大豆子叶节遗传转化受体体系的优化

Optimization of Genetic Transformation System of Soybean Cotyledonary Node

以大豆品种"JN18、JN28、JN54"为试验材料,通过正交试验设计,对种子萌发以及子叶节不定芽诱导阶段的影响因素进行探究,以期对大豆的遗传转化受体体系进行优化。结果表明:选择大豆品种"JN28"经次氯酸钠溶液灭菌10 min,接种于0.2 mg/L 6-BA的MS萌发培养基(pH 6)为大豆种子萌发的最优条件,大豆子叶节不定芽诱导的最佳条件是,将萌发5 d的种子接种于MS+3.0 mg/L 6-BA+0.2 mg/L IBA培养基上进行培养。根据得到的最优条件,利用农杆菌介导法对大豆品种"JN28"进行遗传转化,获得的T_(0)代大豆转化植株,PCR阳性转化率达到5.56%。研究为大豆基因功能的鉴定及自主知识产权的转基因大豆新品系的获得奠定了基础。

In order to optimize the genetic transformation receptor system of soybean,the affecting factors of seed germination and adventitious bud induction were investigated by orthogonal test design on soybean varieties JN18,JN28 and JN54.The results showed that the optimal condition for soybean seed germination is to select JN28 soybean sterilized by sodium hypochlorite solution for 10 min,and inoculated on MS+0.2 mg/L 6-BA germination medium(pH 6).The optimal condition for adventitious bud induction of soybean cotyledon node is to inoculate the seeds germinated for 5 d on MS+3.0 mg/L 6-BA+0.2 mg/L IBA medium.According to the obtained optimal conditions,the Agrobacterium-mediated transformation for JN28 was applied,and the positive transformation rate of PCR was 5.56%.This result lays foundation for the identification of soybean gene function and the acquisition of new genetically modified soybean lines with independent intellectual property rights.