将水稻DADI基因插入Ti质粒,构建成该基因的正反义植物表达载体pWM-DADI,pWM-anrisense DADI,转入根癌农杆菌LBA4404后,利用农杆菌介导的叶盘法转化菊花,经Hyg抗性筛选,并诱导愈伤组织和分化成苗,得到转基因菊花植株,通过PCR检测,从部分转基因植株中检测出预期的目的基因的存在.
We first inserted the DADI gene from rice into Ti plasmid, then constructed sense and antisense DADI gene expression vector which were named as pWM-DADI and pWM-antisense DADI respectively. Afetr that two recombinated plasmids were transformed into Agrobacterium tumefaciens strain LBA4404 and infected chrysanthemum leaves via leaf disc co-cultivation. By the process of Hyg sreceening, the infected leaf discs were induced to form callus and differentiated to shoots further more. At last 28 transgenic chrysanthemum plants were obtained. The PCR results indicated DADI gene exists in part of transgenic plants, which confirmed the fact that the sense and antisense fragment of DADI gene have been integrated into chrysanthemum genome successfully.