作 者 ：何丽,张于光,彭红兰,李迪强,李大全*
期 刊 ：生态学报 2010年 30卷 16期 页码：4340~4350
关键词：川金丝猴(Rhinopithecus roxellana)；神农架国家级自然保护区；非损伤性；遗传多样性；
Keywords:Sichuan snub-nosed monkey (Rhinopithecus roxellana), Shennongjia National Nature Reserve, non-invasive DNA technology, genetic diversity,
摘 要 ：湖北神农架国家级自然保护区是中国川金丝猴（Rhinopithecus roxellana）的重要分布区之一，为了更好地了解神农架川金丝猴种群遗传结构，并制定有效的保护对策，采集了该保护区2个川金丝猴种群126份样品，其中分布在大龙潭的人工补食种群粪便样品60份，分布在千家坪的一个小种群粪便样品63份、肌肉样品3份。通过应用线粒体DNA（mtDNA）引物和筛选出的9对近缘微卫星位点成功地进行了川金丝猴种群的遗传多样性和遗传结构分析，同时使用Y染色体相关标记方法进行性别鉴定。结果显示：在24份川金丝猴mtDNA D-loop区401 bp序列中，共检测出27个变异位点，定义了20个单倍型，单倍型多样性（h）为09820，核苷酸多样性（π）为00129；其中有9对微卫星位点能够在117份粪便样品DNA中稳定扩增，2对位点可能偏离Hardy-Weinberg平衡，平均有效等位基因数（Ne）为3.85，遗传杂合度（Ho）为07450；多态信息含量（PIC）为05950。使用Y染色体标记方法，在58个特有的基因型中检测到18个雄性和40个雌性川金丝猴。微卫星标记和mtDNA D-loop区基因序列均表明神农架川金丝猴存在较丰富的遗传多样性，同时2个取样点川金丝猴种群间出现了明显的遗传分化。
Abstract:Sichuan snub-nosed monkey (Rhinopithecus roxellanae) is an unique primates in china, witch is in severe danger now and is mainly distributed in Shanxi，Hubei, Sichuan and Gansu province. In recent years, since the forest has been cut excessively and human activities have become increasingly frequent, habitat loss and habitat fragmentation have caused the population isolation and differentiation, the reduction of genetic diversity in Sichuan snub-nosed monkey. Hubei Shennongjia National Nature Reserve is one of the major habitating areas for Sichuan snub-nosed monkey in China. In order to understand the genetic structure of the species for effective conservation, a total of 126 samples were collected from two natural populations in the reserved region, including 60 fecal samples from Dalongtan population, and 63 fecal samples plus 3 muscle samples from Qianjiaping population. The genetic structure and diversity of the populations were estimated by analyzing the samples for mitochondrial DNA (mtDNA) gene fragment, and 9 selected microsatellite loci including D6S474, D3S1768, D1S533, D8S505, D15S644, D1S207, D7S1830, D17S1290, D5S820 In addition, sex identification of the samples was performed using Y chromosome-markers. The results of analysis showed that: (1) a total of 20 haplotypes and 27 diversity sites were found in the 24 random samples at a 401 bp region of mtDNA D-loop, with haplotype diversity value equals to 0982, and nucleotide diversity was 00129. (2) All of 9 microsatellite loci were stably detected in 117 of total fecal DNA preparations and genotyping success rate was 87%, two loci showed deviation from Hardy-Weinberg equilibrium. The average effective number of alleles was 3.85, the average observed heterozygosis and polymorphism information content were 0745 and 0595, respectively. By using Y chromosome marker techniques, 18 male and 40 female was identified in 58 unique Sichuan snub-nosed monkey genotypes. This sex identification technique is very simple, reliable, and especially applicable for the fecal DNA test. These results indicated a high level of genetic diversity in Sichuan snub-nosed monkey in Shennongjia National Nature Reserve. All the same time, the two populations we have invested showed a significant genetic differentiation. Some protection advices were proposed for the species-source saving and population rejuvenation of Sichuan snub-nosed monkey according to protection genetics and research achievement above.
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