摘要
以子叶柄为材料，建立了甘蓝型油菜（ＢｒａｓｓｉｃａｎａｐｕｓＬ．）双低品种的再生体系。通过子叶柄与农杆菌（ＡｇｒｏｂａｃｔｅｒｉｕｍｔｕｍｅｆａｃｉｅｎｓＬＢＡ４４０４）共培养，将表达载体ｐＢＴｕ中芜菁花叶病毒外壳蛋白（ＴｕＭＶ－ＣＰ）基因以整合方式导入甘蓝型油菜，然后用卡那霉素进行筛选，获得了油菜再生植株。经ＰＣＲ特异性扩增、点杂交和Ｓｏｕｔｈｅｒｎ印迹分析，证明再生植株基因组ＤＮＡ中整合了ＴｕＭＶ－ＣＰ基因。攻毒实验表明，有ＴｕＭＶ－ＣＰ基因插入的工程油菜对ＴｕＭＶ均有不同程度的抗性。
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.
基金
"八五"攻关经费
院重大项目经费
关键词
甘蓝型油菜
遗传工程
转基因油菜
芜菁花叶病毒
Turnip mosaic viruscoat protein gene, Transgenic Brassica napus, Plant genetic engineering
