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用根癌农杆菌介导的方法 ,将B .t.基因和Cp TI基因分别导入花椰菜“杂交 75天”的父本和母本的无菌苗下胚轴切段细胞 ,都获得了转基因植株。经过预培养的下胚轴切段用根癌农杆菌 (LBA44 0 4/ pG BI4A2B ,含B .t .基因 ;LBA44 0 4/pBRLC ,含CpTI基因 )进行感染后 ,共培养 48h ,继续培养 30d后 ,将分化芽转移至筛选培养基上。 10d后 ,大多数分化芽的顶端变成紫色 ,2 0d后紫色芽逐渐变白死亡 ,而转化芽在选择培养基上长成小植株。小植株移至大田能正常生长、开花、结籽。PCR和Southernblot分析表明B .t和CpTI基因已整合在植物基因组中
The Agrobacterium tumefaciens-mediated method was used to introduce the B.t. gene and the Cp TI gene into the hypocotyl cuttings of the male and female parent plants of “hybridization 75 days” . After pre-cultured hypocotyl cuttings were infected with Agrobacterium tumefaciens (LBA44 0 4 / pG BI4A2B, with Bt. Gene; LBA44 0 4 / pBRLC, CpTI containing gene) for a total of 48 hours and cultured for 30 days Afterwards, the differentiated shoots are transferred to a screening medium. After 10 days, the tops of most of the differentiated shoots became purple, and the purple shoots gradually became white after 20 days, while the transformed shoots grew into small plants on the selective medium. Small plants can be moved to the field to normal growth, flowering, seed. PCR and Southern blot analysis showed that the Bt and CpTI genes were integrated in the plant genome