软枣猕猴桃RAPD分子标记反应体系的优化  

Optimalization of RAPD Molecular Marker Reaction System for Actinidia arguta(Actinidia arguta(Sieb. & Zucc) Planch. ex Miq.)

作　　者：赵成日[1] Zhao Chengri(Agricultural College of Yanbian University,Yanji,133002)

机构地区：[1]延边大学农学院,延吉133002

出　　处：《分子植物育种》2018年第18期6009-6016,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31660319)资助

摘　　要：以长白山地区及韩国等9个采样点采集的软枣猕猴桃新鲜幼叶为试材,进行RAPD分子标记反应扩增体系的优化。对实验结果影响较大的5个因素(模板DNA浓度,引物浓度, MgCl_2浓度, dNTP浓度, Taq DNA聚合酶浓度)进行单因素设计,以寻找最佳反应浓度。结果表明,总反应体积为20μL时,模板DNA 40 ng、引物浓度0.3μmol/L、MgCl_2浓度2.0 mmol/L、dNTP浓度250μmol/L、Taq DNA聚合酶2.0 U。此时扩增条带多、电泳结果清晰稳定,表明该体系是一个适合软枣猕猴桃RAPD分子标记反应的体系。该体系的建立有利于软枣猕猴桃的亲缘关系和遗传多样性分析。Actinidia arguta leaf was used to study optimization of RAPD reaction conditions which was collected in nine place of Changbai mountion and Korea. The five most important influence of template DNA concentration,primer concentration, MgCl2 concentration, dNTP concentration, Taq DNA polymerase concentration amount were analyzed in order to obtain stable and reliable experiment result. The results show that the reaction materials of the optimized RAPD reaction system were as follows： 20 μL total volume of 40 ng DNA, primer at 0.3 μmol/L,MgCl2 at 2.0 mmol/L, dNTP at 250 μmol/L, Taq DNA polymerase at 2.0 U. The results indicate that the reaction system of RAPD could be used in tara vine. This optimized RAPD reaction system would be helpful for analyzing germplasm classification and identification of tara vine.

关 键 词：软枣猕猴桃(Actinidia arguta (Sieb.&Zucc) Planch. EX Miq.) RAPD 反应体系 优化 

分 类 号：S663.4[农业科学—果树学]