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Abstract

The invention provides a method for transforming a plant through a bacterium belonging to genus Agrobacterium, comprising transforming plant cells simultaneously with a first T-DNA (1) and a second T-DNA (2); and selecting the cells which acquired drug resistance; the first T-DNA (1) containing a gene giving the drug resistance, which functions in the plant; the second T-DNA (2) containing a desired DNA fragment to be introduced into the plant, the second T-DNA (2) being contained in a hybrid vector; the hybrid vector being prepared by homologous recombination between an acceptor vector and an intermediate vector in the bacterium belonging to genus Agrobacterium; the acceptor vector containing at least (a) a DNA region having a function to replicate a plasmid in the bacterium belonging to genus Agrobacterium and Escherichia coli, (b) a DNA region containing virB gene and virG gene in virulence region of Ti plasmid pTiBo542 of Agrobacterium tumefaciens, and (c) a DNA region which is homologous with a part of the intermediate vector, which is subjected to homologous recombination in the bacterium belonging to genus Agrobacterium; the intermediate vector containing at least (i) a DNA region having a function to replicate a plasmid in Escherichia coli, which does not function in the bacterium belonging to genus Agrobacterium, (ii) a DNA region which is homologous with a part of the acceptor vector, which is subjected to homologous recombination in the bacterium belonging to genus Agrobacterium, and (iii) a DNA region which constitutes at least a part of the second T-DNA.

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