影响农杆菌介导大豆遗传转化的因素  

Affecting factors of Agrobacterium-mediated transformation of soybean

作　　者：刘银[1] 孔月琴[1] 唐慧[1] 史秀岚[1] 王幼平[1] 

机构地区：[1]扬州大学生物科学与技术学院,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2014年第1期71-75,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家转基因重大专项(201311117091X);江苏省大学生创新训练计划项目(201311117091X)

摘　　要：以大豆子叶节为外植体,采用农杆菌介导的遗传转化方法,探讨农杆菌菌液浓度、侵染时间、抗氧化剂、表面活性剂Silwet L-77及草丁膦(PPT)筛选浓度等因素对大豆转化效率的影响。结果表明:适合大豆转化的农杆菌菌液浓度D600nm为0.8,子叶节外植体侵染时间以30min为宜,外植体侵染后在含有25mg·L-1LA共培养基中暗培养3d有利于提高转化效率。此外,PPT筛选的适宜浓度为5mg·L-1。根据大豆子叶节农杆菌转化特点,研究中采用了延迟筛选的方法。In this study, factors affecting the transformation efficiency of soybean cotyledonary node by Agrobacteriummediated transformation system were assessed, including Agrobacterium concentrations and infection time, antioxidant, Silwet L-77, and PPT selection concentrations. The results indicated that the suitable transformation condition of soybean： 0.8 （D600 ram） for the concentration of Agrobacterium, 30 min for co-infection time, then 3 d dark in the co-cultivation medium added with 25 mg ·L^-1 LA. In addition, the suitable concentration of PPT used in resistant shoots selec tion was 5 mg ·L^-1 , and the late selection was used to overcome the difficulty of adventitious shoot differentiation.

关 键 词：大豆 农杆菌 子叶节 转化 因素 

分 类 号：S565.1[农业科学—作物学]