Genetic Diversity And Phylogeny Studies Of Cabot’s Tragopan By Artificial Breeding

Cabot’s Tragopan(Tragopan caboti)(Galliformes:Phasianidae) is one of the endemic pheasant species in China. Especially, the species is a national first grade protected species in China and one of the most endangered species worldwidely. One captive population has been established in wildlife breeding and ambulance’s center of HuNan Province, and its population has reached80individuals. The population was developed from small number of founders, which lead to loss genetic diversity severely. In this study, genetic diversity of Cabot’s Tragopan from breeding and ambulance’s center of HuNan Province were evaluated with mtDNA sequence analysis and microsatellite markers. Phylogenetic relationships of Tragopan has been controversy. COI gene is used to study phylogenetic relationships of Tragopan. In addition, COI gene, Ctyb gene and ND2gene were used to explore the relationship between species of genus Tragopan. The results were as follows:1.77mtDNA D-loop sequences of Cabot’s Tragopan were sequenced,1164bp in length was eventually determined through analysis of homologous. Based on77sequences obtained,15haplotypes were detected. We also found21variable sites,2Singleton sites,18Parsim-Info sites, and1insertion/deletion. DNAsp (V5.0) analysis showed that haplotype diversity (h) is0.817, Nucleotide diversity (π) is0.462%and average number of nucleotide differences(k) is5.375. Which indicate a low polymorphism, because the data used in this study were collected from a relatively small and closely related population. Phyogenetic trees of mtDNA D-loop sequences for Cabot’s tragopan were made by NJ, ME, ML and UPGMA with Temminck’s Tragopan as outgroup. Topological structures with three branches calculated using the above-mentioned four methods are very similar except bootstrap support values. Kinship network graph and phyogenetic trees demonstrated two basically identical results. Population historical dynamics analysis showed that Cabot’s Tragopan has no obvious population expansion in the distribution area and the group size remained relatively stable state in history.2.Genomic DNA samples from Cabot’s Tragopan were amplified with58pairs of microsatellite primers. Polymorphism was found on3microsatellite loci (TT06、TT07、 TT08) from Tragopan temminckii,2microsatellite loci(BG16、BG18) from Tetrao tetrix,2microsatellite loci (sTuT2、sTuT3) from Tetrao urogallus and1microsatellite loci(Aru-1A1)from Alectoris rufa. Average observed heterozygosity, expected heterozygosity, PIC, effective allelic numbers of eight loci were0.6155,0.6725,0.619,4.116respectively, which indicated genetic polymorphism of Cabot’s Tragopan was poor.3.81COI sequences (690bp) of7species from4genera were obtained, and homologous sequences of46species in21genera were downloaded from Genbank and BOLD. Sequence and phylogenetic analysis, genetic distances were calculated and phyogenetic trees of46species birds in Phasianidae were made by NJ, ME and ML with Mergus squamatus and Cuora amboinensis as outgroup. Genetic distances and phyogenetic trees showed relationship of Tragopan and Lophophorus was very close. Phyogenetic trees showed that Tragopan and Phasianini clustered together, Therefore, this result supports genus Tragopan belong to Phasianini.4. COI, Ctyb and ND2seuqences of species of Tragopan were analyzes, and the nine kinds of phylogenetic tree were made by a variety of methods of three gene sequences. The result showed that Tragopan caboti and Tragopan temminckii clustered together.