| 农杆菌介导玉米H99遗传转化体系的优化 |
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| 引用本文： | 葛敏,张体付,王华,赵涵. 农杆菌介导玉米H99遗传转化体系的优化[J]. 玉米科学, 2013, 21(6): 31-34 |
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| 作者姓名： | 葛敏 张体付 王华 赵涵 |
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| 作者单位： | 江苏省农业科学院农业生物技术研究所/江苏省农业生物学重点实验室, 南京 210014;江苏省农业科学院农业生物技术研究所/江苏省农业生物学重点实验室, 南京 210014;浙江省农业科学院作物与核技术利用研究所, 杭州 310021;江苏省农业科学院农业生物技术研究所/江苏省农业生物学重点实验室, 南京 210014 |
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| 基金项目： | 江苏省农业科技自主创新资金项目[cx（10）435] |
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| 摘 要： | 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%。
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| 关 键 词： | 玉米 农杆菌介导 愈伤组织 转基因 |
| 收稿时间： | 2013-02-14 |
Optimization of Agrobacterium tumefaciens-Mediated Transformation System for Maize Inbred Line H99 |
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| GE Min,ZHANG Ti-fu,WANG Hua and ZHAO Han. Optimization of Agrobacterium tumefaciens-Mediated Transformation System for Maize Inbred Line H99[J]. Journal of Maize Sciences, 2013, 21(6): 31-34 |
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| Authors: | GE Min ZHANG Ti-fu WANG Hua ZHAO Han |
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| Affiliation: | Institute of Agro-biotechnology, Jiangsu Academy of Agricultural Sciences, Provincial Key Laboratory of Agrobiolgy, Nanjing 210014;Institute of Agro-biotechnology, Jiangsu Academy of Agricultural Sciences, Provincial Key Laboratory of Agrobiolgy, Nanjing 210014;Institute of Crop and Nuclear Technology Utilization, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;Institute of Agro-biotechnology, Jiangsu Academy of Agricultural Sciences, Provincial Key Laboratory of Agrobiolgy, Nanjing 210014 |
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| Abstract: | 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 positive 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 Agrobacterium 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%. |
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| Keywords: | Maize Agrobacterium-mediated Callus Transgene |
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