CAJ | 학술논문

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60 h时，GFP瞬时表达率最高，达到23.47%。
유전전화솔저시전기인기술진일보발전적한제인소。우화옥미전화체계적연구대다이유배위수체재료、이GUS기인위보고기인，재일정정도상한제료취재적시간화수량。연구이용휴대pCAMBIA1302(함록색형광단백기인-Green Fluorescent Protein)질립적EHA105균주대옥미자교계H99적유상조직진행침염，차조활체형광검측기술대GFP기인적순시표체진행평개。결과표명，농간균침염액농도OD600위0.7～0.8、침염시간20 min、공배양60 h시，GFP순시표체솔최고，체도23.47%。
The low transformation efficiency limited its large-scale application. Previously, optimization of maize transformation system heavily relied on the GUS staining, which was considered to be time-consuming, high false pos-itive observations and damaging tissues. Using GFP transient expression system, dipped the callus of maize inbred line H99 by EHA105 strain with pCAMBIA1302. The result showed that the concentration of Agrobacterium, the infection time and the co-cultivation time are playing the key roles in determining maize transgenic efficiency. With the A-grobacterium concentration of OD600=0.7-0.8, the infection time of 20 minutes, and the co-cultivation time of 60 hours, the percentage of expressed GFP callus reaches 23.47%.