PLANT PARASITIC NEMATODES ARE HIGHLY EVOLVED OBLIGATE PARASITES THAT THREATEN GLOBAL FOOD SECURITY. THESE NEMATODES HAVE A REMARKABLE ABILITY TO MODIFY HOST CELLS THAT SERVE AS THEIR ONLY SOURCE OF NUTRIENTS THROUGHOUT THEIR LIFE CYCLE. WHILE THESE PESTS HAVE BEEN INVESTIGATED MAINLY BECAUSE THEY ARE MAJOR CONSTRAINTS ON FOOD PRODUCTION GLOBALLY, THEY ALSO PROVIDE AN EXCELLENT SYSTEM TO INVESTIGATE SPECIFIC PROBLEMS IN PLANT AND ANIMAL DEVELOPMENT. HOWEVER, NO METHOD CURRENTLY EXISTS TO EDIT A PLANT-PARASITIC NEMATODE GENE OF INTEREST, CHANGE ITS EXPRESSION LEVEL, OR INCORPORATE NEW GENES. STUDIES ON PLANT-NEMATODE INTERACTIONS THEREFORE LAG THOSE FOR OTHER PATHOSYSTEMS. THE SPECIFIC AIM OF THIS PROPOSAL IS TO DEVELOP FUNCTIONAL GENOMIC TOOLS FOR AN AGRICULTURALLY IMPORTANT PLANT-PARASITIC NEMATODE. BUILDING ON OUR PRELIMINARY RESULTS, WE PROPOSE TO DELIVER A TOOL-KIT FOR THE GENERATION AND IDENTIFICATION OF HERITABLE GENOME EDITING EVENTS IN PLANT-PARASITIC NEMATODES. WE WILL: I) OPTIMIZE TRANSIENT EXPRESSION OF INTRODUCED GENES IN SOMATIC CELLS OF ROOT-KNOT NEMATODES, II) DEVELOP A STRATEGY FOR GENOME EDITING IN MALE GONADS AND IDENTIFY HERITABLE EDITING EVENTS, AND 3) DISSEMINATE THE INFORMATION TO THE SCIENTIFIC COMMUNITY AND THE PUBLIC. ACHIEVEMENT OF THESE GOALS WOULD BE A TRANSFORMATIVE TECHNICAL ADVANCE FOR THE FIELD OF PLANT-NEMATODE INTERACTIONS AND WOULD FACILITATE THE DEVELOPMENT OF MUCH-NEEDED ENVIRONMENTALLY SAFE STRATEGIES TO CONTROL THESE HIGHLY DAMAGING PLANT PATHOGENS. THE GOAL OF PROPOSED RESEARCH IS DIRECTLY RELEVANT TO THE AFRI PROGRAM AREA AGRICULTURAL INNOVATION THROUGH GENE EDITING (A1191), WHICH SUPPORTS THE EXPANSION OF TRANSFORMATION AND GENE-EDITING TECHNOLOGIES FOR USE IN AGRICULTURALLY IMPORTANT MICROORGANISMS.