DOI: 10.3724/SP.J.1245.2014.00228

Asian Herpetological Research 2014/5:4 PP.228-239

A New Species of the Genus Thermophis(Serpentes: Colubridae) from Shangri-La, Northern Yunnan, China, with a Proposal for an Eclectic Rule for Species Delimitation

The hot-spring snakes, Thermophis, were previously known only from the Tibet Autonomous Region and Western Sichuan, China. During the past two years, three adult hot-spring snakes(2 females, 1 male) were sampled in Shangri-La, northern Yunnan, China, thus expanding their known distribution region towards the southeast. This site is the southeastern-most corner of the Tibetan Plateau and the southernmost tip of the Hengduan Mountains(Mts.). Phylogenetic analysis of mitochondrial(mt) and nuclear(n) DNA segments suggested that the three specimens belong to the genus of Thermophis. Morphologically, the new species is more similar to T. zhaoermii. However, it is distinguished from T. zhaoermii in the number of maxillary teeth(15), distance between the two eyes/head width, rostral width/height, mental width/height, in one character limited to female: head width/length, and in four characters restricted to male: occurrence of the reduction from 10 to 8(8 to 6, 6 to 4) scales in each dorsal row on the tail. There are differences in morphology, genetics(mt DNA, n DNA), and geography between the putative new species and T. zhaoermii, the new species meets our proposed eclectic and feasible "four-differences" rule.

Key words:morphology,phylogenetics,"four-differences" rule,Thermophis Shangrila sp.nov.

ReleaseDate:2015-03-27 09:32:50

Arévalo E., Davis S. K., Sites Jr., J. W. 1994. Mitochondrial DNA sequence divergence and phylogenetic relationships among eight chromosome races of the Sceloporus grammicus complex (Phrynosomatidae) in Central Mexico Syst Biol, 43: 387-418

Bininda-Emonds O. R. P., Bryant H. N., Russell A. P. 1998. Supraspecific taxa as terminals in cladistic analysis: implicit assumptions of monophyly and a comparison of methods. Biol J Linn Soc, 64: 101-133

Burbrink F. T., Lawson R., Slowinski J. B. 2000. Mitochondrial DNA phylogeography of the polytypic North American rat snake (Elaphe obsoleta): a critique of the subspecies concept. Evolution, 54: 2107-2118

Che J., Chen H., Yang J., Jin J., Jiang K., Yuan Z., Murphy W., Zhang Y. P. 2012. Universal COI primers for DNA barcoding amphibians. Mol Ecology Resources, 12: 247-258

Chen L., Chen Y., Hu J. J., 2010. Summarization and suggestions for the preference support of NSFC (National Natural Science Foundation of China) to classical animal taxonomy. China Basic Science, 3: 55-57 (In Chinese)

Chen X., Huang S., Guo P., Colli G. R., Montes de Oca A. N., Vitt L. J., Pyron R. A., Burbrink F. T. 2013a. Understanding the formation of ancient intertropical disjunct distributions using Asian and Neotropical hinged-teeth snakes (Sibynophis and Scaphiodontophis: Serpentes: Colubridae). Mol Phylogenet Evol, 66: 254-261

Chen X., Jiang K., Guo P., Huang S., Rao D. Q., Ding L., Takeuchi H., Che J., Zhang Y., Myers E. A., Burbrink F. T. 2013b. Assessing species boundaries and the phylogenetic position of the rare Szechwan ratsnake, Euprepiophis perlacea (Serpentes: Colubridae), using coalescent-based methods, Mol Phylogenet Evol, doi:

Dawson K., Malhotra A., Thorpe R. S., Guo P., Mrinalini, Ziegler T. 2008. Mitochondrial DNA analysis reveals a new member of the Asian pitviper genus Viridovipera (Serpentes: Viperidae: Crotalinae). Mol Phylogenet Evol, 49(1): 356-61

de Queiroz A., Lawson R. 2008. A peninsula as an island: multiple forms of evidence for overwater colonization of Baja California by the gartersnake Thamnophis validus Biol J Linn Soc Lond, 95 (2): 409-424

de Queiroz K. 1998. The general lineage concept of species, species criteria, and the process of speciation. In: Endless Forms: Species and Speciation (eds Howard DJ, Berlocher SH). Oxford University Press, Oxford, UK, pp, 57-75

Dorge T., Hofmann S., Wangdwei M., Duoje L., Solhoy T., Miehe G. 2007. The ecological specialist, Thermophis baileyi (Wall, 1907)-new records, distribution and biogeographic conclusions. Br Herpetol Soc, 101: 8-12

Fu J. Z., Zeng X. M. 2008. How many species are in the genus Batrachuperus? A phylogeographical analysis of the stream salamanders (family Hynobiidae) from southwestern China. Molecular Ecology, 17: 1469-1488

Guo G. Y., Liao Z. J., Liu S. B., Shen M. Z., Tong W., Wang D. X., Zhang B. S., Zhang Z. F., Zhao F. S., Zhu M. X., Wu C. Z., Zhang M. T., Zhou C. J., Kan R. J., Yan F. T., Han Z. S., Bi D. Z., Li G. X., Zhang Z. X., Zhao H., Zhao S. M., Zhang, G. W. 1994. Thermal Springs in Hengduan Mountains. Science Press, Beijing, 1-326 (In Chinese)

Guo P., Liu S. Y., Huang S., He M., Sun Z. Y., Feng J. C., Zhao E. M. 2009. Morphological variation in Thermophis Malnate (Serpentes: Colubridae), with an expanded description of T. zhaoermii. Zootaxa, 1973: 51-60

Guo P., Liu S., Feng J., He M. 2008. The description of a new species of Thermophis (Serpentes: Colubridae). Sichuan J Zool, 27: 321

He M., Feng J., Liu S., Guo P., Zhao E. M. 2009. The phylogenetic position of Thermophis (Serpentes: Colubridae), an endemic snake from the Qinghai-Xizang Plateau, China. J Nat Hist, 43: 479-488

He M., Feng J., Zhao E. M. 2010. The complete mitochondrial genome of the Sichuan hot-spring keel-back (Thermophis zhaoermii; Serpentes: Colubridae) and a mitogenomic phylogeny of the snakes. Mitochondrial DNA, 21: 8-18

Hofmann, S. 2012. Population genetic structure and geographic differentiation in the hot spring snake Thermophis baileyi (Serpentes, Colubridae): Indications for glacial refuges in southern-central Tibet. Mol Phylogenet Evol, 63: 396-406

Huang S., Liu S. Y., Guo P., Zhang Y. P., Zhao, E. M. 2009. What are the closest relatives of the hot-spring snakes (Colubridae, Thermophis), the relict species endemic to the Tibetan Plateau? Mol Phylogenet Evol, 51: 438-446

IUCN. 2010. IUCN Red List of Threatened Animals.

Kelly C. M. R., Barker N. P., Villet M. H. 2003. Phylogenetics of advanced snakes (Caenophidia) based on four mitochondrial genes Syst Biol, 52: 439-459

Kraus F., Brown W. M. 1998. Phylogenetic relationships of colubroid snakes based on mitochondrial DNA sequences. Zool J Linn Soc, 122: 455-487

Lawson R., Slowinski J. B., Crother B. I., Burbrink F. T. 2005. Phylogeny of the Colubroidea (Serpentes): new evidence from mitochondrial and nuclear genes. Mol Phylogenet Evol, 37 (2): 581-601

Li J. J., Wen S. X., Zhang Q. S., Wang F. B., Zheng B. X., Li B. Y. 1979. A discussion on the period, amplitude and type of the uplift of the Qinghai-Xizang Plateau. Scientia Sinica, 22: 608-616 (In Chinese)

Malhotra A., Thorpe R. S. 2004. A phylogeny of four mitochondrial gene regions suggests a revised taxonomy for Asian pitvipers (Trimeresurus and Ovophis) Mol Phylogenet Evol, 32: 83-100

Malnate E.V. 1953. The taxonomic status of the Tibetan colubrid snake Natrix baileyi. Copeia, 2: 92-96

Mayden R. L. 1997. A hierarchy of species concepts: the denouement in the saga of the species problem. In: Species: the Units of Biodiversity (eds Claridge M, Darwah HA, Wilson MR). Chapman & Hall, London, pp, 381-424

Pinou T., Vicarioa S., Marschner M., Caccone A. 2004. Relict snakes of North America and their relationships within Caenophidia, using likelihood-based Bayesian methods on mitochondrial sequences. Mol Phylogenet Evol, 32: 563-574

Pyron R. A., Burbrink F. T., Colli G. R., de Oca A. N., Vitt L. J., Kuczynski C. A., Wiens J. J. 2011. The phylogeny of advanced snakes (Colubroidea), with discovery of a new subfamily and comparison of support methods for likelihood trees. Mol. Phylogenet. Evol., 58 (2): 329-342

Pyron R. A., Kandambi D., Hendry C. R., Pushpamal V., Burbrink F. T., Somaweera R. 2013. Genus-level phylogeny of snakes reveals the origins of species richness in Sri Lanka. Mol Phylogenet Evol, 66(3): 969-78

Sambrook J., Fritsch E. F., Maniatis T. 1989. Molecular cloning: A laboratory manual, seconded. New York: Cold Spring Harbor Laboratory Press

Sun X. Y., Liu S. Y., Huang S. 2011. Tibetan Plateau Relict Snakes of the Genus Thermophis and Their relationship to New World Relict Snakes. Asian Herpetol Res, 2(3): 161-168

Swofford D. L. 2002. PAUP*. Phylogenetic analysis using parsimony (* and other methods), version 4. Sinauer Associates, Sunderland, Massachusetts

Tamura K., Dudley J., Nei M., Kumar S. 2007. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol, 24: 1596-1599

Thompson J. D., Gibson T. J., Plewniak F., Jeanmougin F., Higgins D. G. 1997. The CLUSTAL-X windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucl Acids Res, 25: 4876-488

Tong W., Liao Z. J., Liu S. B., Zhang Z. F., You M. Z., Zhang M. T. 2000. Thermal Springs in Tibet. Science Press, Beijing, 1-300 (In Chinese)

Wall, F. 1907. Some new Asian snakes. J. Bombay Nat. Hist. Soc., 17: 612-618

Yang Z. H., Rannala B. 2010. Bayesian species delimitation using multilocus sequence data. Proc Natl Acad Sci USA, 107: 9264-9269

Zhao E. M. 2006. Snakes of China. Anhui Sciences and Technology Publishing House, Hefei, China, pp, 1-365 (In Chinese)

Zheng D., Li B. Y. 1990. Evolution and differentiation of the physico-geographical environment of Qinghai-xizang Plateau. Geographical research, 9: 1-10 (In Chinese)