农杆菌介导的sbe2a基因RNAi载体对玉米遗传转化的研究

Genetic Transformation with RNAi Vector of sbe2a Gene on Maize via Agrobacterium

将淀粉分支酶基因sbe2a的RNAi表达载体转入玉米自交系H99和丹598胚性愈伤组织。探讨了不同菌浓度、不同侵染时间和不同的共培养时间对玉米转化率的影响,确定了最佳转化条件:菌液浓度OD600值为0.5～0.7,侵染时间为25min,共体培养时间为3d。对转化的愈伤组织分化诱导出苗后进行PCR检测,获得了2株阳性植株,初步证明外源基因已经整合到玉米的基因组中。

Starch branch enzyme sbe2a gene of RNAi expression vector was transformed into embryogenic callus in maize inbred lines of H99 and Dan598 via Agrobacterium transformation system.The different concentration of bacteria,the different time of infection and the different time of co-culture on the infection rate of conversion of corn were determined.The results showed that the bacteria concentration（OD600 0.5~0.7）,infection time（25min） and co-culture time（3d） were optimal.Differentiated transformed callus were detected by PCR,indicating that 2 positive transgenic plants were obtained.