生物工程专业毕业论文CUC3基因RNA干扰载体对烟草的转化及鉴定.docVIP

摘 要 采用叶盘法通过导入了构建好的表达载体根癌农杆菌，对野生型烟草叶片进行转化。烟草叶片在分化培养基MS + 0.1mg/L NAA + 1.0mg/L 6-BA + 50mg/L Hyg + 500mg/L Cb上4周后分化出芽；在生根培养基MS +0.2 mg/L IBA +0mg/L Cb上，2周后形成根；叶片β-葡萄糖甘酸酶GUS）组织化学染色，阳性率为75%；通过PCR 检测潮霉素磷酸转移酶（HPT）基因的DNA序列，获得14个阳性株系。在烟草转化阳性株系中获得了2种表型：叶片融合和叶片缺失。 关键词∶根癌农杆菌；GUS组织化学染色; 聚合酶链式反应( PCR) Abstract Agrobacterium with expression vector transformed the leaves of Nicotiana tabacum. Regenerated shoots were formed in 4 weeks when the leaves of N. tabacum cultured on shoot-inducing medium MS medium supplemented with 0.1mg/L NAA, 1.0mg/L 6-BA, 50mg/L Hyg and 500mg/L Cb. Shoots rooted on the root i