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大白菜小孢子培养获得的子叶期胚状体,经粉碎的玻璃碴摩擦后,与农杆菌共培养,在加筛选剂Basta(含phosphinothricin5mg/L)的培养基上,再生出数株绿苗。自交留种后,对其后代苗期进行的Basta抗性鉴定显示,有4株抗此除草剂。Southern杂交结果显示,抗性植株的基因组中各有一个bar基因插入位点。对转化株的小孢子进行再培养,后代小孢子植株对Basta抗性的分离比显示,此转基因株为杂合体
Cotyledonary embryoid bodies obtained from microspore culture of Chinese cabbage were cocultivated with Agrobacterium by rubbing with crushed glass ballast and several green shoots were regenerated on medium supplemented with screening agent Basta (containing phosphinothricin 5 mg / L). After selfing, the Basta resistance test of the offspring of their progeny showed that there were 4 strains that could resist this herbicide. Southern blot results showed that each of the resistant plants had a bar gene insertion site in its genome. The microspores of the transformants were re-cultured, and segregation ratio of microspore plants to Basta resistance of the offspring showed that the transgenic plants were heterozygous