Advertisement

Accumulation of γ-linolenic acid and stearidonic acid in rapeseeds that heterologously express the Phytophthora citrophthora Δ6 desaturase gene

  • Kyeong-Ryeol LeeEmail author
  • Hami Yu
  • Inhwa Jeon
  • Kyung-Hwan Kim
  • Jong Sug Park
  • Juho Lee
  • Hyun Uk KimEmail author
Original Article
  • 18 Downloads

Abstract

Most oilseeds contain polyunsaturated fatty acids, such as linoleic acid (LA) and α-linolenic acid (ALA) that are important essential fatty acids for human health. γ-Linolenic acid (GLA) and stearidonic acid (SDA), synthesized from LA and ALA by Δ6 desaturase, respectively, are mainly obtained from algae and fish. Given the benefits of GLA and SDA to human health, many researchers have investigated ways to produce these fatty acids in oilseed crops. Canola-type oilseed rape (Brassica napus), widely cultivated in temperate and microthermal zones, accumulates oleic acid to a relatively greater extent (60–70% oleic acid content) than do other oilseed crops. We transformed the Phytophthora citrophthora Δ6 desaturase (PcD6DES) gene under the control of seed-specific vicilin into canola-type oilseed rape ‘Youngsan’. D6DES products (GLA, SDA, and 18:2Δ6,9) accumulated in mature PcD6DES-transformed rapeseeds and leaves. D6DES products of PcD6DES rapeseeds were over 20% in T1 and more than 25% in T2. Expression levels of the PcD6DES gene and the content of D6DES products coincided with each other and were related to expression levels and fatty acid composition in leaves and developing seeds 15, 25, 35, and 45 days after flowering. Seed weights of PcD6DES rapeseeds were not lower than those of Youngsan. In this study, PcD6DES oilseed rapes accumulated GLA, SDA, and putative 18:2Δ6,9 content to a maximum of 25% in the seed oil. Results show that PcD6DES rapeseed oil can potentially be used as a health food to improve human health.

Keywords

Δ6 desaturase D6DES Oilseed rape GLA SDA 

Notes

Acknowledgements

This study was conducted with the support of the Research Program for Agricultural Science & Technology Development (Project no. PJ01257102), the National Institute of Agricultural Science, the Next-Generation BioGreen 21 Program (SSAC, Grant no. PJ01318501), Rural Development Administration, Republic of Korea, and the Mid-Career Researcher Program of the National Research Foundation of Korea (NRF-2017R1A2B4007096).

Supplementary material

11816_2019_547_MOESM1_ESM.pdf (32 kb)
Supplementary material 1 (PDF 31 kb)
11816_2019_547_MOESM2_ESM.xlsx (18 kb)
Supplementary material 2 (XLSX 18 kb)
11816_2019_547_MOESM3_ESM.xlsx (23 kb)
Supplementary material 3 (XLSX 23 kb)
11816_2019_547_MOESM4_ESM.xlsx (17 kb)
Supplementary material 4 (XLSX 17 kb)

References

  1. Chen GQ, Turner C, He X, Nguyen T, McKeon TA, Laudencia-Chingcuanco D (2007) Expression profiles of genes involved in fatty acid and triacylglycerol synthesis in castor bean (Ricinus communis L.). Lipids 42(3):263–274CrossRefGoogle Scholar
  2. Coupland K (2008) Stearidonic acid: a plant produced omega-3 PUFA and a potential alternative for marine oil fatty acids. Lipid Technol 20:152–154CrossRefGoogle Scholar
  3. Guil-Guerrero JL (2007) Stearidonic acid (18: 4n–3): metabolism, nutritional importance, medical uses and natural sources. Eur J Lipid Sci Technol 109:1226–1236CrossRefGoogle Scholar
  4. Guillou H, D’Andrea S, Rioux V, Barnouin R, Dalaine S, Pédrono F, Jan S, Legrand P (2004) Distinct roles of endoplasmic reticulum cytochrome b5 and fused cytochrome b5-like domain for rat Delta6-desaturase activity. J Lipid Res 45:32–40CrossRefGoogle Scholar
  5. Hong H, Datla N, Reed DW, Covello PS, MacKenzie SL, Qiu X (2002) High-level production of gamma-linolenic acid in Brassica juncea using a delta 6 desaturase from Pythium irregulare. Plant Physiol 129:354–362CrossRefGoogle Scholar
  6. Hudson BJF (1984) Evening primrose (Oenothera spp.) oil and seed. J Am Oil Chem Soc 61:540–543CrossRefGoogle Scholar
  7. James MJ, Ursin VM, Cleland LG (2003) Metabolism of stearidonic acid in human subjects: comparison with the metabolism of other n-3 fatty acids. Am J Clin Nutr 77:1140–1145CrossRefGoogle Scholar
  8. Kapoor R, Huang YS (2006) Gamma linolenic acid: an antiinflammatory omega-6 fatty acid. Curr Pharm Biotechnol 7:531–534CrossRefGoogle Scholar
  9. Katavic V, Mietkiewska E, Barton DL, Giblin EM, Reed DW, Taylor DC (2002) Restoring enzyme activity in nonfunctional low erucic acid Brassica napus fatty acid elongase 1 by a single amino acid substitution. Eur J Biochem 269:5625–5631CrossRefGoogle Scholar
  10. Kawamura A, Ooyama K, Kojima K, Kachi H, Abe T, Amano K, Aoyama T (2011) Dietary supplementation of gamma-linolenic acid improves skin parameters in subjects with dry skin and mild atopic dermatitis. J Oleo Sci 60:597–607CrossRefGoogle Scholar
  11. Kim SH, Roh KH, Lee KR, Kang HC, Kim HU, Kim JB (2016) Metabolic engineering to produce γ-linolenic acid in Brassica napus using a Δ6-desaturase from pike eel. Plant Biotechnol Rep 10:475–481CrossRefGoogle Scholar
  12. Larkin PJ, Scowcroft WR (1981) Somaclonal variation—a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60:197CrossRefGoogle Scholar
  13. Lee KR, Kim EH, Roh KH, Kim JB, Kang HC, Go YS, Suh MC, Kim HU (2016) High-oleic oilseed rapes developed with seed-specific suppression of FAD2 gene expression. Appl Biol Chem 59:669–676CrossRefGoogle Scholar
  14. Lee KR, Kim KH, Kim JB, Hong SB, Jeon I, Kim HU, Lee MH, Kim JG (2019) High Accumulation of γ-linolenic acid and stearidonic acid in transgenic perilla (Perilla frutescens var. frutescens) seeds. BMC Plant Biol 19:120.  https://doi.org/10.1186/s12870-019-1713-2 CrossRefGoogle Scholar
  15. Li F, Chen B, Xu K, Wu J, Song W, Bancroft I, Harper AL, Trick M, Liu S, Gao G, Wang N, Yan G, Qiao J, Li J, Li H, Xiao X, Zhang T, Wu X (2014) Genome-wide association study dissects the genetic architecture of seed weight and seed quality in rapeseed (Brassica napus L.). DNA Res 21:355–367.  https://doi.org/10.1093/dnares/dsu002 CrossRefGoogle Scholar
  16. Li N, Peng W, Shi J, Wang X, Liu G, Wang H (2015) The natural variation of seed weight is mainly controlled by maternal genotype in rapeseed (Brassica napus L.). PLoS One 10(4):e0125360CrossRefGoogle Scholar
  17. Liu JW, DeMichele S, Bergana M, Bobik E, Hastilow C, Chuang LT, Mukerji P, Huang YS (2001) Characterization of oil exhibiting high gamma-linolenic acid from a genetically transformed canola strain. J Am Oil Chem Soc 78:489–493CrossRefGoogle Scholar
  18. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plantarum 15(3):473–497.  https://doi.org/10.1111/j.1399-3054.1962.tb08052.x CrossRefGoogle Scholar
  19. Napier JA, Sayanova O, Stobart AK, Shewry PR (1997) A new class of cytochrome b5 fusion proteins. Biochem J 328(Pt 2):717–718Google Scholar
  20. Reddy AS, Nuccio ML, Gross LM, Thomas TL (1993) Isolation of a delta-6-desaturase gene from the Cyanobacterium Synechocystis sp. strain Pcc-6803 by gain-of-function expression in Anabaena sp. strain Pcc-7120. Plant Mol Biol 22:293–300CrossRefGoogle Scholar
  21. Ruiz-Lopez N, Haslam RP, Venegas-Caleron M, Larson TR, Graham IA, Napier JA, Sayanova O (2009) The synthesis and accumulation of stearidonic acid in transgenic plants: a novel source of ‘heart-healthy’ omega-3 fatty acids. Plant Biotechnol J 7:704–716CrossRefGoogle Scholar
  22. Ruiz-López N, Sayanova O, Napier JA, Haslam RP (2012) Metabolic engineering of the omega-3 long chain polyunsaturated fatty acid biosynthetic pathway into transgenic plants. J Exp Bot 63(7):2397–2410.  https://doi.org/10.1093/jxb/err454 CrossRefGoogle Scholar
  23. Ruuska SA, Girke T, Benning C, Ohlrogge JB (2002) Contrapuntal networks of gene expression during arabidopsis seed filling. Plant Cell 14(6):1191–1206.  https://doi.org/10.1105/tpc.000877 CrossRefGoogle Scholar
  24. Sayanova O, Smith MA, Lapinskas P, Stobart AK, Dobson G, Christie WW, Shewry PR, Napier JA (1997) Expression of a borage desaturase cDNA containing an N-terminal cytochrome b5 domain results in the accumulation of high levels of delta6-desaturated fatty acids in transgenic tobacco. Proc Natl Acad Sci USA 94:4211–4216CrossRefGoogle Scholar
  25. Sayanova O, Haslam R, Venegas-Caleron M, Napier JA (2006) Identification of primula “front-end” desaturases with distinct n-6 or n-3 substrate preferences. Planta 224:1269–1277CrossRefGoogle Scholar
  26. Shanklin J, Cahoon EB (1998) Desaturation and related modifications of fatty acids 1. Annu Rev Plant Physiol Plant Mol Biol 49:611–641CrossRefGoogle Scholar
  27. Singh SP, Zhou XR, Liu Q, Stymne S, Green AG (2005) Metabolic engineering of new fatty acids in plants. Curr Opin Plant Biol 8:197–203CrossRefGoogle Scholar
  28. Stefansson BR, Hougen FW, Downey RK (1961) Note on the isolation of rape plants with seed oil free from erucic acid. Can J Plant Sci 41:218–219CrossRefGoogle Scholar
  29. Timoszuk M, Bielawska K, Skrzydlewska E (2018) Evening primrose (Oenothera biennis) biological activity dependent on chemical composition. Antioxidants 7(8):108.  https://doi.org/10.3390/antiox7080108 CrossRefGoogle Scholar
  30. Traitler H, Winter H, Richli U, Ingenbleek Y (1984) Characterization of gamma-linolenic acid in Ribes seed. Lipids 19:923–928CrossRefGoogle Scholar
  31. Vincentz M, Leite A, Neshich G, Vriend G, Mattar C, Barros L, Weinberg D, de Almeida ER, de Carvalho MP, Aragão F, Gander ES (1997) ACGT and vicilin core sequences in a promoter domain required for seed-specific expression of a 2S storage protein gene are recognized by the opaque-2 regulatory protein. Plant Mol Biol 34(6):879–889CrossRefGoogle Scholar

Copyright information

© Korean Society for Plant Biotechnology 2019

Authors and Affiliations

  1. 1.Department of Agricultural BiotechnologyNational Institute of Agricultural Science, Rural Development AdministrationJeonjuRepublic of Korea
  2. 2.Department of Bioindustry and Bioresource Engineering, Plant Engineering Research InstituteSejong UniversitySeoulRepublic of Korea

Personalised recommendations