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Stepwise engineering to produce high yields of very long-chain polyunsaturated fatty acids in plants

Abstract

Very long chain polyunsaturated fatty acids (VLCPUFAs) such as arachidonic acid (AA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are valuable commodities that provide important human health benefits1,2,3,4,5. We report the transgenic production of significant amounts of AA and EPA in Brassica juncea seeds via a stepwise metabolic engineering strategy. Using a series of transformations with increasing numbers of transgenes, we demonstrate the incremental production of VLCPUFAs, achieving AA levels of up to 25% and EPA levels of up to 15% of total seed fatty acids. Both fatty acids were almost exclusively found in triacylglycerols, with AA located preferentially at sn-2 and sn-3 positions and EPA distributed almost equally at all three positions. Moreover, we reconstituted the DHA biosynthetic pathway in plant seeds, demonstrating the practical feasibility of large-scale production of this important ω-3 fatty acid in oilseed crops.

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Figure 1: Simplified maps of the binary vector constructs used for plant transformation.
Figure 2: GC analysis of seed fatty acid methyl esters from wild-type and transgenic B. juncea plants.
Figure 3: Stereospecific analysis of phospholipids and triacylglycerols.

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Acknowledgements

The authors thank Derek Potts for providing B. juncea germplasm, and Darwin Reed and Mike Giblin for assistance with GC and GC/MS analysis. We also thank Jonathan Page for comments on an earlier version of this manuscript.

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Correspondence to Xiao Qiu.

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This research was funded in part by BASF BmbH, Ludwigshafen, Germany.

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Wu, G., Truksa, M., Datla, N. et al. Stepwise engineering to produce high yields of very long-chain polyunsaturated fatty acids in plants. Nat Biotechnol 23, 1013–1017 (2005). https://doi.org/10.1038/nbt1107

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