基于非损伤取样法的林麝微卫星遗传多样性分析

林麝(Moschus berezovskii)为我国一级重点保护野生动物，为了保护野生林麝种群、满足中医药等行业对天然麝香的需求，我国从20世纪50年代开始人工饲养林麝。维持高的遗传多样性是实现饲养林麝种群可持续增长并放归野外的关键因素。本研究旨在筛选可用于粪便DNA扩增的林麝四碱基微卫星位点，并评估陕西凤县2个饲养林麝种群的遗传多样性。通过搜集文献，共获得25个林麝四碱基微卫星位点，其中19个可从粪便DNA中稳定扩增，且基因分型峰型较好，能用于后续分析，与前人研究相比增加了13个粪便DNA微卫星位点。利用19个微卫星位点，对陕西凤县富民和海兴2个饲养林麝种群共计95只林麝进行遗传多样性分析，其中7个位点多态信息含量超过0.5，为高多态性位点，10个位点符合哈迪-温伯格平衡(P > 0.05)。本研究中的95只林麝共存在99个等位基因，有效等位基因总数为43.880 5，平均Shannon’s信息指数为0.930 6，平均多态信息含量为0.428 3，表明陕西饲养林麝种群具有较高的遗传多样性。平均观测杂合度为0.449 4，平均期望杂合度为0.467 5，观测杂合度低于期望杂合度，种群存在近交的趋势。富民麝场饲养林麝遗传多样性高于海兴麝场，2个种群的遗传分化较小，有较大的基因流，所有林麝个体共来源于4个基因簇，富民麝场的林麝主要来自于1和2基因簇，海兴麝场的林麝主要来自于3和4基因簇。

Microsatellite Genetic Diversity Analysis of Forest Musk Deer Moschus berezovskii Based on Noninvasive Sampling Technology

[Objectives] Forest Musk Deer Moschus berezovskii is first class protection wildlife in China. In order to protect the wild Forest Musk Deer population and meet the demand for natural musk in traditional Chinese medicine and other industries, China began to raise Forest Musk Deer artificially in the 1950s. Maintaining high genetic diversity of the population is the key factor to realize the sustainable growth and release of the captive Forest Musk Deer population into the wild. [Methods] In Fengxian County, Shaanxi Province, we collected feces samples from 95 Forest Musk Deer in two captive populations and kept all the samples in a lab refrigerator at﹣20 ℃ until DNA extraction. We selected published tetranucleotide microsatellite loci, then identified microsatellite loci that can be reliably amplified from fecal DNA. The forward primers of these loci were colored with fluorescent dyes and used to analyze the genetic diversity of 95 captive Forest Musk Deer. [Results] Twenty-five tetranucleotide microsatellite loci of Forest Musk Deer were obtained, of which 21 could be steadily amplified from fecal DNA, and two of them had chaotic peaks that were not suitable for analysis (Appendix 1). For the 19 microsatellite loci, there were 7 loci with polymorphism information content > 0.5, which were high polymorphism loci, and 10 loci were consistent with Hardy-Weinberg equilibrium (P > 0.05) (Table 1). Among the two populations studied, 95 individuals had 99 alleles and 43.880 5 effective alleles, the mean of Shannon’s index and polymorphism richness were 0.930 6 and 0.428 3 respectively. The mean observed heterozygosity was 0.449 4, and the mean expected heterozygosity was 0.467 5. The genetic diversity of the Fumin farm population was higher than that of the Haixing farm (Table 3). All the Forest Musk Deer individuals came from four gene clusters. The Forest Musk Deer of the Fumin farm mainly came from gene cluster 1 and 2, and the Forest Musk Deer of the Haixing farm mainly came from gene cluster 3 and 4 (Fig. 2). [Conclusion] The selected microsatellite loci were 13 more than the previous studies, which can provide a reference for future studies on genetic diversity of Forest Musk Deer based on fecal samples. The captive Forest Musk Deer in Fengxian County, Shaanxi Province showed high genetic diversity but a tendency to inbreed. The genetic differentiation of Fumin Forest Musk Deer farm and Haixing Forest Musk Deer farm was small, and there was a large gene flow. It is suggested that the provenance of each captive population can be changed to increase the degree of heterozygosity.