Novel bacterial N-acetyltransferase gene for herbicide detoxification in land plants and selection maker in plant transformation.
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.
Cell-Free Science and Technology Research Center and the Venture Business Laboratory, Ehime University, Ehime, Japan.
Choong-Soo
CS
Yun
Hisakazu
H
Hasegawa
Hideaki
H
Nanamiya
Teruhiko
T
Terakawa
Yuzuru
Y
Tozawa
D000123:Acetyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000613:Aminobutyrates / Q000633:toxicity; D017360:Arabidopsis / Q000166:cytology / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D002736:Chloroplasts / Q000378:metabolism; D003001:Cloning, Molecular; D005818:Genetic Engineering / Q000379:methods*; D005819:Genetic Markers / Q000235:genetics; D053209:Herbicide Resistance / Q000235:genetics; D006540:Herbicides / Q000633:toxicity*; D008715:Methionine / Q000031:analogs & derivatives / Q000378:metabolism; D008969:Molecular Sequence Data; D009615:Nocardia / Q000235:genetics; D012275:Oryza / Q000166:cytology / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D030821:Plants, Genetically Modified; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
D000613:Aminobutyrates; D005819:Genetic Markers; D006540:Herbicides; D011993:Recombinant Fusion Proteins; C018332:methionine sulfone; C003121:phosphinothricin; D008715:Methionine; D000123:Acetyltransferases; C056243:phosphinothricin N-acetyltransferase