Individual Identification And Conservation Gentic Studies In Procapra Przewalskii Using SSR

Procapra przewalskii is endemic to China, and just live around Qinghai Lake region, Qinghai Province, with a population about 300, so it was appointed as CR (Critically Endangered) in the IUCN (The World Conservation Union) Red List in 1996. The conservation genetics research is a key step to conserve the endangered species in individual identification, population size estimation, pedigree analysis and population viability analysis.First of all, we explored a set of techniques for extracting fecal DNA, and established a new extracted DNA method successfully. Compared the extracted fecal DNA with blood DNA and skin DNA by agar electrophoresis, we found this new method is easy operate high yield, practicable and safety to operator because it reduced chance to touch poisonous chemical drugs, and the method was a high veracity and non-invasive DNA technology. Therefore, this new extracted DNA method imply the non-invasive DNA technology can use to study of genetic diversity of endangered species.Microsatellite locus is a highly variable nuclear marker. The two sides of the sequence of microsatellite markers were high conservative, and sequences in the middle part were polymorphic. Thus microsatellite markers are one of the better molecular markers for inter-species genetic diversity study and individual analysis. In our study, 10 polymorphic microsatellite DNA primers which identified in Grant gazelle were amplified in Procapra przewalskii genomes, and we found that five of the loci (TGLA54, BM1392, Oarfcb304, SMHCC, BM1862) were polymorphic. At the same time, we used 10 polymorphic microsatellite DNA primers identified in sheep to amplify in Procapra przewalskii genomes, the amplified results showed that 3 loci were polymorphic (MNS61, MNS64, Oarfel33). We identified individuals and tested the genetic diversity in 39 Procapra przewalskii fecal samples using these eight polymorphic microsatellite loci. The PCR amplified products of microsatellite loci were detected by non-denatured polyacry lamide gel electrophoresis. The results showed these fecal samples belonged to 35 different individuals; we calculated the gene heterozygosity, individual identification potential, polymorphism information content (PIC), and effective number of alleles of each locus. The values of gene heterozygosity was 0.7089-0.8488, the values of effective number of alleles is 3.375-6.592, PIC is 0.657-0.789, TDP =0.999; which showed these loci were high polymorphism and fitted to individual identification. These results implied the microsatellite DNA markers could help study the genetic diversity of endangered species.Finally, We used MtDNA to identify successfully a small captive gazelle (Ling Ling) in the Bird Island in Qinghai National Nature Reserve and a ram that was fed in Qinghai Zoo. The result showed that Ling Ling belonged to Procapra przewalskii i, and ram belonged to Gazella gutturosa.