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Identification of functional BrFAD2-1 gene encoding microsomal delta-12 fatty acid desaturase from Brassica rapa and development of Brassica napus containing high oleic acid contents

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Abstract

Microsomal delta-12 fatty acid desaturase (FAD2) functions in the first committed step of the biosynthesis of polyunsaturated fatty acids via the desaturation of oleic acid to linoleic acid. In this study, two FAD2 genes were identified through genome-wide analysis of Brassica rapa. One BrFAD2-1 gene harbors functional sequence information, but another BrFAD2-2 gene has mutations that generated a premature stop codon, rendering it nonfunctional. From a database of 120,000 B. rapa expressed sequence tags, we determined that all sequences coding for FAD2 corresponded to the BrFAD2-1 gene. The BrFAD2-1 protein was shown to share high sequence homology (71–99%) with FAD2 proteins from other plant species. An intron in the 5′-untranslated region and three histidine boxes in the protein, which are characteristic of plant FAD2 genes, have been well-conserved. BrFAD2-1 transcripts were detected in various organs of B. rapa. When a pBrFAD2-1:mRFP construct was introduced into tobacco epidermal cells, the fluorescent signal was noted in the endoplasmic reticulum. Ectopic expression of BrFAD2-1:mRFP complemented the Arabidopsis fad2-2 mutant. Finally, transgenic Korean rapeseed Tammi containing high oleic acid contents (78 mol%) was developed via the expression of the BrFAD2-1 gene in an antisense orientation. The data demonstrate that B. rapa harbors only one functional FAD2 that can be utilized for the development of the high-oleic acid Korean rapeseed cultivar Tammi, which might be useful for both human consumption and industrial applications.

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Acknowledgments

We thank Beom-Seok Park, RDA, Korea, for providing the B. rapa FAD2-1 gene and John Browse, Washington State University, Pullman, Washington, for providing Arabidopsis fad2-2 seeds. The nucleotide sequence of the BrFAD2-1 gene from B. rapa ssp. pekinensis reported herein has been registered in the GenBankTM/EBI Data Bank under accession number HM189213. This work was supported by grants from Research Programs (PJ007441201008 and PJ0067152010), the Rural Development Administration, Republic of Korea, and Basic Research Program (20090064298) and the World Class University Project (R31-2009-000-20025-0) from the National Research Foundation of Korea.

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Authors and Affiliations

  1. Department of Bioenergy Science and Technology, Chonnam National University, Gwangju, 500-757, Republic of Korea

    Jin Hee Jung, Hyojin Kim, Saet Buyl Lee & Mi Chung Suh

  2. Department of Plant Biotechnology, Chonnam National University, Gwangju, 500-757, Republic of Korea

    Young Sam Go

  3. Genome Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 305-333, Republic of Korea

    Cheol-Goo Hur

  4. Department of Agricultural Bio-resources, National Academy of Agricultural Science, RDA, Suwon, 441-707, Republic of Korea

    Hyun Uk Kim

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  1. Jin Hee Jung
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Correspondence to Hyun Uk Kim or Mi Chung Suh.

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Communicated by J. R. Liu.

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Jung, J.H., Kim, H., Go, Y.S. et al. Identification of functional BrFAD2-1 gene encoding microsomal delta-12 fatty acid desaturase from Brassica rapa and development of Brassica napus containing high oleic acid contents. Plant Cell Rep 30, 1881–1892 (2011). https://doi.org/10.1007/s00299-011-1095-x

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