摘要
采用GUS基因瞬时表达检测的方法,利用正交试验法对玉米根癌农杆菌介导的玉米遗传转化体系的菌液OD值、侵染时间、AS浓度、共培养时间4个因素在3个水平上进行优化试验。通过对试验结果进行直观分析,方差分析和因素内多重比较分析,得到了稳定的转化体系,即菌液OD值0.6,侵染时间30min,AS浓度200μmol/L,共培养时间2d。采用优化后的玉米转化体系进行试验,通过PCR验证,得到了转基因植株,初步建立了农杆菌介导的玉米遗传转化体系。
Using the method of transient expression of GUS gene detection, orthogonal design was appneu to optimize Agrobacterium tumefacie mediated transformation of maize in three levels of four factors （fermenting liquor of OD, infection time, inclusion of acetosyringone and co-cultivation time）. Through the visual analysis, variance analysis and analysis of multiple comparison in factors, the stable transform system that factors were 0.6 of fermenting liquor of OD, 30 minutes of infection time, 200p.mol/L of inclusion of acetosyringone and 2 days of co-cultivation time was built. Using the optimized transformation system, transgenic maize plants were examined by PCR, and the Agrobacterium tumefacie mediated transformation of maize was elementarily built.
出处
《中国农学通报》
CSCD
2007年第11期56-60,共5页
Chinese Agricultural Science Bulletin
基金
国家高技术研究发展计划(863计划)"玉米优质
抗病
对生基因分子标记选择与聚合技术研究"(No.2006AA10Z1B4)
关键词
玉米
正交设计
农杆菌
遗传转化
Zea mays L
orthogonal experiment
agrobacterium tumefacien
genetic transformation
