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以玉米杂交种郑单958为受体材料,建立农杆菌介导的玉米萌动胚转化体系,并获得抗除草剂转基因玉米材料。经过对农杆菌菌液浓度、共培养时间、乙酰丁香酮的浓度的优化,农杆菌介导的玉米萌动胚转化法的最优参数为菌液浓度OD600值为0.8,共培养时间为48 h,乙酰丁香酮的浓度为100μmol/L。获得69株PCR阳性植株,其中54株为bar基因金标免疫试纸条检测阳性植株,29株获得种子;T1代植株具有草丁膦抗性,并显示出胶体金免疫酶联反应阳性,表明目的基因已整合玉米基因组内并有效表达,而且能够稳定遗传给后代。
Using maize hybrid Zhengdan 958 as the receptor material, Agrobacterium -mediated maize embryo transformation system was established, and the herbicide-resistant transgenic maize material was obtained. After optimizing Agrobacterium tumefaciens concentration, co-cultivation time and acetosyringone concentration, the optimum parameters of Agrobacterium -mediated embryo transformation were the OD600 of 0.8, the co-culture time of 48 h, Acetosyringone concentration of 100μmol / L. Sixty-nine PCR-positive plants were obtained, of which 54 were bar gold-labeled immunobanks and 29 were seed; the T1 plants had glufosinate resistance and showed positive colloidal gold immunosorbent assay The target gene has been integrated into the maize genome and efficiently expressed, and can be stably inherited to offspring.