Light-Dark Regulation of Starch Metabolism in Chloroplasts: I. Levels of Metabolites in Chloroplasts and Medium during Light-Dark Transition.

In Spinacia oleracea the kinetics of CO(2) fixation, of starch formation, and of changes in the levels of metabolites in chloroplasts and the surrounding medium has been investigated during light-dark and dark-light transitions with isolated intact chloroplasts.The internal level of orthophosphate stays constant throughout a light-dark-light cycle. The concentration of 3-phosphoglycerate in the chloroplasts is about 4 millimolar in the light and decreases in the dark within 3 minutes to about 1.6 millimolar. The level of the hexose monophosphates shows a reverse trend, increasing from about 2.2 millimolar in the light to 6 millimolar in darkness. In the subsequent light period both compounds reach their original levels within 2 minutes. The chloroplastic concentrations of dihydroxyacetone phosphate, of the pentose monophosphates, and of the hexose- and heptose bisphosphates remain constant at about 0.4 millimolar throughout the light-dark-light cycle.In the medium, the concentration of 3-phosphoglycerate increases and dihydroxyacetone phosphate decreases in the dark phase: this is due to an exchange of internal 3-phosphoglycerate for external dihydroxyacetone phosphate. Part of the reimported dihydroxyacetone phosphate is converted into hexose monophosphates via aldolase and fructose bisphosphatase during the first minutes of darkness. Due to the observed exchange transport reactions, the large difference between the transenvelope concentration gradients of 3-phosphoglycerate, dihydroxyacetone phosphate, and orthophosphate which exist in the light, is completely abolished after 2 to 3 minutes in the dark.The kinetics and the magnitudes of the changes of metabolite concentrations during the light-dark-light cycle are compared to the kinetics of starch formation, and their relevance for a possible light-dark regulation of starch synthesis is discussed.