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基于粪便DNA 的青海雪豹种群遗传结构初步研究

张于光,何丽,朵海瑞,李迪强,金崑   

  1. (中国林业科学研究院森林生态环境与保护研究所,国家林业局森林生态环境重点实验室,北京100091
  • 出版日期:2009-09-03 发布日期:2009-03-08

A preliminary study on the population genetic structure of snow leopard (Unica unica) in Qinghai Province utilizing fecal DNA

ZHANG Yuguang,HE Li,DUO Hairui,LI Diqiang,JIN Kun   

  1. (Institute of ForestEcology,Environment and Protection,and the Key Laboratory of Forest Ecology and Environment of State Forestry Administration,Chinese Academy of Forestry,Beijing 100091,China
  • Online:2009-09-03 Published:2009-03-08

摘要: 雪豹是国际关注的全球濒危物种,由于独特的生活习性,其确切分布区、种群数量和遗传结构信息非常有限,基于粪便DNA 分析技术的发展为该物种的研究提供了新的手段。在青海省都兰县宗加乡、诺木红乡和治多县索加乡收集到106 份疑似雪豹粪便样品,成功地扩增了78 份粪便样品的mtDNA Cyt b 基因片段,并进行测序。经过GenBank 数据库的Blastn 比对,确定了21 份为雪豹粪便,其中宗加乡11 份、诺木红乡5 份、索加乡5份。经过Clustal W 和DNASP 软件比对分析,在21 份雪豹粪便DNA 中,共检测出7 个多态性位点,分为4 个单倍型,单倍型多态性(h)为0. 384,核苷酸多态性(π)为0.35% 。三个样地之间的遗传距离为0.002 ~ 0.005,核苷酸差异为0.200 ~1. 273。结果表明了3 个取样区均有雪豹存在,且存在较丰富的遗传多态性,样地间也存在一定的遗传差异。

关键词: 雪豹, 生物多样性, 遗传结构, Cyt b 基因

Abstract: The snow leopard (Uncia uncia) is a globally endangered and threatened species,and studies on their population
and genetic structure has been hampered due to their rugged and remote habitat. Molecular methods incorporating noninvasive sampling of wildlife populations via the collection of scats have provided a new chance for understanding the mysterious snow leopard. In this study a total of 106 scat samples were collected in the dominant distribution areas of Zongjia Township (ZJ) and Nuomuhong Township (NMH)in Dulan County and Suojia Township (SJ)in Zhiduo County,Qinghai Province. DNA was directly extracted from the scats and the mtDNA Cyt b gene fragment was amplified using PCR. 78 scats were successfully amplified and sequenced. Sequences were aligned with the GenBank by BLAST software,and 21 of the scats were identified utilizing DNA as snow leopards (11,5 and 5 in ZJ,NMH and SJ,respectively. According to the analysis of Clustal W and DNASP software,4 haplotypes and 7 diversity sites were found in the 21 snow leopard scat DNA,and their haplotype diversity was 0.384 and nucleotide diversity was 0.35%. The genetic distance and nucleotide divergence among the 3 snow leopard
populations was 0.002 -0.005 and 0.200 -1.273,respectively. Therefore,the results showed that the snow leopard has high variation in the mt DNA Cyt b gene and significant genetic distance among the three sampling sites.

Key words: Biodiversity, Cyt b gene, Genetic structure, Snow leopard (Unica unica)