Taxonomical Studies On Some Pit-vipers (Gloydius) And Hemiphyllodactylus Yunnanensis From China Using Sequences Of Mitochondrial Genes

The present study discussed the classification of some pit-vipers {Gloydius) from the western plateau and intraspecific classification of Hemiphyllodactylus yunnanensis based on sequences of fragments of Cyt b and ND2-CO I genes, and identified five animal samples by means of molecular identification techniques using 12S rDNA sequences.1. Classification of pit-vipers from the western plateau of ChinaThe sequences of a 319 bp fragment of Cyt b gene were determined for 9 samples of Gloydius strauchii, G liupanensis and G. qinlingensis and the phylogenetic relationships among the 4 pit-vipers were studied. The results obtained do not support the opinion to refer G monticola as a subspecies of G. strauchii, and on the contrary support it to be a separate species; indicate G liupanensis would be a junior synonym of G strauchii, and G qinlingensis is a separate species.2. Intraspecific classification of Hemiphyllodactylus yunnanensisThe sequences of a 385 bp fragment of Cyt b gene and 485 bp fragment of ND2-CO I gene for 20 samples belonging to 4 subspecies of Hemiphyllodactylus yunnanensis were determined. Fifteen haplotypes were defined according to the Cyt b sequences, among which Longling and Baoshan local populations share LLC1, no shared haplotypes is found between local populations. Twelve haplotypes were defined according to the ND2-CO I sequences, among which Kunming and Shilin local populations share KMN1, no shared haplotypes is found between local populations also. The results of AMOVA analysis and phylogenetic analyses indicate both high nucleotide diversity and haplotype diversity for Hemiphyllodactylus yunnanensis. All trees from sequence data derived from molecular phylogenetic analyses strongly support the sister relationship between the Yunnan population and Guizhou population of Hemiphyllodactylus yunnanensis (BCL values at least 95). The Yunnan population contains two clades: Hemiphyllodactylus yunnanensis yunnanensis andHemiphyllodactylus yunnanensis longlingensis. The two individuals of Lvchun local population of Hemiphyllodactylus yunnanensis jinpingensis nested into the nominal subspecies H. y. yunnanensis and H. y. longlingensis respectively. The validity of H. y. yunnanensis, H. y. longlingensis and H. y. dushanensis is supported by both haplotype analysis and phylogenetic analysis. The subspecific status of H. y. jinpingensis is also suppoted by haplotype analysis, but it has not been resolved by phylogenetic analyses.3. DNA molecular identification of 5 animal samplesThe Industry and Commerce Bureau of Yixing City sent us five animal samples for identification. Fragment of 12S rDNA was amplified from the five samples using the primer pair L1091 and H1478. Sequences obtained in the present study, and homologous sequences retrieved from GenBank were aligned by Clustal software. P-distance was assessed by MEGA software version 2.1., and was used as a criterion for identification. The results of the molecular identification indicate: there are two samples of Varanus bengalensis, one sample of Ursus thibetanus, one sample of Manis pentadactyla and one sample of unidentified species of pangolin {Manis sp.).