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Transgenic plants containing the phosphinothricin-N-acetyltransferase gene metabolize the herbicide l-phosphinothricin (glufosinate) differently from untransformed plants

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Abstract

l-Phosphinothricin (l-Pt)-resistant plants were constructed by introducing a modified phosphinothricin-N-acetyl-transferase gene (pat) via Agrobacterium-mediated gene transfer into tobacco (Nicotiana tabacum L), and via direct gene transfer into carrot (Daucus carota L). The metabolism of l-Pt was studied in these transgenic, Pt-resistant plants, as well as in the untransformed species. The degradation of l-Pt, 14C-labeled specifically at different C-atoms, was analysed by measuring the release of 14CO2 and by separating the labeled degradation products on thin-layer-chromatography plates. In untransformed tobacco and carrot plants, l-Pt was deaminated to form its corresponding oxo acid 4-methylphosphinico-2-oxo-butanoic acid (PPO), which subsequently was decarboxylated to form 3-methylphosphinico-propanoic acid (MPP). This compound was stable in plants. A third metabolite remained unidentified. The l-Pt was rapidly N-acetylated in herbicide-resistant tobacco and carrot plants, indicating that the degradation pathway of l-Pt into PPO and MPP was blocked. The N-acetylated product, l-N-acetyl-Pt remained stable with regard to degradation, but was found to exist in a second modified form. In addition, there was a pH-dependent, reversible change in the mobility of l-N-acetyl-Pt thin-layer during chromatography.

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Abbreviations

ac-Pt:

N-acetyl-phosphinothricin

MPE:

2-methylphosphinico-ethanoic acid

MPP:

3-methylphosphinicopropanoic acid

pat :

phosphinothricin-N-acetyltransferase gene

Pat:

phosphinothricin-N-acetyltransferase

pat41:

pat gene modified for expression in plants

PPO:

4-methylphosphinico-2-oxobutanoic acid

Pt:

phosphinothricin

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Authors and Affiliations

  1. Lehrstuhl für Genetik, Fakultät für Biologie, Universität Bielefeld, Postfach 8640, W-4800, Bielefeld 1, Germany

    W. Dröge, I. Broer & A. Pühler

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  1. W. Dröge
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  2. I. Broer
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  3. A. Pühler
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This work was supported in part by a grant from Hoechst AG, Pflanzenschutz, Frankfurt/Main, FRG. The authors wish to thank E. Strauch and W. Arnold (Lehrstuhl f. Genetik, Universität Bielefeld, FRG) for supplying plasmids; R. Eichenlaub (Lehrstuhl f. Mikrobiologie und Gentechnologie, Universität Bielefeld, FRG) for the synthesized oligonucleotides; and I.M. Pretorius-Güth (Lehrstuhl f. Genetik, Universität Bielefeld, FRG) and K. Severin (Lehrstuhl f. Allgemeine Genetik, Universität Tübingen, FRG) for reading the manuscript. W. Droge acknowledges the receipt of a scholarship from DECHEMA, Frankfurt.

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Dröge, W., Broer, I. & Pühler, A. Transgenic plants containing the phosphinothricin-N-acetyltransferase gene metabolize the herbicide l-phosphinothricin (glufosinate) differently from untransformed plants. Planta 187, 142–151 (1992). https://doi.org/10.1007/BF00201636

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