title: Effects of tung oilseed FAD2 and DGAT2 genes on unsaturated fatty acid accumulation in Rhodotorula glutinis and Arabidopsis thaliana.
abstract: Genetic engineering to produce valuable lipids containing unsaturated fatty acids (UFAs) holds great promise for food and industrial applications. Efforts to genetically modify plants to produce desirable UFAs with single enzymes, however, have had modest success. The key enzymes fatty acid desaturase (FAD) and diacylglycerol acyltransferase (DGAT) are responsible for UFA biosynthesis (a push process) and assembling fatty acids into lipids (a pull process) in plants, respectively. To examine their roles in UFA accumulation, VfFAD2 and VfDGAT2 genes cloned from Vernicia fordii (tung tree) oilseeds were conjugated and transformed into Rhodotorula glutinis and Arabidopsis thaliana via Agrobacterium tumefaciens. Real-time quantitative PCR revealed variable gene expression levels in the transformants, with a much higher level of VfDGAT2 than VfFAD2. The relationship between VfFAD2 expression and linoleic acid (C18:2) increases in R. glutinis (R (2) = 0.98) and A. thaliana (R (2) = 0.857) transformants was statistically linear. The VfDGAT2 expression level was statistically correlated with increased total fatty acid content in R. glutinis (R (2) = 0.962) and A. thaliana (R (2) = 0.8157) transformants. With a similar expression level between single- and two-gene transformants, VfFAD2-VfDGAT2 co-transformants showed a higher linolenic acid (C18:3) yield in R. glutinis (174.36 % increase) and A. thaliana (14.61 % increase), and eicosatrienoic acid (C20:3) was enriched (17.10 % increase) in A. thaliana. Our data suggest that VfFAD2-VfDGAT2 had a synergistic effect on UFA metabolism in R. glutinis, and to a lesser extent, A. thaliana. These results show promise for further genetic engineering of plant lipids to produce desirable UFAs.
authors.affiliation: State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing, 100091, People's Republic of China, yicun_chen@163.com.
authors.forename: Yicun
authors.initials: Y
authors.lastname: Chen
authors.forename: Qinqin
authors.initials: Q
authors.lastname: Cui
authors.forename: Yongjie
authors.initials: Y
authors.lastname: Xu
authors.forename: Susu
authors.initials: S
authors.lastname: Yang
authors.forename: Ming
authors.initials: M
authors.lastname: Gao
authors.forename: Yangdong
authors.initials: Y
authors.lastname: Wang
mesh_terms: D015126:8,11,14-Eicosatrienoic Acid / Q000378:metabolism; D031284:Aleurites / Q000201:enzymology* / Q000235:genetics; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D051048:Diacylglycerol O-Acyltransferase / Q000235:genetics / Q000378:metabolism*; D044943:Fatty Acid Desaturases / Q000235:genetics / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D008055:Lipids / Q000032:analysis / Q000096:biosynthesis; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012248:Rhodotorula / Q000235:genetics / Q000378:metabolism*; D012639:Seeds / Q000201:enzymology / Q000235:genetics; D017962:alpha-Linolenic Acid / Q000378:metabolism
chemical_list: D005231:Fatty Acids, Unsaturated; D008055:Lipids; D010940:Plant Proteins; D017962:alpha-Linolenic Acid; D044943:Fatty Acid Desaturases; C040448:delta-12 fatty acid desaturase; D051048:Diacylglycerol O-Acyltransferase; D015126:8,11,14-Eicosatrienoic Acid