真空渗透法转化油菜及转化种子的筛选

真空渗透遗传转化法首先在拟南芥(Arabidopsis thaliana)中获得成功, 是一种简便、快速且无需经过组织培养阶段即可获得大量转化植株的基因转化方法。该研究以不同遗传背景的3个甘蓝型油菜(Brassica napus)品种(系)陕3B、L45和Mg23为材料, 对真空渗透遗传转化方法中真空渗透时间和Silwet L-77浓度与遗传转化效果的关系进行了比较, 同时对转化种子的筛选方法进行了优化。结果表明, 卡那霉素(Km)对油菜种子的萌发影响不显著, 但对其生长发育有明显的抑制作用。不同油菜品种对卡那霉素的敏感性不同, 各自的致死浓度也不一样。在0%-0.05%的浓度范围内, 随着Silwet L-77浓度的增加, 在相同的真空渗透时间内, 3个油菜品种的转化率逐渐升高。当Silwet L-77浓度为0.05%时, 10分钟的真空渗透时间可获得最高转化效率, 此时陕3B、L45和Mg23的转化率分别达到1.97%、2.09%和2.30%。

Transformation of Brassica napus by Vacuum Infiltration

We studied the effect of vacuum infiltration time and Silwet L-77 concentration on transformation of different rapeseed cultivars (Brassica napus) Shaan 3B, L45 and Mg23. The selection methods for transformed seeds were optimized. Seed germination rates were not affected significantly by kanamycin, but the shoot growth was inhibited. The higher the concentration of kanamycin, the stronger the inhibition. The lethal concentration of kanamycin differed among different B. napus cultivars. The transformation frequency increased when 0%-0.05% Silwet L-77 was used but declined with 0.05%-0.10% Silwet L-77. The transformation frequency was highest with 0.05% Silwet L-77 for 10-min vacuum infiltration: as high as 1.97%, 2.09% and 2.30% for Shaan 3B, L45 and Mg23, respectively.