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刘正华, 陈金辉, 黄明敏, 郑康, 罗琛. 草鱼基因组DNA一些RAPD位点的遗传分析及分子标记筛选[J]. 水生生物学报, 2006, 30(3): 292-297.
引用本文: 刘正华, 陈金辉, 黄明敏, 郑康, 罗琛. 草鱼基因组DNA一些RAPD位点的遗传分析及分子标记筛选[J]. 水生生物学报, 2006, 30(3): 292-297.
LIU Zheng-Hua, CHEN Jin-Hui, HUANG Ming-Min, ZHENG Kang, LUO Chen. GENETIC ANALYSIS ON SOME RAPD LOCI FOR IDENTIFICATION OF MOLECULAR MARKERS IN GRASS CARP GENOMIC DNA[J]. ACTA HYDROBIOLOGICA SINICA, 2006, 30(3): 292-297.
Citation: LIU Zheng-Hua, CHEN Jin-Hui, HUANG Ming-Min, ZHENG Kang, LUO Chen. GENETIC ANALYSIS ON SOME RAPD LOCI FOR IDENTIFICATION OF MOLECULAR MARKERS IN GRASS CARP GENOMIC DNA[J]. ACTA HYDROBIOLOGICA SINICA, 2006, 30(3): 292-297.

草鱼基因组DNA一些RAPD位点的遗传分析及分子标记筛选

GENETIC ANALYSIS ON SOME RAPD LOCI FOR IDENTIFICATION OF MOLECULAR MARKERS IN GRASS CARP GENOMIC DNA

  • 摘要: 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。为此,本实验选择具有遗传多态性的湘江流域草鱼群体和经连续两代人工诱导雌核发育获得的雌核发育草鱼品系,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条多态性随机引物共检测到30个多态性RAPD位点。两个不同遗传背景群体的遗传统计对比分析结果表明:这30个多态位性位点在湘江流域草鱼群体和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。本实验的观察结果还表明: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17个多态性位点。因此,加强对自然水体中草鱼种质资源多样性的保护和利用各种现代生物学技术纯化、筛选和组合优良性状基因,是草鱼遗传育种中同样重要和不可或缺的两个方面。

     

    Abstract: The products amplified by Random Amplified Polymorphism DNA(RAPD) techniques 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. In this experiment, some polymorphic RAPD loci that could be employed as molecular markers for grass carp genomic DNA analysis have been examined by comparatively analyzing the genomes of theXiangjiang River grass carp group(Xiangjiang group) and a two-generation artificial induced meio-gynogenetic grass carp group(meio-gynogenetic-2 group). The Xiangjiang group can provide distinct genetic backgrounds samples andmay include all the genotype of grass carp,while the meiogynogenetic-2 group can provide clear genetic background samples. The different characterizations of the two groups can help identify the polymorphic RAPD loci in grass carp. Total 36 individuals of the Xiangjiang group and total 29 individuals of meiogynogenetic-2 group were examined with 10 polymorphism random primers in this experiment.With RAPD-PCR method, total 30 polymorphic loci have been identified in Xiangjiang group. In meiogynogenetic- 2 group, 17 of the polymorphic loci identified in Xiangjiang group were not detected in all the examined individuals. Only 13 of the 30 polymorphic loci in Xiangjiang group were identified in meio-gynogenetic-2 group.However only 7 of the 13 detected loci showed polymorphism, the other 6 lociwere identified in all the individuals in meio-gynogenetic-2 group and no polymorphism was observed. Because the meio-gynogenetic-2 individuals are highly purified during the gynogenesis and artificial selection for two generations, obviously, the 17 polymorphic loci that were undetected in meio-gynogenetic-2 group but in Xiangjiang group and the 6 polymorphic loci that detected in all the examined individuals of the meio-gynogenetic-2 group are the real polymorphic loci in grass carp.A chi-squared test was done on the 7 loci that show polymorphism to determine if the ratio of the detected and undetected individualswere consistent with the 1B1 expected ratio. The P values of the 7 polymorphic loci are from 0. 1936 to 0. 5775. This genetic statistical analysis indicated that the 7 polymorphic loci in Xiangjiang group and meio- gynogenetic-2 group are also consistent with the law of Mendelian genetics well. The 30 polymorphic RAPD loci, therefore, could be used as reliable molecular markers for the grass carp genomic DNA analysis. The observat ion that 17 polymorphic loci have been lost during the process of the two-generation artificial gynogenesis strongly suggested that protection of the diversity of natural grass carp resource and selection of homozygous traits with desired economic genotypes are both very important for grass carp breeding.

     

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