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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (36): 71-75.

所属专题： 玉米

• 农学 农业基础科学 • 上一篇    下一篇

农杆菌介导玉米遗传转化体系的研究

孙传波 郭嘉 陶蕊 李海华 刘文国 袁英   

  • 收稿日期:2012-04-24 修回日期:2012-06-22 出版日期:2012-12-25 发布日期:2012-12-25

Study on Genetic Transformation System of Maize Mediated by Agrobacterium tumefaciens

  • Received:2012-04-24 Revised:2012-06-22 Online:2012-12-25 Published:2012-12-25

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摘要：

为了提高玉米自交系的遗传转化率，建立农杆菌介导玉米遗传转化体系。通过农杆菌菌株EHA105侵染玉米自交系7922的胚性愈伤组织，以β-葡萄糖苷酶基因(GUS)为报告基因研究几种因素对遗传转化体系的影响。结果表明：（1）当农杆菌浓度OD600=0.6时，GUS瞬时表达率最高为35.3%；（2）当侵染时间为15 min时，GUS瞬时表达率最高为37.3%；（3）当共培养时间为3天和恢复培养时间为4天时，GUS瞬时表达率最高为29.1%。因此选择农杆菌浓度OD600=0.6，侵染时间15 min，共培养3天和恢复培养4天，为最佳遗传转化条件。通过优化影响遗传转化体系的因素，初步建立了农杆菌介导的玉米遗传转化体系。

关键词: 对策, 对策

Abstract:

In order to improve transformation efficiency of maize inbred lines and establish genetic transformation system of maize mediated by Agrobacterium tumefaciens, Agrobacterium strains EHA105 were used to inoculate maize embryonic calli of maize inbred lines 7922 and factors influencing genetic transformation system were studied with GUS as reporter gene. The results showed that: (1) with OD600=0.6 of bacterial concentration, the highest GUS transient expression efficiency was 35.3%. (2) with 15 min of infection time, the highest GUS transient expression efficiency was 37.3%. (3) with 3 days of co-cultivation time and 4 days of resting, the highest GUS transient expression efficiency was highest 29.1%. So OD600=0.6 for bacterium concentration, 15 minutes for infection, 3 days for co-cultivation and 4 days for resting were the best transformation conditions. Genetic transformation system of maize mediated by Agrobacterium tumefaciens was established by optimizing factors influencing this system.