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Beside RFOs, which are commonly present in legume seeds, seeds of some species contain galactosyl pinitols (GPs). These carbohydrates, like RFOs, have been hypothesized to constitute an important energy and carbon skeletal source during germination. To test this hypothesis we have applied a specific α-galactosidase inhibitor (1-deoxygalactonojirimycin, DGJ) to germinating winter vetch (Vicia villosa Roth.) seeds, containing more galactosyl pinitols than RFOs. The breakdown of RFOs but not that of GPs was completely blocked in both embryonic axes and cotyledons tissues, during the first 18 h of imbibition in DGJ. The inhibitor only decreased the rate of GPs degradation. The inhibitory effect of DGJ on GPs degradation was partially alleviated by addition of sucrose or galactose to DGJ solutions. After three days of germination in water, RFOs and GPs disappeared in axial tissues of seeds imbibed in water, galactose or sucrose. Eighteen-hour imbibition of seeds in DGJ drastically reduced germination, by ca 50%, during the first three days. The inhibitory effect of DGJ decreased during the next seven days of germination. The presence of galactose or sucrose in imbibition solution initially stimulated seed germination, but later this effect was not statistically significant. Our study provides clear evidence that galactosyl pinitols play an important role in early winter vetch seeds germination. Additionally, we suggest that galactosyl pinitols can replace RFOs as reserve material necessary for early germination.
Changes in the accumulation of two types of α-D-galactosides: raffinose family oligosaccharides and galactosyl pinitols were compared with changes in the activities of galactosyltransferases during winter vetch (Vicia villosa Roth.) seed development and maturation. Occurrence of galactinol and raffinose in young seeds and changes in activities of galactinol synthase and raffinose synthase during seed development indicated that formation of raffinose oligosaccharides (RFOs) preceded synthesis of galactopinitols. Although transfer of galactose residues into raffinose oligosaccharides increased as seeds were maturing, at late stages of seed maturation the accumulation of galactopinitols was preferred to that of RFOs. In the present study, activities of enzymes transferring galactose moieties from galactinol to D-pinitol forming galactopinitol A, and further transfer of galactose moieties from galactinol to mono- and di-galactopinitol A were detected throughout seed development and maturation. This is a new observation, indicating biological potential of winter vetch seeds to synthesize mono-, di- and tri-galactosides of D-pinitol in a pathway similar to RFOs. The pattern of changes in activities of stachyose synthase and enzymes synthesizing galactopinitols (named galactopinitol A synthase and ciceritol synthase) suggests that formation of stachyose, mono- and di-galactopinitol A (ciceritol) is catalyzed by one enzyme. High correlation between activities of verbascose synthase and enzyme catalyzing synthesis of tri-galactopinitol A from galactinol and ciceritol (named tri-galactopinitol A synthase) also suggests that biosynthesis of both types of tri-galactosides was catalyzed by one enzyme, but distinct from stachyose synthase. Changes in concentrations of galactosyl acceptors (sucrose and D-pinitol) can be a factor which regulates splitting of galactose moieties between both types of galactosides in winter vetch seeds.
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