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根癌农杆菌介导的水稻高效转化系统的建立

刘巧泉, 张景六, 王宗阳, 洪孟民, 顾铭洪

扬州大学农学院农学系.江苏,扬州 225009；中科院上海植物生理研究所.上海 200032

摘要

应用国内的7个栽培品种和3个广亲和品种,详细比较了根癌农杆菌转化水稻的各种条件,建立了优化的转化系统,我们用这一系统转化水稻获得了比前述报告更高的转化频率,并得到了大量转基因可育植株。

关键词： 水稻; 根癌农杆菌; 基因转化; 转基因植株; 幼胚

A highly efficient transformation system mediated by Agrobacterium tumefaciens in rice ( Oryza sativa L.)

Liu Qiaoquan, Zhang Jingliu, Wang Zongyang, Hong Mengmin, Gu Minghong

Department of Agronomy,Agricultural College,Yangzhou University.Yangzhou 225009,Jiangsu；China

Abstract

After the factors influencing transformation were carefully investigated, a highly efficient Agrobacterium-mediated gene transfer system has been developed for rice. Using this protocol, immature embryos of rice variety Zhonghua No. 11 cultured for 4 days were infected and transformed by A. tumefaciens strain EHA105 harboring Ti plasmid pCAMBIA1301. The results showed that infected embryos gave a high frequency (50%-90%) of GUS gene expression after co-cultivation. The transformation afficiency of stable integration of T-DNA reached 87.6% measured as hygromycin-resistant calli produced and 64.6% measured as number of transgenic plants regenerated, respectively. Southern blotting analysis of the total DNA extracted from transgenic plants indicated that the T-DNA had been integrated into the rice genome, and most of transgenic plants had one copy of T-DNA insert.

Key words: Rice;Agrobacterium tumefaciens;Transformation;Transgenic plant;Immature embryo

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引用本文：

刘巧泉, 张景六, 王宗阳, 洪孟民, 顾铭洪. 根癌农杆菌介导的水稻高效转化系统的建立[J]. 生理学报 1998; 50 (3): .

Liu Qiaoquan, Zhang Jingliu, Wang Zongyang, Hong Mengmin, Gu Minghong. A highly efficient transformation system mediated by Agrobacterium tumefaciens in rice ( Oryza sativa L.). Acta Physiol Sin 1998; 50 (3): (in Chinese with English abstract).