The major goal of this project is to develop plant-derived nanovesicles (PDNVs) as a new nanomaterial platform for studying and controlling devastating plant pathogens. Importantly, this will enable expansion of spray-induced gene silencing to previously untreatable microbes with limited RNA uptake and provide a new transformation method for investigating microbial genetics. PDNVs can be isolated from various plant sources, making them easy to scale, environmentally-friendly, and economical relative to other nanomaterials. Completion of this work will lead to a new suite of nanomaterials for nucleic acid delivery to microbes in agriculture and human health and increased knowledge on the nanoscale features and biological pathways controlling microbial nanoparticle uptake. Objective 1: Evaluate the use of PDNVs for dsRNA delivery and disease control against multiple microbial pathogens on pre- and post-harvest plant materials. These results will serve as the basis for identifying PDNVs with low or high microbial uptake. Objective 2: Optimize plasmid and protein loading in PDNVs to facilitate microbial transformation and genetic engineering. Objective 3: Determine the key features that dictate microbial uptake of PDNVs through multi-omics analysis of PDNV compositions and experimental confirmation using artificial PDNVs and chemical inhibitors.