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A New Species of Kurixalus (Anura, Rhacophoridae) from Guizhou, China

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A New Species of Kurixalus (Anura, Rhacophoridae) from Guizhou, China ZOOLOGICAL RESEARCH A new species of Kurixalus (Anura, Rhacophoridae) from Guizhou, China DEAR EDITOR, collected several small-sized tree frog specimens resembling members of Kurixalus based on various characters, notably We describe a new species of the genus Kurixalus, Kurixalus presence of serrated dermal fringes along outer edge of raoi sp. nov., from Guizhou Province, China, based on forearm and tarsus and saddle-shaped mark on dorsum. morphological and molecular evidence. Phylogenetically, the Phylogenetically, these specimens represent a new lineage, new species is sister to K. idiootocus, but is distinguishable which is sister to K. idiootocus (currently endemic in Taiwan), from all known congeners by a combination of the following but morphologically distinguishable from K. idiootocus and characters: small body size (snout-vent length 28.2‒32.2 mm other members of Kurixalus by a series of characters. in males; 38.6 mm in female); snout rounded, with no prominence on tip; single internal vocal sac; dorsal surface Therefore, we formally describe this lineage as a new species brown, rough, scattered with several small warts; chin clouded of Kurixalus. with blackish marking; pair of large symmetrical dark blotches Field surveys were conducted in May 2019 at Qingshuihe on chest; vomerine teeth present; iris brown; tibiotarsal Nature Reserve, Southwest Guizhou Autonomous Prefecture, articulation reaching center of eye; nuptial pad slight; flank Guizhou, China (Figure 1A). Specimens were photographed, rough; mandibular symphysis weak; throat skin granular; and euthanized, fixed, and then stored in 75% ethanol. Liver toes moderately webbed, formula I2 ‒2II1.5 ‒3III2 ‒3IV3 ‒2V. tissues were preserved in 99% ethanol. Specimens were The genetic distances between the new species and K. deposited at Guangxi Normal University (GXNU), Guangxi, idiootocus were 2.9% and 5.4% for 16S rRNA and COI, China. respectively. Sequences encoding three mitochondrial genes (12S rRNA, Kurixalus Ye, Fei & Dubois, a genus within the family 16S rRNA, and COI) were amplified and sequenced using the Rhacophoridae, is widely distributed from northeastern India, primers and experimental protocols of Yu et al. (2020). eastwards to Indochina, southern mainland China, adjacent Homologous sequences of other Kurixalus species were continental islands (Hainan, Taiwan), and the Ryukyu obtained from GenBank. All new sequences were deposited in archipelago, and southwards to the Sunda and Philippine GenBank under Accession Nos. MW345613‒MW345628 and archipelagos (Frost, 2020). Currently, a total of 19 species are MW346332 ‒MW346339 (Supplementary Table S1). recognized in the genus Kurixalus, nine of which have been Phylogenetic relationships among the genus Kurixalus were recorded from China, namely: K. berylliniris Wu, Huang, Tsai, inferred using Bayesian inference (Supplementary Materials Li, Jhang & Wu, K. eiffingeri (Boettger), K. hainanus (Zhao, and Methods). Uncorrected pairwise distances between Wang & Shi), K. idiootocus (Kuramoto & Wang), K. species were calculated in MEGA 7 (Kumar et al., 2016). lenquanensis Yu, Wang, Hou, Rao & Yang, K. naso Morphometric data were taken using digital calipers to the (Annandale), K. odontotarsus (Ye & Fei), K. wangi Wu, nearest 0.1 mm. Morphological terminology followed Fei et al. Huang, Tsai, Li, Jhang & Wu, and K. yangi Yu, Hui, Rao & (2017) (Supplementary Materials and Methods). Comparative Yang (Frost, 2020; Yu et al., 2017a, 2018). morphological data of other Kurixalus species were taken from At present, a single species of the genus (K. hainanus) is previous publications (Annandale, 1912; Bossuyt & Dubois, known from Guizhou Province, China (Yu et al., 2017a). During our recent field surveys in southwest Guizhou, we Received: 22 October 2020; Accepted: 06 January 2021; Online: 18 February 2021 Open Access Foundation items: This work was supported by the National Natural This is an open-access article distributed under the terms of the Science Foundation of China (No. 32060114), Key Laboratory of Creative Commons Attribution Non-Commercial License (http:// Ecology of Rare and Endangered Species and Environmental creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted Protection (Guangxi Normal University), Ministry of Education non-commercial use, distribution, and reproduction in any medium, (ERESEP2020Z22), and Guangxi Key Laboratory of Rare and provided the original work is properly cited. Endangered Animal Ecology, Guangxi Normal University (18-A-01-08 Copyright ©2021 Editorial Office of Zoological Research, Kunming and 19-A-01-06) Institute of Zoology, Chinese Academy of Sciences DOI: 10.24272/j.issn.2095-8137.2020.311 Science Press Zoological Research 42(2): 227−233, 2021 227 Figure 1 Collection site (A), habitat (B), phylogenetic position (C), and scatterplot of morphometric data of Kurixalus raoi sp. nov. (D) 2001; Boulenger, 1893; Günther, 1858; Inger et al., 1999; supported the new specimens as a distinct lineage of the Kuramoto & Wang, 1987; Mediyansyah et al., 2019; Nguyen genus Kurixalus and sister to K. idiootocus (Figure 1C). The et al., 2014a, 2014b; Smith, 1924; Taylor, 1962; Van Nguyen genetic distances between the new lineage and K. idiootocus et al., 2020; Wu et al., 2016; Yu et al., 2017b, 2018; Zhao et were 2.9% and 5.4% for 16S rRNA and COI, respectively al., 2005). Multivariate principal component analysis (PCA) (Supplementary Table S2). was conducted using SPSS 17.0 (SPSS Inc., USA) based on Morphometric data are summarized in Table 1. We retained a correlation matrix of size-standardized measurements (all the first two principal components, which accounted for measurements divided by SVL). Scatter plots of the first two 65.97% of total variance and had eigenvalues above 1.0. PCA factors were used to examine the differentiation between Loadings for PC1 were positive, except for distance between the new species and K. idiootocus. Specimens of K. nostril and eye (DNE), and were most heavily loaded on head idiootocus were collected from Xinbei, Taiwan, China, and can length (HL), upper eyelid width (UEW), and eye diameter (ED) be easily distinguished from other Kurixalus species in Taiwan (Table 2). Differentiation was found along the PC1 axis by the presence of two large brownish rounded blotches on between the new lineage and K. idiootocus (Figure 1D). The the chest (Wu et al., 2016). PCA results indicated that the new lineage differs The obtained sequence alignments were 401 bp, 873 bp, morphometrically from its sister species (K. idiootocus) by and 807 bp for 12S rRNA, 16S rRNA, and COI genes, shorter head, narrower UEW, and smaller eye. In addition, the respectively. Phylogenetic analysis of the three genes strongly new lineage possesses a combination of morphological 228 www.zoores.ac.cn Table 1 Measurements (in mm) of Kurixalus raoi sp. nov. and K. idiootocus collected from Xinbei, Taiwan, China Species Voucher No. Sex SVL HL HW SL IND IOD GXNU YU140141 M 28.2 8.5 10.4 4.1 2.6 3.2 GXNU YU140143 M 31.1 9.5 10.9 4.5 2.6 3.3 GXNU YU140144 M 30.2 9.2 10.6 4.4 2.6 3.1 GXNU YU140145* M 30.8 9.4 10.8 4.5 2.7 3.0 Kurixalus raoi sp. nov. GXNU YU140146 M 29.6 9.2 10.2 4.2 2.7 3.1 GXNU YU140147 M 32.2 9.8 11.7 4.8 2.9 3.4 GXNU YU140148 M 29.6 9.3 10.4 4.1 2.3 2.8 GXNU YU20160078 F 38.6 11.9 13.3 5.5 3.7 4.2 GXNU YU1406033 M 30.8 9.4 10.2 4.4 2.6 3.4 Kurixalus idiootocus GXNU YU140220 M 27.5 9.2 9.7 3.9 2.8 3.0 GXNU YU140217 M 29.3 9.5 10.2 4.3 2.7 3.0 GXNU YU140219 M 26.6 9.3 9.8 3.9 2.6 2.9 GXNU YU140215 M 25.7 8.8 9.5 3.8 2.7 2.9 GXNU YU140216 F 37.5 12.0 12.9 5.5 3.9 4.1 GXNU YU140218 M 28.9 9.3 10.0 4.1 3.0 3.3 Species Voucher No. UEW ED TD FHL TL FL DNE Kurixalus raoi sp. nov. GXNU YU140141 2.4 3.9 1.5 13.9 12.9 12.2 2.4 GXNU YU140143 2.4 4.0 1.8 14.5 13.8 12.6 2.8 GXNU YU140144 2.5 4.2 1.7 14.2 13.6 12.5 2.4 GXNU YU140145* 2.5 3.9 1.7 14.3 13.7 12.6 2.6 GXNU YU140146 2.6 3.8 1.6 14.0 13.4 12.2 2.5 GXNU YU140147 2.8 4.1 1.8 16.0 15.5 14.2 2.8 GXNU YU140148 2.6 4.2 1.7 14.1 13.5 12.8 2.3 GXNU YU20160078 3.3 4.7 2.0 19.0 17.6 16.7 3.1 GXNU YU1406033 2.5 3.9 1.6 14.6 13.7 12.7 2.5 Kurixalus idiootocus GXNU YU140220 2.8 4.3 1.6 13.1 13.1 11.3 2.0 GXNU YU140217 3.1 4.4 1.9 13.4 12.7 11.9 2.1 GXNU YU140219 2.9 4.3 1.7 13.1 12.2 11.3 1.9 GXNU YU140215 2.8 4.2 1.6 12.6 11.9 10.9 2.1 GXNU YU140216 3.4 5.3 1.9 18.2 17.3 15.3 3.0 GXNU YU140218 2.7 4.4 1.6 13.8 12.4 11.4 2.2 Abbreviations are defined in Table 2 and Supplementary Materials and Methods. Holotype is marked with asterisk. M: Male; F: Female. characters different from all known congeners (Supplementary Etymology: The species name is dedicated to Dr. Ding-Qi Table S3).
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