Abstract
Triacylglycerol (TAG) is the major carbon storage reserve in oilseeds such as Arabidopsis. Acyl-CoA:diacylglycerol acyltransferase (DGAT) catalyses the final step of the TAG synthesis pathway. Although TAG is mainly accumulated during seed development, and DGAT has presumably the highest activity in developing seeds, we show here that TAG synthesis is also actively taking place during germination and seedling development in Arabidopsis. The expression pattern of the DGAT1 gene was studied in transgenic plants containing the reporter gene β-glucuronidase (GUS) fused with DNA sequences flanking the DGAT1coding region. GUS activity was not only detected in developing seeds and pollen, which normally accumulate storage TAG, but also in germinating seeds and seedlings. Western blots showed that DGAT1 protein is present in several tissues, though is most abundant in developing seeds. In seedlings, DGAT1 is expressed in shoot and root apical regions, correlating with rapid cell division and growth. The expression of GUS in seedlings was consistent with the results of RNA gel blot analyses, precursor feeding and DGAT assay. In addition, DGAT1gene expression is up-regulated by glucose and associated with glucose-induced changes in seedling development.
Similar content being viewed by others
References
Baumlein, H., Nagy, I., Villarroel, R., Inzé D. and Wobus U. 1992. Cis-analysis of a seed protein gene promoter: the conservative RY repeat CATGCATG within the legumin box is essential for tissue-specific expression of a legumin gene. Plant J. 2: 233–239.
Bevan, M. 1984. Binary Agrobacterium vectors for plant transformation. Nucl. Acids Res. 12: 8711–8721.
Bouvier-Nave, P., Benveniste, P., Oelkers, P., Sturley, S.L. and Schaller, H. 2000. Expression in yeast and tobacco of plant cDNAs encoding acyl-CoA:diacylglycerol acyltransferase. Eur. J. Biochem. 267: 85–96.
Brown, A.P., Johnson, P.E., Rawsthorne, S. and Hills, M.J. 1998. Expression and properties of acyl-CoA binding protein from Brassica napus. Plant Physiol. Biochem. 36: 629–635.
Bradford, M.M. 1976. A rapid and sensitive method for the detection of microgram quantities of protein utilising the principle of protein-dye binding. Anal. Biochem. 72: 248–254.
Cases, S., Stone, S.J., Zhou, P., Yen, E., Tow, B., Lardizabal, K.D., Voelker, T. and Farese, R.V., Jr. 2001. Cloning of DGAT2, a second mammalian diacylglycerol acyltransferase, and related family members. J. Biol. Chem. 276: 38870–38876.
Clough, S.J. and Bent A.F. 1998. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 16: 735–743.
Dahlqvist, A., Stahl, U., Lenman, M., Banas, A., Lee, M., Sandager, L., Ronne, H. and Stymne, S. 2000. Phospholipid: diacylglycerol acyltransferase: an enzyme that catalyzes the acyl-CoA-independent formation of triacylglycerol in yeast and plants. Proc. Natl. Acad. Sci. USA 97: 6487–6492.
Dickinson, C.D., Evans, R.P. and Nielsen, N.C. 1988. RY repeats are conserved in the 5?-flanking regions of legume seed-protein genes. Nucl. Acids Res. 16: 371.
Eastmond, P.J., Germain, V., Lange, P.R., Bryce, J.H., Smith, S.M. and Graham, I.A. 2000. Postgerminative growth and lipid catabolism in oilseeds lacking the glyoxylate cycle. Proc. Natl. Acad. Sci. USA 97: 5669–5674.
Ellerstrom, M., Stalberg, K., Ezcurra, I. and Rask L. 1996. Functional dissection of a napin gene promoter: identification of promoter elements required for embryo and endosperm-specific transcription. Plant Mol. Biol. 32: 1019–1027.
Ericson, M.L., Muren, E., Gustavsson, H.O., Josefsson, L.G. and Rask, L. 1991. Analysis of the promoter region of napin genes from Brassica napus demonstrates binding of nuclear protein in vitro to a conserved sequence motif. Eur. J. Biochem. 197: 741–746.
Ezcurra, I., Ellerstrom, M., Wycliffe, P., Stalberg, K. and Rask, L. 1999. Interaction between composite elements in the napA promoter: both the B-box ABA-responsive complex and the RY/G complex are necessary for seed-specific expression. Plant Mol. Biol. 40: 699–709.
Farese, R.V., Jr., Cases, S. and Smith, S.J. 2000. Triglyceride synthesis: insights from the cloning of diacylglycerol acyltransferase. Curr. Opin. Lipidol. 11: 229–234.
Fujiwara, T. and Beachy, R.N. 1994. Tissue-specific and temporal regulation of a β-conglycinin gene: roles of the RY repeat and other cis-acting elements. Plant Mol. Biol. 24: 261–272.
Gibson, S.I. 2000. Plant sugar-response pathways. Part of a complex regulatory web. Plant Physiol. 124: 1532–1539.
Goldberg, R.B., De Paiva, G. and Yadegari, R. 1994. Plant embryogenesis: zygote to seed. Science 266: 605–614.
Hobbs, D.H., Lu, C. and Hills, M.J. 1999. Cloning of a cDNA encoding diacylglycerol acyltransferase from Arabidopsis thaliana and its functional expression. FEBS Lett. 452: 145–149.
Itzhaki, H., Maxson, J.M. and Woodson, W.R. 1994. An ethyleneresponsive enhancer element is involved in the senescencerelated expression of the carnation glutathione-S-transferase (GST1) gene. Proc. Natl. Acad. Sci. USA 91: 8925–8929.
Jang, J.C. and Sheen, J. 1994. Sugar sensing in higher plants. Plant Cell 6: 1665–1679.
Jefferson, R.A., Kavanagh, T.A. and Bevan, M.W. 1987. GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6: 3901–3907.
Katavic, V., Reed, D.W., Taylor, D.C., Giblin, E.M., Barton, D.L., Zou, J., Mackenzie, S.L., Covello, P.S. and Kunst, L. 1995. Alteration of seed fatty acid composition by an ethyl methanesulfonate-induced mutation in Arabidopsis thaliana affecting diacylglycerol acyltransferase activity. Plant Physiol. 108: 399–409.
Kaup, M.T., Froese, C.D. and Thompson, J.E. 2002. A role for diacylglycerol acyltransferase during leaf senescence. Plant Physiol. 129: 1616–1626.
Keddie, J.S., Tsiantis, M., Piffanelli, P., Cella, R., Hatzopoulos, P. and Murphy D.J. 1994. A seed-specific Brassica napus oleosin promoter interacts with a G-box-specific protein and may be bi-directional. Plant Mol. Biol. 24: 327–340.
Laby, R.J., Kincaid, M.S., Kim, D. and Gibson, S.I. 2000. The Arabidopsis sugar-insensitive mutants sis4 and sis5 are defective in abscisic acid synthesis and response. Plant J. 23: 587–596.
Lardizabal, K.D., Mai, J.T., Wagner, N.W., Wyrick, A., Voelker, T. and Hawkins, D.J. 2001. DGAT2 is a new diacylglycerol acyltransferase gene family. Purification, cloning, and expression in insect cells of two polypeptides from Mortierella ramanniana with diacylglycerol acyltransferase activity. J. Biol. Chem. 276: 38862–38869.
Lu, C. and Hills, M.J. 2002. Arabidopsis mutants deficient in diacylglycerol acyltransferase display increased sensitivity to abscisic acid, sugars, and osmotic stress during germination and seedling development. Plant Physiol. 129: 1352–1358.
Mansfield S.G., Briarty L.G. 1991. Cotyledon cell development in Arabidopsis thaliana during reserve deposition. Can. J. Bot. 70: 151–164.
Martin, B.A. and Wilson, R.F. 1983. Sub-cellular location of triacylglycerol synthesis in spinach leaves. Lipids 19: 117–121.
Martin, T., Oswald, O. and Graham, I.A. 2002. Arabidopsis seedling growth, storage lipid mobilization, and photosynthetic gene expression are regulated by carbon:nitrogen availability. Plant Physiol. 128: 472–481.
Montgomery, J., Goldman, S., Deikman, J., Margossian, L. and Fischer R.L. 1993. Identification of an ethylene-responsive region in the promoter of a fruit ripening gene. Proc. Natl. Acad. Sci. USA 90: 5939–5943.
Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Plant Physiol. 15: 473–497.
Murphy, D.J. 2001. The biogenesis and functions of lipid bodies in animals, plants and microorganisms. Prog. Lipid Res. 40: 325–438.
Murphy, D.J., Rawsthorne, S. and Hills M.J. 1993. Storage lipid formation in seeds. Seed Sci. Res. 3: 79–95.
Nishizuka, Y. 1992. Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. Science 258: 607–614.
Nykiforuk, C.L., Furukawa-Stoffer, T.L., Huff, P.W., Sarna, M., Laroche, A. Moloney, M.M. and Weselake R.J. 2002. Characterisation of cDNAs encoding diacylglycerol acyltransferase from cultures of Brassica napus and sucrose-mediated induction of enzyme biosynthesis. Biochim. Biophys. Acta. 1580: 95–109.
Ohlrogge, J. and Browse, J. 1995. Lipid biosynthesis. Plant Cell 7: 957–970.
Pongdontri, P. and Hills, M.J. 2001. Characterization of a novel plant acyl-CoA synthetase that is expressed in lipogenic tissues of Brassica napus L. Plant Mol. Biol. 47: 717–726.
Reidt, W., Wohlfarth, T., Ellerstrom, M., Czihal, A., Tewes, A., Ezcurra, I., Rask, L. and Baumlein, H. 2000. Gene regulation during late embryogenesis: the RY motif of maturation-specific gene promoters is a direct target of the FUS3 gene product. Plant J. 21: 401–408.
Riggs, C.D., Voelker, T.A. and Chrispeels M.J. 1989. Cotyledon nuclear proteins bind to DNA fragments harboring regulatory elements of phytohemagglutinin genes. Plant Cell 1: 609–621.
Rook, F., Corke, F., Card, R., Munz, G., Smith, C. and Bevan, M.W. 2001. Impaired sucrose-induction mutants reveal the modulation of sugar-induced starch biosynthetic gene expression by abscisic acid signalling. Plant J. 26: 421–433.
Routaboul, J., Benning, C., Bechtold, N., Caboche, M. and Lepiniec, L. 1999. The TAG1 locus of Arabidopsis encodes for a diacylglycerol acyltransferase. Plant Physiol. Biochem. 37: 831–840.
Sakaki, T., Saito, K., Kawaguchi, A., Kondo, N. and Yamada M. 1990. Conversion of monogalactosyldiacylglycerols to triacylglycerols in ozone-fumigated spinach leaves. Plant Physiol. 94: 766–772.
Stalberg, K., Ellerstom, M., Ezcurra, I., Ablov, S. and Rask, L. 1996. Disruption of an overlapping E-box/ABRE motif abolished high transcription of the napA storage-protein promoter in transgenic Brassica napus seeds. Planta 199: 515–519.
Weterings, K., Schrauwen, J., Wullems, G. and Twell, D. 1995. Functional dissection of the promoter of the pollen-specific gene NTP303 reveals a novel pollen-specific, and conserved cis-regulatory element. Plant J. 8: 55–63.
Wilson, R.F. and Kwanyuen P. 1986. Triacylglycerol synthesis and metabolism in germinating soybean cotyledons. Biochim. Biophys. Acta 877: 231–237.
Zhou, L., Jang, J.C., Jones, T.L. and Sheen J. 1998. Glucose and ethylene signal transduction crosstalk revealed by an Arabidopsis glucose-insensitive mutant. Proc. Natl. Acad. Sci. USA 95: 10294–10299.
Zou, J., Wei, Y., Jako, C., Kumar, A., Selvaraj, G. and Taylor, D.C. 1999. The Arabidopsis thaliana TAG1 mutant has a mutation in a diacylglycerol acyltransferase gene. Plant J. 19: 645–653.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lu, C.L., de Noyer, S.B., Hobbs, D.H. et al. Expression pattern of diacylglycerol acyltransferase-1, an enzyme involved in triacylglycerol biosynthesis, in Arabidopsis thaliana . Plant Mol Biol 52, 31–41 (2003). https://doi.org/10.1023/A:1023935605864
Issue Date:
DOI: https://doi.org/10.1023/A:1023935605864