Biologia plantarum 2014, 58:348-354 | DOI: 10.1007/s10535-014-0406-9
Transgenic barley producing essential polyunsaturated fatty acids
- 1 Plant Production Research Center, Piešťany, Slovak Republic
- 2 Faculty of Chemical and Food Technology, Slovak University of Technology, Bratislava, Slovak Republic
Polyunsaturated fatty acids (PUFAs) affect diverse physiological processes and human health. Most cereals are poor in n-3 and n-6 PUFAs. Using biolistics, barley (Hordeum vulgare L. cv. Golden Promise) was transformed with an artificial gene encoding Δ6-desaturase (D6D) under an endosperm-specific promoter. This artificial gene was designed from the sequence of D6D of the filamentous fungus Thamnidium elegans, but codon usage was optimised for cereals. A signal sequence from the gene encoding for high molecular mass glutenin Dx5 was added to a destinate mature protein. Successful transformation was confirmed in T0 plants at the genomic level and in T1 seeds at the transcriptomic and metabolomic levels. Transformed plants produced up to 0.141 % of γ-linolenic acid (GLA) and 0.294 % of stearidonic acid (SDA) of the total amount of fatty acids in their grains. Although the content of these fatty acids was relatively low, the current study provides the first evidence that transgenic barley can be a source of GLA/SDA.
Keywords: biolistic transformation; Δ6-desaturase; γ-linolenic acid; Hordeum vulgare; stearidonic acid
Subjects: transgenic plants; polyunsaturated fatty acids; biolistic transformation; barley
Species: Hordeum vulgare
Received: April 8, 2013; Revised: September 5, 2013; Accepted: September 16, 2013; Published: June 1, 2014Show citation
ACS | AIP | APA | ASA | Harvard | Chicago | IEEE | ISO690 | MLA | NLM | Turabian | Vancouver |
References
- Anderson, O.D., Greene, F.C., Anderson, O.D., Greene, F.C., Yip, R.E., Halford, N.G., Shewry, P.R., Malpica-Romero, J.M.: Nucleotide sequences of the two high-molecularweight glutenin genes from the D-genome of a hexaploid bread wheat, Triticum aestivum L. cv Cheyenne. - Nucl. Acids Res. 17: 461-462, 1989.
Go to original source...
- Brenner, R.R.: Regulatory function of D6-desaturase - a key enzyme of polyunsaturated fatty acid synthesis. - Adv. exp. Med. Biol. 83: 85-101, 1976.
Go to original source...
- Certík, M., Adamechova, Z.: Cereal-based bioproducts containing polyunsaturated fatty acids. - Lipid Technol. 21: 250-253, 2009.
Go to original source...
- Certik, M., Adamechova, Z., Slavikova, L.: Biotechnological enrichment of cereals with polyunsaturated fatty acids. - In: Hou, C.T., Shaw, J.F. (ed.): Biocatalysis and Biomolecular Engineering. Pp. 175-193. John Wiley & Sons, Hoboken 2010.
- Certik, M., Sakuradani, E., Shimizu, S.: Desaturase-defective fungal mutants: useful tools for the regulation and overproduction of polyunsaturated fatty acids. - Trends Biotechnol. 16: 500-505, 1998.
Go to original source...
- Certik, M., Shimizu, S.: Biosynthesis and regulation of microbial polyunsaturated fatty acid production. - J. Biosci. Bioeng. 87: 1-14, 1999.
Go to original source...
- Cheawchanlertfa, P., Cheevadhanarak, S., Tanticharoen, M., Maresca, B., Laoteng, K.: Up-regulated expression of desaturase genes of Mucor rouxii in response to low temperature associates with pre-existing cellular fatty acid constituents. - Mol. Biol. Rep. 38: 3455-3462, 2011.
Go to original source...
- Chen, R., Tsuda, S., Matsui, K., Mizutani, M.F., Ochiai, M., Shimizu, S., Sakuradani, E., Aoki, T., Imaizumi, R., Ayabe, S., Tanaka, Y.: Production of γ-linolenic acid in Lotus japonicus and Vigna angularis by expression of the Δ-6-fatty-acid desaturase gene isolated from Mortirella alpina. - Plant Sci. 169: 599-605, 2005.
Go to original source...
- Christensen, A.H., Quail, P.H.: Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants. - Transgenic Res. 5: 213-218, 1996.
Go to original source...
- Dahleen, L.S., Manoharan, M.: Recent advances in barley transformation. - In Vitro cell. dev. Biol. Plant 43: 493-506, 2007.
Go to original source...
- Flider, F.J.: GLA: uses and new sources. - Inform 16: 279-282, 2005.
- Furtado, A., Henry, R.J., Pellegrineschi, A.: Analysis of promoters in transgenic barley and wheat. - Plant Biotechnol. J. 7: 240-253, 2009.
Go to original source...
- García-Maroto, F., Garrido-Cárdenas, J.A., Rodríguez-Ruiz, J., Vilches-Ferrón, M., Adam, A.C., Polaina, J., López Alonso, D.: Cloning and molecular characterization of the Δ6-desaturase from two Echium plant species: production of GLA by heterologous expression in yeast and tobacco. - Lipids 37: 417-426, 2002.
Go to original source...
- Gill, I., Valivety, R.: Polyunsaturated fatty acids, part 1: occurrence, biological activities and applications. - Trends Biotechnol. 15: 401-409, 1997.
Go to original source...
- Gregova, E., Mihalik, D., Slikova, S., Sramkova, Z.: Allelic variation of HMW glutenin subunits and 1BL. 1RS translocation in Slovak common wheats. - Cereal Res. Commun. 35: 1675-1683, 2007.
Go to original source...
- Gubisova, M., Mihalik, D., Konopkova, L.: Regeneration efficiency of Slovak spring barley cultivars and winter wheat cultivars. - Agriculture 57: 76-83, 2011.
- Gunstone, F.D.: Gamma linolenic acid - occurrence and physical and chemical properties. - Progr. Lipid Res. 31: 145-161, 1992.
Go to original source...
- Harwood, W.A., Smedley, M.A.: Barley transformation using biolistic techniques. - In: Jones, H.D., Shewry, P.R. (ed.): Methods in Molecular Biology. Transgenic Wheat, Barley and Oats. Pp. 125-136. Humana Press, New York 2009.
- He, G.Y., Jones, H.D., D'Ovidio, R., Masci, S., Chen, M., West, J., Butow, B., Anderson, O.D., Lazzeri, P., Fido, R., Shewry, P.R.: Expression of an extended HMW subunit in transgenic wheat and the effect on dough mixing properties. - J. Cereal Sci. 42: 225-231, 2005.
Go to original source...
- Hong, H., Datla, N., Reed, D.W., Covello, P.S., MacKenzie S.L., Qiu, X.: High-level production of γ-linolenic acid in Brassica juncea using a D6 desaturase from Pythium irregulare. - Plant Physiol. 129: 354-362, 2002.
Go to original source...
- Hudson, B.J.: Evening primrose (Oenothera spp.) oil and seed. - J. amer. Oil Chem. Soc. 61: 540-542, 1984.
Go to original source...
- Liu, J.W., DeMichele, S., Bergana, M., Bobik, E., Hastilow, C., Chuang, L.T., Mukerji, P, Huang, Y.S.: Characterization of oil exhibiting high gamma-linolenic acid from a genetically transformed canola strain. - J. amer. Oil Chem. Soc. 78: 489-493, 2001.
Go to original source...
- Matsumoto, T., Tanaka, T., Sakai, H., Amano, N., Kanamori, H., Kurita, K., Kikuta, A., Kamiya, K., Yamamoto, M., Ikawa, H., Fujii, N., Hori, K., Itoh, T., Sato, K.: Comprehensive sequence analysis of 24,783 barley fulllength cDNAs derived from 12 clone libraries. - Plant Physiol. 156: 20-28, 2011.
Go to original source...
- Na-Ranong, S., Laoteng, K., Kittakoop, P., Tanticharoen, M., Cheevadhanarak, S.: Substrate specificity and preference of delta-6-desaturase of Mucor rouxii. - FEBS Lett. 579: 2744-2748, 2005.
Go to original source...
- Nykiforuk, C.L., Shewmaker, C., Harry, I., Yurchenko, O.P., Zhang, M., Reed, C., Oinam, G.S., Zaplachinski, S., Fidantsef, A., Boothe, J.G., Moloney, M.M.: High level accumulation of gamma linolenic acid (C18:3Δ6.9,12 cis) in transgenic safflower (Carthamus tinctorius) seeds. - Transgenic Res. 21: 367-381, 2012.
Go to original source...
- Puigbó, P., Guzmán, E., Romeu, A., Garcia-Vallvé, S.: OPTIMIZER: a web server for optimizing the codon usage of DNA sequences. - Nucl. Acids Res. 35: 126-131, 2007.
Go to original source...
- Ratledge, C.: Fatty acid biosynthesis in microorganisms being used for single cell oil production. - Biochimie 86: 807-815, 2004.
Go to original source...
- Reddy, A.S., Nuccio, M.L., Gross, L.M.: Isolation of a D6-desaturase gene from the cyanobacterium Synechocystis sp. strain PCC6803 by gain-of-function expression in Anabaena sp. strain PCC7120. - Plant mol. Biol. 22: 293-300, 1993.
Go to original source...
- Reddy, A.S., Thomas, T.L.: Expression of a cyanobacterial D6-desaturase gene results in Γ-linolenic acid production in transgenic plants. - Nat. Biotech. 14: 639-642, 1996.
Go to original source...
- Sato, S., Xing, A., Ye, X., Schweiger, B., Kinney, A., Graef, G., Clemente, T.: Production of Γ-linolenic acid and stearidoinic acid in seeds of marker-free transgenic soybean. - Crop Sci. 44: 646-652, 2004.
Go to original source...
- Sayanova, O., Smith, M.A., Lapinskas, P., Stobart, A.K., Dobson, G., Christie, W.W., Shewry, P.R., Napier, J.A.: Expression of borage desaturase cDNA containing an Nterminal cytochrom b5 domain results in the accumulation of high levels of D6-desaturated fatty acids in transgenic tobacco. - Proc. nat. Acad. Sci. USA 94: 4211-4216, 1997.
Go to original source...
- Sood, P., Bhattacharya, A., Sood, A.: Problems and possibilities of monocot transformation. - Biol. Plant. 55: 1-15, 2011.
Go to original source...
- Stymne, S., Stobart, A. K.: Biosynthesis of γ-linolenic acid in cotyledons and microsomal preparation of the developing seeds of common borage (Borago officinalis). - Biochem. J. 24: 385-393, 1986.
Go to original source...
- Tang, G.Y., Wei, L.Q, Liu, Z.J., Bi, Y.P., Shan, L.: Ectopic expression of peanut acyl carrier protein in tobacco alters fatty acid composition in the leaf and resistance to cold stress. - Biol. Plant. 56: 493-501, 2012.
Go to original source...
- Traitler, H., Wille, H.J., Studer, A.: Fractionation of black currant seed oil. - J. amer. Oil Chem. Soc. 65: 755-760, 1988.
Go to original source...
- Ucciani, E.: Potential sources of gamma-linolenic acid. - OCL. Oleagineux Corps Gras Lipides 2: 319-322, 1995.
- Wang, D., Li, M., Wei, D., Cai, Y., Zhang, Y., Xing, L.: Identification and functional characterization of the delta 6-fatty acid desaturase gene from Thamnidium elegans. - J. Eukaryot. Microbiol. 54: 110-117, 2007.
Go to original source...
- Zhao, X., Kong, X., Hua, Y., Feng, B., Zhao, Z.K.: Medium optimization for lipid production through co-fermentation of glucose and xylose by oleaginous yeast Lipomyces starkeyi. - Eur. J. Lipid Sci. Technol. 110: 405-412, 2008.
Go to original source...
- Zhou, X., R., Robert, S., Singh, S., Green, A.: Heterelogous production of GLA and SDA by expression of an Echium plantagineum Δ-6 desaturase gene. - Plant Sci. 170: 665-673, 2006.
Go to original source...