession numbers of all sequences are shown in Table 1. Genomic DNA was extracted from liver tissue sam- ples, using Trelief Hi-Pure Animal Genomic DNA Kit from Beying Tsingke Biotechnology Co., Ltd. We ampli- fied and sequenced the complete mitochondrial NADH dehydrogenase subunit 2 gene (ND2) of three specimens, totaling 1,038 bp using the primers L4437b and H5934 (Macey etal. 1997). The protocol for polymerase chain re- action (PCR) amplifications followed Agung et al. (2021). DNA sequences were aligned in MEGA 11 (Tamura et al. 2021) by the Clustal W algorithm with default pa- rameters (Thompson et al. 1994). The dataset was tested Table 1. List of specimens used for phylogenetic analyses in this study. Species GenBank no. Locality Voucher information H. harterti KF219760 Bukit Larut, Malaysia LSUHC 10383 H. harterti KF219761 Bukit Larut, Malaysia LSUHC 10384 H. zhutangxiangensis MW962150 Zhutangxiang town, Lancang Lahu, Yunnan, China KIZ061163 H. zhutangxiangensis MW962151 Zhutangxiang town, Lancang Lahu, Yunnan, China KIZ061164 H. zhutangxiangensis MW962152 Zhutangxiang town, Lancang Lahu, Yunnan, China KIZ061165 H. zhutangxiangensis MW962153 Zhutangxiang town, Lancang Lahu, Yunnan, China KIZ061166 H. zhutangxiangensis MW962154 Zhutangxiang town, Lancang Lahu, Yunnan, China KIZ061167 H. longlingensis FJ971045 Longyang District, Baoshan, Yunnan, China isolate N30 H. longlingensis FJ971046 Longyang District, Baoshan, Yunnan, China NJNUhOO104 H. longlingensis FJ971047 Longyang District, Baoshan, Yunnan, China isolate N32 H. longlingensis FJ971048 Longyang District, Baoshan, Yunnan, China isolate N33 H. zalonicus MW039150 Zalon Taung National Forest, Ban Mauk, Sagaing, Myanmar ZMMU R 16635 H. changningensis ON676073 Yongde County, Yunnan, China KIZ 061990 H. changningensis ON676074 Yongde County, Yunnan, China KIZ 061991 H. changningensis ON676075 Yongde County, Yunnan, China KIZ 061992 H. changningensis ON676076 Yongde County, Yunnan, China KIZ 061993 H. changningensis ON676077 Yongde County, Yunnan, China KIZ 061994 H. changningensis ON676078 Yongde County, Yunnan, China KIZ 061995 H. changningensis ON676079 Yongde County, Yunnan, China KIZ 061996 H. changningensis ON676080 Yongde County, Yunnan, China KIZ 061997 H. gengmaensis PP540023 Gengma Dai and Wa Autonomous County, Yunnan, China 2014002297 H. gengmaensis PP540024 Gengma Dai and Wa Autonomous County, Yunnan, China 2014002298 H. gengmaensis PP540022 Gengma Dai and Wa Autonomous County, Yunnan, China 2014002299 H. gengmaensis PP540021 Gengma Dai and Wa Autonomous County, Yunnan, China 2014002300 H. gengmaensis PP540025 Gengma Dai and Wa Autonomous County, Yunnan, China 2014002302 H. laowozhenensis sp. nov. PQ819178 Laowo town, Lushui City, Yunnan, China LW2024R004 H. laowozhenensis sp. nov. PQ819179 Laowo town, Lushui City, Yunnan, China LW2024R005 H. laowozhenensis sp. nov. PQ819180 Laowo town, Lushui City, Yunnan, China LW2024R006 zse.pensoft.net 248 for the best-fit substitution model of ML and BI respec- tively in ModelFinder v. 2.2.0 (Kalyaanamoorthy et al. 2017) with Akaike and Bayesian Information Criteria. A maximum-likelihood (ML) analysis was conducted using TN+F+Gé4 as the best-fit substitution model. We applied 1,000 bootstrap pseudoreplicates with the ultrafast boot- strap approximation algorithm (UFBoot), where nodes having values of 95 and above were considered highly supported (Minh et al. 2013). A Bayesian-inference (BI) analysis was conducted using GTR+F+G4 model follow- ing the methods by Agung et al. (2021). We ran two inde- pendent Markov Chain Monte Carlo (MCMC) analyses with four chains in each analysis (three hot and one cold). The MCMC analyses were implemented with 10 million generations, sampled every 5000 generations with the first 10% sampled discarded as burn-in. Nodes with Bayesian posterior probabilities (BPP) of 0.95 and above were con- sidered highly supported (Huelsenbeck et al. 2001; Wil- cox et al. 2002). Uncorrected pairwise divergences were calculated using MEGA 11 (Tamura et al. 2021). Species delineation Our species definition and species delimitation methods are based on Agung et al. (2022): first, new lineages not clustered within the named species lineages in phyloge- netic topology were marked as potential new species; sec- ond, uncorrected pairwise genetic distances among the new lineages with either known species or other putative species lineages were measured, with a 3.0% difference in mtDNA ND2 considered minimal to define a potential new species; third, those lineages showing >3.0% genetic Zhou, H. et al.: Hemiphyllodactylus laowozhenensis sp. nov. from Yunnan difference from their nearest relatives were examined for morphological distinctiveness from closely related spe- cies. If this third criterion was also met, the lineage was considered a confirmed new species. Results Our phylogenetic topology is consistent with previously published studies (Grismer et al. 2013; Grismer et al. 2020; Agung et al. 2021, 2022; Yushchenko et al. 2023). The ML and BI analyses (Fig. 2) recovered the Laowo town popu- lations to be nested within clade 3 of Agung et al. (2022). The phylogenetic analyses recovered a strongly-supported monophyletic lineage containing all Hemiphyllodactylus samples used in this study (ML 100/BI 1.00). Uncorrect- ed genetic P-distances among and within the ND2 gene of the newly recovered OTUs (within clades 3) are presented in Table 2. The mean percentage of uncorrected pairwise distance between the Laowo Town specimens and its sister species H. longlingensis is 4.8% (Table 2). The clustering of species in the PCA of Clade 3 showed the first two principal components (PC1 and PC2) recovered 65.45% of the vari- ation in the normalized morphometric and meristic dataset (Fig. 3, Suppl. material 1). PC1 represented 50.65% of the dataset variability, with the heaviest loadings for DS, VS, Chin and CN, while PC2 represented 14.80% of the dataset variability, with the heaviest loadings for HW, Chin, DS and CN. The PCA recovered Hemiphyllodactylus laowozhenen- sis Sp. nov. to be widely separated from other species. Based on the molecular and morphological evidence, we hereby describe these specimens from Laowo Town as a new species, Hemiphyllodactylus laowozhenensis sp. nov. Hemiphyllodactylus longlingensis FJ971045 95/1 Hemiphyllodactylus longlingensis FJ971046 Hemiphyllodactylus longlingensis FJ971047 100/1 Hemiphyllodactylus longlingensis FJ97 1048 LW2024R004 Lw2024R005 HT Jao} an 100/1 LL w2024R006 Hemiphyllodactylus zhutangxiangensis MW962150 Hemiphyllodactylus zhutangxiangensis MW962153 99/1 | Heniphylodactylus zhutangxiangensis MW962152 Hemiphyllodactylus zhutangxiangensis MW962151 Hemiphylodactylus zhutangxiangensis MW962 154 100/1 Clade 3 0.09 Hemiphyllodactylus gengmaensis PP540024 Hemiphylodactylus zalonicus MW039150 Hemiphyllodactylus changningensis ON676073 Hemiphyllodactylus changningensis ON676079 Hemiphyllodactylus changningensis ON676076 Hemiphyltlodactylus changningensis ON676077 Hemiphyllodactylus changningensis ON676074 Hemiphylodactylus changningensis ON676075 Hemiphyllodactylus changningensis ON676078 Hemiphyllodactylus changningensis ON676080 Hemiphyliodactylus gengmaensis PP540021 Hemiphyllodactylus gengmaensis PP540023 Hemiphyllodactylus gengmaensis PP540022 Hemiphyllodactylus gengmaensis PP540025 Hemiphylodactylus harterti KF219760 Aemiphyllodactylus harterti KF219761 Figure 2. Maximum-likelihood consensus tree mitochondrial ND2 gene. Numbers by the nodes indicate ML bootstrap support values and posterior probability values of the BI. zse.pensoft.net Zoosyst. Evol. 101 (1) 2025, 245-255 249 Table 2. The mean percentage of the uncorrected p—distance among the Hemiphyllodactylus species studied based on mitochondrial ND2 gene fragments. Species name (no.) 1 1. H. gengmaensis (5) 2. H. longlingensis (4) 24.4 3. H. zalonicus (2) De? 4. H. zhutangxiangensis (5) 20.3 5. H. changningensis (8) 9.6 6. H. laowozhenensis (3) PC2 (14.80%) PC1 (50.65%) 2 3 4 ) 6 26.9 225 24.2 2a, 19.0 22.0 4.8 27.5 21.8 25.5 H. changningensis H. gengmaensis H. jinghongensis H. laowozhenensis sp. nov. H. zhutangxiangensis El + Ee + Te) Figure 3. PCA of Hemiphyllodactylus species between new species with their related species showing their morphospatial relation- ships along the first two principal components based on their morphological traits (Suppl. material 2). Taxonomic account Hemiphyllodactylus laowozhenensis sp. nov. https://zoobank.org/DE9C8C 1E-CAF 1-4280-BE45-359157B79B39 Fig. 4 Type material. Holotype. * LW2024R006, adult male, collected by Hongxin Zhou, Xiuyan Li, Jiazhong Wang on August 2024 from Laowo Town, Lushui City, Yunnan, China (25.865°N, 99.038°E, at an elevation of 1511 m). Paratypes. * LW2024R004, LW2024R005, adult males, collected at the same locality as the holotype on August 2024. Diagnosis. Hemiphyllodactylus laowozhenensis sp. nov. can be distinguished from its congeners by the com- bination of the following characters: maximum SVL of 38.5 mm; 10 or 11 chin scales; enlarged postmentals; 6 circumnasal scales; 2 or 3 internasal scales; 8 or 9 su- pralabial scales; 8 or 9 infralabial scales; 15 or 16 dorsal scales; 9 ventral scales; a manual lamellar formula of 3-4- 4-4 and a pedal lamellar formula of 3(4)-4-4-4; 26-28 precloacal and femoral pore-bearing scales contiguous in males; 1—3 cloacal spur on each side; a dark postorbit- al stripe extending to at least base of neck; dorsolateral light-colored spots on trunk present; dark ventrolateral stripe on trunk present; dark dorsal transverse blotches present; and a dark and brown postsacral mark bearing anteriorly projecting arms. Description of holotype. Adult male, one longitu- dinal incision on ventral surface used for liver tissue sampling, small-sized (SVL 38.30 mm), and somewhat flattened in body shape; head triangular, elongated (HL/ SVL = 0.25); dorsum of head covered in granular scales, which are relatively small; six circumnasal scales, with the lower two being the rostral and the largest upper la- bial, while the other three are circular; two internasal scales; circular mental scale; ten chin scales touching the internal edges of the infralabials, extending from the juncture of the 2"! and 3” infralabial scales on the left of the mental scale to the same juncture on the right (Chin); scales in the gular region are rounded, non—over- lapping, becoming larger and more ovoid on the venter; snout short and narrow (SnW = 1.57 mm; SnW/HL = 0.16); small eyes (ED = 2.48 mm; ED/HL = 0.25); robust body shape (TrunkL/SVL=0.54); granular scales on the dorsum, with 16 scales within one eye diameter; ventral scales are flattened, with 9 scales within one eye diam- eter; granular scales on the limbs; Finger I is vestigial, zse.pensoft.net 250 Zhou, H. et al.: Hemiphyllodactylus laowozhenensis sp. nov. from Yunnan B F Figure 4. Hemiphyllodactylus laowozhenensis sp. nov. A. Holotype, LW2024R006, dorsal view in alcohol; B. Holotype, LW2024R006, ventral view in alcohol; C. The ventral view of the head; black lines indicate chin scales; D, E. In view of lamellae formula counting on fingers and feet I to V; F. Lateral view of basal tail. clawless, and with rectangular subdigital lamellae, while Fingers II—V are well—developed; the proximal subdigi- tal lamellae are undivided and rectangular, while the dis- tal subdigital lamellae are divided, angular, U-shaped, except for the terminal lamellae, which are rounded and undivided; subdigital lamellae count 3-4-4-4 (hand) and 4-4-4-4 (foot); femoral pores and precloacal pores continuous, with a total count 28; Tail long (TL/SVL = 0.90), with dorsal scales larger than those on the body and head, and smaller than the subcaudals; subcaudals are large and flat. Coloration in ethanol. The dorsal surface of head and body is dark-gray; dark stripes extend from the posterior corner of the eye socket to the neck; the back 1s covered with dark and light-colored dorsal transverse blotches; the dorsal surfaces of the limbs are dark; the dorsal surface of tail is brown-gray, with several dark-brown transverse stripes and its ventral surface is orange-red; the ventral surfaces of the head are gray with dark spots; the ventral surfaces of the body are dark. Variation. The primary variations observed in this species pertain to scale counts and morphometric data, which are comprehensively documented in Table 3. Distribution. This species is currently known to be distributed at the type locality Laowo Town, Lushui City of Yunnan Province in China (Fig. 1). Natural history. Hemiphyllodactylus laowozhenensis Sp. nov. was discovered on the walls of houses situated alongside the streets of Laowo Town, where its popula- zse.pensoft.net tion is scant. It coexists in the same region with the Hemi- dactylus aquilonius. Etymology. The scientific name “/aowozhenensis” 1s derived from its type locality Laowo Town, Lushui City in Yunnan province. We suggest Laowo Town Slender Gecko in English and “% 5 #4-F M1 HLEE (Lao Wo Zhén Ban Ye Zhi Ht)” in Chinese. Comparisons. We compared the morphology of Hemiphyllodactylus laowozhenensis against its closely related species, specifically species from clade 3, as in- ferred from the phylogeny we constructed (Table 4). H. laowozhenensis sp. nov. differences from H. geng- maensis by shorter head (HL = 9.71—9.96 versus 10.80- 11.24; HL/SVL = 0.25-0.26 versus 0.26—0.31); shorter SnEye distance (SnEye = 3.43—3.64 versus 3.88—4.52; SnEye/HL = 0.35—0.37 versus 0.36—0.41); thinner SnW distance (SnW = 1.60—1.57 versus 1.60—2.72); smaller eyes (ED/HL=0.22—0.24 versus 0.24—0.25); more chin scales (10 or 11 versus 8 or 9); few supralabial scales (SL=8 or 9 versus 9-11); few lamelar fomnlae hands and foot II-V (hands = 3-4-4-4 versus 5-5(6)-5-4; foot = 3(4)- 4-4-4 versus 5-5-6-5); few SLIF (SLIF = 4 versus 5); few SLIT (SLIT = 4 versus 6); more femoroprecloacal pores (26—28 versus 10-25); dorsolateral light-colored spots on trunk present (versus absent); dark ventrolater- al stripe on trunk present (versus absent); dark reticulate pattern on dorsum absent (versus Yes or Indistinct). H. laowozhenensis sp. nov. differences from H. longlingensis by shorter head (HL/SVL = 0.25-0.26 Zoosyst. Evol. 101 (1) 2025, 245-255 251 Table 3. Mensural (in mm), meristic, color pattern, and proportions of the type series of Hemiphyllodactylus laowozhenensis sp. nov. Character Holotype Paratype Paratype LW2024R006 LW2024R004 LW2024RO005 Sex Male male male SVL 38.30 36.74 38.50 He 34.58 31.90 32.30 TrunkL 20.5 18.76 19.84 HL 9.73 9.71 9.96 HW 6.66 7.41 Ti22 ED 2.48 2.42 2.4 SnEye 3.64 3.43 3.56 NarEye 2.88 2.94 2.92 SnW 152 1.6 1.58 TrunkL/SVL 0.54 0.51 0.52 HL/SVL 0.25 0.26 0.26 HW/SVL Onl? 0.20 0.19 HW/HL 0.68 0.76 0.72 SnEye/HL 0.37 0.35 0.36 NarEye/HL 0.30 0.30 0.29 ED/HL 0.25 0.25 0.24 SnW/HL 0.16 0.16 0.16 ED/NarEye 0.86 0.82 0.82 Snw/HW 0.24 0.22 0.22 Chin 10 11 11 CN 6 6 IS 2 S| 2 SL (left/right) 8/8 8/8 8/9 IL (left/right) 8/8 8/8 9/9 VS 9 9 9 DS 16 15 15 Lamelar formulae hands II-V (left) 3-4-4-4 3-4-4-4 3-4-4-4 Lamelar formulae hands II-V (right) 3-4-4-4 3-4-4-4 3-4-4-4 Lamelar formulae foot II-V (left) 4-4-4-4 3-4-4-4 3-4-4-4 Lamelar formulae foot II-V (right) 4-4-4-4 3-4-4-4 3-4-4-4 SLIF 4/4 4/4 4/4 SLIT 4/4 4/4 4/4 Precloacal and femoral pore series separate (1) or 0 0 0 continuous (0) Total femoroprecloacal pores 28 26 28 CloacS on each side 3/2 1/1 1/1 Subcaudals enlarged, plate-like No No No Dark postorbital stripe Yes Yes Yes Dorsolateral light-colored spots on trunk Yes Yes Yes Dark dorsolateral stripe on trunk No No No Dark ventrolateral stripe on trunk Yes Yes Yes Dark dorsal transverse blotches Yes Yes Yes Dark reticulate pattern on dorsum No No No Postsacral marking anteriorly projecting arms Yes Yes Yes versus 0.22—0.24); longer SnEye distance (SnEye/HL = 0.35—0.37 versus 0.42—0.45); smaller eyes (ED/HL=0.22- 0.24 versus 0.15—0.18); more chin scales (10 or 11 versus 7-9); more circumnasal scales (CN = 6 versus 5); more ventral scales contained with one eye diameter (VS = 8 or 9 versus 6-7); more dorsal scales contained with one eye diameter (DS = 15 or 16 versus 10—14); dorsolateral light-colored spots on trunk present (versus absent); dark ventrolateral stripe on trunk present (versus absent). H. laowozhenensis sp. nov. differences from H. zalon- icus by longer head (HL = 9.71—9.96 versus 8.4—8.5; HL/ SVL = 0.25-0.26 versus 0.22—0.23); wider head (HW = 6.66—-7.41 versus 5.7—5.8; HW/SVL = 0.17—20 versus 0.15; HW/HL = 0.68-0.76 versus 0.68); longer NarEye zse.pensoft.net 252 Zhou, H. et al.: Hemiphyllodactylus laowozhenensis sp. nov. from Yunnan Table 4. Diagnostic characters separating Hemiphyllodactylus laowozhenensis sp. nov. from other nominal taxa of Hemiphyl- lodactylus within clade 3 of Agung et al. (2022). (/) = data unavailable. Mensural characters are in mm. Data for H. zalonicus and H. longlingensis were obtained from Grismer et al. (2020). Data for H. changningensis were obtained from Guo et al. (2015). Data for H. zhutangxiangensis were obtained from Agung et al. (2021). Data for H. gengmaensis were obtained from Zhou et al. (2024a). Character H. H. H. zalonicus H. H. H. laowozhenensis gengmaensis __ longlingensis changningensis zhutangxiangensis sp. nov. Max SVL 43.24 45.7 Se 43.8 44.42 38.5 n 6 2 10 13 3 TrunkL 17.46-23.22 / 18.1-18.9 17.4-22.5 16.1-23.1 18.76-20.5 HL 10.8-11.24 / 8.4-8.5 8.2-10.1 6.2-7.6 9.71-9.96 HW 6.88-8.2 3 5.7-5.8 6.1-7.5 6.5-8.2 6.66-7.41 ED 2.12-2.62 a 2.0-2.6 1.7-2.3 2.1-2.7 2.40-2.48 SnEye 3.88-4.52 / 3.4-3.5 3.5-4.5 3.4-4.4 3.43-3.64 NarEye 2.12-3.32 i 2.4-2.7 2.8-3.4 2.6-3.2 2.88-2.94 SnW 1.6-2.72 / re 1.1-1.4 1.3-1.6 1.57-1.60 TrunkL/SVL 0.45-0.54 0.47-0.52 0.49-0.50 0.46-0.51 0.48-0.52 0.51-0.54 HL/SVL 0.26-0.31 0.22-0.24 0.22-0.23 0.22-0.25 0.17-0.20 0.25-0.26 HW/SVL 0.19-0.2 0.17-0.19 OS 0.17-0.18 0.18-0.20 0.17-0.20 HW/HL 0.64-0.74 0.75-0.80 0.68 0.72-0.80 1.03-1.13 0.68-0.76 SnEye/HL 0.36-0.41 0.42-0.45 0.40-0.42 0.41-0.49 0.53-0.60 0.35-0.37 NarEye/HL 0.24-0.30 0.29-0.34 0.28-0.31 0.30-0.37 0.39-0.44 0.29-0.30 ED/HL 0.20-0.24 0.22-0.25 0.23-0.30 0.21-0.25 0.30-0.36 0.24-0.25 SnW/HL 0.15-0.24 0.12-0.14 0.14 0.12-0.16 0.19-0.22 0.16 ED/NarEye 0.73-0.94 0.66-0.82 0.74-1.08 0.61-0.77 0.70-0.91 0.82-0.86 Snw/HW 0.22-0.33 0.15-0.18 0.21 0.16-0.20 0.16-0.21 0.22-0.24 Chin 8-9 7-9 8-10 7-8 7-9 10 or 11 CN 6 4-5 5 3-4 5 6 IS 2-3 1-3 3-4 2-3 2-4 2or3 SL 9-11 9-10 10 8-11 8-11 8or9 IL 8-9 8-10 8-9 8-10 8-11 8 or9 VS 8-10 6-7 9-10 6-8 5-7 8or9 DS 11-18 10-14 17-18 11-15 11-15 15 or 16 Lamelar formulae 5-5(6)-5-4 3-4-4-4(3) 3-3-3-3 3-3(4)-3(4)-3 (3-5){4-6)(4 or 544 3-4-4-4 hands |I-V 4-4-A(5)-4 or 5) Lamelar formulae 5-5-6-5 4-4(5)-4(5)-4 3-4-4-4 3-3-3-3 (4 or 5)}(4 or 5)-4-6-(4 3(4)-4-4-4 foot II-LV 34-4-4 or 5) SLIF 5 4-5 3 3-4 4-5 4 SLIT 6 4-6 4 3-4 4-5 4 Precloacal and 0 0 0 0 0 0 femoral pore series separate (1) or continuous (0) Total 10-25 16-27 16-20 19-22 20-23 26-28 femoroprecloacal pores CloacS on each 1 or 2 Loer2 1 lor 2 1 or 2 1-3 Side Subcaudals No No No No No No enlarged, plate- like Dark postorbital Yes Yes Yes Yes Yes Yes stripe Dorsolateral light- No No No No No Yes colored spots on trunk Dark dorsolateral No No No No No No stripe on trunk zse.pensoft.net Zoosyst. Evol. 101 (1) 2025, 245-255 293 Character H. H. H. zalonicus H. H. H. laowozhenensis gengmaensis __longlingensis changningensis zhutangxiangensis sp. nov. Dark ventrolateral No No No No No Yes stripe on trunk Dark dorsal Yes Variable No Yes Yes Yes transverse blotches Dark reticulate Yes or Indistinct Variable No Yes No No pattern on dorsum Postsacral Indistinct or Light-colored Indistinct or No Fork-like dark-colored Yes marking anteriorly — Fork-like dark- not projecting arms colored distance (NarEye = 2.88—2.94 versus 2.4—2.7); wider SnW distance (SnW = 1.57—1.60 versus 1.20; SnW/HL = 0.16 versus 0.14; SnW/HW = 0.22-0.24 versus 0.21); more chin scales (10 or 11 versus 8-10); more circum- nasal scales (CN = 6 versus 5); few supralabial scales (SL = 8 or 9 versus 10); few dorsal scales contained with one eye diameter (DS = 15 or 16 versus 17—18); more SLIF (SLIF = 4 versus 3); more femoroprecloacal pores (26-28 versus 16-20); dorsolateral light-colored spots on trunk present (versus absent); dark ventrolateral stripe on trunk present (versus absent); dark dorsal transverse blotches present (absent). H. laowozhenensis sp. nov. differences from H. changningensis by greater eyes (ED = 2.40—2.48 versus 1.7—2.3); wider SnW distance (SnW = 1.57—1.60 versus 1.10-1.40; SnW/HL = 0.16 versus 0.12—0.16); more chin scales (10 or 11 versus 7 or 8); more circumnasal scales (CN = 6 versus 3 or 4); more femoroprecloacal pores (26—28 versus 19-22); dorsolateral light-colored spots on trunk present (versus absent); dark ventrolateral stripe on trunk present (versus absent); dark reticulate pattern on dorsum absent (versus present); postsacral marking ante- riorly projecting arms present (versus absent). H. laowozhenensis sp. nov. differences from H. zhutangxiangensis by longer head (HL = 9.71—9.96 ver- sus 6.2—7.6; HL/SVL = 0.25-0.26 versus 0.17—0.20); more chin scales (10 or 11 versus 7-9); more circumnasal scales (CN = 6 versus 5); more ventral scales contained with one eye diameter (VS = 8 or 9 versus 5—7); more dorsal scales contained with one eye diameter (DS = 15 or 16 versus 11—15); more femoroprecloacal pores (26—28 versus 20—23); dorsolateral light-colored spots on trunk present (versus absent); dark ventrolateral stripe on trunk present (versus absent). Discussion Our research supports the recognition of the Hemiphyl- lodactylus laowozhenensis sp. nov. as a new species, be- longing to clade 3 of Agung et al. (2022). It is sister taxa to H. longlingensis, H. zalonicus, H. changningensis, H. zhutangxiangensis and H. gengmaensis. Our research team had overlooked species of this genus during our more than 10 years of surveys in the region. The discov- ery of this new species further emphasizes the importance of repeated surveys in determining true local species di- versity and highlights the diversity of Hemiphyllodacty- Jus species in Yunnan Province, China. In Yunnan Province, our decade-long surveys have found that all habitats of Hemiphyllodactylus are adjacent to human settlements, which are precisely areas that are not protected. Most Hemiphyllodactylus species in Yun- nan have narrow distributions, occurring in only one or several limited number of localities. Therefore, the con- servation of Hemiphyllodactylus species in the future may face significant challenges. Ethical statement The Ethics Committee of Kunming Institute of Zoology, Chinese Academy of Sciences, approved the study and provided ethics permission (no. SMKX-20191221-216). Funding This work was supported by Science Technology Basic Condition Platform from the Ministry of Science and Technology of the People’s Republic of China (grant no. 2005DKA21402), National Natural Science Foundation Project: Investigation and Classificatory and Phylogenet- ic Studies on the Lizards of Gekkonidae of China (grant no. NSFC-—31970404). Acknowledgements We thank Jiazhong Wang for assistance in the field. We also thank the editors and reviews for their comments on the manuscript. References Agarwal I, Khandekar A, Giri VB, Ramakrishnan U, Karanth KP (2019) The hills are alive with geckos! A radiation of a dozen species on sky islands across peninsular India (Squamata: Gekkonidae, Hemi- Phyllodactylus) with the description of three new species. 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Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons. org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow us- ers to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://do1.org/10.3897/zse.101.138964.suppl1 fas) Supplementary material 2 Morphometric and meristic data used in the analyses from specimens of Hemiphyllodactylus members within clade 3 Authors: Hongxin Zhou, Xiaohan Wang, Liangwei Cuil, Dongru Zhang, Ziqi Shen, Xiuyan Li, Shuo Liu, Dingqi Rao Data type: xlsx Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons. org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow us- ers to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/zse.101.138964.suppl2 zse.pensoft.net