Research On Germplasm Resource Of Panthera Tigris Altaica And Phylogeny Of Several Felidae Species

Panthera tigris altaica is mainly distributed over the northeasten of China, Russion and North Korea. There are fewer wild Panthera tigris altaica in our country now, about 12 to 16. It has been the most speeding of animals extinction. So it is very important to do some researches on the genetic resource of the Panthera tigris altaica. And then some methods can be taken to protect and exploit the resource of Panthera tigris altaica. According to its categorization, Panthera tigris altaica belongs to Mammalia, Carnivora, Felidae, a subspecies of the Panthera tigris. But there is still some dispute about whether it belongs to Panthera pardus. Research on the genome DNA diversity of the Panthera tigris altaica was did using 15 microsatellites. The germplasm resource diversity was evaluated. And then the genetic distance of the Panthera tigris altaica population was calculated. On the other hand, part of mtDNA Cytb gene of the Panthera tigris altaica Persia cat, African lion and Panthera tigris Leopard were sequencd Then all these sequences were compared and analysised. The results showed that:1. Fifteen pairs of microsatellite primers were successfully amplified in the Panthera tigris altaica genome. The number of alleles ranged from 3 to 7 for each locus. The average number of alleles was 5.53 and effective number of alleles was 4.3664. The gene heterozygosity (He) ranged from 0.5850 to 0.8413, and the average value was 0.7140; the polymorphism information content (PIC) ranged from 0.5129 to 0.8217, and the average value was 0.7504, which implied that the 15 microsatellite loci were high polymorphism in these Panthera tigris altaica.2. The cluster analysis for 20 tigers was carried out with SPSS11.5 software and the 20 tigers were divided into three categories at 5.988. Except for few tigers, the blood relationship among the individuals was clear. Three categories were (A, B , C), (D), and (E , F, G).3. According to the results, 138 nucleotide sites substituted in Cytb gene(24.86% of the total sequenced sites in Cytb gene) were obtained. Among these nucleotide sites, there were 18 singleton variable sites, 118 parsimony informative sites, 2 gaps/missing data. Five types of mutation were found in these nucleotide sites: transition(85.5%), transversion(6.25%), transition and transversion(6.25%), gaps/missing data(0.73%), transition and gaps/missing data(0.73%). On the other hand, 22 haplotypes were detected, which were no shared haplotype. The diversity of haplotype within populations ranged from 0.7636 to 1.000, and the total diversity of haplotype was 0.9571±0.017, which showed high diversity.4 Among the four felidae species, the ramification degree of nucleotide(Dxy) ranged from 0.05286% to 0.14261%, the net genetic distance of nucleotide(Da) ranged from 0.02456% to 0.12955%, the value of kimura ranged from 0.056 to 0.164, which all showed the genetic distance of the Persia cat and the other three felidae species was further. And the value of Fst showed the genetic relationship of Panthera tigris altaica and Panthera tigris Leopard was the nearest.5. According to the 22 haplotypes, molecule phylogenetic trees(NJ, ME, UPGMA) were constructed. Thus, the opinion that Panthera tigris altaica, African lion and Panthera tigris Leopard should be grouped into the Panthera pardus were supported.