Abstract
The canola industry generates more than $11 billion of yearly income to the Canadian economy. One problem of meal quality is the dark polyphenolic pigments that accumulate in the seed coat. Seed coat-specific promoters are a pre-requisite to regulate the genes involved in seed coat development and metabolism. The β-glucuronidase (GUS) reporter gene was used to test an Arabidopsis promoter in developing and mature seeds of canola (Brassica napus). The promoter tested is the regulatory region of the laccase gene (AtLAC15) from Arabidopsis thaliana. The AtLAC15 promoter::GUS construct was inserted into canola double haploid line DH12075 using Agrobacterium-mediated transformation. Southern blot analysis using a 536 bp GUS probe showed variation among the transformed plants in the T-DNA copy numbers and the position of the insertion in their genomes. Histochemical assay of the GUS enzyme in different tissues (roots, leaves, stem, pollen grains, flowers, siliques, embryos and seed coats) showed ascending GUS activity only in the seed coat from 10 days after pollination (DAP) to the fully mature stage (35 DAP). GUS stain was observed in the mucilage cell layer, in the outer integument layer of the seed coat but not in the inner integument. The AtLAC15 promoter exhibited a specificity and expression level that is useful as a seed coat-specific promoter for canola.
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Acknowledgements
This research was funded by Genome Canada in the project “Designing oilseeds for Tomorrow’s Markets”. We wish to thank Dr. Yajun Wu, Utah State University, USA for providing the AtLAC15 promoter construct, Dr. Gerhard Rakow, Agriculture and Agri-Food Canada, Canada, for providing DH12075 canola seeds and Dr. Michael Deyholos, University of Alberta, Canada, for his support and advice.
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El-Mezawy, A., Wu, L. & Shah, S. A seed coat-specific promoter for canola. Biotechnol Lett 31, 1961–1965 (2009). https://doi.org/10.1007/s10529-009-0098-y
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DOI: https://doi.org/10.1007/s10529-009-0098-y