Metabolomic Profiling of Lipids and Their Compositional Dynamics in Plant Stress Responses

该项目汇集了具有互补专业知识的研究人员，以开发代谢组能力，以定量描述植物中的脂质分子种类及其变化。该项目的长期目标是了解细胞脂类及其代谢产物如何相互作用，以产生和维持复杂的细胞膜，并调节细胞功能。该项目的目标有两个：一是利用一个基于质谱学的通用平台，发展一种全面的能力，以充分描述细胞内的脂质种类。另一种是利用谱分析能力来确定神秘的、帕特丁样酰基水解酶和几种可能参与植物胁迫反应的脂肪酶的代谢功能。本应用程序的具体目标是：(1)剖析和定量调节脂类：游离脂肪酸、选定的脂肪酸衍生物、溶菌脂种类和磷脂酰肌醇；(2)剖析和定量中性甘油酯：二酰甘油和三酰甘油；(3)剖析和定量鞘磷脂分子种类；(4)发现新的脂类和代谢物；(5)利用剖析能力来确定可能的脂解酶的代谢功能：类Patatin蛋白、RL-PLA、AAM10310、AtSABP2、PRLIP1、PAD4和EDS1。利用电喷雾电离串联质谱仪(ESI-MS/MS)灵敏、高效地进行脂类分析，有可能实现对细胞脂类的全面表征。这一能力的发展将极大地促进新兴的、全面的研究战略和代谢组学。脂类分析将为解决涉及脂类功能的生物学问题提供一个强有力的策略。这些功能研究将深入了解拟南芥脂水解酶的体内底物和产物，酶被激活的条件，以及活性的空间分布。该项目将产生更广泛的影响。除了通过研究活动培训研究生外，该项目还将把代谢图谱和功能基因组学的最新知识带到课堂上。它还将提供一个机会，扩大代表不足的群体对植物科学的参与。堪萨斯脂质组学研究中心提供的能力和服务对许多研究人员来说将是重要的。该项目将确定调节植物生长和胁迫反应的重要代谢和调节步骤。这些步骤的操作可能会导致作物的抗逆性增强，产品质量和/或生产率提高。

This project assembles researchers with complementary expertise to develop the metabolomic capacity to quantitatively profile lipid molecular species and their changes in plants. The long-term goal of this project is to understand how cellular lipids and their metabolites interact to produce and maintain complex cell membranes and to regulate cell functions. The goal of this project is two-fold: One is to develop a comprehensive capacity to fully profile cellular lipid species, using a mass spectrometry-based, common platform. The other is to use the profiling capability to determine the metabolic function of enigmatic, patatin-like acyl hydrolases and several putative lipases involved in plant stress responses. The specific objectives of this application are to: (1) profile and quantify regulatory lipids: free fatty acids, selected fatty acid derivatives, lysolipid species, and phosphoinositides; (2) profile and quantify neutral glycerides: diacylglycerol and triacylglycerol; (3) profile and quantify sphingolipid molecular species; (4) discover new lipid species and metabolites; (5) use the profiling capability to determine the metabolic functions of putative lipolytic enzymes: the patatin-like proteins, RL-PLA, AAM10310, AtSABP2, PRLIP1, PAD4, and EDS1. Sensitive and efficient lipid profiling by electrospray ionization tandem mass spectrometry (ESI-MS/MS) has the potential to achieve full characterization of cellular lipids. The development of this capability will contribute greatly to the emerging, comprehensive research strategy, and metabolomics. Lipid profiling will provide a powerful strategy to address biological questions that involve the function of lipids. The functional studies will produce insights into in vivo substrates and products of Arabidopsis lipid-hydrolyzing enzymes, the conditions under which the enzymes are activated, and the spatial distribution of the activity.The project will have broader impacts. In addition to training graduate students through research activities, this project will bring current knowledge of metabolic profiling and functional genomics to the classroom. It will also provide an opportunity to broaden participation of underrepresented groups in the plant sciences. The capability and service provided by the Kansas Lipidomics Research Center will be important to many researchers. The project will identify important metabolic and regulatory steps mediating plant growth and stress responses. Manipulation of these steps may lead to production of crop plants with enhanced stress tolerance and increased product quality and/or productivity.