大豆外源基因转化体系建立及条件优化研究  

Establishment of the genetic transformation system of soybean and its optimum conditions

作　　者：姬丹丹[1] 王鹏[1] 向凤宁[1] 

机构地区：[1]山东大学生命科学学院,山东济南250100

出　　处：《山东大学学报（理学版）》2011年第5期116-122,共7页Journal of Shandong University(Natural Science)

基　　金：国家自然科学基金资助项目(30970243);山东省杰出青年基金资助项目(QG200810);山东省公关项目(2009GG10002001)

摘　　要：建立了适用于多个大豆品种的萌动胚真空渗透辅助的外源基因转化方法,对影响农杆菌介导转化的有关参数进行了比较研究。确定的优化条件为:预培养3d,在菌种活化液的OD600值为0.6并加以200μmol.L-1乙酰丁香酮的农杆菌菌种活化液,侵染时间为6h,共培养3d。在优化条件下,将携带GUS基因的35S启动子驱动的表达载体pCAMBIA1304的根癌农杆菌菌株GV3101分别转入鲁豆11和潍6823中,获得510株鲁豆11再生植株和591株潍6823再生植株,其中,抗性再生植株分别为444株和462株。通过PCR和GUS检测,证明分别获得了13株和15株转基因植株,转化率分别为2.2%和2.5%。An exogenous gene transformation method used by germinating embryo co-culture with Agrobacterium tumefaciens GV3101 under vacuum permeation conditions was established in different soybean varieties.Several parameters of Agrobacterium-mediated gene transformation were compared to determine the optimum conditions.The results indicated that the optimal pre-training time,infection time,and co-culture time are respective 3 days,6 hours,and 3 days, the optimal concentration of Agrobacterium is OD600=0.6,the optimal concentration of acetosyringone is 200 μmol·L-1.Under optimized conditions,the pCAMBIA1304 vector was transferred by GV3101 into Soybean cv.Ludou 11 and Wei 6823,obtaining 510 Ludou 11 and 591 Wei 6823 regenerated plants,respectively.Among them,444 and 462 plants had the resistance on MB＋0.6 mg·L-1 IAA medium with 25 mg·L-1 kanamycin.13 and 15 transgenic plants were identified from the resistance plants by GUS and PCR analysis.Their transformation efficiencies were 2.2% and 2.5% respectively.

关 键 词：农杆菌介导 pCAMBIA1304 遗传转化 

分 类 号：S565.1[农业科学—作物学]