title: Biosynthesis of Essential Polyunsaturated Fatty Acids in Wheat Triggered by Expression of Artificial Gene.
abstract: The artificial gene D6D encoding the enzyme ∆⁶desaturase was designed and synthesized using the sequence of the same gene from the fungus Thamnidium elegans. The original start codon was replaced by the signal sequence derived from the wheat gene for high-molecular-weight glutenin subunit and the codon usage was completely changed for optimal expression in wheat. Synthesized artificial D6D gene was delivered into plants of the spring wheat line CY-45 and the gene itself, as well as transcribed D6D mRNA were confirmed in plants of T₀ and T₁ generations. The desired product of the wheat genetic modification by artificial D6D gene was the γ-linolenic acid. Its presence was confirmed in mature grains of transgenic wheat plants in the amount 0.04%-0.32% (v/v) of the total amount of fatty acids. Both newly synthesized γ-linolenic acid and stearidonic acid have been detected also in leaves, stems, roots, awns, paleas, rachillas, and immature grains of the T₁ generation as well as in immature and mature grains of the T₂ generation. Contents of γ-linolenic acid and stearidonic acid varied in range 0%-1.40% (v/v) and 0%-1.53% (v/v) from the total amount of fatty acids, respectively. This approach has opened the pathway of desaturation of fatty acids and production of essential polyunsaturated fatty acids in wheat.
authors.affiliation: Research Institute of Plant Production, National Agricultural and Food Center, 921 68 Piešťany, Slovakia. mihalik@vurv.sk.|Department of Biotechnology, Faculty of Natural Sciences, University of SS, Cyril and Methodius in Trnava, 917 01 Trnava, Slovakia. mihalik@vurv.sk.
authors.forename: Daniel
authors.initials: D
authors.lastname: Mihálik
authors.affiliation: Research Institute of Plant Production, National Agricultural and Food Center, 921 68 Piešťany, Slovakia. klcova.l@vurv.sk.|Department of Botany and Genetics, Faculty of Natural Sciences, Constantine the Philosopher University in Nitra, 949 74 Nitra, Slovakia. klcova.l@vurv.sk.
authors.forename: Lenka
authors.initials: L
authors.lastname: Klčová
authors.affiliation: Research Institute of Plant Production, National Agricultural and Food Center, 921 68 Piešťany, Slovakia. ondreickova@vurv.sk.
authors.forename: Katarína
authors.initials: K
authors.lastname: Ondreičková
authors.affiliation: Research Institute of Plant Production, National Agricultural and Food Center, 921 68 Piešťany, Slovakia. hudcovicova@vurv.sk.
authors.forename: Martina
authors.initials: M
authors.lastname: Hudcovicová
authors.affiliation: Research Institute of Plant Production, National Agricultural and Food Center, 921 68 Piešťany, Slovakia. gubisova@vurv.sk.
authors.forename: Marcela
authors.initials: M
authors.lastname: Gubišová
authors.affiliation: Faculty of Chemical and Food Technology, Slovak University of Technology, 812 37 Bratislava, Slovakia. tatiana.klempova@stuba.sk.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Klempová
authors.affiliation: Faculty of Chemical and Food Technology, Slovak University of Technology, 812 37 Bratislava, Slovakia. milan.certik@stuba.sk.
authors.forename: Milan
authors.initials: M
authors.lastname: Čertík
authors.affiliation: Cereal Research Non-profit Ltd., Szeged, Alsó kikötö sor 9, H-6726 Szeged, Hungary. janos.pauk@gk-szeged.hu.
authors.forename: János
authors.initials: J
authors.lastname: Pauk
authors.affiliation: Research Institute of Plant Production, National Agricultural and Food Center, 921 68 Piešťany, Slovakia. kraic@vurv.sk.|Department of Biotechnology, Faculty of Natural Sciences, University of SS, Cyril and Methodius in Trnava, 917 01 Trnava, Slovakia. kraic@vurv.sk.
authors.forename: Ján
authors.initials: J
authors.lastname: Kraic
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D019470:Biolistics; D053898:Biosynthetic Pathways; D002849:Chromatography, Gas; D005231:Fatty Acids, Unsaturated / Q000096:biosynthesis*; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant; D005813:Genes, Synthetic; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D012639:Seeds / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D014170:Transformation, Genetic; D019076:Transgenes; D014908:Triticum / Q000235:genetics* / Q000382:microbiology
chemical_list: D005231:Fatty Acids, Unsaturated; D010940:Plant Proteins
keywords: artificial gene; essential fatty acid; transgene; wheat; γ-linolenic acid; ∆6desaturase