Parentage And Genetic Diversity Studies In Yanbian Captive Black Bear Using Microsatellite DNA Markers

Asiatic black bear(Ursus thibetanus), which have several alternative names like "black bear" or "dog hunchback" or "black blind animal" or "crescent moon bear", is the most heavyweight land carnivores nowadays. In the past, black bear was not included within the national aegis animals. In the around of mountain, black bear has been considered as a harmfuL animal. It has always been killed or captured to be raised artificially for gaining its economic value and medicinal value. With the sharp decline of wild black bear’s popuLation, our country has already listed it into the national Ⅱ aegis animals. Many scholars all over the world have started to launch more and more research works about black bear. Acording to the increasing scales from breeding industry of the black bear, it was necessary to further investigate the genetic diversity of black bear subspecies in our country for more scientific promotion of the bear breeding with standardized management. Therefore, in this study we take nineteen adult black bears as a representative animal, contrasting the results of the microsatellite amplification reaction influenced by different quality and quantity of template DNA extracted from three body samples including hair or blood or tissue, try to find out the best template DNA concentration for appropriate reaction of microsatellite amplification. This study also discussed the feasibility of making the genetic diversity research by using the hair of black bear as a testing material. Using microsatellite markers, this study screened one hundred and fifteen black bears specimens in the area of Yanbian to analyze the microsatellite diversity of them statistically. Father-son Parentage of black bear in Yanbian region has also been clearly identified. The results are in the follwing:1. The PCR products using template DNA extracted from hair or blood or tissue had no obvious differences in its quality and quantity. The noninvasive sampling method collecting one hundred hairs with hair follicles was good for extracting enough template DNA for microsatellite amplification.50ng/μL was the best template DNA concentration for efficient microsatellite amplification reactions. The results of this study have further proved that the template DNA extracted from hair samples could be taken for black bear genetic diversity research as good testing materials.2. In this study, fifteen loci screened among nineteen different microsatellite sites of black bear genomic DNA were taken to be done genetic diversity analysis. Total111loci and alleles were investigated in this study. It was calculated that each microsatellite locus has almost4-11allele types. Average number of each allele types investigated in this study was7.40, and the allele frequency was between0-1. These results showed there was relatively rich genetic diversities in black bear population farming at north-east area of china.3. The PIC value of fifteen microsatellite loci invesigated in this study was between0.4533-0.7422and average number was0.6133. The PIC value of ABB3, ABB7, ABB12, UT36was between0.25to0.5, which were belonging to the moderate polymorphism loci and the others were over0.5. Average expectation heterozygosis of fifteen loci was0.6213. It can be explained that the selected markers have relatively rich polymorphism, which could reflect the genetic relationship of black bear accurately.4. Fourteen little bears were confirmed its parents through paternity test using fifteen microsatellite loci mentioned above. Cumulative chance of exclusion (CCE) and paternity index between the parents and their off-springs were high. The CCE was0.999985and the RCP range was99.9473%-99.9993%respectively between the parents and their off-springs investigated in this study.