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Increase of the tocochromanol content in transgenic Brassica napus seeds by overexpression of key enzymes involved in prenylquinone biosynthesis

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Abstract

Lipid soluble tocochromanols, only synthesised in photosynthetic organisms, are industrially interesting compounds because of their antioxidative properties and their essential function in nutrition. In order to increase the tocochromanol content in the seed oil of transgenic plants, approaches were undertaken to engineer the flux of substrates and intermediates through the pathway. To this end, we overexpressed genes encoding hydroxyphenylpyruvate dioxygenases, alone or in combination with chimeric homogentisate phytyltransferase and tocopherol cyclase genes, in seeds of transgenic Brassica napus plants and analysed total tocochromanol content and composition. Overexpression of chimeric hydroxyphenylpyruvate dioxygenase genes, both in the cytosol or in the plastids of developing seeds, yielded a slight although significant increase in total tocochromanol level. Coexpression of a hydroxyphenylpyruvate dioxygenase gene with both a homogentisate phytyltransferase gene and a tocopherol cyclase gene elevated this effect with maximum increases of up to two-fold in individual lines and this phenotype was found to be stably inherited. These data showed that the three enzymes are critical in determining the total tocochromanol content in the seed oil of Brassica napus plants, while the tocopherol cyclase, unlike hydroxyphenylpyruvate dioxygenase and homogentisate phytyltransferase, had additionally an effect on the relative abundance of individual tocochromanol species and resulted in an increase of δ-tocopherol and plastochromanol-8 in the seeds.

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Abbreviations

DMGGBQ:

2,3-dimethyl-5-geranylgeranyl-1,4-benzoquinol

DMPBQ:

2,3-dimethyl-5-phytyl-1,4- benzoquinol

DMSBQ:

2,3-dimethyl-5-solanesyl-1,4-benzoquinol

HPPD:

4-hydroxyphenylpyruvate dioxygenase

HGGT:

homogentisate geranylgeranyltransferase

HPT:

homogentisate phytyltransferase

HST:

homogentisate solanesyl transferase

MPBQ:

2-methyl-6-phytyl-1,4-benzoquinol

MGGBQ:

2-methyl-6-geranylgeranyl-1,4-benzoquinol

MSBQ:

2-methyl-6-solanesyl-1,4-benzoquinol

MT1:

MPBQ/MGGBQ/MSBQ methyltransferase

MT2:

γ-tocopherol methyltransferase

P-8:

plastochromanol-8

TC:

tocopherol cyclase

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Acknowledgements

The authors thank Christof Stoll for performing statistical analysis and Ingrid Klawiter, Marion Heitmann, Petra Degen, Gabi Ciobanu, Ingrid Frantz, Violeta Cadar, Sonja Sahner and Stavros Tzigos for technical assistance.

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Authors and Affiliations

  1. Plant Breeding Department, IFZ Research Centre for Biosystems, Land Use and Nutrition, Justus-Liebig-University Giessen, Heinrich-Buff-Ring 26-32, 35392, Giessen, Germany

    Mirela Raclaru, Wilfried Lühs, M. Karim Zarhloul & Wolfgang Friedt

  2. Institute for Biology I—Botany, RWTH Aachen University, Worringerweg 1, 52074, Aachen, Germany

    Jens Gruber, Rajeev Kumar, Radin Sadre, Margrit Frentzen & Dagmar Weier

Authors
  1. Mirela Raclaru
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  2. Jens Gruber
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  3. Rajeev Kumar
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  4. Radin Sadre
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  5. Wilfried Lühs
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  6. M. Karim Zarhloul
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  7. Wolfgang Friedt
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  8. Margrit Frentzen
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  9. Dagmar Weier
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Corresponding author

Correspondence to Dagmar Weier.

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Mirela Raclaru and Jens Gruber contributed equally

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Raclaru, M., Gruber, J., Kumar, R. et al. Increase of the tocochromanol content in transgenic Brassica napus seeds by overexpression of key enzymes involved in prenylquinone biosynthesis. Mol Breeding 18, 93–107 (2006). https://doi.org/10.1007/s11032-006-9014-5

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  • DOI: https://doi.org/10.1007/s11032-006-9014-5

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