【作者】 刘正华；

【导师】 罗琛；

【作者基本信息】 湖南师范大学 ， 发育生物学， 2005， 硕士

【摘要】 本实验利用随机扩增多态性DNA技术和微卫星DNA分析技术对湘江流域普通草鱼群体、一代及连续两代人工诱导雌核发育获得的极体型雌核发育草鱼群体的基因纯合性及遗传多样性进行了分析。 主要研究内容和结果包括: 由于RAPD技术的实验结果很容易因实验条件和反应参数的不同而造成差异。为了建立能通用的草鱼基因组多态性分析RAPD分子标记体系,需要利用RAPD位点按照孟德尔共显性规律遗传的特点,用不同遗传背景的材料对多态性的RAPD位点进行统计遗传学比较分析以判断其真实性。本实验选择具有遗传多态性的湘江流域草鱼群体、一代及连续两代人工诱导雌核发育获得的极体型雌核发育草鱼品系,对一些可能作为草鱼基因组DNA分子标记的RAPD位点进行了遗传学比较分析。所用的10条多态性随机引物在三个群体中共检测到116个RAPD位点和30个多态性RAPD位点。三个不同遗传背景群体的遗传统计对比分析结果表明:30个在湘江流域普通草鱼群体中存在的多态位性位点,有23个在两个极体型雌核发育草鱼群体中的分布符合孟德尔遗传规律,可以作为草鱼基因组DNA分析的可靠分子标记。通过对这23个位点的检测结果分析表明:在人工诱导雌核发育过程中,存在RAPD位点的快速丢失现象,两次人工诱导雌核发育过程中共丢失了14个多态性RAPD位点。 采用微卫星DNA技术,对湘江流域普通草鱼群体,一代及连续两代极体型人工诱导雌核发育草鱼群体基因组DNA进行统计分析。结果发现,湘江流域草鱼群体存在一定程度的遗传多态性;大多数被检测的微卫星位点存在两个以上的等位基因,但遗传多样性程度低。一代人工诱导雌核发育草鱼群体中个体的基因位点已经基本纯合,但就整个群体而言,个体之间的基因型还不完全一致,表现出一定的多态性。连续两代人工诱导雌核发育草鱼群体中不仅所检测个体的基因位点已经完全纯合,并且各个体的基因型也完全相同。这些观察结果说明所检测的两代人工诱导雌核发育草鱼群体是纯合的,经连续两代人工诱导雌核发育可能建立起草鱼纯系。本实验结果还发现不仅草鱼的微更多还原

【Abstract】 Random Amplified Polymorphism DNA (RAPD) and microsatellite were employed for analysis on the gene gynozygosity and genetic diversity of a Xiangjiang river grass carp group(xiangjiang group), a one-generation artificial induced meio-gynogenetic grass carp group (meio-gynogenetic-1 group) and a two-generation artificial induced meio-gynogenetic grass carp group (meio-gynogenetic-2 group).The main contents and results were as follows:The products amplified by RAPD are always variable in different experiments. In order to establish a universal RAPD molecular marker system for grass carp genomic DNA polymorphism analysis, selected polymorphism RAPD loci should be checked by using different strains with distinct genetic backgrounds according to the law of Mendelian genetics. The Xiangjiang group with genetic diversity, meio-gynogenetic-1 group and meio-gyno-genetic-2 group with distinct genetic background and high homozygous were employed for RAPD analysis. Some polymorphic RAPD loci that could be the molecular markers of grass carp genomic DNA were comparatively analyzed. With 10 polymorphism random primers, total 116 distinct loci and 30 polymorphic loci were detected in the three groups by the RAPD-PCR. Among the 30 polymorphism loci, 23 of loci distributs accorded with the law of Mendelian genetics judged by genetic statistical analysis with the two gynogenetic grass carp groups. Therefore, the 23 polymorphic loci were the authentic molecular markers for grass carp genomic DNA. Some of the RAPD polymorphic loci have been lost rapidly during the process of the artificial gynogenesis. 14 loci of 23 polymorphic loci have been lost after two-generation artificial gynogenesis.The genomes of the Xiangjiang group, meio- gynogenetic-1更多还原