TUNABLE RELEASE OF DSRNA MOLECULES INTO PLANTS FROM SUSTAINABLE NANOCARRIERS: A NOVEL MANAGEMENT TOOL FOR VIRAL PATHOGENS

Short-term goal:synthesize three different nanoparticle platforms (chitosan, silica, and carbon dots) and test their capacity to provide stable and sustained delivery of dsRNA molecules into potato plant tissue.Long-term goal:develop a tunable platform for dsRNA delivery (nanoparticle-dsRNA complex) that can be translated to a range of viral-crop systems.Objective 1 - Synthesis and screening of nanoparticles and loading of target dsRNAs:Objective 1a.Synthesis of chitosan-based nanoparticles.Objective 1b. Synthesis of silica NPs.Objective 1c.Synthesis ofCarbon dots.Objective 2. Selection of the optimum nucleic acid delivery system:To choose the ideal dsRNA delivery system, the different nanoparticles will be tested for their loading capacity and potential leaching of dsRNA, and their ability to protect dsRNA from degradation.Objective 3 - Bioassays to evaluate dsRNA-nanoparticles in planta and their efficacy to control virus infections.Besides testing the potential use of the combinations dsRNA-NP (dsRNA loaded into nanoparticles) to control PVY infections, experiments will be designed to evaluate the release and degradation rates of dsRNAs and the systemic movement of dsRNAsin plantato aid the creation of a sustainable PVY control strategy.Objective 3a.Inoculation assays.The treatments will be:1 - mock (plants treated with water solution), 2 - NP (plant treated with nanoparticles only), 3 - dsRNA (plants treated with dsRNA only), and 4 - NP-dsRNA (plants treated with nanoparticle-dsRNA complex).Objective 3b.NP-dsRNA effect on the host.We will monitor plant health and yield in response to each nanocarrier complex. Plant growth measurements (height, canopy, and biomass) will be recorded and compared between treatments. Objective 3c.NP-dsRNA effect on the microbiome.We will also investigate if the treatments proposed (more specifically the dsRNAs and NPs used) have any effect on the plant and rhizosphere microbiomes.The microbiomes in plants and rhizospheres play an important role in plant nutrition, development, and help plants to fight pest and diseases; therefore any technology that affects these microbiomes will ultimate affect the overall plant health.