Abstract
In addition to the normal-type sunflower (Helianthus annuus L.) where linoleic acid is the major seed fatty acid, a dominant negative high-oleic mutant with oleic acid as the predominant fatty acid was previously obtained. We report the isolation and characterization of three different cDNA sequences, designated Ha89FAD2-1, Ha89FAD2-2, and Ha89FAD2-3, encoding sunflower microsomal oleate desaturases (FAD2), using a PCR strategy. All three deduced amino acid sequences showed significant homology to the known plant FAD2 sequences. Genomic Southern blot analysis revealed that at least one copy of each of these genes is present in the sunflower genome, except for the FAD2-1 gene from the high-oleic mutant, which might be duplicated. The FAD2-2 and FAD2-3 genes were weakly expressed in all tissues studied from both varieties. In contrast, the FAD2-1 gene was expressed strongly and exclusively in developing embryos of normal-type sunflower, whereas its expression in high-oleic developing embryos was drastically reduced. Functional expression of the corresponding cDNAs in yeast confirmed that they encode microsomal oleate desaturases. Furthermore, the FAD2-1 gene from the high-oleic variety also expresses a fully active enzyme. These results suggest that the high-oleic mutation in sunflower interferes with the regulation of the transcription of the seed-specific FAD2 gene.
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Ausubel F.M., Brent R., Kingston R.E., Moore D.D., Seidman J.G., Smith J.A., Struhl K., Albright L.M., Coen D.M. and Varki A. 1995. Current Protocols in Molecular Biology. John Wiley, New York.
Brown A.P., Dann R., Bowra S. and Hills M. 1998. Characterization of expression of a plant oleate desaturase in yeast. J. Am. Oil. Chem. Soc. 75: 77–82.
Covello P.S. and Reed D.W. 1996. Functional expression of the extraplastidial Arabidopsis thaliana oleate desaturase gene (FAD2) in Saccharomyces cerevisiae. Plant Physiol. 111: 223–226.
Fernández-Martínez J., Jiménez A., Domínguez J., García J.M., Garcés R. and Mancha M. 1989. Genetic analysis of the high oleic acid content in cultivated sunflower (Helianthus annuus L.). Euphytica 41: 39–51.
Garcés R. and Mancha M. 1989. Oleate desaturation in seeds of two genotypes of sunflower. Phytochemistry 28: 2593–2595.
Garcés R. and Mancha M. 1991. In vitro oleate desaturase in developing sunflower seeds. Phytochemistry 30: 2127–2130.
Garcés R., García J.M. and Mancha M. 1989. Lipid characterization in seeds of a high oleic acid sunflower mutant. Phytochemistry 28: 2597–2600.
Garcés R., Sarmiento C. and Mancha M. 1992. Temperature regulation of oleate desaturase in sunflower (Helianthus annuus L.) seeds. Planta 186: 461–465.
Girke T., Schmidt H., Zähringer U., Reski R. and Heinz E. 1998. Identification of a novel Δ6-acyl-group desaturase by targeted gene disruption in Physcomitrella patens. Plant J. 15: 39–48.
Harris H.C., McWilliam J.R. and Mason W.K. 1978. Influence of temperature on oil content and composition of sunflower seed. Aust. J. Agric. Res. 29: 1203–1212.
Heppard E.P., Kinney A.J., Stecca K.L. and Miao G.-H. 1996. Developmental and growth temperature regulation of two different microsomal ω-6 desaturase genes in soybeans. Plant Physiol. 110: 311–319.
Hongtrakul V., Slabaugh M.B. and Knapp S.J. 1998. A seed specific Δ-12 oleate desaturase gene is duplicated, rearranged, and weakly expressed in high oleic acid sunflower lines. Crop Sci. 38: 1245–1249.
Jung S., Powell G., Moore K. and Abbott A. 2000. The high oleate trait in the cultivated peanut (Arachis hypogaea L.). II. Molecular basis and genetics of the trait. Mol. Gen. Genet. 263: 806–811.
Kajiwara S., Shirai A., Fujii T., Toguri T., Nakamura K. and Ohtaguchi K. 1996. Polyunsaturated fatty acid biosynthesis in Saccharomyces cerevisiae: Expression of ethanol tolerance and the FAD2 gene from Arabidopsis thaliana. Appl. Environ. Microbiol. 62: 4309–4313.
Liu Q., Singh S.P., Brubaker C.L., Sharp P.J., Green A.G. and Marshall D.R. 1997. Isolation and characterization of two different microsomal ω-6 desaturase genes in cotton (Gossypium hirsutum L.). In: Williams J.P., Khan M.U. and Lem N.W. (eds), Physiology, Biochemistry and Molecular Biology of Plant Lipids. Kluwer Academic Publishers, Dordrecht, Netherlands, pp. 383–385.
Miller J.F., Zimmerman D.C. and Vick B.A. 1987. Genetic control of high oleic acid content in sunflower oil. Crop Sci. 27: 923–926.
Murphy D.J. and Piffanelli P. 1998. Fatty acid desaturases: structure, mechanism and regulation. In: Harwood J.L. (ed.), Plant Lipid Biosynthesis. Fundamentals and Agricultural Applications, Seminar Series 67. Cambridge University Press, Cambridge, UK, pp. 95–130.
Okuley J., Lightner J., Feldmann K., Yadav N., Lark E. and Browse J. 1994. Arabidopsis FAD2 gene encodes the enzyme that is essential for polyunsaturated lipid synthesis. Plant Cell 6: 147–158.
Shanklin J. and Cahoon E.B. 1998. Desaturation and related modifications of fatty acids. Annu. Rev. Plant Physiol. Plant Mol. Biol. 49: 611–641.
Smith M.A., Cross A.R., Jones O.T.G., Griffiths W.T., Stymne S. and Stobart K. 1990. Electron-transport components of the 1-acyl-2-oleoyl-sn-glycero-3-phosphocholine Δ12-desaturase (Δ12-desaturase) in microsomal preparations from developing safflower (Carthamus tinctorius L.) cotyledons. Biochem. J. 272: 23–29.
Soldatov K.I. 1976. Chemical mutagenesis in sunflower breeding. In: Proceedings of the 7th International Sunflower Conference. Internatiional Sunflower Association, Vlaardingen, Netherlands, pp. 352–357.
Sperling P., Hammer U., Friedt W. and Heinz E. 1990. High oleic sunflower: Studies on composition and desaturation of acyl groups in different lipids and organs. Z. Naturforsch. 45c: 166–172.
Thompson J.D., Gibson T.J., Plewniak F., Jeanmougin F. and Higgins D.G. 1997. The CLUSTALX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucl. Acid Res. 25: 4876–4882.
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Martínez-Rivas, J.M., Sperling, P., Lühs, W. et al. Spatial and temporal regulation of three different microsomal oleate desaturase genes (FAD2) from normal-type and high-oleic varieties of sunflower (Helianthus annuus L.). Molecular Breeding 8, 159–168 (2001). https://doi.org/10.1023/A:1013324329322
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DOI: https://doi.org/10.1023/A:1013324329322