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Functional characterization of β-ketoacyl-CoA synthase genes from Brassica napus L.

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Abstract

Seed-specifically expressed β-ketoacyl-CoA synthase genes of Brassica napus (Bn-FAE1.1 genes) were cloned from two cultivars, namely Askari, a high-erucic-acid type, and Drakkar, a low-erucic-acid type. The genes from the two cultivars were found to be nearly identical. They encode proteins of 507 amino acids, the sequences of which differ only at position 282. The Bn-FAE1.1 gene of Askari, unlike that of Drakkar, was functionally expressed in yeast cells suggesting that the single amino acid exchange effects the low erucic acid phenotype at the E1 gene locus. In yeast cells the β-ketoacyl-CoA synthase of Askari elongated not only oleoyl but also palmitoleoyl groups as well as saturated acyl groups in such a way that monounsaturated acyl groups of 22 carbons and saturated ones of 26 carbons were formed as main products. A reporter gene fused to the promoter region of the Bn-FAE1.1 gene from Askari showed seed-specific expression in transgenic rapeseed plants. Over-expression of the coding region of the Askari gene in developing seeds of transgenic Drakkar plants resulted in a significant increase in the levels of eicosenoic acid and erucic acid esterified in the seed oil. On the other hand, in transgenic high-erucic-acid rapeseed plants the increase in erucic acid level was at most 60% although the chimeric Bn-FAE1.1 gene was co-expressed with an erucoyl-CoA-specific lysophosphatidate acyltransferase gene enabling trierucoyl glycerol to accumulate in the seed oil.

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Authors and Affiliations

  1. Institut für Allgemeine Botanik, Universität Hamburg, Ohnhorststrasse 18, 22609, Hamburg, Germany

    Jixiang Han, Frank P. Wolter, Ernst Heinz & Margrit Frentzen

  2. Lehrstuhl für Pflanzenzüchtung, Universität Giessen, Heinrich-Buff-Ring 26–32, 35392, Giessen, Germany

    Wilfried Lühs

  3. BAZ-Institut für Züchtung landwirtschaftlicher Kulturpflanzen, Institutsplatz, 18190, Gross Lüsewitz, Germany

    Karin Sonntag

  4. LG Immunchemie, Parkallee 1-40, 23845, Borstel, Germany

    Ulrich Zähringer

  5. KWS Saat AG, Grimselstrasse 31, 37555, Einbeck, Germany

    Dorothea S. Borchardt

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  1. Jixiang Han
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Han, J., Lühs, W., Sonntag, K. et al. Functional characterization of β-ketoacyl-CoA synthase genes from Brassica napus L.. Plant Mol Biol 46, 229–239 (2001). https://doi.org/10.1023/A:1010665121980

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