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Genomic microstructure and differential expression of the genes encoding UDP-glucose:sinapate glucosyltransferase (UGT84A9) in oilseed rape (Brassica napus)

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Abstract

In oilseed rape (Brassica napus), the glucosyltransferase UGT84A9 catalyzes the formation of 1-O-sinapoyl-β-glucose, which feeds as acyl donor into a broad range of accumulating sinapate esters, including the major antinutritive seed component sinapoylcholine (sinapine). Since down-regulation of UGT84A9 was highly efficient in decreasing the sinapate ester content, the genes encoding this enzyme were considered as potential targets for molecular breeding of low sinapine oilseed rape. B. napus harbors two distinguishable sequence types of the UGT84A9 gene designated as UGT84A9-1 and UGT84A9-2. UGT84A9-1 is the predominantly expressed variant, which is significantly up-regulated during the seed filling phase, when sinapate ester biosynthesis exhibits strongest activity. In the allotetraploid genome of B. napus, UGT84A9-1 is represented by two loci, one derived from the Brassica C-genome (UGT84A9a) and one from the Brassica A-genome (UGT84A9b). Likewise, for UGT84A9-2 two loci were identified in B. napus originating from both diploid ancestor genomes (UGT84A9c, Brassica C-genome; UGT84A9d, Brassica A-genome). The distinct UGT84A9 loci were genetically mapped to linkage groups N15 (UGT84A9a), N05 (UGT84A9b), N11 (UGT84A9c) and N01 (UGT84A9d). All four UGT84A9 genomic loci from B. napus display a remarkably low micro-collinearity with the homologous genomic region of Arabidopsis thaliana chromosome III, but exhibit a high density of transposon-derived sequence elements. Expression patterns indicate that the orthologous genes UGT84A9a and UGT84A9b should be considered for mutagenesis inactivation to introduce the low sinapine trait into oilseed rape.

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Acknowledgments

The authors thank Rod Snowdon (University of Giessen, Germany) for providing the genomic BAC library from B. napus, for help with the library screen and for supplying the identified positive clones. BAC end sequencing by Prisca Viehöver and Bernd Weisshaar (University of Bielefeld, Germany) is greatly acknowledged. Seeds of B. napus, B. oleracea and B. rapa were kindly provided by Norddeutsche Pflanzenzucht (Holtsee, Germany). Excellent technical assistance was given by Sylvia Vetter, Claudia Horn, Anja Henning and Alexandra Jestadt. This work was part of the research project “YelLowSin Rapeseed: Functional genomics approaches for the development of yellow-seeded, low sinapine (“YelLowSin”) oilseed rape/canola (Brassica napus)”, financially supported by the Bundesministerium für Bildung und Forschung.

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  1. Carsten Milkowski

    Present address: Institute of General Botany and Plant Physiology, Friedrich Schiller University Jena, Am Planetarium 1, 07743, Jena, Germany

Authors and Affiliations

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

    Juliane Mittasch, Dieter Strack & Carsten Milkowski

  2. Institute for Crop Science and Plant Breeding, Bavarian State Research Center for Agriculture, Am Gereuth 8, 85354, Freising, Germany

    Sabine Mikolajewski

  3. Molecular Breeding of Oilseed Rape, KWS Saat AG, Grimsehlstraße 31, 37555, Einbeck, Germany

    Frank Breuer

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  1. Juliane Mittasch
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Correspondence to Carsten Milkowski.

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Mittasch, J., Mikolajewski, S., Breuer, F. et al. Genomic microstructure and differential expression of the genes encoding UDP-glucose:sinapate glucosyltransferase (UGT84A9) in oilseed rape (Brassica napus). Theor Appl Genet 120, 1485–1500 (2010). https://doi.org/10.1007/s00122-010-1270-4

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