THE LONG-TERM GOAL OF THIS PROPOSED PROJECT IS TO PROVIDE AGRICULTURE SECTORS IN THE USA WITH ENVIRONMENTALLY FRIENDLY APPROACHES FOR BETTER PEST AND DISEASE CONTROL, CROP PROTECTION, AND AGRICULTURAL PRODUCTION. THE GOAL WILL BE ACHIEVED BY DESIGN AND SYNTHESIS OF MULTIFUNCTIONAL NANOPARTICLES INCORPORATING BIODEGARABLE POLYSACCHARIDE, GLUCOSINOLATE, BIOTIN-STREPTAVIDIN COMPLEX, AND MYROSINASE. THE GLUCOSINOLATE-MYROSINASE CHEMICAL DEFENSE SYSTEM IS BASED ON THE ACCUMULATION OF INERT COMPONENTS IN SEPARATE COMPARTMENTS AND RAPID RELEASE OF LARGE AMOUNTS OF DEFENSIVE ISOTHIOCYANATES THROUGH MIXING OF THESE COMPONENTS AND THEIR REACTION WITH EACH OTHER UPON TISSUE DISRUPTION. THE PROPOSED STUDY INTENDS TO HARNESS CHITOSAN NANOPARTICLES TO HOLD GLUCOSINOLATE AND MYROSINASE IN TWO SEPARATE LAYERS, AND THEN TRIGGER THE FORMATION OF ISOTHIOCYANATE BY SLOW RELEASE OF GLUCOSINATE. THE SPECIFIC AIMS OF THIS SEED GRANT PROJECT ARE TO 1) CONSTRUCT PLASMIDS CONTAINING A MYROSINASE-STREPTAVIDIN FUSION GENE FOR BACTERIA TRANSFORMATION, PROTEIN EXPRESSION AND PURIFICATION, 2) FABRICATE BIOTIN-COATED CHITOSAN NANOPARTICLES PRE-LOADED WITH ANIONIC GLUCOSINATE VIA PROTEASE-CLEAVABLE LINKERS, 3) ASSEMBLE MULTIFUNCTIONAL CHITOSAN NANOPARTICLES BY BIOTIN-STREPTAVIDIN AFFINITY, WITH GLUCOSINOLATE AND MYROSINASE CONFINED IN SEGREGATED TWO DIFFERENT COMPARTMENTS, 4) ELICIT THE FORMATION/RELEASE OF ISOTHIOCYANATE FROM CHITOSAN NANOPARTICLES. THE SUCCESSFUL OUTCOME OF THIS PROJECT WILL LAY THE FRAMEWORK FOR PD TO BECOME COMPETITIVE FOR FUTURE GRANTS FROM AFRI.