Breeding structure in fish population by RFLP analysis of mitochondrial DNA

线粒体DNA（MTDNA）基因组的限制性片段长度多态性（RFLP）分析是估计养殖种群母系数量的一种合适的方法，因为：母系遗传，在动物中是一个非重组基因组（约16-18Kbp），单倍型的数量直接代表母系的数量等。在本研究中，I:(1)对鲤鱼和马苏鲑鱼复合体mtDNA的快速分离方法进行了改进，用玻璃丝碱法处理成功。(2)利用15种6碱基识别限制性内切酶在两种养殖鱼类中检测到5个单倍型。(3)利用内切酶双酶切法绘制单倍型酶切图谱，确定了33个酶切位点。(4)从限制性片段的结果来看，可以认为mtDNA的变异是由碱基的简单替换引起的。II：利用6个6碱基识别的限制性内切酶对马苏鲑鱼复合体养殖种群的mtDNA进行RFLP分析，(5)在4个种群的59个个体中观察到13个mtDNA单倍型。(6)非选择群体的核苷酸多样性在0.00170 ~ 0.00253之间，而选择的amago-parr群体的核苷酸多样性没有变化。(7)结果表明，amago-parr种群由一个母系组成，而其他种群由多个母系组成。

The RFLP(restriction fragment length polymorphism)analysis of the mitochondrial DNA(MTDNA)genome is a suitable method for estimating the number of maternal lines in cultured stocks because : maternal inheritance, it is a nonrecombining genome(apploximately 16-18Kbp)in animals, the number of haplotypes represents the number of maternal lines directly and so on.In the present study, I : (l)modification of the method of rapid isolation of mtDNA from carp and masu salmon complex is successfully carried out by alkaline treatment with glassailk. (2)The 5 haplotypes were detected in the two cultured stocks of carp by using 15 kinds of 6-base recognizing restriction endonucleases. (3)Restriction maps of the haplotypes were made up by double-digestion of endonucleases and then the 33 restriction sites were defined in the carp. (4)From the results of restriction naps, the variation of mtDNA is considered to be occurred by the simple substitution of bases. II : Based on the results of RFLP analysis of mtDNA using six 6-base recognized restriction endonuclease in the cultured stocks of masu salmon complex, (5)13 mtDNA haplotypes were observed among 59 individuals of the 4 stocks. (6)The nucleotide diversity within the non-selected stocks ranged from 0.00170 to 0.00253, while no variation was observed in selected amago-parr stock. (7)The results suggest that the amago-parr stock consists of one maternal line while the other stocks consist of several maternal lines.