Novel Bacterial N-Acetyltransferase Gene for Herbicide Detoxification in Land Plants and Selection Maker in Plant Transformation

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  • YUN Choong-Soo
    Cell-Free Science and Technology Research Center and the Venture Business Laboratory, Ehime University
  • HASEGAWA Hisakazu
    Hokko Chemical Industry, Ltd.
  • NANAMIYA Hideaki
    Cell-Free Science and Technology Research Center and the Venture Business Laboratory, Ehime University
  • TERAKAWA Teruhiko
    Hokko Chemical Industry, Ltd.
  • TOZAWA Yuzuru
    Cell-Free Science and Technology Research Center and the Venture Business Laboratory, Ehime University

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  • Novel Bacterial<i>N</i>-Acetyltransferase Gene for Herbicide Detoxification in Land Plants and Selection Maker in Plant Transformation

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Abstract

Phosphinothricin (PPT) is the active ingredient in bialaphos, which specifically inhibits glutamine synthetase in land plants. We isolated a novel PPT-resistant gene from a soil bacterium, Nocardia sp., and characterized it. The encoded protein, consisting of 177 amino acids, showed significant similarity to bacterial N-acetyltransferases, and we originally designated the gene MAT (methionine sulfone N-acetyltransferase). The recombinant MAT protein exhibited functions as a methionine sulfone and PPT N-acetyltransferase in vitro. The PPT N-acetyltransferase activity reached the maximum at pH 8–8.5, indicating that the protein might optimally function in chloroplasts. We therefore constructed a MAT gene, encoding the enzyme with a chloroplast-localizing signal in its amino-terminus. Plant transformation with the construct resulted in the generation of PPT-resistant rice and Arabidopsis. Furthermore, the transformed Arabidopsis was selectable in a synthetic medium containing PPT. The MAT gene thus facilitated establishment of herbicide-resistant plants, and as a new selectable gene marker.

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