摘要
通过油菜子叶外植体-农杆菌共培养法将苏云金杆菌杀虫蛋白基因导入甘蓝型油菜，获得抗虫的转基因植株。带有1～2mm子叶柄的油菜子叶经农杆菌感染后，共培养2～3天，然后转移到附加15mg／L卡那霉素的MS选择培养基上筛选转化愈伤组织及不定芽。卡那霉素抗性苗相继在含20～50mg／L卡那霉素的选择培养基上继代培养，再转移到合25mg／L卡那霉素的生根培养基上诱导生根。以苏云金杆菌杀虫蛋白基因为探针，进行Southernblot分析，证明外源基因已整合到油菜基因组中。部分转基因植株具有明显的杀虫活力，用转基因植株叶片饲喂甜菜夜蛾等幼虫后，幼虫出现中毒症状，导致幼虫发育受阻和死亡。转基因植株的抗虫性和卡那霉素抗性在自交后代中得到保持，外源基因星核基因单一位点显性遗传。
Cotyledons, each with a 1-2mm petiole at its base, were cut from axenic seedlings and infected with Arobacterium tumefaciens. After 2-3 days of cocultivation, the cotyledon explants were transferred to MS selection medium containing 15mg / L kanamycin and 4.5ing / L 6-BA to induce shoot differentiation. Kanamycin-resistant shoots were subeultured on selection medium with 20-50mg / L kanamycin for 3-6 months for eliminating escaped non-transformants, and then rooted on MS medium containing 25mg/ L kanamycin and 0.1mg/ L NAA. Whole Plants were transplanted into soil and grew in the field. DNA Southem blot hybridization and polymerase chain reaction showed that some of the plants were positive when probed with the insecticidal crystal protein gene. The transgenic plants exhibited tolerant to pest insects such as lerhygma exigua and Pieris rapae in leaf feeding experiments Kanamycin-resistance and insect-resistance were maintained in the progeny. The foreign genes were delivered to the progeny according to Mendelian Law of single gene segregation.
基金
国家自然科学基金
湖北省自然科学基金
关键词
甘蓝型油菜
杀虫蛋白基因
转基因植株
抗虫性
Brassica napus, Insecticidal crystal protein gene, Transgenic plants, Explanation of Plate
