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遗传转化率低是转基因技术进一步发展的限制因素。优化玉米转化体系的研究大多以幼胚为受体材料、以GUS基因为报告基因,在一定程度上限制了取材的时间和数量。研究利用携带pCAMBIA1302(含绿色荧光蛋白基因-Green Fluorescent Protein)质粒的EHA105菌株对玉米自交系H99的愈伤组织进行侵染,借助活体荧光检测技术对GFP基因的瞬时表达进行评价。结果表明,农杆菌侵染液浓度OD600为0.7～0.8、侵染时间20 min、共培养60h时,GFP瞬时表达率最高,达到23.47%。
Low genetic conversion is a limiting factor in the further development of transgenic technology. Most of the studies on the optimization of maize transformation system use the immature embryo as the receptor material, and the GUS gene as the reporter gene, to a certain extent, limits the time and amount of the material taken. In this study, the EHA105 strain harboring pCAMBIA1302 (containing green fluorescent protein) was used to infect the callus of maize inbred line H99. The transient expression of GFP gene was evaluated by real-time fluorescence detection. The results showed that the Agrobacterium tumefaciens concentration of OD600 was 0.7-0.8 and the infection time was 20 min. The highest transient expression rate of GFP reached 23.47% at 60 h after co-culture.