Lu A, Chen Z, Kong L, Fang R, Cun S, Mang K
Institute of Genetics, Academia Sinica, Beijing.
Yi Chuan Xue Bao. 1996;23(1):77-83.
A system for obtaining regenerated plantlets of "double low" Brassica napus by using cotyledonary petioles as material was established. The turnip mosaic virus coat protein (TuMV-CP) gene inserted in the binary vector pBTu in the Agrobacterium tumefaciens LBA 4404 was integrated into Brassica napus through co-culture of cotyledonary petioles with LBA4404, and the material in co-culture was selected under the stress of Kanamycin. Regenerated plantlets were obtained, and the specific TuMV-CP gene was proved to be integrated into the genomic DNA of the regenerated plants by the specific PCR amplification, dot hybridization and Southern blot. All these transgenic plants were proved to be resistant to virulent TuMV by virus challenge in different degree.
建立了一种以子叶柄为材料获得“双低”甘蓝型油菜再生植株的体系。农杆菌LBA 4404中二元载体pBTu上插入的芜菁花叶病毒外壳蛋白（TuMV-CP）基因，通过子叶柄与LBA4404共培养整合到甘蓝型油菜中，共培养材料在卡那霉素胁迫下进行筛选。获得了再生植株，通过特异性PCR扩增、斑点杂交和Southern杂交证明特异性TuMV-CP基因已整合到再生植株的基因组DNA中。通过病毒挑战试验证明所有这些转基因植株都不同程度地对强毒力TuMV具有抗性。
