甘蓝型油菜抗病虫双价基因转化体系的建立

被引:4
作者
刘宏波
郭翔
崔鹏
刘旦
许玲
唐桂香
周伟军
机构
[1] 浙江大学作物科学研究所
基金
浙江省自然科学基金;
关键词
甘蓝型油菜; 转化体系; PCR; Southern杂交; 病虫害抗性;
D O I
暂无
中图分类号
S565.4 [油菜籽(芸薹)];
学科分类号
摘要
以甘蓝型油菜(Brassica napus L.)品种浙双758子叶柄外植体为受体,建立农杆菌介导转化体系,研究乙酰丁香酮、羧苄青霉素浓度和潮霉素筛选浓度等对农杆菌遗传转化效果的影响。结果发现,羧苄青霉素对农杆菌的抑制效果以500mg/L最佳,且对子叶柄外植体愈伤组织诱导和分化的影响最小,愈伤组织诱导率和芽再生率分别为75.2%和65.1%。5.0mg/L的潮霉素能完全抑制未转化再生植株的生长,使其最终褐化死亡,在此潮霉素筛选浓度下,获得了32株转化再生植株。共培养时培养基中添加100μmol/L AS的芽再生率为3.9%,显著高于未添加AS的芽再生率(2.0%),说明共培养阶段添加酚类物质乙酰丁香酮有利于转化载体T-DNA的转化。部分转化再生植株经PCR和Southern杂交检测呈阳性,病虫害接种试验表明转化植株对菌核病和小菜蛾有较好的抗性。
引用
收藏
页码:26 / 31
页数:6
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