勐养保护区亚洲象微卫星位点筛选及种群遗传多样性分析  

Screening microsatellite DNA markers and genetic variation analysis of wild Asian elephant population in Mengyang Nature Reserve

作　　者：蔡清秀[1] 林柳[1] 潘文婧[1] 罗述金 张立[1] 

机构地区：[1]教育部生物多样性与生态工程重点实验室,北京师范大学生态研究所,北京100875 [2]Laboratory of Genomic Diversity,National Cancer Institute,Frederick,MD 21702-1201,USA

出　　处：《兽类学报》2008年第2期126-134,共9页Acta Theriologica Sinica

基　　金：美国内政部鱼和野生动物管理局亚洲象保护基金资助项目;国际爱护动物基金会资助项目

摘　　要：本文以亚洲象肌肉样品提出的DNA为模板,从非洲象31个微卫星位点和5个已知亚洲象微卫星位点中筛选勐养亚洲象的微卫星位点,进而对在西双版纳勐养保护区3年采集到的191份亚洲象粪便样品中提出的DNA进行特异性PCR扩增、基因分型、检测位点信息和遗传多样性分析。结果表明:36个位点中有14个位点能在亚洲象肌肉样品提出的DNA中成功扩增,且经测序证实为微卫星位点。其中9个多态位点能在185份粪便样品DNA中稳定扩增。勐养种群中,3个位点可能偏离Hardy-Weubberg平衡,至少8个位点间无明显连锁存在,平均等位基因数3.78±1.72,平均期望杂和度0.32±0.06,平均观察杂和度0.36±0.02,平均多态信息含量0.28,说明这9个位点适用于勐养亚洲象的遗传学研究。根据微卫星位点的杂合度和等位基因频率,相比于其他亚洲象种群,勐养亚洲象种群遗传多样性较低且等位基因频率具有特异性。To get the appropriate microsatellite markers, we first screened 31 loci originally developed in the African elephant and five in the Asian elephant （Elephas maximua） using one muscle sample and then test the locus polymorphism and analyze genetic variation by the total 191 faecal elephant samples collected in Mengyang Nature Reserve through 4 -8 repeat PCR and genoiyping. The results showed 14 loci from the 36 can amplify in the target muscle DNA sample and verified to be microsatellite after sequencing. Nine microsatellite loci were optimized for faecal DNA amplification in the targeted Asian elephant population. For Mengyang population, we obtained amplified product from 185 of the total 191 samples （96.9%） and 55 unique genotypes. 3 loci deviated from Hardy-Weinberg equilibrium and at least 8 loci lacked linkage. The average number of alleles is 3.78 ：t： 1.72, the average expected and observed heterozygosity and PIC were 0. 32 ±0. 06, 0. 36 ±0. 02 and 0. 28, respectively, which indicated that these loci were suitable to genetic researches for Asian elephant. The relatively low heterozygosity （0. 32 ± 0. 06 ） and allele frequencies compared with other populations showed that the genetic variation was low and genetic structure was potentially skewed in the Mengyang population.

关 键 词：亚洲象 近缘种 微卫星位点 遗传多样性 粪便样品 

分 类 号：Q786[生物学—分子生物学]