农杆菌介导玉米遗传转化影响因子的研究(精选).docxVIP

分子植物育种，2006 年，第 4 卷，第 2 期，第 228-232 页Molecular Plant Breeding, 2006, Vol.4, No.2, 228-232研究报告Research Report农杆菌介导玉米遗传转化影响因子的研究袁鹰1李启云1郝文媛1谭化1孔祥梅1张光弟2刘德璞1*1 吉林省农业科学院生物技术研究中心, 公主岭, 136100; 2 吉林省梨树县农业技术推广站, 四平, 136500* 通讯作者, liudp@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 OD600 0.5~0.6、侵染时间 5~10min、共培养时间 3d 最佳。关键词　农杆菌介导法, 转化频率, 影响因素, 抗性愈伤组织Studies on Influencing Factors of Agrobacterium tumefaciens MediatedMaize TransformationYuan Ying1Li Qiyun1Hao Wenyuan1Tan Hua1Kong Xiangmei1Zhang Guangdi2Liu Depu1*1 Jilin Academy of Agricultural Sciences, Gongzhuling, 136100; 2 Agriculture Generalize Station of Lishu County , Siping, 136500* Corresponding author, liudp@Abstract Maize inbred lines in Northeast (7922、340、78599 etc.) immature embryos calli were transformed byAgrobacterium tumefaciens (CryIA 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 showedthat callus subculture for 3~5 days was suitable for transformation and that transformation efficiency of 7922 callusreached the highest of 46.6% after 5 days subculture. Dominant difference existed among different maize genotypeon infection sensitivity and H99 was the most sensitive genotype with 34.57% resistant callus. Transformation effi-ciency was improved obviously with acetosyringone (AS) (100~200mg/L) appended in medium. In addition, bac-terium concentration (OD600 0.5~0.6), 5~10 minutes infection time and co-culture 3 days were the optimal condi-tions for Agrobacterium tumefaciens mediated maize transformation.Keywords Agrobacterium tumefaciens mediated transformation, Transf