Preliminary Studies Of Molecular Phylogeography In The Chinese Blue Sheep(Pseudois Nayaur)

Blue sheep or bharal(Pseudois nayaur) is one of wild ungulates of the subfamily Cparinae genus Pseudois (Artiodactyla, Bovidae), and is listed in the second grade nationally protected animals of China. Traditionally, the blue sheep was divided into two subspecies, the Tibetan subspecies (Pseudois nayaur nayaur) and the Sichuan subspecies (Pseudois nayaur szechuanensis).Two kinds of molecular makers, the complete Cyt b gene sequence of mtDNA and the hypervariable region in the5’end of control region,was used in this study to research the Genetic structure of geographic populations, genetic diversity, systematic geography, gene flow level of the Chinese blue sheep, explore the taxonomic status of Chinese blue sheep subspecies, as well as the differentiation of the Sichuan subspecies at the molecular level. Combined with historical events, preliminary study on the main reasons and processes of formation of Chinese blue sheep in the geographic distribution pattern. The results show that:1. The combination of the Cyt b and D-loop region sequences downloaded from GenBank of blue sheep identified45Cyt b single haplotype and27haplotype of the D-loop region. These two sets of data were displayed:the Tibetan population and the Ningxia population had shared haplotype, and other populations did not. A and C was higher than the proportion of G and T, the base content had a certain partial difference.2. The calculation of the haplotype diversity (Cyt b:0.911±0.027; D-loop region of gene:0.924±0.027) and nucleotide diversity (Cyt b:0.02821±0.03443; D-loop region of gene:0.11412±0.0064) of the different geographic populations were higher, indicate that the Chinese blue sheep had a wealth of genetic diversity. The haplotype diversity and nucleotide diversity of Ningxia population were the lowest; the haplotype diversity and nucleotide diversity of the Sichuan and Tibet populations were relatively highest, implying a relatively high genetic diversity.3. Calculation of Chinese blue sheep in different geographic genetic distance between populations found that,the geographic populations genetic distance values of the Tibetan subspecies and the Sichuan subspecies were (Cyt b:0.035-0.050; D-loop:0.209-0.280).averages were (Cyt b:0.042±0.007; D-loop:0.243±0.032), Values between the the various groups within Sichuan population were (Cyt b:0.002-0.047; D-loop:0.006-0.299), averages were (Cyt b:0.033±0.011; D-loop:0.200±0.083).Ningxia, Tibetan and other populations had larger genetic distance. Population distances between Qinghai,Sichuan and Gansu populations were relatively smaller.4. Calculated the Sichuan subspecies’ gene flow among the various geographic populations based on Cyt b and D-loop region found that:There was almost no gene flow between the Ningxia populations and other geographic populations of Sichuan subspecies,implying that a there was a significant genetic differentiation between the Ningxia populations and other geographic populations of Sichuan subspecies.5. ML and Bayesian phylogenetic tree constructed based on Cyt b and D-loop region found that:Tibet subspecies alone clustered into one, Sichuan subspecies divided into two branches, namely:the Sichuan, Gansu and Qinghai branch and the Helan Mountain branch.6. Calculated the Tibet subspecies and Sichuan subspecies divergence time based on Cyt b and D-loop region was1.05-1.14million years ago, that the late period of the new generation Early Pleistocene of geological age——Middle Pleistocene early.7. Two sets of genetic distance datas from Cyt b and D-loop region for the t-test both showed that the genetic distance differences between species of various geographic populations of Sichuan subspecies and Tibet subspecies was not significant, indicated that the internal of Sichuan subspecies generated great genetic differentiation.8. The Chinese blue sheep should be devided into three evolutionary significant units,Tibet subspecies, Sichuan-Gansu-Qinghai branch, and the Helan Mountain branch,to be managed and protected. In particular, the Helan Mountain branch should be highly concerned about.