Obata Toshihiro, Rosado-Souza Laise, Fernie Alisdair R
Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476, Potsdam-Golm, Germany.
Department of Biochemistry, University of Nebraska Lincoln, 1901 Vine St, 68588, Lincoln, NE, USA.
Methods Mol Biol. 2017;1670:17-30. doi: 10.1007/978-1-4939-7292-0_2.
Carbohydrates catabolized via respiratory processes are not only used for energy production but also for biosynthesis of cellular components including soluble molecules (sugars, amino acids, organic acids, and their derivatives) and insoluble macromolecules (proteins, starch, and cell wall). Radiotracer experiments using C-labeled glucose provide a global picture of the fate of respired carbon in the metabolic network. This method is based on a chemical fractionation of biomolecules in C-glucose fed plant materials and the subsequent determination of radioactivity in each fraction. Metabolic flux into each fraction can be estimated from the specific activity of the hexose phosphate pool. Here, we describe the procedure for glucose metabolism in potato tuber but similar protocols can be adopted for various plant organs and substrates.
通过呼吸过程分解代谢的碳水化合物不仅用于能量产生，还用于细胞成分的生物合成，包括可溶性分子（糖、氨基酸、有机酸及其衍生物）和不溶性大分子（蛋白质、淀粉和细胞壁）。使用碳标记葡萄糖的放射性示踪实验提供了代谢网络中呼吸碳命运的全局图景。该方法基于对喂食含碳葡萄糖的植物材料中生物分子的化学分级分离以及随后对每个级分中放射性的测定。进入每个级分的代谢通量可以根据磷酸己糖池的比活性来估计。在这里，我们描述了马铃薯块茎中葡萄糖代谢的程序，但类似的方案可用于各种植物器官和底物。
