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.
