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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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Citing This Article：

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).