非损伤采样提取DNA技术在猫科动物调查中的应用

The application of extracting DNA from noninvasive samples in feline species identification

猫科动物广泛分布于世界各地（Johnson et al.,2006）,全球现存37种猫科动物,其中25种已被国际自然保护联盟（IUCN）列为濒危和易危,86%以上的种群数量处于下降或未知状态（IUCN Red List,2011）。我国有13种猫科动物,占世界种类的35%（王应祥,2003）,其中属于国家一级保护的有4种,二级保护的有8种,然而,对这些珍稀物种的生态学研究并不充分（高耀亭,1987；国家林业局，2009）。

Field surveys utilizing indirect indicators are often adopted in population ecology studies of wild cats in China.However,research using molecular biology techniques are still relatively rare. Confirmation of molecular techniques as an informative method for wide use in population analysis is required. Therefore,in this paper,molecular markers（ mt DNA,sex gene and microsatellites） were used for determining applicability in conservation genetic research by comparing fecal samples from 4 feline species. Nine of 11 fecal samples suspected for Siberian tiger were identified as Panthera tigris altaica and16 of 30 fecal samples suspected as Eurasian lynx were confirmed as Lynx lynx. Additionally five tigers（ four males and one female） and five Eurasian lynx（ three males and two females） were identified. Genetic diversity analysis indicated that the average number of allele of microsatellite loci of the Siberian tiger was 2. 64; the average observed heterozygosity was 0. 489 and the average polymorphism information content was 0. 301,while these values for Eurasian lynx were 2. 88,0. 575,and0. 428,respectively,indicating a moderate level of genetic diversity for the two species. Analysis of Eurasian lynx fecal samples preserved in pure ethanol were as follows： species identification rate of 69%; sex identification rate of 100%; microsatellite amplification rate 85%. By comparison analysis of air dried samples were 41%,71%,and 25% for species identification,sex identification and microsatellite amplification,respectively. The species identification rate for fecal samples preserved for one year was 80%,the sex identification rate was 75% and the microsatellite amplification rate was 28%. Our results provide a reliable method for high success in DNA extraction,from noninvasive fecal samples,for identifying wild cats and determining population genetic structure.