植物遗传转化中农杆菌抑制的研究

Study on the Inhibition of Agrbacterium in Plant Genetic Transformation

以农杆菌GV3101侵染预培养的无菌烟草叶方块，共培养2d后，外植体通过5种表面脱菌处理后转接在培养基上进行抑菌和筛选培养。结果表明，共培养后的材料先用无菌水冲洗6～8遍，以脱去材料表面大部分农杆菌，再用2％（V/V）NaClO漂洗10min，然后无菌水冲洗5遍，捞出后放在无菌滤纸上吸干多余水分，转接在抑菌和筛选培养基Ms＋BA2．0mg／L＋NAA0．1mg／L＋Cef500mg／L＋HygB20mg／L上的抑菌效果良好。

Agrobacterium tumefaciens strain GV3101 was used to infect pre-cultured sterile tobacco leaf squares. After they were co-cultured together for 2 d, the explants were transplanted in medium for bacterial-restraining and selective culture, when they were made by 5 kinds of eliminating bacteria treatments. The results showed that the co-cultured materials was washed with aseptic water for 6 - 8 times at In,st to eliminate most agrobacterium on the surfaces of materials, and bathed with 2% （V/V） NaClO for 10 min, and then washed with aseptic water for 5 times. They were put on the aseptic filter paper after fished out to sop up the superfluous water. Transplanting them in the bacterial-restraining and selective medium of MS ＋ BA 2.0 mg/L ＋ NAA 0.1 mg/L ＋ Cef 500 mg/L ＋ HygB 20 mg/L could get better bacterial-restraining effect.