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探讨植物激素、外植体类型、抗生素、外植体预培养时间、共培养时间等多因素对小白菜再生体系和转化体系建立的影响。结果表明,基本(播种)培养基中添加6-BA3mg·L-1+NAA0.3mg·L-1,有助于提高不定芽的诱导频率;带柄子叶的不定芽再生频率显著地高于下胚轴;将外植体预培养2d后,再感染,转化频率较高;共培养2d有助于提高转化效果。经优化组合这些技术参数,初步建立了小白菜农杆菌介导的转化体系,获得了转抗虫基因sck的小白菜植株。PCR检测结果证明,sck基因已整合到小白菜的基因组中。
The effects of plant hormones, explant type, antibiotics, explant pre-culture time and co-culture time on the regeneration system and transformation system of Chinese cabbage were discussed. The results showed that the addition of 6-BA3mg · L-1 + NAA0.3mg · L-1 to the basic (seeding) medium was helpful to improve the induction frequency of adventitious buds; Hypocotyls. After explants were pre-cultured for 2 days, the explants were infected again and the transformation frequency was higher. Co-cultivation for 2 days helped to improve the transformation efficiency. After optimizing the combination of these technical parameters, the Agrobacterium-mediated transformation system of Brassica campestris was initially established and the Brassica campestris plants transformed with insect sck gene sck were obtained. PCR test results show that the sck gene has been integrated into the genome of Chinese cabbage.