双丙氨磷在橡胶树遗传转化中的应用  

Application of Bialaphos in Hevea Genetic Transformation

作　　者：谢黎黎[1,2] 安泽伟[1,2] 姜泽海 李雅超[1,2] 翟琪麟[1,2] 黄华孙[2] 

机构地区：[1]海南大学农学院,海口570228 [2]国家橡胶树育种中心,中国热带农业科学院橡胶研究所,儋州571737 [3]广东农垦热带作物研究所,化州525145

出　　处：《分子植物育种》2013年第4期545-550,共6页Molecular Plant Breeding

基　　金：国家天然橡胶产业技术体系(CARS-34);国家自然科学基金项目(30860221;30960319);中国热带农业科学院橡胶研究所基本科研业务费专项(1630022011002)共同资助

摘　　要：本研究以GV3101::pMP90RK为工程菌株,bar基因作为筛选标记基因,研究了双丙氨磷在橡胶树根癌农杆菌介导法转基因体系中对转基因愈伤组织的筛选效果,并利用该体系将橡胶树转录因子HbWRKY5转入橡胶树无性系热研8-79中。实验结果表明,3mg/L双丙氨磷可作为橡胶树转基因体系中抗性愈伤组织筛选的最佳浓度。本研究中共获得了3个抗性愈伤组织系,抗性愈伤经过体细胞胚发生共获得234个体细胞胚,经植株再生培养后共获得22株再生植株。对再生植株进行的PCR鉴定结果表明,所获得的再生植株均为转基因植株。因此,bar基因结合双丙氨磷的筛选体系可以作为橡胶树转化体系中的有效筛选体系。In this study, bar gene was used as a selecting marker, and the effects of bialaphos on selection of the transgenic calli were investigated. The results showed that 3 mg/L of bialaphos was the optimal concentration for embryogenic calli screening. Using this selecting system, HbWRKY5 was transformed into embryogenic calli of rubber tree clone CATAS 8-79 mediated by Agrobacterium strain GV3101：：pMP90RK. Two hundred and thirty-four somatic embryo and 22 regeneration plantlets were obtained from three resistant callus lines, and the PCR analysis showed that all of the regeneration plantlets were positive individuals. So bar gene and bialaphos can be used as an effective screening system in Hevea genetic transformation.

关 键 词：双丙氨磷 巴西橡胶树 BAR基因 抗性筛选 再生植株 

分 类 号：S794.1[农业科学—林木遗传育种]