摘要
根据甘蓝型油菜不同品系下胚轴分化能力的不同，选用再生率及苗质均较好的特选4号作为受体材料，研究了各因素对其再生及转化效率的影响。结果表明，6d苗龄的下胚轴分化能力较强。以MS＋6-BA（3mg·L^-1）＋NAA（0．1mg·L^-1）＋AgNO3（6mg·L^-1）作为分化培养基诱导频率最高。培养基中添加5mg·L^-1Kan（卡那霉素）就能筛选出转化芽苗；顸培3d的下胚轴在稀释10倍的农杆菌菌液（OD0．4～0．5）侵染30s后共培2（1获得的转化率最高。根据上述条件的优化，建立了以常规种特选4号下胚轴为转化受体的良好的再生及转化体系，转化率可达8％。
According to the different differentiating frequency of hypocotyls of rapeseed （ Brassica napus） , the Texuan 4 with high regeneration frequency and good seedling quality was used as the recipient of transformation, and the factors affecting plant regeneration and genetic transformation were researched. The results were as follows ： 6 - day hypocotyls segments had better differentiating ability. And using MS ＋ 6 - BA （3 mg·L^-1 ） ＋ NAA （0.1 mg·L^-1 ） ＋ AgNO3 （ 6mg·L^-1 ） as differetiating medium could get higher induced frequence. Medium with 5mg·L^-1Kan could select transgened plants. The hypocotyl segments precultured on preculture medium for 3 days, then submerged into 10 - fold diluted Agrobacterium suspension （ OD 0.4 - 0.5 ） for 30 second, and placed on co-cultivation medium for 2 days could result in the highest transformation efficiency. On the basis of all factors mentioned above, a better system using the Texuan 4 hypocotyls as the recipient of regeneration and transformation, was set up and the transformation frequency could reach 8%.
出处
《山地农业生物学报》
2006年第5期377-381,433,共6页
Journal of Mountain Agriculture and Biology
基金
贵州省优秀青年科技人才培养计划资助项目[黔科合人字(2003)0312]
贵州省国际合作计划资助项目[黔科合国字(2004)11001]
关键词
甘蓝型油菜
植株再生
遗传转化
农杆菌介导法
影响因素
rapeseed （ Brassica napus）
plant regeneration
genetic transformation
Agrobacterium mediated method
influence factors
