林麝微卫星标记开发与圈养种群遗传多样性分析

发布时间：2018-02-01 00:50

  本文关键词： 微卫星 林麝 遗传多样性 磁珠富集法　出处：《西北农林科技大学》2017年硕士论文　论文类型：学位论文

【摘要】：林麝(Moschus berezovskii)是我国重要的经济动物,因其所产麝香有极高的经济价值而被大量捕杀,导致种群数量锐减,2002年被列为我国Ⅰ级重点保护的珍稀濒危野生动物。目前,开展人工养麝是保护林麝野生资源的重要手段之一,但圈养林麝又带来了新的问题,遗传相关问题尤为突出。因林麝保护级别高,分布区域有限,样品获取较难,目前有关林麝的研究报道较少。为丰富林麝遗传学有关数据,同时为麝场的养殖管理提供资料,本文开展了以下研究:运用磁珠富集法构建林麝微卫星富集文库,从文库中筛选多态微卫星位点;用8个林麝多态微卫星位点分析陕西某麝场圈养林麝群体的遗传多样性,并对所采40个林麝样进行UPGMA聚类分析。所得主要结果如下:1.采用磁珠富集法构建林麝(AC)n微卫星文库,获得16个微卫星位点,其中完全型微卫星8个,占50%,不完全型微卫星7个,占43.75%,复合型微卫星1个,占6.25%。2.设计14个林麝微卫星位点引物,经筛选发现3个多态的林麝微卫星位点,可用于林麝群体遗传多样性分析;1个林麝和马麝的种间鉴别位点。3.圈养林麝种群的遗传多样性分析结果显示,8个林麝微卫星位点的多态信息含量在0.679~0.870之间,平均多态信息含量为0.798;共检测到68个等位基因,平均等位基因数为8.5,平均有效等位基因数为5.8770;圈养林麝种群的平均观测杂合度为0.5428,平均期望杂合度为0.8337,平均Shannon’s信息指数为1.8971。卡方检验结果显示,除Mber76C位点外,其余7个微卫星位点偏离Hardy-Weinberg平衡,其中2个位点杂合子过剩,5个位点杂合子缺失。8个微卫星位点共检测到14个稀有等位基因,占等位基因总数的20.59%。4.40个圈养林麝样品的聚类分析结果表明,群体的相似系数在0.72~0.97之间,整个群体可分为9类,同一类群个体数最多为22个,最少为1个。本研究丰富了林麝微卫星标记数据,分析发现陕西麝场圈养林麝群体的遗传多样水平较高,但为了麝场长远的发展,仍需不断提高麝场的养殖管理水平,建立完善的群体遗传谱系,实时进行群体遗传监测,避免种群退化的风险。
[Abstract]:Moschus berezovskii (Moschus berezovskii) is an important economic animal in China. Because of its high economic value, Moschus berezovskii has been killed in large numbers, resulting in a sharp decrease in population. In 2002, it was listed as a rare and endangered wild animal in China. At present, artificial breeding of musk deer is one of the important means to protect the wild resources of forest musk deer, but the captive forest musk deer has brought new problems. Because of the high protection level and limited distribution area, it is difficult to obtain samples, so there are few reports about forest musk deer. It is to enrich the genetic data of forest musk deer. At the same time, the following studies were carried out for the culture management of musk deer farm: the microsatellite enrichment library of forest musk deer was constructed by magnetic bead enrichment method, and the polymorphic microsatellite loci were screened from the library; The genetic diversity of captive forest musk deer populations in a musk deer farm in Shaanxi Province was analyzed by using 8 loci of microsatellite polymorphism of forest musk deer. The main results were as follows: 1. The microsatellite library of ACN was constructed by magnetic bead enrichment method, and 16 microsatellite loci were obtained. There were 8 complete microsatellites (50 cents), 7 incomplete microsatellites (43.75) and 1 compound microsatellite (6.25.2.). Fourteen primers were designed for the microsatellite loci of forest musk deer. Three polymorphic microsatellite loci were identified, which can be used to analyze the genetic diversity of forest musk deer populations. The results of genetic diversity analysis showed that the polymorphic information content of microsatellite loci of 8 forest musk deer was between 0.679 and 0.870. The average polymorphic information content was 0.798; 68 alleles were detected, the average number of alleles was 8.5 and the number of effective alleles was 5.8770; The average observed heterozygosity, expected heterozygosity and Shannon's information index were 0.5428, 0.8337 and 1.8971 respectively. The chi-square test results showed that the observed heterozygosity was 0.5428, the average expected heterozygosity was 0.8337, and the average Shannon's information index was 1.8971. Except for Mber76C, the other 7 microsatellite sites deviated from the Hardy-Weinberg equilibrium, and 2 of them had excess heterozygotes. A total of 14 rare alleles were detected in 8 microsatellite loci, accounting for 20.59% of the total alleles, and the cluster analysis of 4.40 captive forest musk deer samples showed that. The similarity coefficient of the population was between 0.72 and 0.97, the whole population could be divided into 9 categories, the number of individuals in the same group was 22 at most, and at least one. This study enriched the microsatellite marker data of forest musk deer. It was found that the genetic diversity of the population in the captive forest of Shaanxi musk deer farm was high, but in order to develop the musk deer farm for a long time, it was still necessary to improve the management level of the musk deer farm and establish a complete genetic pedigree of the population. Population genetic monitoring is carried out in real time to avoid the risk of population degradation.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S865.41
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本文编号：1480461