Journal of the American Oil Chemists' Society

, Volume 78, Issue 9, pp 941–947

Transgenic cotton plants with increased seed oleic acid content

  • Kent D. Chapman
  • Shea Austin-Brown
  • Salvatore A. Sparace
  • Anthony J. Kinney
  • Kevin G. Ripp
  • Irma L. Pirtle
  • Robert M. Pirtle
Article

Abstract

Cottonseed typically contains about 15% oleic acid. Here we report the development of transgenic cotton plants with higher seed oleic acid levels. Plants were generated by Agrobacterium-mediated transformation. A binary vector was designed to suppress expression of the endogenous cottonseed †-12 desaturase (fad2) by subcloning a mutant allele of a rapeseed fad2 gene downstream from a heterologous, seedspecific promoter (phaseolin). Fatty acid profiles of total seed lipids from 43 independent transgenic lines were analyzed by gas chromatography. Increased seed oleic acid content ranged from 21 to 30% (by weight) of total fatty acid content in 22 of the primary transformants. The increase in oleic acid content was at the expense of linoleic acid, consistent with reduced activity of cottonseed FAD2. Progeny of some lines yielded oleic acid content as high as 47% (three times that of standard cottonseed oil). Molecular analyses of nuclear DNA from transgenics confirmed the integration of the canola transgene into the cotton genome. Collectively, our results extend the metabolic engineering of vegetable oils to cottonseed and should provide the basis for the development of a family of novel cottonseed oils.

Key Words

Cottonseed oil fatty acid metabolism metabolic engineering oilseeds 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Cottonseed: World's No. 3 Oilseed Yields Diverse Products, inform 11:820–839 (2000).Google Scholar
  2. 2.
    Jones, L.A., and C.C. King, Cottonseed Oil, in Bailey's Industrial Oil and Fat Products, Vol. 2 Edible Oil and Fat Products: Oils and Oil Seeds, 5th edn. edited by Y. Hui, John Wiley & Sons, New York, 1996, pp. 159–240.Google Scholar
  3. 3.
    Kinney, A.L., and S. Knowlton, Designer Oils:: The High-Oleic Soybean, in Genetic Modification in the Food Industry, edited by S. Roller and S. Harlander, Blackie Academic and Professional, New York, 1998, pp. 193–213.Google Scholar
  4. 4.
    Kinney, A.J., Genetic Engineering of the Storage Lipids of Plants, Curr. Opinion Biotechnol. 5:144–151 (1994).CrossRefGoogle Scholar
  5. 5.
    Browse, J., J. Spychalla, J. Okuley, and J. Lightner, Altering the Fatty Acid Composition of Vegetable Oils, in Plant Lipid Biosynthesis: Fundamentals and Agricultural Applications, edited by J.L. Harwood, Society for Experimental Biology Seminar Series 67, Cambridge University Press, Cambridge, 1998, pp. 131–153.Google Scholar
  6. 6.
    Downey, R.K., and G. Röbbelen, Brassica Species, in Oil Crops of the World: Their Breeding and Utilization, edited by G. Röbbelen, R.K. Downey, and A. Ashri, McGraw Hill, New York, 1989, pp. 339–362.Google Scholar
  7. 7.
    Green, A.G., and D.R. Marshall, Isolation of Induced Mutants in Linseed (Linum usitatissimum) Having Reduced Linolenic Acid Content, Euphytica 33:321–328 (1984).CrossRefGoogle Scholar
  8. 8.
    Kinney, A.J., Genetic Engineering of Oilseeds for Desired Traits, in Genetic Engineering, edited by J.K. Setlow, Plenum Press, New York 1997, Vol. 19, pp. 149–166.Google Scholar
  9. 9.
    Del Vecchio, A.J., High-Laurate Canola, inform 7:230–243 (1996).Google Scholar
  10. 10.
    Kinney, A.J., Production of Specialised Oils for Industry, in Plant Lipid Biosynthesis: Fundamentals and Agricultural Applications, edited by J.L. Harwood, Society for Experimental Biology Seminar Series 67, Cambridge University Press, Cambridge, 1998, pp. 273–285.Google Scholar
  11. 11.
    Mazur, B., E. Krebbers, and S. Tingey, Gene Discovery and Product Development for Grain Quality Traits, Science 285: 372–375 (1999).CrossRefGoogle Scholar
  12. 12.
    Ohlrogge, J.B., Design of New Plant Products: Engineering of Fatty Acid Metabolism, Plant Physiol. 104:821–826 (1994).Google Scholar
  13. 13.
    Voelker, T., Plant Acyl-ACP Thioesterases: Chain-Length Determining Enzymes in Plant Fatty Acid Biosynthesis, in Genetic Engineering, edited by J.K. Setlow, Plenum Press, New York, 1996, Vol. 18, pp. 111–133.Google Scholar
  14. 14.
    Somerville, C., J. Browse, J. Jaworski, and J.B. Ohlrogge, Lipids, in Biochemistry and Molecular Biology of Plants, edited by B. Buchanan, W. Gruissem, and R. Jones, American Society of Plant Physiologists, Rockville, MD, 2000, pp. 456–527.Google Scholar
  15. 15.
    Harwood, J.L., Recent Advances in the Biosynthesis of Plant Fatty Acids, Biochim. Biophys. Acta 1301:7–56 (1996).Google Scholar
  16. 16.
    Mekhedov, S., O. Martínez de Ilárduya, and J.B. Ohlrogge, Toward a Functional Catalog of the Plant Genome. A Survey of Genes for Lipid Biosynthesis, Plant Physiol. 122:389–402 (2000).CrossRefGoogle Scholar
  17. 17.
    Voelker, T.A., A.C. Worrell, L. Anderson, J. Bleibaum, C. Fan, D.J. Hawkins, S.E. Radke, and H.M. Davies, Fatty Acid Biosynthesis Redirected to Medium Chains in Transgenic Oilseed Plants, Science 257:72–74 (1992).CrossRefGoogle Scholar
  18. 18.
    Shanklin, J., and E.B. Cahoon, Desaturation and Related Modifications of Fatty Acids, Annu. Rev. Plant Physiol. Plant Mol. Biol. 49:611–642 (1998).CrossRefGoogle Scholar
  19. 19.
    Okuley, J., J. Lightner, K. Feldmann, N. Yadav, E. Lark, and J. Browse, Arabidopsis FAD2 Gene Encodes the Enzyme That Is Essential for Polyunsaturated Lipid Synthesis, Plant Cell 6:147–158 (1994).CrossRefGoogle Scholar
  20. 20.
    Burow, M.D., P. Sen, C.A. Chlan, and N. Murai, Developmental Control of the beta-Phaseolin Gene Requires Positive, Negative, and Temporal Seed-Specific Transcriptional Regulatory Elements and a Negative Element for Stem and Root Expression, Plant J. 2:537–548 (1992).CrossRefGoogle Scholar
  21. 21.
    Thomas, J.C., D.G. Adams, V.D. Keppenne, C.C. Wasmann, J.K. Brown, M.R. Kanost, and H.J. Bohnert, Protease Inhibitors of Madhuca sexta Expressed in Transgenic Cotton, Plant Cell Rep. 14:758–762 (1995).CrossRefGoogle Scholar
  22. 22.
    Trolinder, N.L., and J.R. Goodin, Somatic Embryogenesis in Cotton (Gossypium). II. Requirements for Embryo Development and Plant Regeneration, Plant Cell Tissue Org. Cult. 12:43–53 (1988).CrossRefGoogle Scholar
  23. 23.
    Umbeck, P., G. Johnson, K. Baron, and W. Swain, Genetically Transformed Cotton (Gossypium hirsutum L.), Plants Biotechnol. 5:263–266 (1987).Google Scholar
  24. 24.
    Firoozabady, E., D.L. DeBoer, D.J. Merlo, E.L. Halk, L.N. Amerson, K.E. Rashka, and E.E. Murray, Transformation of Cotton (Gossypium hirsutum L.) by Agrobacterium tumefaciens and Regeneration of Transgenic Plants, Plant Mol. Biol. 10:105–116 (1987).CrossRefGoogle Scholar
  25. 25.
    Hemphill, J.K., C.G.A. Maier, and K.D. Chapman, Rapid in vitro Regeneration of Cotton (Gossypium hirsutum L.), Plant Cell Rep. 17:273–278 (1998).CrossRefGoogle Scholar
  26. 26.
    Christie, W.W., Lipid Analysis, 2nd edn, Pergamon Press, New York, 1982, pp. 52–54.Google Scholar
  27. 27.
    Chapman, K.D., and R.N. Trelease, Acquisition of Membrane Lipids by Differentiating Glyoxysomes: Role of Lipid Bodies, J. Cell Biol. 115:995–1007 (1991).CrossRefGoogle Scholar
  28. 28.
    Paterson, A.H., C.L. Brubaker, and J.F. Wendel, A Rapid Method for Extraction of Cotton (Gossypium spp.) Genomic DNA Suitable for RFLP or PCR Analysis, Plant Mol. Biol. Reporter 11:122–127 (1993).CrossRefGoogle Scholar
  29. 29.
    Sambrook, J., E.F. Fritsch, and T. Maniatis, Molecular Cloning: A Laboratory Manual 2nd edn., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989.Google Scholar
  30. 30.
    Feinberg, A.P., and B. Vogelstein, A Technique for Radiolabeling DNA Restriction Endonuclease Fragments to High Specific Activity, Anal. Biochem. 132:6–13 (1983).CrossRefGoogle Scholar
  31. 31.
    Reed, K.C., and D.A. Mann, Rapid Transfer of DNA from Agarose Gels to Nylon Membranes, Nucl. Acids Res. 13:7207–7221 (1985).Google Scholar
  32. 32.
    Trelease, R.N., J.A. Miernyk, J.S. Choinski, Jr., and S.J. Bortman, Synthesis and Compartmentation of Enzymes During Cottonseed Maturation, in Cotton Physiology, edited by J.M. Stewart, and J.R. Mauney, Cotton Foundation, Memphis, TN, 1986.Google Scholar
  33. 33.
    Croteau, R., T.M. Kutchan, and N.G. Lewis, Natural Products (secondary metabolites), in Biochemistry and Molecular Biology of Plants, edited by B. Buchannan, W. Gruissem, and R. Jones, American Society of Plant Physiologists, Rockville, MD, 2000, pp. 1250–1318.Google Scholar
  34. 34.
    Wood, R., Comparison of the Cyclopropene Fatty Acid Content of Cottoneed Varieties, Glanded and Glandless Seeds, and Various Seed Structures, Biochem. Arch. 2:73–80 (1986).Google Scholar

Copyright information

© AOCS Press 2001

Authors and Affiliations

  • Kent D. Chapman
    • 1
  • Shea Austin-Brown
    • 1
  • Salvatore A. Sparace
    • 2
  • Anthony J. Kinney
    • 3
  • Kevin G. Ripp
    • 3
  • Irma L. Pirtle
    • 1
  • Robert M. Pirtle
    • 1
  1. 1.Division of Biochemistry and Molecular Biology, Cottonseed Development GroupUniversity of North Texas Department of Biological SciencesDentoh
  2. 2.Department of Plant SciencesMcGill University, McDonald CampusQuébecCanada
  3. 3.DuPont Experimental StationWilmington

Personalised recommendations