Starch is the major storage compound in plants. The biochemical and genetic characterization of mutants established maize as a model system to investigate the biosynthesis of starch. In this study, a combination of stable isotope labeling and NMR-spectroscopy was used to study the starch biosynthetic pathway and the primary carbohydrate metabolism. Maize kernels were grown in sterile culture and labeled in steady state experiments with [U-13C6]glucose or [U-13C12]sucrose. From starch and phytoglycogen hydrolysates the concentrations of all biochemical relevant glucose isotopologs were estimated by 13C-NMR-spectroscopy and in this way the isotopolog patterns of several inbred lines (W64A, W22, Fa56) and starch-deficient mutants (wx, ae, su1, bt1, bt2, sh2, Sh2-Rev6, sh1, sh1 sus1, mn1) were determined. Three labeling experiments at different developmental stages show on the one hand that fluxes through the glycolysis, the pentose-phosphate pathway, and the tricarboxylic acid cycle are flexible and change with kernel development. On the other hand, the similar isotopolog patterns of the majority of the mutants studied here, support the rigidity of central metabolism. Solely the isotopolog patterns of su1-phytoglycogen, mn1-starch, and bt2-starch differ significantly from the profiles of according wild type starches. Based on these results, a new model of the biosynthesis of starch and phytoglycogen in su1 endosperm was proposed.
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Starch is the major storage compound in plants. The biochemical and genetic characterization of mutants established maize as a model system to investigate the biosynthesis of starch. In this study, a combination of stable isotope labeling and NMR-spectroscopy was used to study the starch biosynthetic pathway and the primary carbohydrate metabolism. Maize kernels were grown in sterile culture and labeled in steady state experiments with [U-13C6]glucose or [U-13C12]sucrose. From starch and phytogl...
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