【作者】 葛学亮；

【导师】 孙中武；

【作者基本信息】 东北林业大学 ， 细胞生物学， 2007， 硕士

【摘要】 唇(鱼骨)(Hemibarbus labeo)是黑龙江优质名贵淡水鱼，2004年在黑龙江省首次成功实现人工繁育。本研究通过同工酶和RAPD分析方法系统研究黑龙江、乌苏里江和长江野生唇(鱼骨)与养殖唇(鱼骨)群体的遗传多样性，为唇(鱼骨)种质保护提供科学依据，为唇(鱼骨)的资源开发利用及其选育与推广提供遗传学基础资料。首先，采用聚丙烯酰胺凝胶电泳对野生和养殖唇(鱼骨)群体进行同工酶组织特异性和遗传多样性分析，结果显示，大部分的同工酶在组织间呈现较强的特异性，其中以EST、GDH、ME、G-6-PDH、CAT、ADH和POD较为明显。α-AMY、POD在雌雄个体间差异性较强：EST、G-6-PDH与COX在野生和养殖唇(鱼骨)间差异性较大。选取10种同工酶的23个位点用于群体遗传多样性分析，其中Amy-1、Amy-2、Est-2、G-6-pdh-2、Ldh-1、Mdh-1、Mdh-2、Me-2、Sdh-2共8个座位表现为多态。在群体水平上，各个群体的多态座位比例差异很小，平均值为38.26％，平均每个位点的有效等位基因数为1.3129，平均杂合度预期值为0.1588，各群体的Shannon多样性指数为0.2267。Hardy-Weinberg遗传平衡符合度检验表明唇(鱼骨)遗传变异性比较高，处于杂合子过剩状态。其次，运用RAPD技术对野生和养殖唇(鱼骨)群体进行遗传多样性分析。60个随机引物中筛选出6个有效引物进行遗传学分析。在群体水平上，各个群体的多态座位比例差异不大，平均值为43.10％，平均每个位点的有效等位基因数为1.4274，Nei’s基因多样度为0.3223，Shannon’s多样性指数为0.2111。各项遗传学指标与同工酶数据趋势一致。无论从蛋白质水平还是从DNA水平来看，唇(鱼骨)遗传多样性水平较高，逊克群体最高，涪陵群体最低。各群体的遗传变异主要存在于群体内，群体间几乎无遗传分化，群体内的遗传分化大于群体间的分化。基因流程度较高，有利于保持良好的遗传多样性水平。更多还原

【Abstract】 Hemibarbus labeo is a high quality valuable fresh water fish in Heilongjiang, and wascultrued for the first time in the province of Heilongjiang in 2004. By using allozyme andRAPD analysis, the genetic diversity of 3 wild populations in Heilongjiang River, WusuliRiver, Changjiang River and a hatchery H. labeo population were studied. Those providedscientific basis of germ plasm protection, resources development, seed selection and geneticsfoundation data.Firstly, vertical PAGE was used to investigate the tissue-specific and genetic diversity ofisozymic systems in wild and breeding H. Iabeo populations. The results showed that theexpression of all the isozymes exhibited a tissue specific expression apparently. Especiallyamong EST, GDH, ME, G-6-PDH, CAT, ADH and POD.α-AMY and POD showed greatdifference between male and female individuals. While, EST, G-6-PDH and COX had distinctdifference among wild and hatchery populations. Among the 10 enzymes which were studied,23 genetic loci were detected. Amy-1, Amy-2, Est-2, G-6-pdh-2, Ldh-1, Mdh-1, Mdh-2, Me-2.Sdh-2 were polymorphic loci. The percentage of polymorphic loci had small difference amongpopulations. Mean value was 38.26%, the average effective number of alleles was 1.3129, theNei’s expected heterozygosity was 0.1588, and Shannon’s Information index was 0.2267.Hardy-Weinberg heredity equilibrium proof-test indicated that H. labeo resource quality wasgood.Secondly, RAPD method was used in analyzing the genetic diversity of wild and breedingH. labeo populations. 60 random primers were used for the analysis of nuclear DNApolymorphism of 5 populations, and 6 primers were effective.The percentage of polymorphicloci also had small difference among populations, mean value was 43.10%, the averageeffective number of alleles was 1.4274, the Nei’s gene diversity was 0.3223. and Shannon’sInformation index was 0.2111. Genetics index was consistent with isozyme data.Regardless of the DNA level or protein level, H. labeo resource quality was stillgood.With the highest level in Xunke population and the lowest level in Fuling population.Inherit variation were detected within population level. The gene differentiation coefficient indicated that there was almost no genetic differentiation among them, while the geneticdifferentiation degree within population level was stronger than that was among populationlevel.These results implied that various genetic indexes in different populations would give usthe theoretical guidance in breeding and genetic improvement.更多还原