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高效的遗传转化体系对植物的转基因有重要意义,除菌是遗传转化工作中的重要环节.头孢除菌剂在蓖麻子叶节部位能完全去除农杆菌,并保证外植体子叶节的生长状况不受损伤.本研究按不同头孢浓度、不同处理时间设置梯度摸索最佳除菌条件.结果表明:选择头孢浓度在4000mg/L、处理时间为10min时处理子叶节,用灭菌的1/8MS液体培养基涮洗3次,并用无菌滤纸吸干表面后再接种到新培养基内,用这种办法除菌效果较好,并且可以保证外植体的长势不受太大影响.该浓度和处理时间对保证农杆菌最小程度的污染率起到了关键作用.
Efficient genetic transformation system is of great significance to plant genetically modified, and sterilization is an important part of genetic transformation.Cephalosporins can completely remove Agrobacterium in the castor leaves and ensure the growth of explants cotyledonary node The results showed that when the concentration of cefotaxime was 4000mg / L and the treatment time was 10min, the cotyledonary node was treated with sterile 1/8 MS The liquid medium was rinsed three times, and the surface was inoculated with sterilized filter paper and then inoculated into the new medium. In this way, the sterilization effect is better and the growth of the explants is not affected much. And treatment time play a crucial role in ensuring the minimum contamination rate of Agrobacterium.