Fettke Joerg, Nunes-Nesi Adriano, Alpers Jessica, Szkop Michal, Fernie Alisdair R, Steup Martin
Department of Plant Physiology, University of Potsdam, 14476 Potsdam-Golm, Germany.
Plant Physiol. 2008 Nov;148(3):1614-29. doi: 10.1104/pp.108.127969. Epub 2008 Sep 19.
The cytosolic pools of glucose-1-phosphate (Glc-1-P) and glucose-6-phosphate are essential intermediates in several biosynthetic paths, including the formation of sucrose and cell wall constituents, and they are also linked to the cytosolic starch-related heteroglycans. In this work, structural features and biochemical properties of starch-related heteroglycans were analyzed as affected by the cytosolic glucose monophosphate metabolism using both source and sink organs from wild-type and various transgenic potato (Solanum tuberosum) plants. In leaves, increased levels of the cytosolic phosphoglucomutase (cPGM) did affect the cytosolic heteroglycans, as both the glucosyl content and the size distribution were diminished. By contrast, underexpression of cPGM resulted in an unchanged size distribution and an unaltered or even increased glucosyl content of the heteroglycans. Heteroglycans prepared from potato tubers were found to be similar to those from leaves but were not significantly affected by the level of cPGM activity. However, external glucose or Glc-1-P exerted entirely different effects on the cytosolic heteroglycans when added to tuber discs. Glucose was directed mainly toward starch and cell wall material, but incorporation into the constituents of the cytosolic heteroglycans was very low and roughly reflected the relative monomeric abundance. By contrast, Glc-1-P was selectively taken up by the tuber discs and resulted in a fast increase in the glucosyl content of the heteroglycans that quantitatively reflected the level of the cytosolic phosphorylase activity. Based on (14)C labeling experiments, we propose that in the cytosol, glucose and Glc-1-P are metabolized by largely separated paths.
葡萄糖-1-磷酸(Glc-1-P)和葡萄糖-6-磷酸的胞质池是多种生物合成途径中的关键中间体,包括蔗糖和细胞壁成分的形成,并且它们还与胞质淀粉相关杂聚糖有关。在这项研究中,利用野生型和各种转基因马铃薯(Solanum tuberosum)植株的源器官和库器官,分析了胞质单磷酸葡萄糖代谢对淀粉相关杂聚糖的结构特征和生化特性的影响。在叶片中,胞质磷酸葡萄糖变位酶(cPGM)水平的升高确实影响了胞质杂聚糖,因为葡萄糖基含量和大小分布均降低。相比之下,cPGM的低表达导致杂聚糖的大小分布不变,葡萄糖基含量未改变甚至增加。发现从马铃薯块茎制备的杂聚糖与叶片中的杂聚糖相似,但不受cPGM活性水平的显著影响。然而,当向块茎切片中添加外源葡萄糖或Glc-1-P时,它们对胞质杂聚糖产生了完全不同的影响。葡萄糖主要用于淀粉和细胞壁物质,但掺入胞质杂聚糖成分中的量非常低,大致反映了相对单体丰度。相比之下,Glc-1-P被块茎切片选择性吸收,并导致杂聚糖的葡萄糖基含量快速增加,这在数量上反映了胞质磷酸化酶活性水平。基于¹⁴C标记实验,我们提出在胞质溶胶中,葡萄糖和Glc-1-P通过基本分离的途径进行代谢。