Abstract
Previous studies have identified two tissue- and cell-specific, yet functionally redundant, sucrose synthase (SuSy) genes, Sh1 and Sus1, which encode biochemically similar isozymes, SH1 and SUS1 (previously referred to as SS1 and SS2, respectively). Here we report evidence for a third SuSy gene in maize, Sus3, which is more similar to dicot than to monocot SuSys. RNA and/or protein blot analyses on developing kernels and other tissues show evidence of expression of Sus3, although at the lowest steady-state levels of the three SuSy gene products and without a unique pattern of tissue specificity. Immunoblots of sh1sus1-1 embryos that are either lacking or deficient for the embryo-specific SUS1 protein have shown a protein band which we attribute to the Sus3 gene, and may contribute to the residual enzyme activity seen in embryos of the double mutant. We also studied developing seeds of the double mutant sh1sus1-1, which is missing 99.5% of SuSy enzyme activity, for evidence of co-regulation of several genes of sugar metabolism. We found a significant reduction in the steady-state levels of Miniature-1 encoded cell wall invertase2, and Sucrose transporter (Sut) mRNAs in the double mutant, relative to the lineage-related sh1Sus1 and sh1Sus1 kernels. Down-regulation of the Mn1 gene was also reflected in significant reductions in cell wall invertase activity. Co-regulatory changes were not seen in the expression of Sucrose phosphate synthase, UDP-glucose pyrophosphorylase, and ADP-glucose pyrophosphorylase.
Similar content being viewed by others
References
Altschul, S.F., Madden, T.L., Schäffer, A., Zhang, J., Zhang, Z., Miller, W. and Lipman, D.J. 1997. Gapped BLAST and PSIBLAST: a new generation of protein database search programs. Nucl. Acids Res. 25: 3389-3402.
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. Proc. Natl. Acad. Sci. USA 92: 9353-9357.
Aoki, N., Hirose, T., Takahashi, S., Ono, K., Ishimaru, K. and Ohsugi, R. 1999. Molecular cloning and expression analysis of a gene for a sucrose transporter in maize (Zea mays L.). Plant Cell Physiol. 40: 1072-1078.
Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struhl, K. 1993. Current Protocols in Molecular Biology. John Wiley, New York.
Buczynski, S.R., Thom, M., Chourey, P.S. and Maretzki, A. 1993. Tissue distribution and characterization of sucrose synthase isozymes in sugarcane. J. Plant Physiol. 142: 641-646.
Carlson, S.J. and Chourey, P.S. 1996. Evidence of plasma membrane-associated forms of sucrose synthase in maize. Mol. Gen. Genet. 252: 303-310.
Carlson, S.J. and Chourey, P.S. 1999. A re-evaluation of the relative roles of two invertases, Incw2 and IVR1, in developing maize kernels and other tissues. Plant Physiol. 121: 1025-1035.
Carlson, S.J., Shanker, S. and Chourey, P.S. 2000. A point mutation at the Miniature1 seed locus reduces levels of the encoded protein, but not its mRNA, in maize. Mol. Gen. Genet. 263: 367-373.
Cheng, W.-H. and Chourey, P.S. 1999. Genetic evidence that invertase mediated release of hexoses is critical for appropriate carbon partitioning and normal seed development in maize. Theor. Appl. Genet. 98: 485-495.
Cheng, W.-H., Taliercio, E.W. and Chourey, P.S. 1996. The Miniature1 seed locus of maize encodes a cell wall invertase required for normal development of endosperm and maternal cells in the pedicel. Plant Cell 8: 971-983.
Chengappa, S., Guilleroux, M., Phillips, W. and Shields, R. 1999. Transgenic tomato plants with decreased sucrose synthase are unaltered in starch and sugar accumulation in the fruit. Plant Mol. Biol. 40: 213-221.
Chourey, P.S. 1981. Genetic control of sucrose synthetase in maize endosperm. Mol. Gen. Genet. 184: 372-376.
Chourey, P.S. and Nelson, O.E. 1976. The enzymatic deficiency conditioned by the shrunken-1 mutations in maize. Biochem. Genet. 14: 1041-1054.
Chourey, P.S. and Nelson, O.E. 1979. Interallelic complementation at the sh locus in maize at the enzyme level. Genetics 91: 317-325.
Chourey, P.S. and Taliercio, E.W. 1994. Epistatic interaction and functional compensation between the two tissue-and cell-specific sucrose synthase genes in maize. Proc. Natl. Acad. Sci. USA 91: 7917-7921.
Chourey, P.S., DeRobertis, G.A. and Still, P.E. 1988. Altered tissue specificity of the revertant shrunken allele upon DS excision is associated with loss of expression and molecular rearrangement at the corresponding non-allelic isozyme locus in maize. Mol. Gen. Genet. 214: 300-306.
Chourey, P.S., Chen, Y.-C. and Miller, M.E. 1991a. Early cell degeneration in developing endosperm is unique to the shrunken mutation in maize. Maydica 36: 141-146.
Chourey, P.S., Taliercio, E.W. and Kane, E.J. 1991b. Tissue specific expression and anaerobically induced posttranscriptional modulation of sucrose synthase genes in Sorghum bicolor M. Plant Physiol. 96: 485-490.
Chourey, P.S., Taliercio, E.W., Carlson, S.J. and Ruan, Y.-L. 1998. Genetic evidence that the two isozymes of sucrose synthase present in developing maize endosperm are critical, one for cell wall integrity and the other for starch biosynthesis. Mol. Gen. Genet. 259: 88-96.
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: 2407-2418.
Doehlert, D.C. and Kuo, T.M. 1990. Sugar metabolism in developing kernels of starch-deficient endosperm mutants of maize. Plant Physiol. 92: 990-994.
Fu, H. and Park, W.D. 1995. Sink-and vascular-associated sucrose synthase functions are encoded by different gene classes in potato. Plant Cell 7: 1369-1385.
Gai, X., Lal, S., Xing, L., Brendel, V. and Walbot, V. 2000. Gene discovery using the maize genome database ZmDB. Nucl. Acids Res. 28: 94-96.
Giroux, M.J. and Hannah, L.C. 1994. ADP-glucopyrophosphorylase in shrunken-2 and brittle-2 mutants of maize. Mol. Gen. Genet. 243: 400-408.
Guerin, J. and Carbonero, P. 1997. The spatial distribution of sucrose synthase isozymes in barley. Plant Physiol. 114: 55-62.
Gupta, M., Chourey, P.S., Burr, B. and Still, P.E. 1988. cDNAs of two non-allelic sucrose synthase genes in maize: cloning, expression, characterization and molecular mapping of sucrose synthase-2 gene. Plant Mol. Biol. 10: 215-224.
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: 394-401.
Hesse, H. and Willmitzer, L. 1996. Expression analysis of a sucrose synthase gene from sugar beet (Beta vulgaris L.). Plant Mol. Biol. 30: 863-872.
Hofmann, K. and Stoffel, W. 1993. Tmbase: a database of membrane spanning proteins segments. Biol. Chem. Hoppe-Seyler 347: 166.
Huber, S.C., Huber, J.L., Liao, P.-C., Gage, D.A., McMichael, R.J. Jr., Chourey, P.S., Hannah, L.C. and Koch, K. 1996. Phosphorylation of serine-15 of maize leaf sucrose synthase: occurrence in vivo and possible regulatory significance. Plant Physiol 112: 793-802.
Im, K.H. 1996. Regulation of sucrose phosphate synthase expression in maize. Ph.D. thesis, University of Florida, Gainesville.
Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680.
Marana, C., Gracia-Olmedo, F. and Carbonero, P. 1988. Linked sucrose synthase genes in group-7 chromosomes in hexaploid wheat (Triticum aestivum L.). Gene 63: 253-260.
Martin, T., Frommer, W.B., Salanonbat, M. and Willmitzer, L. 1993. Expression of an Arabidopsis sucrose synthase gene indicates a role in metabolization of sucrose both during phloem loading and in sink organs. Plant J 4: 367-377.
McCarty D.R., Shaw J.R. and Hannah L.C. 1986. The cloning, genetic mapping, and expression of the constitutive sucrose synthase locus of maize. Proc. Natl. Acad. Sci. USA 83: 9099-9103.
McElfresh, K.C. and Chourey, P.S. 1988. Anaerobiosis induces transcription but not translation of sucrose synthase in maize. Plant Physiol 87: 542-546.
Miller M.E. and Chourey, P.S. 1992. The maize invertase-deficient minature-1 seed mutation is associated with aberrant pedicel and endosperm development. Plant Cell 4: 297-305.
Page, R.D.M. 1996. TreeView for Macintosh, version 1.3. Published electronically on the Internet, available at http://taxonomy.zoology.gla.ac.uk/rod/treeview.html
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: 399-406.
Scott, P. and Chourey, P.S. 2000. Metabolic analyses of a double mutant of sucrose synthase (SuSy) genes in developing endosperm. Maize Genetics Conference Abstracts, Coeur d'Alene, Idaho.
Sebkova, V., Unger, C., Hardegger, M. and Sturm, A. 1995. Biochemical, physiological and molecular characterization of sucrose synthase from Daucus carota. Plant Physiol. 108: 75-83.
Shaw, J.R., Ferl, R.J., Baier, J., St. Clair, D., Carson, C., McCarty, D.R. and Hannah, L.C. 1994. Structural features of the maize sus1 gene and protein. Plant Physiol. 106: 1659-1663.
Sowokinos, J.R., Thomas, C. and Burrell, M.M. 1997. Pyrophosphorylases in potato. V. Allelic polymorphism of UGP-glucose pyrophosphorylase in potato cultivars and its association with tuber resistance to sweetening in the cold. Plant Physiol 113: 511-517.
Springer, B., Werr, W., Starlinger, P., Bennett, D.C., Zokolica, M. and Freeling, M. 1986. The shrunken gene on chromosome 9 of Zea mays L. is expressed in various plant tissues and encodes an anaerobic protein. Mol. Gen. Genet. 205: 461-468.
Tang, Q.-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: 465-479.
Thompson, J.D., Higgins, D.G. and Gibson, T.J. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucl. Acids Res. 22: 4673-4680.
Tsai, C.Y., Salamini, F. and Nelson, O.E. 1970. Enzymes of carbohydrate metabolism in the developing endosperm of maize. Plant Physiol. 46: 299-306.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Carlson, S.J., Chourey, P.S., Helentjaris, T. et al. Gene expression studies on developing kernels of maize sucrose synthase (SuSy) mutants show evidence for a third SuSy gene. Plant Mol Biol 49, 15–29 (2002). https://doi.org/10.1023/A:1014457901992
Issue Date:
DOI: https://doi.org/10.1023/A:1014457901992