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Gene targeting in plants using theAgrobacterium vector system

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Abstract

In the past decade several methods have been developed for the introduction of foreign DNA into plant cells to obtain transgenic plants. In some of these methods, purified DNA is directly introduced into protoplasts that for some species can be regenerated into mature plants. The more commonly used protocols, however, employ the natural capacity ofAgrobacterium tumefaciens to transfer a defined peice of DNa, called T-DNA, to the nucleus of plant cells that are more easy to regenerate than protoplasts. In plant cells, like in animal cells, foreign DNA (including T-DNA) is readily inserted into the genome via illegitimates recombination. In contrast, targeted integration via homologous recombination, referred to as ‘gene targeting’, can only be obtained at relatively low frequencies. Nevertheless, gene targeting has become a standard strategy for reverse genetics studies in animals. In plants, the occurrence of gene targeting was only reported recently. This review focuses on the use of theAgrobacterium vector system to achieve gene targeting in plants. Recent experimental data concerning gene targeting in plants are presented and the overall suitability ofAgrobacterium T-DNA transfer for this purpose is assessed in light of contemporary views on the mechanism of T-DNA transfer.

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  1. MOGEN International Einsteinweg 97, 2333 CB, Leiden, The Netherlands

    Remko Offringa & Peter J. M. van den Elzen

  2. Institute for Molecular Plant Sciences Molecular Biology, Leiden University, Clusius Laboratories, Wassenaarseweg 64, 2333 AL, Leiden, The Netherlands

    Remko Offringa & Paul J. J. Hooykaas

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  1. Remko Offringa
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Offringa, R., van den Elzen, P.J.M. & Hooykaas, P.J.J. Gene targeting in plants using theAgrobacterium vector system. Transgenic Research 1, 114–123 (1992). https://doi.org/10.1007/BF02528776

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