Transgenic Research

, Volume 19, Issue 2, pp 221–229

Towards the production of high levels of eicosapentaenoic acid in transgenic plants: the effects of different host species, genes and promoters

  • Bifang Cheng
  • Guohai Wu
  • Patricia Vrinten
  • Kevin Falk
  • Joerg Bauer
  • Xiao Qiu
Original Paper

Abstract

Eicosapentaenoic acid (EPA, 20:5n-3) plays an important role in many aspects of human health. In our efforts towards producing high levels of EPA in plants, we investigated the effects of different host species, genes and promoters on EPA biosynthesis. Zero-erucic acid Brassica carinata appeared to be an outstanding host species for EPA production, with EPA levels in transgenic seed of this line reaching up to 25%. Two novel genes, an 18-carbon ω3 desaturase (CpDesX) from Claviceps purpurea and a 20-carbon ω3 desaturase (Pir-ω3) from Pythium irregulare, proved to be very effective in increasing EPA levels in high-erucic acid B. carinata. The conlinin1 promoter from flax functioned reasonably well in B. carinata, and can serve as an alternative to the napin promoter from B. napus. In summary, the judicious selection of host species and promoters, together with the inclusion of genes that enhance the basic very long chain polyunsaturated fatty acid biosynthetic pathway, can greatly influence the production of EPA in plants.

Keywords

Eicosapentaenoic acid Brassica carinata B. juncea Transformation 

Supplementary material

11248_2009_9302_MOESM_ESM.doc (310 kb)
(DOC 310 kb)

References

  1. Abbadi A, Domergue F, Bauer J, Napier JA, Welti R, Zähringer U, Cirpus P, Heinz E (2004) Biosynthesis of very-long-chain polyunsaturated fatty acids in transgenic oilseeds: constraints on their accumulation. Plant Cell 16:1–15CrossRefGoogle Scholar
  2. Babic V, Datla RS, Scoles GJ, Keller WA (1998) Development of an efficient Agrobacterium-mediated transformation system for Brassica carinata. Plant Cell Rep 17:183–188CrossRefGoogle Scholar
  3. Bauer J, Wu G, Qiu X (2008) Isolation and characterization of a novel Pythium omega-3 desaturase with specificity to all omega-6 fatty acids longer than 18 carbon chains. WO/2008/022963Google Scholar
  4. Hong HP, Datla N, Reed DW, Covello PS, MacKenzie SL, Qiu X (2002) High-level production of γ-linolenic acid in Brassica juncea using a Δ6 desaturase from Pythium irregulare. Plant Physiol 129:354–362CrossRefPubMedGoogle Scholar
  5. Kinney AJ, Cahoon EB, Damude HG, Hitz WD, Kolar CW, Liu ZB (2004) Production of very long chain polyunsaturated fatty acids in oilseed plants. Patent WO/2004/071467 A2Google Scholar
  6. Kooter JM, Matzke MA, Meyer P (1999) Listening to the silent genes: transgene silencing, gene regulation and pathogen control. Trends in Plant Sci 4:340–347CrossRefGoogle Scholar
  7. Matzke AJM, Matzke MA (1998) Position effects and epigenetic silencing of plant transgenes. Curr Opin Plant Biol 1:142–148CrossRefPubMedGoogle Scholar
  8. Meesapyodsuk D, Reed DW, Covello PS, Qiu X (2007) Primary structure, regioselectivity, and evoloution of the membrane-bound fatty acid desaturases of Claviceps purpurea. J Biol Chem 282:20191–20199CrossRefPubMedGoogle Scholar
  9. Meyer A, Kirsch H, Domergue F, Abbadi A, Sperling P, Bauer J, Cirpus P, Zank TK, Moreau H, Roscoe TJ, Zähringer U, Heinz E (2004) Novel fatty acid elongases and their use for the reconstitution of docosahexaenoic acid biosynthesis. J Lipid Res 45:1899–1909CrossRefPubMedGoogle Scholar
  10. Myers RA, Worm B (2003) Rapid worldwide depletion of predatory fish communities. Nature 423:280–283CrossRefPubMedGoogle Scholar
  11. Naylor RL, Goldburg RJ, Primavera JH, Kautsky N, Beveridge MCM, Clay J, Folke C, Lubchenco J, Mooney H, Troell M (2000) Effect of aquaculture on world fish supplies. Nature 405:1017–1024CrossRefPubMedGoogle Scholar
  12. Qi BX, Fraser T, Mugford S, Dobson G, Sayanova O, Butler J, Napier JA, Stobart AK, Lazarus CM (2004) Production of very long chain polyunsaturated omega-3 and omega-6 fatty acids in plants. Nat Biotechnol 22:739–745CrossRefPubMedGoogle Scholar
  13. Qiu X, Hong HP, MacKenzie SL (2001a) Identification of a △4 fatty acid desaturase from Thraustochytrium sp. involved in the biosynthesis of docosahexanoic acid by heterologous expression in Saccharomyces cerevisiae and Brassica juncea. J Biol Chem 276:31561–31566CrossRefPubMedGoogle Scholar
  14. Qiu X, Reed DR, Hong HP, MacKenzie SL, Covello PS (2001b) Identification and analysis of a gene from Calendula officinalis encoding a fatty acid conjugase. Plant Physiol 125:847–855CrossRefPubMedGoogle Scholar
  15. Radke SE, Turner JC, Facciotti D (1992) Transformation and regeneration of Brassica rapa using Agrobacterium tumefaciens. Plant Cell Rep 11:499–505CrossRefGoogle Scholar
  16. Robert SS, Singh SP, Zhou XR, Petrie JR, Blackburn SI, Mansour PM, Nichols PD, Liu Q, Green AG (2005) Metabolic engineering of Arabidopsis to produce nutritionally important DHA in seed oil. Funct Plant Biol 32:473–479CrossRefGoogle Scholar
  17. Simopoulos AP (2003) Importance of the ratio of omega-6/omega-3 essential fatty acids: evolutionary aspects. World Rev Nutr Diet 92:1–22CrossRefPubMedGoogle Scholar
  18. Truksa T, MacKenzie SL, Qiu X (2003) Molecular analysis of flax 2S storage protein conlinin and seed specific activity of its promoter. Plant Physiol Biochem 41:141–147CrossRefGoogle Scholar
  19. Wahlroos T, Susi P, Solovyev A, Dorokhov Y, Morozov S, Atabekov J, Korpela T (2004) Increase of histidine content in Brassica rapa subsp. oleifera by over-expression of histidine-rich fusion proteins. Mol Breed 14:455–462CrossRefGoogle Scholar
  20. Wang YP, Sonntag K, Rudloff E, Han J (2005) Production of fertile transgenic Brassica napus by Agrobacterium-mediated transformation of protoplasts. Plant Breed 124:1–4CrossRefGoogle Scholar
  21. Wu G, Truksa M, Datla N, Vrinten P, Bauer J, Zank T, Cirpus P, Heinz E, Qiu X (2005) Stepwise engineering to produce high yields of very long-chain polyunsaturated fatty acids in plants. Nat Biotechnol 23:1013–1017CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Bifang Cheng
    • 1
    • 2
  • Guohai Wu
    • 1
  • Patricia Vrinten
    • 1
  • Kevin Falk
    • 2
  • Joerg Bauer
    • 3
  • Xiao Qiu
    • 4
  1. 1.Bioriginal Food and Science CorporationSaskatoonCanada
  2. 2.Agriculture and Agri-Food CanadaSaskatoon Research CentreSaskatoonCanada
  3. 3.BASF Plant Science GmbHLimburgerhofGermany
  4. 4.Department of Food and Bioproduct SciencesUniversity of SaskatchewanSaskatoonCanada

Personalised recommendations