D-A-CH Project:Regulation of primary metabolism under oxidative stress

由于其固着的生活方式，植物在其分布和性能受到许多非生物因素的显着影响。供水、温度和土壤质量对植物生长的影响最为相关，并且具有相似的作用。在细胞水平上，这些应激源可导致膜和蛋白质的损伤，并且在许多情况下，这是由氧化应激引起的，由细胞内产生活性氧引起。以往的研究表明，模式植物拟南芥的胁迫耐受性的显着差异与可溶性碳水化合物的积累和中心碳水化合物代谢酶活性的增加。然而，研究也表明，有必要分析整个初级代谢的亚细胞组织，以允许绘制一个有代表性的图片复杂的机制参与植物胁迫responses.In拟议中的项目，我们的目标是解剖在亚细胞水平上的氧化胁迫引起的初级代谢代谢和信号传导的影响。我们的项目是阐明昼夜调节和室的相互作用，有助于重新调整氧化还原稳态后，非生物胁迫的干扰。将拟南芥（Arabidopsis thaliana）登录号哥伦比亚-0的植物以及己糖激酶1和其己糖感应功能的S177 A互补物缺陷的gin-2突变体暴露于光、热和冷胁迫条件。对于初级代谢的亚细胞分析，非水分馏（NAF）技术将与代谢组和蛋白质组的高通量测量相结合。提出的新组合的NAF与鸟枪蛋白质组学，代谢产物分析和精细的数据处理技术的目的是提出一个更精确的图片的植物细胞作为一个系统协调氧化还原和代谢活动的子系统。这将涉及对低分子量ROS清除剂如抗坏血酸盐以及氧化损伤指标如马龙和保护性分子如糖或脯氨酸的隔室特异性分析。叶片初级代谢的昼夜动态和亚细胞代谢相互作用的数学建模将产生代谢适应非生物胁迫的瓶颈信息。除了植物与环境相互作用的生物化学分析和综合系统生物学表征之外，该项目还旨在提供一种增强的方法，以了解植物细胞内不同反应空间的相互作用。我们打算模拟整个细胞的氧化还原调节，从而证明或证伪细胞氧化还原状态的概念。这反过来又会打破旨在提高植物非生物胁迫耐受性的育种概念。

Due to their sessile lifestyle plants are significantly affected in their distribution and performance by many abiotic factors. Water supply, temperature and soil quality are most relevant and share similar effects on plant growth. At the cellular level, these stressors can cause damage to membranes and proteins, and in many cases this is entailed by oxidative stress, resulting from intracellular production of reactive oxygen species. Previous studies revealed significant differences in stress tolerance of the model plant Arabidopsis thaliana to be correlated with accumulation of soluble carbohydrates and increased enzyme activities of the central carbohydrate metabolism. Yet, studies also indicated the necessity of analysing subcellular organisation of the whole primary metabolism to allow drawing a representative picture of the complex mechanisms involved in plant stress response.In the proposed project, we aim to dissect metabolic and signalling effects on primary metabolism caused by oxidative stress at the subcellular level. Our project is set out to elucidate diurnal regulation and compartment interactions contributing to the re-adjustement of redox homeostasis after a disturbance by abiotic stresses. Plants of the Arabidopsis thaliana accession Columbia-0 as well as the gin-2 mutant defective in hexokinase 1 and the S177A complement of its hexose sensing function will be exposed to conditions of light, heat and cold stress. For subcellular analysis of primary metabolism, the non-aqueous fractionation (NAF) technique will be applied in combination with high-throughput measurement of the metabolome and proteome. The proposed novel combination of NAF with shotgun proteomics, metabolite profiling and refined data processing techniques are intended to present a more precise picture of the plant cell as a system co-ordinating redox and metabolic activities of its subsystems. This shall involve the compartment-specific analysis of low molecular weight ROS scavengers like ascorbate as well as indicators of oxidative damage, e.g. malone dialdehyde and protective molecules like sugars or proline. Mathematical modelling of both diurnal dynamics and subcellular metabolic interactions of leaf primary metabolism shall yield information about bottlenecks for metabolic adaptation to abiotic stress. Beyond biochemical analysis and comprehensive systems biological characterization of plant-environment interactions, this project intends to contribute an enhanced methodology that allows understanding of the interplay of the different reaction spaces within a plant cell. We intend to simulate whole cell redox adjustments, thus proving or disproving the concept of a cellular redox state. This should, in turn, burst breeding concepts aimed at improving plant abiotic stress tolerance.