J Integr Plant Biol. ›› 2005, Vol. 47 ›› Issue (2): -.DOI: 10.1111/j.1744-7909.2005.00015.x

• Research Articles •    

Agrobacterium-Mediated Multiple Gene Transformation in Rice Using a Single Vector

Ming-Xia CAO, Jian-Qiu HUANG, Zhi-Ming WEI, Quan-Hong YAO, Chang-Zhao WAN and Jia-An LU   

Abstract: The homodimeric hemoglobin gene (VHb), the trans-zeatin synthetase gene (tzs), the modified 5-enolpyruvylshikimate-3-phosphate synthase gene (EPSPS), a selectable marker gene (hpt), and a reporter gene (gus), as linked expression cassettes, were stacked into the T-DNA region of a binary vector and introduced simultaneously into immature embryos of the rice (Oryza sativa L.) varieties Xiushui-11, Qiufeng, Youfeng, and Hanfeng by Agrobacterium tumefaciens. A total of 1 153 transgenic lines was obtained through selection for hygromycin B resistance. Approximately 90.2% of the transgenic lines harbored all the transgenes. Integration of multiple transgenes occurred at one to three genetic loci. Expression analysis revealed that the transgenes were coexpressed and inherited in a simple Mendelian fashion in transgenic plants and the frequency of coexpression was approximately 85%. On the basis of the cointegration and coexpression of the transgenes, most transgenic families were considered to be useful in a breeding program.

Key words: Agrobacterium tumefaciens, linked expression cassettes, multiple genes, rice, transformation.

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