基于RAPD和SRAP分子标记的柑桔大实蝇种群多态性及其亲缘关系研究

DNA Polymorphism and Genetic Relationship of Bactrocera(Tetradacus) minax(Enderlein) Populations Based on RAPD and SRAP Molecular Markers

目的：利用RAPD（Random Amplified polymorphic DNA）和SRAP（Sequence-Related Amplified Polymorphism）分子标记技术,研究来自重庆、湖北、四川和陕西4省市10个地理种群柑桔大实蝇的亲缘关系.方法：利用10条RAPD引物和5对SRAP引物对10个不同地理种群柑桔大实蝇的基因组DNA进行扩增,统计多态性谱带,然后运用DPS统计分析软件进行遗传多态性及聚类分析.结果：10条RAPD引物扩增产生152条清晰稳定的谱带,其中多态性谱带132条,多态性谱带的比例为86.84%;不同地理种群间的遗传距离在0.250 0~0.625 0之间,10个地理种群的柑桔大实蝇被划分为4个大类群.5对SRAP引物扩增共获得66条重复性好的清晰谱带,其中多态性谱带60条,多态性谱带比例为90.91%,不同地理种群间的遗传距离在0.200 0~0.789 5之间,10个地理种群的柑桔大实蝇被分为5个大的类群.将RAPD和SRAP两种标记的数据整合,聚类结果显示,10个地理种群的柑桔大实蝇被划分为6个大类群.结论：柑桔大实蝇不同地理种群间具有丰富的遗传多样性,两种方法结合分析更能准确地反映不同地理种群间的亲缘关系.

Objective：Two molecular marker techniques ,RAPD （random amplified poloymorphic DNA ） and SRAP （sequence‐related aAmplified polymorphism ） ,were used to investigate the genetic relationship among 10 geographic populations of Bactrocera （Tetradacus） minax （Enderlein） from Chongqing ,Hu‐bei ,Sichuan and Shaanxi .Methods ：Ten RAPD primers and five pairs of SRAP primers were used to am‐plify the genomic DNA of 10 different geographic populations of B. （Tetradacus） minax （Enderlein） ,and polymorphic bands were counted .Then the DPS （data processing system） software was used to obtain the information of genetic polymorphism and clustering .Result ：One hundred and fifty‐two clear and stable bands were amplified by PCR with the 10 RAPD primers ,of which 132 were polymorphic ,the proportion of polymorphic bands being 86.84% .The genetic distance among the 10 different geographic populations ranged from 0.250 0 to 0.625 0 .The 10 different geographic populations were clustered into four groups . Meanwhile ,66 clear and repeatable bands were obtained by PCR with the 5 pairs of SRAP primers ,of which 60 were polymorphic ,the proportion of polymorphic bands being 90.91% .The genetic distance a‐mong the 10 different geographic populations ranged from 0.200 0 to 0.789 5 .Clustering analysis divided the 10 different geographic populations into five groups .However ,integration of the RAPD data and the SRAP data clustered the 10 different geographic populations into six groups .Conclusion：There is abun‐dant genetic polymorphism among different geographic populations of B. （Tetradacus） minax （Ender‐lein） .The clustering result based on both RAPD and SRAP data can more accurately reflect the genetic re‐lationship between the different geographic populations .