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A New Species of the Genus Elaphe (Squamata: Colubridae) from Zoige County, Sichuan, China

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A New Species of the Genus Elaphe (Squamata: Colubridae) from Zoige County, Sichuan, China Asian Herpetological Research 2012, 3(1): 38–45 DOI: 10.3724/SP.J.1245.2012.00038 A New Species of the Genus Elaphe (Squamata: Colubridae) from Zoige County, Sichuan, China Song HUANG1, 2, 3*, Li DING4*, Frank T. BURBRINK5, 6, Jun YANG7, Jietang HUANG3, Chen LING8, Xin CHEN5, 6 and Yaping ZHANG2** 1 College of Life and Environment Sciences, Huangshan University, Huangshan 245041, Anhui, China 2 State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, Yunnan, China 3 Huangshan Institute of Ophiology, Huangshan 245000, Anhui, China 4 Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, Sichuan, China 5 Department of Biology, the College of Staten Island, the City University of New York, 2800 Victory Blvd, Staten Island, NY 10314, USA 6 Department of Biology, the Graduate Center, the City University of New York, 365 Fifth Avenue, New York, NY 10016- 4309, USA 7 School of Life Sciences, Sichuan University, Chengdu 610064, Sichuan, China 8 College of Forestry, Southwest Forestry University, Kunming 650224, Yunnan, China Abstract Previous phylogenetic analyses of DNA sequence data reveal a likely new species of Elaphe Fitzinger from China, which forms the sister group of all extant Elaphe. One of the three original specimens is crushed and the other two are neonates, which precludes a morphological analysis. Three adult snakes (2 females, 1 male) collected from Jiangzha Hot Spring in Zoige County, Sichuan, China on 26 July 2010 now facilitate the species’ description. Mitochondrial DNA nucleotide sequences of cytochrome b (cyt b), NADH dehydrogenase subunit 4 (ND4), 12S ribosomal RNA (12S rRNA), and cytochrome oxidase subunit 1 (CO1) from the adults are identical to sequences from the crashed specimen. Interspecific uncorrected p-distances of partial CO1 gene of 26 sequences from all 11 species of Elaphe, including this new species, are large (8.5–15.2%). This new species is morphologically distinguished from all other species of Elaphe by several characters: “M”-shaped marking on the supraoculars and adjacent frontal; four parallel series of red-brown spots on the dorsum extending from the neck to the vent; four longitudinal stripes associated with the spots; each spot is usually composed of a whole (or incomplete) red-brown scale with puce borders; spots of each stripe are similar in shape, they occur at relatively regular intervals of one or two scales, and they are slightly red-brown in color; and it has three preoculars, whereas all other species have one or two. Now Elaphe contains 11 species and this discovery highlights the need for continued exploration of temperate regions. Keywords new species, mitochondrial DNA, DNA barcoding, Zoige ratsnake 1. Introduction molecular phylogenetics and increased taxon sampling. These changes are important not only for classification, Nowadays, taxonomic revisions of major groups of but also for comparative biology, which requires credible vertebrates are common due, in part, to the revolution in phylogenetic estimates and accurate assessments of extant taxa for hypothesis testing (Felsenstein, 1988; * Both authors contribute equally to this work. ** Corresponding author: Prof. Yaping ZHANG, from Kunming Institute Díaz-Uriarte and Garland, 1996, 1998). Advanced snakes of Zoology, Chinese Academy of Sciences, with his research focusing (Caenophidia), representing about 3 000 species, are the on molecular phylogeny and evolution of vertebrates. E-mail: [email protected] subject of several recent massive phylogenetic studies Received: 5 January 2012 Accepted: 27 Feburary 2012 that propose numerous changes in classification (Pyron No. 1 Song HUANG et al. New Species of the Genus Elaphe from Zoige, China 39 et al., 2011; Burbrink and Crother, 2011) as well as members from all genera of Old World ratsnakes and provide important tests of major hypotheses in evolution representatives from the New World strongly indicates (Burbrink and Pyron, 2010). Within the advanced snakes, that the three specimens represent the sister species to all the ratsnakes are one of the most-well-studied groups. other taxa in the genus Elaphe. This relationship suggests Recent major taxonomic changes occur at generic and an undescribed species of Elaphe but it is not described species levels (Utiger et al., 2002; Burbrink and Lawson, because the absence of adult specimens precluded a 2007; Pyron and Burbrink, 2009a). morphological description. Several studies demonstrate that the ratsnake genus On 26 July 2010, we captured three adult snakes (2 Elaphe (sensu lato) is likely paraphyletic, particularly females, 1 male) at Jiangzha Hot Spring, Zoige County, with respect to New World taxa that are more closely Sichuan, China. Based on molecular data from cyt b, related to kingsnakes, pinesnakes and their allies (e. g., ND4 and 12S rRNA, the new specimens were determined Lampropeltis, Pituophis, Rhinocheilus, Cemophora). to belong to Elaphe and to be conspecific with those The genus Elaphe, with the type species Coluber reported by Ling et al. (2010). sauromates Pallas 1811 from Europe, is now restricted to the following ten taxa found throughout Eurasia: 2. Materials and Methods E. anomala (Boulenger), E. bimaculata Schmidt, E. carinata (Günther), E. climacophora (Boie), E. davidi Three adult snakes (2 females, 1 male) were sampled at (Sauvage), E. dione (Pallas), E. quadrivirgata (Boie), the side of a stream (Figure 1) by Li DING within 40 m E. quatuorlineata (Lacépède), E. sauromates (Pallas) of the mouth of Jiangzha Hot Spring at 12:00 hr on 26 and E. schrenckii (Strauch) (Schulz, 1996; Helfenberger, July 2010. Several days later, two females laid 11 eggs, 2001; Utiger et al., 2002; Burbrink and Lawson, 2007). respectively (Figure 2) in field terrariums. The eggs were Phylogenetic support for the monophyly of Elaphe is smooth-shelled and adhesive. The holotype (Collection strong and placement of species within in this genus is number: HS201014) and paratypes (HS201015–16) were confirmed by both nuclear and mtDNA genes (Utiger et preserved and deposited in the Museum of Huangshan al., 2002; Burbrink and Lawson, 2007). Geographically, University (Voucher numbers: HUM20110001–3). the highest diversity is centered in East Asia, although the Tissues from the distal ends of ventral scales were range of the genus extends as in the far north to Russia sampled from all the new specimens. Partial mtDNA (E. dione, E. schrenckii), in the south to Vietnam (E. sequences were obtained by polymerase chain reaction carinata), east to the China Sea, Korean Peninsula and (PCR) and direct sequencing using the primers and Japan (E. anomala, E. bimaculata, E. carinata, E. davidi, methods described in Arévalo et al. (1994), Burbrink et al. E. climacophora, E. quadrivirgata), and west to Italy (E. (2000), and Huang et al. (2009) for cyt b (1 015 bp) and quatuorlineata). ND4 (696 bp), and Utiger et al. (2002) for 12S rRNA Molecular data provide a powerful means of (428 bp) and CO1 (513 bp). The new cyt b, ND4 and identifying cryptic diversity, such as that within Elaphe 12S sequences were identical to those reported by Ling (E. quatuorlineata) and the related genera Zamenis (Z. et al. (2010) (GenBank accession number: HQ330525, longissimus), Pantherophis (P. guttatus, P. obsoletus), and HQ330527 and HQ330523, respectively). The absence of Lampropeltis (L. getula), (Lenk and Joger, 1994; Lenk et al., variation indicated that the six snakes were conspecific. 2001; Burbrink et al., 2000; Burbrink, 2001, 2002; Pyron Because sequences from COI—DNA barcoding— and Burbrink, 2009b, c). The rapid radiation of these have enabled the discrimination of closely allied species advanced snakes may be responsible for the absence of (Hebert et al., 2003) and provided a standardized tool morphological differentiation. For example, both E. dione to inventory biodiversity (Naro-Maciel et al., 2010), and E. bimaculata are morphologically quite similar with we sequenced the gene for the three new specimens respect to scutellation and color pattern, and both occur in (GenBank accession numbers: JF510474–76). The China. uncorrected p-distances of partial CO1 gene of 26 Based on molecular phylogenetic analyses of sequences from all the 11 species of Elaphe (23 from nucleotide sequences from three mitochondrial DNA Utiger et al., 2002; 3 from this study) were calculated genes (cyt b, ND4 and 12S rRNA), Ling et al. (2010) using the program MAGA 4 (Tamura et al., 2007). COI suggest that three specimens collected from Zoige p-distances ranged between 8.2–15.0 among species and County, Sichuan, China likely represent a new species between 0–2.5 within species (Table 1). of Elaphe. The matrilineal genealogy, which contains All measurements and observations were conducted 40 Table 1 Matrix of uncorrected P-distances of partial CO1 gene (513bp) for 26 sequences from 11 species of Elaphe. GenBank accession numbers follow species names. [ 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 ] [ 1] HUM20110001 JF510474 0 [ 2] HUM20110003 JF510475 0.4 0 [ 3] HUM20110002 JF510476 0.4 0.4 0 [ 4] E. climacophora AY122687 14.0 13.6 13.6 0 [ 5] E. climacophora AY122686 14.0 13.6 13.6 0.0 0 [ 6] E. climacophora AY122688 13.5 13.1 13.1 0.6 0.6 0 [ 7] E. bimaculata AY122753 13.5 13.5 13.1 13.8 13.8 13.5 0 [ 8] E. bimaculata AY122683 13.5 13.5 13.1 13.5 13.5 13.1 0.4 0 [ 9] E. bimaculata AY122684 13.5 13.5 13.1 13.1 13.1 12.7 1.8 1.4 0 [10] E. dione AY122750 12.9 12.9 12.5 11.5 11.5 11.3 9.6 9.6 9.6 0 Asian HerpetologicalResearch [11] E. dione AY122748 12.9 12.9 12.5 11.5 11.5 11.3 9.6 9.6 9.6 0.0 0 [12] E.
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