| The coat color is one of important morphological characteristics of animals and is mainly decided by melanin. The melanin consists of eumelanin and pheomelanin. At present, there has been an impressive accumulation of researches on the coat color diversity and adaptive evolution in animals with the color related genes. Colobinae is one of the Old World monkeys, and is distributed in various forests ranging from Africa to Southeast Asia. There are distinctive features in Colobine coat color, but very little quantitative information is available concerning differences in their coat color. One of main objectives in this thesis is to explore the difference of several genes related to coat color in Colobine, and provide further understanding on its adaptive evolution in Colobinae. The preliminary results are as follows:With PCR amplification and molecular cloning methods, we successfully got seven genes’ exons sequencing related to coat color (namely MC1R, ASIP, MITF, MLPH, TYR, TYRP1 and SLC45A2) in Presbytis melalophos, Colobus guereza, Nasalis larvatus, Semnopithecus entellus, Pygathrix nigripes, Rhinopithecus roxellanae and Trachypithecus framcoisi. Based on these seven genes sequencing, we got the complete coding sequences by the software of DNAstar and MEGA, and analysized the difference in bases and amino acids of coding sequences in seven Colobine species. We found that the 272th codon in MLPH was under positive selection when we used the site model of PAML to compute the related color genes. Thus, we suggest that the mutation of 272th codon could be one of the factors accounting for the coat color differentiation in Colobinae.The Rhinopithecus roxellana, belonging to Rhinopithecus genus of Colobinae, is one of endangered primate species endemic to China. It is alsothe flagship species in biological conservation. Human activities and illegal hunting have been the greatest threat to the long-term survival of this species. Numerous molecular markers, such as mitochondria marker, microsatellite marker, are used in conservation genetics of R. roxellana. In this thesis, we used a new molecular diagnosis method (HRM) for genotyping. Another main objective in this thesis is to select a set of SNP by HRM, which was used to individual identification in R. roxellana population. The preliminary results are as follows:We designed and synthesized 85 primer pairs based on the upstream and downstream sequence of 124 candidate SNPs. Of the 85 primer pairs,37 SNPs were succeed in PCR, and were selected for HRM. The genotypes of 37 SNPs by HRM were the same as the results by sequencing in the blood samples from seven individuals of R. roxellana. The accuracy rate of genotyping by HRM is 100%.15 feces samples were used for genotyping by HRM and individual identification. Of selected 37 SNPs,34 were success in genotyping in the 15 feces samples. The successful rate was 91.89%. We used the genotypes of 34 SNPs by cervus 3.0 for individual identification. The result indicated that the 15 feces samples came from seven individuals, which was in according with the number of sampling individuals. We estimated the power of the 34 SNPs in individual identification. The discrimination power (DP) ranged from 0.0364 to 0.4993, and the cumulative discrimination power (CDP) is 99.99%. We reconstructed the model in Heaton’s study and confirmed that the 34 SNP markers in our study had enough power to identify 40,000 individuals. At present, the population of R. roxellana is approximately 22,000. So we can used the 34 SNP markers for individual identification in the R. roxellana population. |
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