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，以开发代谢组学能力，定量描述脂质分子物种及其在植物中的变化。该项目的长期目标是了解细胞脂质及其代谢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方面：一是利用基于质谱的通用平台，开发全面分析细胞脂质种类的综合能力。另一个是利用分析能力来确定神秘的，类似patatin的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)描述和量化调节脂质：游离脂肪酸、选定的脂肪酸衍生物、溶脂种类和磷酸肌苷；(2)描述和定量中性甘油酯：二酰基甘油和三酰基甘油；(3)鞘脂分子种类分析与定量；(4)发现新的脂质种类和代谢产物；(5)利用分析能力确定假定的脂溶酶的代谢功能：patatin样蛋白，RL-PLA, AAM10310, AtSABP2, PRLIP1， PAD4和EDS1。灵敏、高效的电喷雾电离串联质谱（ESI-MS/MS）脂质谱分析有可能实现细胞脂质的全面表征。这种能力的发展将对新兴的综合研究策略和代谢组学做出巨大贡献。脂质分析将为解决涉及脂质功能的生物学问题提供强有力的策略。这些功能研究将深入了解拟南芥脂质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养研究生外，该项目还将把代谢分析和功能基因组学的最新知识带入课堂。它还将提供机会，扩大代表性不足的群体在植物科学领域的参与。堪萨斯脂质组学研究中心提供的能力和服务将对许多研究人员很重要。该项目将确定介导植物生长和胁迫反应的重要代谢和调控步骤。这些步骤的操作可能导致作物的生产具有更强的抗逆性，提高产品质量和/或生产力。

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.