Hostname: page-component-586b7cd67f-2plfb Total loading time: 0 Render date: 2024-11-23T20:09:29.488Z Has data issue: false hasContentIssue false
  • English
  • Français

Soybean galactinol synthase forms fagopyritol B1 but not galactopinitols: substrate feeding of isolated embryos and heterologous expression

Published online by Cambridge University Press:  22 February 2007

Ralph L. Obendorf ,
Silvia Odorcic ,
Takashi Ueda ,
Mark P. Coseo  and
Elizabeth Vassallo
Ralph L. Obendorf*
Affiliation:
Seed Biology, Department of Crop and Soil Sciences, Cornell University, Cornell University Agricultural Experiment Station, 617 Bradfield Hall, Ithaca, NY, 14853-1901
Silvia Odorcic
Affiliation:
Seed Biology, Department of Crop and Soil Sciences, Cornell University, Cornell University Agricultural Experiment Station, 617 Bradfield Hall, Ithaca, NY, 14853-1901
Takashi Ueda
Affiliation:
Seed Biology, Department of Crop and Soil Sciences, Cornell University, Cornell University Agricultural Experiment Station, 617 Bradfield Hall, Ithaca, NY, 14853-1901 College of Arts and Sciences, Florida Gulf Coast University, 1051 FGCU Boulevard South, Fort Myers, FL, 33965-6565, USA
Mark P. Coseo
Affiliation:
Seed Biology, Department of Crop and Soil Sciences, Cornell University, Cornell University Agricultural Experiment Station, 617 Bradfield Hall, Ithaca, NY, 14853-1901
Elizabeth Vassallo
Affiliation:
Seed Biology, Department of Crop and Soil Sciences, Cornell University, Cornell University Agricultural Experiment Station, 617 Bradfield Hall, Ithaca, NY, 14853-1901
*
*Corresponding author: Fax: +1 607 255 2644, Email: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Soybean (Glycine max (L.) Merrill) seeds accumulate sucrose, raffinose, stachyose and lesser amounts of galactopinitol A, galactopinitol B and fagopyritol B1 in axis and cotyledon tissues as part of the seed maturation process. Somatic embryos appear to be deficient in D-pinitol and galactopinitols, indicating a lack of synthesis by embryo tissues in vitro. Isolated immature soybean zygotic embryos were fed myo-inositol, D-pinitol, D-chiro-inositol and sucrose, individually and in combination, to evaluate the role of substrate availability on galactosyl cyclitol accumulation during precocious maturation. Feeding myo-inositol transiently doubled galactinol accumulation with little effect on other soluble carbohydrates. Feeding D-pinitol increased free D-pinitol 8-fold, galactopinitol A 4.5-fold and galactopinitol B 4.2-fold. Stachyose concentration was 2-fold higher in cotyledons after feeding D-pinitol than after feeding D-chiro-inositol. Feeding D-chiro-inositol increased fagopyritol B1 17-fold in the axis and 7-fold in the cotyledons, but did not increase other soluble carbohydrates. Feeding D-pinitol and D-chiro-inositol together reduced uptake of D-chiro-inositol and steady-state accumulation of galactinol and galactopinitols by 50%, compared to feeding D-pinitol alone. Increasing sucrose concentration from 0 to 200 mM had no effect. Recombinant soybean galactinol synthase, heterologously expressed in Escherichia coli, catalysed the synthesis of fagopyritol B1 and galactinol, but not galactopinitols. These results were consistent with the following interpretations: D-pinitol and D-chiro-inositol were transported from maternal tissues and not synthesized in the embryo, D-chiro-inositol uptake into embryos may be reduced by D-pinitol, fagopyritol B1 was synthesized by galactinol synthase while galactopinitols were not, and fagopyritol B1 and galactopinitols accumulated in response to the supply of free D-chiro-inositol and D-pinitol to embryos.

Keywords

Glycine maxD-chiro-inositolcyclitol substratefagopyritol synthesisgalactinol synthaseheterologous expressionsoybean embryo
Type
Research Article
Information
Seed Science Research , Volume 14 , Issue 4 , December 2004 , pp. 321 - 333
Copyright
Copyright © Cambridge University Press 2004

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Blackman, S.A., Obendorf, R.L. and Leopold, A.C. (1992) Maturation proteins and sugars in desiccation tolerance of developing soybean seeds. Plant Physiology 100, 225–230.Google Scholar
Carmi, N., Zhang, G.F., Petreikov, M., Gao, Z.F., Eyal, Y., Granot, D. and Schaffer, A.A. (2003) Cloning and functional expression of alkaline α-galactosidase from melon fruit: similarity to plant SIP proteins uncovers a novel family of plant glycosyl hydrolases. Plant Journal 33, 97–106.CrossRefGoogle ScholarPubMed
Chanprame, S., Kuo, T.M. and Widholm, J.M. (1998) Soluble carbohydrate content of soybean [ Glycine max (L.) Merr.] somatic and zygotic embryos during development. In Vitro Cellular and Developmental Biology–Plant 34, 64–68.CrossRefGoogle Scholar
Dittrich, P. and Brandl, A. (1987) Revision of the pathway of D -pinitol formation in Leguminosae. Phytochemistry 26, 1925–1926.Google Scholar
Frydman, R.B. and Neufeld, E.F. (1963) Synthesis of galactosylinositol by extracts from peas. Biochemical and Biophysical Research Communications 12, 121–125.Google Scholar
Handley, L.W., Pharr, D.M. and McFeeters, R.F. (1983) Relationship between galactinol synthase activity and sugar composition of leaves and seeds of several crop species. Journal of the American Society for Horticultural Science 108, 600–605.CrossRefGoogle Scholar
Hegeman, C.E., Good, L.L. and Grabau, E.A. (2001) Expression of D - myo -inositol-3-phosphate synthase in soybean. Implications for phytic acid biosynthesis. Plant Physiology 125, 1941–1948.CrossRefGoogle ScholarPubMed
Hitz, W.D., Carlson, T.J., Kerr, P.S. and Sebastian, S.A. (2002) Biochemical and molecular characterization of a mutation that confers a decreased raffinosaccharide and phytic acid phenotype on soybean seeds. Plant Physiology 128, 650–660.Google Scholar
Hoch, G., Peterbauer, T. and Richter, A. (1999) Purification and characterization of stachyose synthase from lentil ( Lens culinaris ) seeds: Galactopinitol and stachyose synthesis. Archives of Biochemistry and Biophysics 366, 75–81.CrossRefGoogle ScholarPubMed
Horbowicz, M. and Obendorf, R.L. (1994) Seed desiccation tolerance and storability: Dependence on flatulence-producing oligosaccharides and cyclitols – review and survey. Seed Science Research 4, 385–405.CrossRefGoogle Scholar
Horbowicz, M., Obendorf, R.L., McKersie, B.D. and Viands, D.R. (1995) Soluble saccharides and cyclitols in alfalfa ( Medicago sativa L.) somatic embryos, leaflets, and mature seeds. Plant Science 109, 191–198.Google Scholar
Horbowicz, M., Brenac, P. and Obendorf, R.L. (1998) Fagopyritol B1, O -α- D -galactopyranosyl-(1→2)- D - chiro -inositol, a galactosyl cyclitol in maturing buckwheat seeds associated with desiccation tolerance. Planta 205, 1–11.Google Scholar
Kerr, P.S., Pearlstein, R.W., Schweiger, B.J., Becker-Manley, M.F. and Pierce, J.W. (inventors) (1997) Nucleotide sequences of galactinol synthase from zucchini and soybean. United States Patent Number 5648210, 15 July 1997.Google Scholar
Kuo, T.M., Lowell, C.A. and Nelsen, T.C. (1997a) Occurrence of pinitol in developing soybean seed tissues. Phytochemistry 45, 29–35.CrossRefGoogle Scholar
Kuo, T.M., Lowell, C.A. and Smith, P.T. (1997b) Changes in soluble carbohydrates and enzymatic activities in maturing soybean seed tissues. Plant Science 125, 1–11.Google Scholar
Loewus, F.A., Murthy, P.P.N. (2000) myo -Inositol metabolism in plants. Plant Science 150, 1–19.Google Scholar
Lowell, C.A. and Kuo, T.M. (1989) Oligosaccharide metabolism and accumulation in developing soybean seeds. Crop Science 29, 459–465.CrossRefGoogle Scholar
Obendorf, R.L. (1997) Oligosaccharides and galactosyl cyclitols in seed desiccation tolerance (review update). Seed Science Research 7, 63–74.Google Scholar
Obendorf, R.L., Ashworth, E.N. and Rytko, G.T. (1980) Influence of seed maturation on germinability in soybean. Crop Science 20, 483–486.CrossRefGoogle Scholar
Obendorf, R.L., Moon, H., Hildebrand, D.F., Torisky, R. and Collins, G.B. (1996) A comparison of pinitols in somatic and zygotic soybean embryos. Molecular and Cellular Biology of the Soybean 6, 40Google Scholar
Obendorf, R.L., Dickerman, A.M., Pflum, T.M., Kacalanos, M.A. and Smith, M.E. (1998a) Drying rate alters soluble carbohydrates, desiccation tolerance, and subsequent seedling growth of soybean ( Glycine max L. Merrill) zygotic embryos during in vitro maturation. Plant Science 132, 1–12.CrossRefGoogle Scholar
Obendorf, R.L., Horbowicz, M., Dickerman, A.M., Brenac, P. and Smith, M.E. (1998b) Soluble oligosaccharides and galactosyl cyclitols in maturing soybean seeds in planta and in vitro. Crop Science 38, 78–84.CrossRefGoogle Scholar
Odorcic, S. (2003) The anabolic and catabolic pathways of soluble α-galactosides in soybean ( Glycine max (L.) Merrill) seeds. Senior Biology Research Honors Thesis, Cornell University, Ithaca New York.Google Scholar
Odorcic, S. and Obendorf, R.L. (2003) Galactosyl cyclitol accumulation enhanced by substrate feeding of soybean embryos. pp. 51–60. in Nicolás, G.;, Bradford, K.J.;, Côme, D.;, Pritchard, H. (Eds) The biology of seeds: Recent research advances. Wallingford, CABI PublishingGoogle Scholar
Peterbauer, T. and Richter, A. (1998) Galactosylononitol and stachyose synthesis in seeds of adzuki bean: Purification and characterization of stachyose synthase. Plant Physiology 117, 165–172.CrossRefGoogle ScholarPubMed
Peterbauer, T. and Richter, A. (2001) Biochemistry and physiology of raffinose family oligosaccharides and galactosyl cyclitols in seeds. Seed Science Research 11, 185–198.Google Scholar
Peterbauer, T., Mach, L., Mucha, J. and Richter, A. (2002a) Functional expression of a cDNA encoding pea ( Pisum sativum L.) raffinose synthase, partial purification of the enzyme from maturing seeds, and steady-state kinetic analysis of raffinose synthesis. Planta 215, 839–846.CrossRefGoogle ScholarPubMed
Peterbauer, T., Mucha, J., Mach, L. and Richter, A. (2002b) Chain-elongation of raffinose in pea seeds: Isolation, characterization and molecular cloning of a multifunctional enzyme catalyzing the synthesis of stachyose and verbascose. Journal of Biological Chemistry 277, 194–200.CrossRefGoogle ScholarPubMed
Saravitz, D.M., Pharr, D.M. and Carter, T.E. (1987) Galactinol synthase activity and soluble sugars in developing seeds of four soybean genotypes. Plant Physiology 83, 185–189.CrossRefGoogle ScholarPubMed
Schweizer, T.F. and Horman, I. (1981) Purification and structure determination of three α- D -galactopyranosylcyclitols from soya beans. Carbohydrate Research 95, 61–71.CrossRefGoogle Scholar
Shoemaker, R., Keim, P., Vodkin, L., Erpelding, J., Coryell, V., Khanna, A., Bolla, B., Marra, M., Hillier, L., Kucaba, T., Martin, J., Beck, C., Wylie, T., Underwood, K., Steptoe, M., Theising, B., Allen, M., Bowers, Y., Person, B., Swaller, T., Gibbons, M., Pape, D., Harvey, N., Schurk, R., Ritter, E., Kohn, S., Shin, T., Jackson, Y., Cardenas, M., McCann, R., Waterson, R. and Wilson, R. (1999) Public soybean EST project. (unpublished); GenBank BE33077; Genome Systems Clone ID: Gm-c1041–80 (5'), cDNA clone from mature senescing soybean leaf, Genome Systems Inc., 4633 World Parkway Circle, St. Louis, Missouri 63134.Google Scholar
Streeter, J.G., Lohnes, D.G. and Fioritto, R.J. (2001) Patterns of pinitol accumulation in soybean plants and relationships to drought tolerance. Plant, Cell and Environment 24, 429–438.CrossRefGoogle Scholar
Volk, G.M. (1998) Plasmodesmatal formation and galactinol synthase expression in melon. PhD dissertation, Cornell University, Ithaca, New York, pp. 176–187.Google Scholar
Wanek, W. and Richter, A. (1997) Biosynthesis and accumulation of D -ononitol in Vigna umbellata in response to drought stress. Physiologia Plantarum 101, 416–424.CrossRefGoogle Scholar
Whistler, R.L. and Durso, D.F. (1950) Chromatographic separation of sugars on charcoal. Journal of the American Chemical Society 72, 677–679.Google Scholar