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BIOCHEMICAL PROCESSES AND MACROMOLECULAR STRUCTURES

Engineering Plant Shikimate Pathway for Production of Tocotrienol and Improving Herbicide Resistance¹

Laboratoire Mixte Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Bayer CropScience (Unité Mixte de Recherche 1932), 14-20 rue Pierre Baizet, Boite Postale 9163, 69263 Lyon cedex 09, France (P.R., C.S., M.M.); and Laboratoire de Biotechnologie, Bayer CropScience, 14-20 rue Pierre Baizet, Boite Postale 9163, 69263 Lyon cedex 09, France (M.D.)

Tocochromanols (tocopherols and tocotrienols), collectively known as vitamin E, are essential antioxidant components of both human and animal diets. Because of their potential health benefits, there is a considerable interest in plants with increased or customized vitamin E content. Here, we have explored a new strategy to reach this goal. In plants, phenylalanine is the precursor of a myriad of secondary compounds termed phenylpropanoids. In contrast, much less carbon is incorporated into tyrosine that provides p-hydroxyphenylpyruvate and homogentisate, the aromatic precursors of vitamin E. Therefore, we intended to increase the flux of these two compounds by deriving their synthesis directly at the level of prephenate. This was achieved by the expression of the yeast (Saccharomyces cerevisiae) prephenate dehydrogenase gene in tobacco (Nicotiana tabacum) plants that already overexpress the Arabidopsis p-hydroxyphenylpyruvate dioxygenase coding sequence. A massive accumulation of tocotrienols was observed in leaves. These molecules, which were undetectable in wild-type leaves, became the major forms of vitamin E in the leaves of the transgenic lines. An increased resistance of the transgenic plants toward the herbicidal p-hydroxyphenylpyruvate dioxygenase inhibitor diketonitril was also observed. This work demonstrates that the synthesis of p-hydroxyphenylpyruvate is a limiting step for the accumulation of vitamin E in plants.

¹ This work was supported by the Centre National de la Recherche Scientifique, by the Institut National de la Recherche Agronomique, and by Bayer CropScience, formerly Aventis CropScience.

² Present address: Plant Biochemistry and Physiology Group, Institute of Plant Sciences, ETH-Zurich, Universitastrasse 2, CH-8092 Zurich, Switzerland.

³ Laboratoire de Physiologie Cellulaire Végétale, Unité Mixte de Recherche 5168, Centre National de la Recherche Scientifique/Commissariat à l'Energie Atomique/Institut National de la Recherche Agronomique/Université Joseph Fourier, Commissariat à l'Energie Atomique-Grenoble, 38054 Grenoble cedex 9, France.

^* Corresponding author; e-mail mmatringe{at}cea.fr; fax 33-438-78-50-91.

Received August 27, 2003; returned for revision September 16, 2003; accepted October 20, 2003.

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