ABB  Vol.1 No.5 , December 2010
Overexpression of aspen sucrose synthase gene promotes growth and development of transgenic Arabidopsis plants
Abstract: In plants, sucrose synthase (SUS) enzymes catalyze conversion of sucrose into fructose and UDP-glucose in the presence of UDP. To investigate the impact of overexpression of heterologous SUS on the growth and development of Arabidopsis, we transformed Arabidopsis plants with an overexpression vector containing an aspen SUS gene (PtrSUS1). The genomic PCR confirmed the successful integration of PtrSUS1 transgene in the Arabidopsis genome. PtrSUS1 expression in transgenic Arabidopsis plants was confirmed by RT-PCR. The SUS activity was dramatically increased in all transgenic lines examined. The three selected transgenic PtrSUS1 lines exhibited faster growth and flowered about 10 days earlier compared to untransformed controls, and also possessed 133%, 139%, and 143% SUS activity compared to controls. Both fresh weights and dry biomass yields of the whole plants from these three selected transgenic lines were significantly increased to 125% of the controls. Transgenic PtrSUS1 lines also had a higher tolerance to higher concentration of sucrose which was reflective of the increased SUS activity in transgenic versus wild-type plants. The growth differences between wild-type and transgenic plants, either in root and hypocotyl length or in fresh and dry weight of whole plant, became more pronounced on the media containing higher sucrose concentrations. Taken together, these results showed that the early flowering, faster growth and increased tolerance to higher sucrose in transgenic lines were caused by the genome integration and constitutive expression of the aspen PtrSUS1 gene in transgenic Arabidopsis.
Cite this paper: nullXu, F. and Joshi, C. (2010) Overexpression of aspen sucrose synthase gene promotes growth and development of transgenic Arabidopsis plants. Advances in Bioscience and Biotechnology, 1, 426-438. doi: 10.4236/abb.2010.15056.

[1]   Munoz, F.J., Baroja-Fernandez, E., Moran-Zorzano, M.T., Viale, A.M., Etxeberria, E., Alonso-Casajus, N., et al. (2005) Sucrose synthase controls both intracellular ADP glucose levels and transitory starch biosynthesis in source leaves. Plant Cell Physiol, 46(8), 1366-1376.

[2]   Baroja-Fernandez, E., Munoz, F.J., Saikusa, T., Rodriguez-Lopez, M., Akazawa, T. and Pozueta-Romero, J. (2003) Sucrose synthase catalyzes the de novo production of ADPglucose linked to starch biosynthesis in heterotrophic tissues of plants. Plant Cell Physiol, 44(5), 500-509.

[3]   Konishi, T., Ohmiya, Y. and Hayashi, T. (2004) Evidence that sucrose loaded into the phloem of a poplar leaf is used directly by sucrose synthase associated with various beta-glucan synthases in the stem. Plant Physiol, 134(3), 1146-1152.

[4]   Klotz, K.L. and Haagenson, D.M. (2008) Wounding, anoxia and cold induce sugarbeet sucrose synthase transcriptional changes that are unrelated to protein expression and activity. J Plant Physiol, 165(4), 423-434.

[5]   Fallahi, H., Scofield, G.N., Badger, M.R., Chow, W.S., Furbank, R.T. and Ruan, Y.L. (2008) Localization of sucrose synthase in developing seed and siliques of Arabidopsis thaliana reveals diverse roles for SUS during development. J Exp Bot, 59(12), 3283-3295.

[6]   Winter, H., Huber, J.L. and Huber, S.C. (1997) Membrane association of sucrose synthase: changes during the graviresponse and possible control by protein phosphorylation. Febs Letters, 420(2-3), 151-155.

[7]   Matic, S., Akerlund, H.E., Everitt, E. and Widell, S. (2004) Sucrose synthase isoforms in cultured tobacco cells. Plant Physiology and Biochemistry, 42(4), 299-306.

[8]   Winter, H., Huber, J.L. and Huber, S.C. (1998) Identification of sucrose synthase as an actin-binding protein. Febs Letters, 430(3), 205-208.

[9]   Etxeberria, E. and Gonzalez, P. (2003) Evidence for a tonoplast-associated form of sucrose synthase and its potential involvement in sucrose mobilization from the vacuole. Journal of Experimental Botany, 54(386), 1407-1414.

[10]   Carlson, S.J. and Chourey, P.S. (1996) Evidence for plasma membrane-associated forms of sucrose synthase in maize. Molecular & General Genetics, 252(3), 303-10.

[11]   Hardin, S.C., Duncan, K.A. and Huber, S.C. (2006) Determination of structural requirements and probable regulatory effectors for membrane association of maize sucrose synthase. Plant Physiology, 141(3), 1106-1119.

[12]   Amor, Y., Haigler, C.H., Johnson, S., Wainscott, M. and Delmer, D.P. (1995) A Membrane-Associated Form of Sucrose Synthase and Its Potential Role in Synthesis of Cellulose and Callose in Plants. Proceedings of the National Academy of Sciences of the United States of America, 92(20), 9353-9357.

[13]   Salnikov, V.V., Grimson, M.J., Delmer, D.P. and Haigler, C.H. (2001) Sucrose synthase localizes to cellulose synthesis sites in tracheary elements. Phytochemistry, 57(6), 823-833.

[14]   Salnikov, V.V., Grimson, M.J., Seagull, R.W. and Haigler, C.H. (2003) Localization of sucrose synthase and callose in freeze-substituted secondary-wall-stage cotton fibers. Protoplasma, 221(3-4), 175-184.

[15]   Albrecht, G. and Mustroph, A. (2003) Localization of sucrose synthase in wheat roots: increased in situ activity of sucrose synthase correlates with cell wall thickening by cellulose deposition under hypoxia. Planta, 217(2), 252-260.

[16]   Albrecht, G. and Mustroph, A. (2003) Sucrose utilization via invertase and sucrose synthase with respect to accumulation of cellulose and callose synthesis in wheat roots under oxygen deficiency. Russian Journal of Plant Physiology, 50(6), 813-820.

[17]   Zrenner, R., Salanoubat, M., Willmitzer, L. and Sonnewald, U. (1995) Evidence of the crucial role of sucrose synthase for sink strength using transgenic potato plants (Solanum tuberosum L.). Plant J, 7(1), 97-107.

[18]   Sun, J.D., Loboda, T., Sung, S.J.S. and Black, C.C. (1992) Sucrose Synthase in Wild Tomato, Lycopersicon-Chmielewskii, and Tomato Fruit Sink Strength. Plant Physiology, 98(3), 1163-1169.

[19]   Ruan, Y.L., Chourey, P.S., Delmer, D.P. and Perezgrau, L. (1997) The differential expression of sucrose synthase in relation to diverse patterns of carbon partitioning in developing cotton seed. Plant Physiology, 115(2), 375-85.

[20]   Heinlein, M. and Starlinger, P. (1989) Tissue-Specific and Cell-Specific Expression of the 2 Sucrose Synthase Isoenzymes in Developing Maize Kernels. Molecular & General Genetics, 215(3), 441-446.

[21]   Wittich, P.E. and Vreugdenhil, D. (1998) Localization of sucrose synthase activity in developing maize kernels by in situ enzyme histochemistry. Journal of Experimental Botany, 49(324), 1163-1171.

[22]   Dejardin, A., Rochat, C., Wuilleme, S. and Boutin, J.P. (1997) Contribution of sucrose synthase, ADP-glucose pyrophosphorylase and starch synthase to starch synthesis in developing pea seeds. Plant Cell and Environment, 20(11), 1421-1430.

[23]   Dejardin, A., Rochat, C., Maugenest, S. and Boutin, J.P. (1997) Purification, characterization and physiological role of sucrose synthase in the pea seed coat (Pisum sativum L). Planta, 201(2), 128-137.

[24]   Coleman, H.D., Ellis, D.D., Gilbert, M. and Mansfield, S.D. (2006) Up-regulation of sucrose synthase and UDP-glucose pyrophosphorylase impacts plant growth and metabolism. Plant Biotechnol J, 4(1), 87-101.

[25]   Chopra, J., Kaur, N. and Gupta, A.K. (2007) Sustained activities of carbon metabolizing enzymes determine seed size in Vigna radiata (mungbean). Current Science, 92(10), 1420-1424.

[26]   Chopra, J., Kaur, N. and Gupta, A. (2005) Role of enzymes of sucrose-starch conversion in seed sink strength in mung bean. Biologia Plantarum, 49(4), 561-566.

[27]   Chopra, J., Kaur, N. and Gupta, A.K. (2000) Ontogenic changes in enzymes of carbon metabolism in relation to carbohydrate status in developing mungbean reproductive structures. Phytochemistry, 53(5), 539-548.

[28]   Ito, A., Hayama, H. and Kashimura, Y. (2002) Sugar metabolism in buds during flower bud formation: a comparison of two Japanese pear [Pyrus pyrifolia (Burm.) Nak.] cultivars possessing different flowering habits. Scientia Horticulturae, 96(1-4), 163-175.

[29]   Coleman, H.D., Yan, J. and Mansfield, S.D. (2009) Sucrose synthase affects carbon partitioning to increase cellulose production and altered cell wall ultrastructure. Proc Natl Acad Sci U S A, 106(31), 13118-13123.

[30]   D’aoust, M.A., Yelle, S. and Nguyen-Quoc, B. (1999) Antisense inhibition of tomato fruit sucrose synthase decreases fruit setting and the sucrose unloading capacity of young fruit. Plant Cell, 11(12), 2407-2418.

[31]   Ruan, Y.L., Llewellyn, D.J. and Furbank, R.T. (2003) Suppression of sucrose synthase gene expression represses cotton fiber cell initiation, elongation, and seed development. Plant Cell, 15(4), 952-964.

[32]   Meng M., Geisler M., Johansson H., Mellerowicz E.J., Karpinski S., and Kleczkowski LA. (2007) Differential tissue/organ-dependent expression of two sucrose- and cold-responsive genes for UDP-glucose pyrophosphorylase in Populus. Gene 389(2), 186-195.

[33]   Clough, S.J. and Bent, A.F. (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J, 16(6), 735-743.

[34]   Sergeeva, L.I., Vonk, J., Keurentjes, J.J., Van Der Plas, L.H., Koornneef, M. and Vreugdenhil, D. (2004) Histochemical analysis reveals organ-specific quantitative trait loci for enzyme activities in Arabidopsis. Plant Physiol, 134(1), 237-245.

[35]   Xu, D.P., Sung, S.J., Loboda, T., Kormanik, P.P. and Black, C.C. (1989) Characterization of Sucrolysis via the Uridine Diphosphate and Pyrophosphate-Dependent Sucrose Synthase Pathway. Plant Physiol, 90(2), 635-642.

[36]   Sergeeva, L.I. and Vreugdenhil, D. (2002) In situ staining of activities of enzymes involved in carbohydrate metabolism in plant tissues. J Exp Bot, 53(367), 361-370.

[37]   Ruan, Y.L., Xu, S.M., White, R. and Furbank, R.T. (2004) Genotypic and developmental evidence for the role of plasmodesmatal regulation in cotton fiber elongation mediated by callose turnover. Plant Physiology, 136(4), 4104-4113.

[38]   Chourey, P.S. and Nelson, O.E. (1976) The enzymatic deficiency conditioned by the shrunken-1 mutations in maize. Biochem Genet, 14(11-12), 1041-1055.

[39]   Tang, G.Q. and Sturm, A. (1999) Antisense repression of sucrose synthase in carrot (Daucus carota L.) affects growth rather than sucrose partitioning. Plant Mol Biol, 41(4), 465-479.

[40]   Sturm, A. and Tang, G.Q. (1999) The sucrose-cleaving enzymes of plants are crucial for development, growth and carbon partitioning. Trends Plant Sci, 4(10), 401-407.

[41]   Barratt, D.H., Barber, L., Kruger, N.J., Smith, A.M., Wang, T.L. and Martin, C. (2001) Multiple, distinct isoforms of sucrose synthase in pea. Plant Physiol, 127(2), 655-664.

[42]   Bieniawska, Z., Paul Barratt, D.H., Garlick, A.P., Thole, V., Kruger, N.J., Martin, C., et al. (2007) Analysis of the sucrose synthase gene family in Arabidopsis. Plant J, 49(5), 810-828.

[43]   Tymowska-Lalanne, Z. and Kreis, M. (1998) Expression of the Arabidopsis thaliana invertase gene family. Planta, 207(2), 259-265.

[44]   Ruan, Y.L., Llewellyn, D.J., Liu, Q., Xu, S.M., Wu, L.M., Wang, L., et al. (2008) Expression of sucrose synthase in the developing endosperm is essential for early seed development in cotton. Functional Plant Biology, 35(5), 382-293.

[45]   Komatsu, A., Moriguchi, T., Koyama, K., Omura, M. and Akihama, T. (2002) Analysis of sucrose synthase genes in citrus suggests different roles and phylogenetic relationships. J Exp Bot, 53(366), 61-71.

[46]   Heim, U., Weber, H., Baumlein, H. and Wobus, U. (1993) A sucrose-synthase gene of Vicia faba L.: expression pattern in developing seeds in relation to starch synthesis and metabolic regulation. Planta, 191(3), 394-401.

[47]   Ren, G., Healy, R.A., Horner, H.T., James, M.G. and Thornburg, R.W. (2007) Expression of starch metabolic genes in the developing nectaries of ornamental tobacco plants. Plant Science, 173(6), 621-637.

[48]   Ruan, Y.L., Llewellyn, D.J. and Furbank, R.T. (2000) Pathway and control of sucrose import into initiating cotton fibre cells. Australian Journal of Plant Physiology, 27(8-9), 795-800.

[49]   King, S.P., Lunn, J.E. and Furbank, R.T. (1997) Carbohydrate Content and Enzyme Metabolism in Developing Canola Siliques. Plant Physiol, 114(1), 153-160.

[50]   Ruan, Y.L. and Chourey, P.S. (1998) A fiberless seed mutation in cotton is associated with lack of fiber cell initiation in ovule epidermis and alterations in sucrose synthase expression and carbon partitioning in developing seeds. Plant Physiol, 118(2), 399-406.

[51]   Dale, E.M. and Housley, T.L. (1986) Sucrose Synthase Activity in Developing Wheat Endosperms Differing in Maximum Weight. Plant Physiol, 82(1), 7-10.

[52]   Chan, H.Y., Ling, T.Y., Juang, R.H., Ting, I.N., Sung, H.Y. and Su, J.C. (1990) Sucrose Synthase in Rice Plants: Growth-Associated Changes in Tissue Specific Distributions. Plant Physiol, 94(3), 1456-1461.

[53]   Baud, S., Vaultier, M.N. and Rochat, C. (2004) Structure and expression profile of the sucrose synthase multigene family in Arabidopsis. J Exp Bot, 55(396), 397-409.

[54]   Dejardin, A., Sokolov, L.N. and Kleczkowski, L.A. (1999) Sugar/osmoticum levels modulate differential abscisic acid-independent expression of two stress-responsive sucrose synthase genes in Arabidopsis. Biochem J, 344(2), 503-509.

[55]   Ciereszkoa, I. and Kleczkowski, L.A. (2002) Glucose and mannose regulate the expression of a major sucrose synthase gene in Arabidopsis via hexokinase-dependent mechanisms. Plant Physiology and Biochemistry, 40(11), 907-911.

[56]   Koch, K.E. (1996) Carbohydrate-Modulated Gene Expression in Plants. Annu Rev Plant Physiol Plant Mol Biol, 47, 509-540.

[57]   Pego, J.V., Kortstee, A.J., Huijser, C. and Smeekens, S.C. (2000) Photosynthesis, sugars and the regulation of gene expression. J Exp Bot, 51 Spec No, 407-416.