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利用根癌农杆菌(Agrobacterium tumefaciens)介导,将豇豆胰蛋白酶抑制剂(CpTI)基因导入甘蓝(Brassica oleracea var.capitata L.)栽培品种“京丰”和“迎春”,并分别获得了转基因再生植株。对NPTⅡ的ELISA检测表明,标记基因NPTⅡ已在再生植株中得到有效表达。对转化植株进行Southern blot分子杂交实验进一步证明CpTI基因已整合到甘蓝的基因组。实验室内的叶片离体饲虫及盆栽饲虫试验表明,转基因甘蓝对菜青虫具有一定抗性。实验中还观察到,采用愈伤组织分化途径可以获得较高的转化频率,但嵌合体较多;而直接分芽的途径转化率低,但嵌合体较少。
The gene of cowpea trypsin inhibitor (CpTI) was introduced into “Jingfeng” and “Yingchun” cultivars of Brassica oleracea var.capitata L. mediated by Agrobacterium tumefaciens, Plant. ELISA detection of NPTII showed that the marker gene NPTII has been effectively expressed in the regenerated plants. Southern blot analysis of transformed plants further confirmed that the CpTI gene has been integrated into the genome of cabbage. Experiments in vitro showed that the transgenic cabbage was resistant to Pieris rapae. The experiment also observed that the use of callus differentiation approach can get a higher frequency of transformation, but more chimeras; and direct sub-bud pathway transformation rate is low, but fewer chimeras.