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基于粪便DNA 的九龙山黑麂种群的遗传多样性

王艳妮,鲍毅新,刘军,林杰君,张旭,郑伟成,潘成椿   

  1. 浙江师范大学生态研究所
  • 出版日期:2012-05-24 发布日期:2012-05-20

Genetic diversity of black muntjac (Muntiacus crinifrons)in Jiulongshan National Nature Reserve by feces DNA

  • Online:2012-05-24 Published:2012-05-20

摘要: 近年来,由于人类活动的干扰使黑麂的栖息地明显减少并呈斑块状分布。因此,深入了解该物种各地理分布种群的遗传现状是制定保护及管理策略的重要科学依据。本研究运用非损伤取样技术在浙江省九龙山国家级自然保护区的黑麂分布区共采集61 份粪便样品、2 份肌肉样品和2 份皮张样品,采用8 个特异微卫星位点对61 份粪便样品中提取的DNA 进行检测,肌肉和皮张样品DNA 的检测结果作为对照,分析了九龙山国家级自然保护区黑麂种群的遗传多样性。经多次重复提取和PCR 扩增后,发现61 份粪便样本中有38 份可被稳定扩增,且不同样本间的条带大小有较大差异,因此我们认为38 份粪便样本来源于38 只不同的黑麂个体。8 个微卫星位点共检测到50 个等位基因,平均等位基因数(A)为6 (5 ~ 8);平均有效等位基因数(Ne)为5. 297 (4.031 ~6.353);平均期望杂合度(He) 为0.817 (0.763 ~ 0.855);平均多态信息含量(PIC ) 为0. 775 (0.709 ~0.822)。8 个位点的平均个体识别率(DP)为0.931,累积个体识别率(CDP)为0.999 9。有1 个微卫星位点(BM1706)极显著地偏离了Hardy-Weinberg 平衡(P < 0.01)。以上结果显示该分布区黑麂种群具有较高的遗传多样性。

关键词: 遗传多样性, 粪便DNA分析, 黑麂, 微卫星分子标记, 九龙山国家级自然保护区

Abstract: Black muntjac (Muntiacus crinifrons)is an important protected animal in China. Recently,the habitat of black muntjac has been seriously damaged and fragmented because of human activities. Thus,it is very important to fully understand the genetic status of black muntjac,providing the scientific foundation on which to develop protective and management
measures. We collected 61 feces samples,2 muscle samples and 2 skin samples of black muntjac in Jiulongshan National Nature Reserves using non-invasive sampling techniques. Eight specific microsatellite loci were selected for analysis of genetic diversity. The skin and muscle DNA samples served as controls. The results showed that in 61 feces DNA samples, 38 were amplified stably and showed different PCR bands. Thus,we considered the 38 faces samples to be from 38 different individuals. Fifty alleles were detected in 8 microsatellite loci,and the mean number of alleles (A)was 6,ranging from 5 to 8. The effective number of alleles (Ne)between 4.031 and 6.353,with an average of
5. 297. The expected heterozygosity (He) ranged from 0.763 to 0. 855,with an average of 0.817,and the polymorphism information content (PIC)ranged from 0. 709 to 0. 822,with an average of 0. 775. The average power of discrimination (DP )of 8 loci was 0.931,and the cumulative DP reached to 0.9999. One microsatellite locus (BM1706)very significantly deviated from Hardy-Weinberg equilibrium (P < 0. 01). According to the results,we can see that the black muntjac population in Jiulongshan National Nature Reserves has a high level of genetic diversity.