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The genes BnSCT1 and BnSCT2 from Brassica napus encoding the final enzyme of sinapine biosynthesis: molecular characterization and suppression

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Abstract

This study describes the molecular characterization of the genes BnSCT1 and BnSCT2 from oilseed rape (Brassica napus) encoding the enzyme 1-O-sinapoyl-β-glucose:choline sinapoyltransferase (SCT; EC 2.3.1.91). SCT catalyzes the 1-O-β-acetal ester-dependent biosynthesis of sinapoylcholine (sinapine), the most abundant phenolic compound in seeds of B. napus. GUS fusion experiments indicated that seed specificity of BnSCT1 expression is caused by an inducible promoter confining transcription to embryo tissues and the aleurone layer. A dsRNAi construct designed to silence seed-specifically the BnSCT1 gene was effective in reducing the sinapine content of Arabidopsis seeds thus defining SCT genes as targets for molecular breeding of low sinapine cultivars of B. napus. Sequence analyses revealed that in the allotetraploid genome of B. napus the gene BnSCT1 represents the C genome homologue from the B. oleracea progenitor whereas BnSCT2 was derived from the Brassica A genome of B. rapa. The BnSCT1 and BnSCT2 loci showed colinearity with the homologous Arabidopsis SNG2 gene locus although the genomic microstructure revealed the deletion of a cluster of three genes and several coding regions in the B. napus genome.

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Abbreviations

SCT:

1-O-sinapoyl-β-glucose:choline sinapoyltransferase

BAC:

Bacterial artificial chromosome

SE:

Sinapate esters

SC:

Sinapoylcholine

SG:

1-O-sinapoyl-β-glucose

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Acknowledgments

The authors thank the Norddeutsche Pflanzenzucht Hans-Georg Lembke KG (NPZ), Hohenlieth, for supply of seed samples. This study was part of the research project “NAPUS 2000—Healthy Food from Transgenic Rape Seeds” and was financially supported by the Bundesministerium für Bildung und Forschung (BMBF). We thank Bettina Hause for helpful discussions during the course of this work and Claudia Horn and Sylvia Vetter for excellent technical assistance.

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  1. Diana Weier

    Present address: Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, 06466, Gatersleben, Germany

Authors and Affiliations

  1. Department of Secondary Metabolism, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle (Saale), Germany

    Diana Weier, Juliane Mittasch, Dieter Strack & Carsten Milkowski

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  1. Diana Weier
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Correspondence to Carsten Milkowski.

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Weier, D., Mittasch, J., Strack, D. et al. The genes BnSCT1 and BnSCT2 from Brassica napus encoding the final enzyme of sinapine biosynthesis: molecular characterization and suppression. Planta 227, 375–385 (2008). https://doi.org/10.1007/s00425-007-0624-x

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