Effect of Overexpression of PDAT Gene on Arabidopsis Growth Rate and Seed Oil Content

Arabidopsis (ecotype Columbia 0) plants was transformed with AtPDAT (phospholipid: diacylglycerol acyltransferase) gene behind the cauliflower mosaic virus 35S promoter. Transformed plants were selected by exposure to kanamycin and further grown on peat-based soil media in growth chamber. The seeds of T1 plants (T2 seeds) segregated and containing homozygotes, heterozygotes and null segregants (without transgene). The T3 seeds of selected null segregants and transformed homozygotes were further grown on peat-based soil media in growth chamber (T3 plants) and the lipid content in their seeds (T4 seeds) were evaluated. Additionally T4 seeds of selected null segregants and selected transformed plants (homozygotes) were used for evaluation of AtPDAT overexpression effect on seedlings growth rate, measurement of PDAT activity and for greenhouse experiments. Results show that PDAT overexpression increased the growth rate of Arabidopsis plants. This phenomenon was observed both during germination and further growth of the seedlings on agar plate as well as during the germination and further growth of the plant on peat-based soil media during the greenhouse experiment. The positive effect on oil production in transformed plants was also observed, however, it was probably not directly attributable to PDAT action but rather an indirect cause of growth stimulation.

[1]  Uday K. Divi,et al.  Metabolic engineering of biomass for high energy density: oilseed-like triacylglycerol yields from plant leaves , 2013, Plant biotechnology journal.

[2]  Changcheng Xu,et al.  Phospholipid:diacylglycerol acyltransferase-mediated triacylglycerol biosynthesis is crucial for protection against fatty acid-induced cell death in growing tissues of Arabidopsis. , 2013, The Plant journal : for cell and molecular biology.

[3]  Changcheng Xu,et al.  Dual Role for Phospholipid:Diacylglycerol Acyltransferase: Enhancing Fatty Acid Synthesis and Diverting Fatty Acids from Membrane Lipids to Triacylglycerol in Arabidopsis Leaves[C][W] , 2013, Plant Cell.

[4]  A. Kelly,et al.  The SUGAR-DEPENDENT1 Lipase Limits Triacylglycerol Accumulation in Vegetative Tissues of Arabidopsis1[W] , 2013, Plant Physiology.

[5]  S. Stymne,et al.  Activities of acyl-CoA:diacylglycerol acyltransferase (DGAT) and phospholipid:diacylglycerol acyltransferase (PDAT) in microsomal preparations of developing sunflower and safflower seeds , 2013, Planta.

[6]  A. Banaś,et al.  Lipids in hairy roots and non-Agrobacterium induced roots of Crambe abyssinica , 2013, Acta Physiologiae Plantarum.

[7]  Rodrigo M. P. Siloto,et al.  Acyl-CoA:diacylglycerol acyltransferase: molecular biology, biochemistry and biotechnology. , 2012, Progress in lipid research.

[8]  I. Graham,et al.  A Cytosolic Acyltransferase Contributes to Triacylglycerol Synthesis in Sucrose-Rescued Arabidopsis Seed Oil Catabolism Mutants1[W][OA] , 2012, Plant Physiology.

[9]  K. Chapman,et al.  Biogenesis and functions of lipid droplets in plants , 2012, Journal of Lipid Research.

[10]  J. Ohlrogge,et al.  DGAT1 and PDAT1 Acyltransferases Have Overlapping Functions in Arabidopsis Triacylglycerol Biosynthesis and Are Essential for Normal Pollen and Seed Development[W][OA] , 2009, The Plant Cell Online.

[11]  J. Kroon,et al.  Identification and functional expression of a type 2 acyl-CoA:diacylglycerol acyltransferase (DGAT2) in developing castor bean seeds which has high homology to the major triglyceride biosynthetic enzyme of fungi and animals. , 2006, Phytochemistry.

[12]  J. Shockey,et al.  Tung Tree DGAT1 and DGAT2 Have Nonredundant Functions in Triacylglycerol Biosynthesis and Are Localized to Different Subdomains of the Endoplasmic Reticulum[W] , 2006, The Plant Cell Online.

[13]  R. Rajasekharan,et al.  Cytosolic Triacylglycerol Biosynthetic Pathway in Oilseeds. Molecular Cloning and Expression of Peanut Cytosolic Diacylglycerol Acyltransferase1[W] , 2006, Plant Physiology.

[14]  Anders S Carlsson,et al.  Cloning and Functional Characterization of a Phospholipid:Diacylglycerol Acyltransferase from Arabidopsis1 , 2004, Plant Physiology.

[15]  S. Stymne,et al.  The regulation of the fatty-acid composition of the triacylglycerols in microsomal preparations from avocado mesocarp and the developing cotyledons of safflower , 2004, Planta.

[16]  T. Voelker,et al.  DGAT2 Is a New Diacylglycerol Acyltransferase Gene Family , 2001, The Journal of Biological Chemistry.

[17]  P. Covello,et al.  Seed-specific over-expression of an Arabidopsis cDNA encoding a diacylglycerol acyltransferase enhances seed oil content and seed weight. , 2001, Plant physiology.

[18]  S. Stymne,et al.  The involvement of phospholipid:diacylglycerol acyltransferases in triacylglycerol production. , 2000, Biochemical Society transactions.

[19]  S. Stymne,et al.  Phospholipid:diacylglycerol acyltransferase: an enzyme that catalyzes the acyl-CoA-independent formation of triacylglycerol in yeast and plants. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[20]  A. Tinkelenberg,et al.  A Lecithin Cholesterol Acyltransferase-like Gene Mediates Diacylglycerol Esterification in Yeast* , 2000, The Journal of Biological Chemistry.

[21]  H. Schaller,et al.  Expression in yeast and tobacco of plant cDNAs encoding acyl CoA:diacylglycerol acyltransferase. , 2000, European journal of biochemistry.

[22]  J. Harwood,et al.  Changes in Kennedy pathway intermediates associated with increased triacylglycerol synthesis in oil-seed rape , 1999 .

[23]  A. Kumar,et al.  The Arabidopsis thaliana TAG1 mutant has a mutation in a diacylglycerol acyltransferase gene. , 1999, The Plant journal : for cell and molecular biology.

[24]  Chaofu Lu,et al.  Cloning of a cDNA encoding diacylglycerol acyltransferase from Arabidopsis thaliana and its functional expression , 1999, FEBS letters.

[25]  Robert V Farese,et al.  Identification of a gene encoding an acyl CoA:diacylglycerol acyltransferase, a key enzyme in triacylglycerol synthesis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[26]  J. Ohlrogge,et al.  Lipid biosynthesis. , 1995, The Plant cell.

[27]  S. B. Settlage,et al.  Localization of diacylglycerol acyltransferase to oil body associated endoplasmic reticulum , 1995 .

[28]  J. Harwood,et al.  Use of [2-3H]glycerol precursor in radiolabelling studies of acyl lipids in developing seeds of Brassica napus☆ , 1993 .

[29]  J. Harwood,et al.  Changes in the lipid content of developing seeds of Brassica napus , 1993 .

[30]  S. Stymne,et al.  8 – Triacylglycerol Biosynthesis , 1987 .

[31]  E. P. Kennedy.  Biosynthesis of complex lipids. , 1961, Federation proceedings.

[32]  W. J. Dyer,et al.  A rapid method of total lipid extraction and purification. , 1959, Canadian journal of biochemistry and physiology.

[33]  C. H..  Oleosins and Oil Bodies in Seeds and Other Organs ' , 2022 .