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经比较影响农杆菌介导Barnase嵌合基因转化甘蓝型油菜的各种因素后 ,建立了高效稳定的转基因实验体系。按该体系 ,甘蓝型油菜“湘油 15”子叶柄预培养 2 0h ,OD60 0 为 0 5农杆菌菌液感染后共培养 3d ,在MS +2mg·L-1AgNO3 +4 5mg·L-1BA +10mg·L-1Km +30 0mg·L-1Carb培养基上进行选择 ,转化率为 8 8% ,PCR检测和Southern杂交检测证明外源基因已整合了甘蓝型油菜基因组中。
After comparing the various factors that affect the transformation of Barnase chimeric gene into Brassica napus by Agrobacterium tumefaciens, a highly efficient and stable transgenic experiment system was established. According to this system, the petiole of “Xiang You 15” was pre-cultured for 20 h and the OD60 0 was 0 5. After co-culture with Agrobacterium tumefaciens for 3 days, MSCs were cultured in the medium of MS +2 mg · L-1AgNO3 +4 5 mg · L -1 BA + 10mg · L-1Km +30 0mg · L-1Carb medium selection, the conversion rate of 8 8%, PCR test and Southern blot showed that foreign genes have been integrated into the Brassica napus genome.