UNDERSTANDING THE FUNCTIONS OF CROP GENES IS KEY FOR MOLECULAR BREEDING AND CROP ENGINEERING, AND THUS IT IS AN IMPORTANT STEP TOWARDS MAINTAINING NATIONAL FOOD SECURITY. HOWEVER, THE PRIMARY, SEQUENCE-SIMILARITY-BASED METHOD OF GENE FUNCTION PREDICTION IS OF LIMITED USE AND ERROR-PRONE, ESPECIALLY FOR ENZYMES THAT INFLUENCE AGRONOMICALLY IMPORTANT METABOLIC TRAITS. THIS PROJECT SEEKS TO IMPROVE THE ANNOTATION OF METABOLIC GENES IN GRASSES BY INTEGRATING HIGH-RESOLUTION UNTARGETED METABOLOMICS, TRANSCRIPTOMICS AND REVERSE GENETICS METHODS USING STATE-OF-THE-ART COMPUTATIONAL FRAMEWORKS. BRACHYPODIUM DISTACHYON - A MODEL SPECIES IN THE POACEAE (GRASS) FAMILY CLOSELY RELATED TO IMPORTANT CEREAL CROPS - WILL BE USED AS A MODEL FOR THIS PROJECT. THE NECESSARY OMICS DATA FOR METHOD DEVELOPMENT WAS GENERATED - IN COLLABORATION WITH THE JOINT GENOME INSTITUTE - FROM PLANTS GROWN UNDER VEGETATIVE GROWTH, LOW COPPER STRESS, HEAT STRESS, AND MYCORRHIZAL SYMBIOSIS. THESE DATASETS WILL BE ANALYZED TO CONSTRUCT METABOLITE AND TRANSCRIPT CO-REGULATION NETWORKS, AND FURTHER FILTERED TO IDENTIFY HIGH-CONFIDENCE METABOLITE-TRANSCRIPT PAIRS. THE FOCUS OF THIS PROJECT WILL BE TO IDENTIFY AND EXPERIMENTALLY VALIDATE ENZYMES IMPORTANT FOR BIOSYNTHESIS OF BLUMENOLS, A CLASS OF CAROTENOID-DERIVED COMPOUNDS THAT ARE BIOMARKERS FOR MYCORRHIZAL SYMBIOSIS. DATA GENERATED HEREIN WILL FACILITATE IDENTIFICATION OF GENETIC AND METABOLIC MARKERS FOR THE TESTED STRESSES, WHICH WILL PINPOINT BREEDING TARGETS FOR INCREASING CEREAL SURVIVAL AND YIELD. THROUGH INCREASING OUR KNOWLEDGE OF PLANT PRODUCTIVITY, THIS PROJECT ALIGNS WITH THE PLANT HEALTH AND PRODUCTION AND PLANT PRODUCTS PRIORITY AREA. THIS PROJECT ARMS THE PD WITH SKILLS HIGHLY SOUGHT AFTER IN AGRICULTURAL RESEARCH, THUS FULFILLING THE AFRI-EWD OBJECTIVE OF DEVELOPING FUTURE AGRICULTURAL LEADERS.