基于粪便DNA的雪豹种群调查和遗传多样性

Population survey and genetic diversity of snow leopards Panthera uncia as revealed by fecal DNA

雪豹(Pantherauncia)是仅分布于亚洲高海拔山区的珍稀濒危猫科动物。本研究在印度西南部(Ladakh)、中国青海和蒙古国的南部(南Gobi)3个独立的雪豹分布区共采集109份粪便样品。应用线粒体DNA(mtDNA)cytb基因特异性引物对109份粪便样品进行鉴定,发现有31份粪便来自雪豹,其中印度Ladakh、我国青海和蒙古国南Gobi的雪豹样品分别为17份、3份和11份。利用重新筛选设计的7对家猫(Feliscatus)微卫星引物,对雪豹粪便样品进行了基因分型分析,结果发现在Ladakh和南Gobi检测到的雪豹粪便样品分别来自4只和5只不同的雪豹个体,而青海的样品则来自同一只雪豹;遗传多样性统计分析表明,蒙古国南Gobi的雪豹微卫星遗传多样性水平低于印度的Ladakh。研究结果表明了粪便DNA在雪豹种群监测和遗传多样性研究中的可行性。

Snow leopards Panthera uncia occur in rugged, high altitude regions of Central Asia, where they are endangered as a result of human induced factors including low prey densities and poaching. Information on the status of this felid is limited in many regions. We examined the feasibility of using noninvasive genetic methods to monitor snow leopard populations and to study their genetic diversity. We observed snow leopard scats in three separate geographic regions during brief surveys and collected 109 scats from southwestern India （Ladakh）, western China （Qinghai）, and southern Mongolia （South Gobi ）. We used a panel of mitochondrial DNA （mtDNA） primers and seven domestic cat （Fells catus） microsatellite loci to survey the snow leopard population and to examine their genetic diversity. Of the 32 scat samples found in Ladakh, 50 found in Qinghai, and 27 found in Mongolia, 80% produced positive species identification. Positive snow leopard identification was made in 17 of 32 scats collected in Ladakh, where 6 red fox and 2 wolf/dog scats were also detected. In Qinghai, 36 scats were identified by mtDNA analysis, including 3 snow leopard, 10 lynx, 21 red fox, and 2 wolf/dog. Only snow leopard （11） and red fox （13） scats were observed in South Gobi. We genotyped 31 confirmed snow leopard scats with the 7 microsatellites selected. There were a total of 9 unique genotypes detected： 4 in Ladakh and 5 in South Gobi. Using the Y-linked AMELY marker, we identified 2 males and 2 females in Ladakh, and 3 males and 2 females in South Gobi. All 7 loci were polymorphic in both Ladakh and South Gobi with numbers of alleles ranging from 2 to d. No loci were found to be out of HW equilibrium after sequential Bonferroni correction of significance levels. The numbers of alleles and heterozygosity were lower in South Gobi, although our sample sizes were too small to test for significance. The Fsy estimated between the two areas was 0. 171. Our findings highlight the efficacy of noninvasive genetic surveys for monitoring snow leopards and the feasibility of large-scale studies that will provide critical information for conservation of snow leopards throughout Central Asia