Functional analysis of the starch related glucose 1-phosphate metabolism in higher plants

高等植物淀粉相关葡萄糖1-磷酸代谢的功能分析

• 批准号: 264514754
• 负责人: Professor Dr. Joerg Fettke
• 金额: --
• 依托单位: Institut für Biochemie und Biologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/264514754?language=en
• 关键词: Functional; analysis; starch; related; glucose

Over the last years the biochemistry of starch metabolism is massively increased. The complexity is related to a concerted action of more than 30 evolutionary conserved enzymes and of various carbon fluxes related to starch syntheses as well as degradation. Recently, a direct transport of glucose 1-phosphate into Arabidopsis mesophyll cells and potato tuber cells and further into plastids (chloroplasts and amyloplast) was identified using in situ experiments. Moreover, a direct link of the up taken glucose 1-phosphate and starch turn-over was demonstrated. The molecular nature of the involved transporters as well as the detailed importance for starch metabolism is so far unclear. This proposal uses various experimental approaches and focuses on the following aspects:a) The identification of the involved glucose 1-phosphate transporters in Arabidopsis as well as in potato, including a detailed analysis of the specificity of the potentially transporters. Therefore, the transporters will be analysed expressed in yeast and/or Xenopus oocytes.b) The importance of the glucose 1-phosphate transport for starch metabolism will be analyzed by generating various Arabidopsis mutants lacking starch related enzymes in addition to the potentially glucose 1-phosphate transporters. The various mutants will be analyzed in respect to carbon fluxes and starch metabolism, including starch granule morphology, starch internal structure and starch related enzymes.c) In the various mutants the distribution of glucose 1-phosphate in different organs, tissues, and compartments will be analyzed using a highly sensitive assay for glucose 1-phosphate and non-aqueous fractionation.

在过去的几年中，淀粉代谢的生物化学大大增加。复杂性与30多种进化保守酶以及与淀粉合成和降解有关的各种碳通量的协同作用有关。最近，使用原位实验鉴定了葡萄糖1-磷酸盐直接转运到拟南芥叶肉细胞和马铃薯块茎细胞中，然后进一步转移到质体（叶绿体和淀粉样体）中。此外，证明了UP葡萄糖1-磷酸盐和淀粉转换的直接连接。到目前为止，涉及转运蛋白的分子性质以及对淀粉代谢的详细重要性尚不清楚。该建议使用各种实验方法，并侧重于以下方面：a）鉴定拟南芥和马铃薯中所涉及的葡萄糖1-磷酸转运蛋白，包括对潜在转运蛋白的特异性的详细分析。因此，将在酵母和/或异武卵母细胞中分析转运蛋白。B）b）葡萄糖1-磷酸转运对淀粉代谢的重要性，通过产生缺乏淀粉相关的各种拟南芥突变体，除了具有潜在的1-葡萄糖1-磷酸盐迁移蛋白外，还将分析。各种突变体将在碳通量和淀粉代谢方面进行分析，包括淀粉颗粒形态，淀粉内部结构和相关的淀粉相关酶.c）在各种突变体中，使用高度敏感的测定法对不同的器官，组织和腔室中的不同器官，组织和腔室中的葡萄糖分布进行葡萄糖和裂缝的分析。