Possible Roles of Calcium and Calmodulin in the Biosynthesis and Secretion of α-Amylase in Rice Seed Scutellar Epithelium

The scutellar epithelial cells of rice (Oryza sativa L. cv Kimmazé) seeds actively secrete alpha-amylase in an early stage of germination. Employing an in vivo system of freshly dissected scutellar tissues, effect of Ca(2+) on the biosynthesis and the secretion of alpha-amylase have been studied. The maximum biosynthetic rate was saturated at about 0.5 mm external Ca(2+) concentrations, whereas the secretion continued to increase to concentrations above 10 mm Ca(2+). In the presence of 1 mm Ca(2+), 0.01 mum A-23187 significantly increased both the biosynthesis and the secretion of alpha-amylase.A cation-specific requirement for Ca(2+) was apparent, since both biosynthesis and extracellular secretion of alpha-amylase were inhibited by 0.1 mm EGTA but were increased above basal rate only with Ca(2+) and Sr(2+); K(+), Mg(2+), and Ba(2+) being ineffective.La(3+) and ruthenium red (selective inhibitors of [Ca(2+) + Mg(2+)]-ATPase) were found to profoundly inhibit the secretion of alpha-amylase. A calmodulin antagonist, W-7, also inhibited the secretion of alpha-amylase at concentrations where the enzyme synthesis was not much affected. Overall data indicate that Ca(2+) movement and secretion of alpha-amylase are tightly linked and it is likely that they are regulated by the cytoplasmic Ca(2+) concentration under possible control by calmodulin.