摘要
以东北春玉米自交系7922、340、78599、921及美国杂交种H99等幼胚愈伤组织为受体,利用农杆菌介导法转化Bt(CryIA.)基因,研究菌液预处理、愈伤组织状态、侵染培养基、侵染时间及共培养条件等因素对转化频率的影响。根据转化愈伤组织的除草剂(Basta)抗性筛选结果评估转化率,表明继代培养3￣5d的愈伤组织适于转化,其中以继代5d的7922愈伤组织转化率最高,为46.6%;不同基因型对农杆菌转化率存在差异,H99的抗性愈伤组织率为34.57%,7922为26.27%、340为21.36%、78599和921抗性愈伤组织率仅为8.02%和6.78%。基因型间差异显著;浸染培养基中加入(AS100￣200mg/L)抗性愈伤转化率明显提高;菌液浓度OD6000.5￣0.6、侵染时间5￣10min、共培养时间3d最佳。
Maize inbred lines in Northeast （7922,340,78599 etc.） immature embryos calli were transformed by Agrobacterium tumefaciens （CrylA Bt.） in this report. Factors influencing transformation efficiency were studied, including pre-treatment, callus state, infection medium, infection time and co-cultivation et al.. Results showed that callus subculture for 3-5 days was suitable for transformation and that transformation efficiency of 7922 callus reached the highest of 46.6% after 5 days subculture. Dominant difference existed among different maize genotype on infection sensitivity and H99 was the most sensitive genotype with 34.57% resistant callus. Transformation efficiency was improved obviously with acetosyringone （AS） （100-200mg/L） appended in medium. In addition, bacterium concentration （OD600 0.5～0.6）, 5～10 minutes infection time and co-culture 3 days were the optimal conditions for Agrobacterium tumefaciens mediated maize transformation.
出处
《分子植物育种》
CAS
CSCD
2006年第2期228-232,共5页
Molecular Plant Breeding
基金
国家转基因植物中试及产业化基地（吉林）专项（JY03-17-1）资助.
关键词
农杆菌介导法
转化频率
影响因素
抗性愈伤组织
Agrobacterium tumefaciens mediated transformation, Transformation efficiency, Influencing factors,Resistant calli
