Research Assistant Carnegie Institution for Science Burlingame, California
Body of Abstract: Sorghum bicolor is a heat and drought tolerant, globally important C4 grain crop that is commonly used as a food source and biofuel. Despite increasing scientific interest in sorghum metabolism, the availability of experimentally validated enzyme localization data is limited. In order to predict and design desirable metabolic traits to increase plant productivity, it is crucial to understand where in the cell Sorghum bicolor enzymes are active, and which reactions they catalyze. Enzyme localization is important for its function. The goal of this project is to create an image-based map of the sorghum metabolic enzymes to systematically improve our understanding of the sorghum metabolic network. Our enzyme localization pipeline currently consists of 387 biologically relevant enzymes spanning 165 metabolic pathways in Sorghum bicolor. We are using two approaches to identify enzyme localization: 1) Heterologous expression of sorghum enzymes into Egeria densa via particle bombardment; and 2) transient expression of enzymes into sorghum protoplasts. The localization is identified by visualizing fluorescently tagged enzymes under a confocal microscope. The localization of 71 of the proposed enzymes have been identified to date. The dataset generated through this project will be publicly available through a searchable and browsable database online. The successful completion of this project will provide a large-scale, image-based localization dataset of sorghum enzymes, help decipher the complexity of sorghum metabolic networks, and provide improved metabolic engineering strategies.