双齿围沙蚕遗传多样性的随机扩增多态DNA(RAPD)分析  

Genetic Diversity in Perineeris aibuhitensis by RAPD Analysis

作　　者：蒋霞敏[1] 柳敏海 符方尧[1] 李正[1] 

机构地区：[1]宁波大学生命科学与生物工程学院,浙江宁波315211 [2]舟山市水产研究所,浙江舟山316000

出　　处：《宁波大学学报（理工版）》2006年第2期166-172,共7页Journal of Ningbo University：Natural Science and Engineering Edition

基　　金：浙江省科技厅重点项目(2005C22043);浙江省教育厅科研基金(20030463);宁波市农办项目(2004703)

摘　　要：通过实验比较获得了一种高效、简便的双齿围沙蚕基因组DNA的提取方法,以该法提取的基因组DNA为模板,对双齿围沙蚕RAPD分析中的DNA模板浓度、镁离子浓度、dNTPs浓度、引物浓度和Taq酶浓度进行了研究,确定了适于双齿围沙蚕RAPD分析的最佳反应体系:25μL反应体系中,含模板DNA 25 ng,10×PCR缓冲液2.5μL,MgCl2 1.5 mmol/L,dNTPs 0.15 mmol/L,引物24 ng,Taq DNA聚合酶1.5 U.对浙江温岭18个双齿围沙蚕个体进行了RAPD分析,从100个随机引物中筛选的26个引物检测出229个位点(400～4 200 bp),群体的多态位点比例为80.78%;遗传多态度在0.225 9;Shannon指数在0.353 2.个体间遗传距离最大的为0.461 0,最小的为0.160 9,18个个体的平均遗传距离为0.342 1±0.045 13;同时用UPGMA对18个个体进行聚类分析,构建谱系关系图,表明双齿围沙蚕的遗传多样性水平较高,为双齿围沙蚕的种质保护、合理开发利用以及选择育种提供了新的分析参数.The nereidid polychaete Penineresis aibuhitensis （Grube, 1878） is a marine euryhaline species found in marine habitats. Its habitation covers a wide range from the Bo Sea to South China Sea, Korea, Thailand, Philippines India （Andamanese）, Indonesia （Java and Sumatra）. However, the genetic background of Penineresis aibuhitensis largely remains unclear. To investigate its population structure and the diversity for the conservation of its germ plasm and prevention of gene loss, in this research genetic diversity analysis is conducted using RAPD-PCR technique. The simple and effective methods of DNA extraction of Perineris aibihitensis are studied, and then applied to template DNA extraction. This paper optimizes the reaction factors. Amplifications are performed in the volume of 25 μl. It contains 2.5 μl 10×PCR buffer, 25 ng template DNA, 0.2 mmol/L dNTPs, 1.5 mmol/L MgCl2, 24 ng primer, 1.5 U Taq DNA polymerase. Twenty-seven 10 bp RAPD primers are used for DNA amplification. Totally 229 markers are detected ranging 400-4 200 bp. Percentage of polymorphic loci of this stock is 80.78% and Genetic diversity is 0.225 9 and Shannon index is 0.353 2. The genetic distance between two individuals is found to be from 0.1609 to 0.4610. The average genetic distance is 0.342 1± 0.045 13 among the 18 individuals. The phylogenetic tree of 18 individuals is obtained by UPGMA analysis. In conclusion, genetic diversity of Perineris aibihitensis appears significant.

关 键 词：双齿围沙蚕 DNA提取 RAPD 遗传多样性 

分 类 号：S963[农业科学—水产养殖] S953[农业科学—水产科学]