应用RAPD技术对6种玻璃鳗的种质鉴定和遗传多样性分析

Identification of 6 Populations of Glass Eeland Genetic Diversity Analysis by Means of Random DNA(RAPD)

运用随机多态性扩增DNA(random amplified polymorphic DNA,RAPD)技术对日本鳗鲡(Anguilla japoni-ca)、欧洲鳗鲡(A.anguilla)、美洲鳗鲡(A.rostrata)、花鳗鲡(A.marmorata)、莫桑比克鳗鲡(A.mossambica)和西里伯斯鳗鲡(A.celebesensis)的遗传多样性进行研究,分析比较6种鳗鲡的种间特异性、种间相似性和遗传距离。筛选出14个重复性好的RAPD随机引物,分别用于6种鳗鲡基因组DNA的分析,其中,引物S21、S28、S42在6种鳗鲡中所产生的条带均有所差异,可单独或组合作为6个鳗鲡品种鉴别的分子标记。应用DPS数据处理系统中的聚类分析软件包分析6种鳗鲡的遗传差异和亲缘关系,结果表明,6种鳗鲡群体间的遗传相似系数在0.0769～0.4615,差异显著(P<0.05),美洲鳗鲡与欧洲鳗鲡之间的遗传距离最小,为0.5385。依据群体间的遗传距离,绘制鳗鲡遗传关系图谱,6种鳗鲡分为2类,其中,美洲鳗鲡与欧洲鳗鲡首先聚类,西里伯斯鳗鲡与这两种鳗鲡聚为一类;同时,莫桑比克鳗鲡与花鳗鲡的亲缘关系较近,而与日本鳗鲡的亲缘关系渐远,这3种鳗鲡聚为一类。结果证实这6种鳗鲡进化分布主要受到地理结构和洋流的影响,而并非只是单纯的地理位置上远近关系的影响,为鳗鲡品种的快速鉴定和种质资源的研究奠定基础。

RAPD（random amplified polymorphic DNA） markers were applied to detect the genetic diversity and molecular evolution of 6 populations of glass ell included Anguilla mossambica,A.japonica,A.marmorata,A.rostrata,A.anguilla and A.celebesensis.14 arbitrary primers screened out of 32 random primers were adopted to analyze the polymorphism in RAPD profiles,the random primers S21,S28 and S42 produced different bands in the 6 populations glass eel,which could be used to identify the stock.Total 89 markers were amplified in a range of 300-2 200 bp,which were distinguishable and reproducible,87 bands of them were polymorphic,about 97.75% of the total bands.Genetic distance coefficient analysis of different bands was carried out by statistical data software （DPS） .The un-weighted pair-group method using arithmetic average （UPGMA） cluster analysis was used and dendrograms was protracted.The results showed as the following： the polymorphic loci ratio in A.mossambica,A.japonica,A.marmorata,A.rostrata,A.Anguilla and A.celebesensis were 34.83% ,26.96% ,22.47% ,26.96% ,13.48% and 31.46% ,respectively.The mean genetic similarity indexes of intra-species （S） were 0.901 1,0.889 9,0.803 8,0.701 1 and 0.788 7,respectively.The mean genetic distances of intra-species（D） were 0.098 9,0.110 1,0.196 2,0.298 9 and 0.211 3,respectively.The mean genetic distances of inter-species （Dxy） were in a range of 0.538 5-0.923 3.The 6 populations glass ell were divided into 2 groups,the results confirmed the geographical distribution of the 6 glass eel,this can compensate the deficiency of the identification of the glass eel by shape.