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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]
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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

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