DESIGN AND SYNTHESIS OF MYROSINASE-GLUCOSINOLATE MAGNETIC NANOPARTICLES FOR AGRI-FOOD APPLICATIONS

Long-term Goal and ObjectivesThe long-term goal of this proposed project is to provide agriculture in the USA with environmentally friendly approaches for better pest and disease control, crop protection, and agricultural production. The goal will be achieved through (1) design and synthesis of multifunctional nanomaterials incorporating chitosan, glucosinolate, biotin-streptavidin complex, and myrosinase, and (2) study of the release efficacy of these compounds and their byproducts in the defined agricultural systems. Accomplishment of this goal would greatly contribute to the rural economy, Idaho's potato industry, the welfare of farmers, and our society in general.Objectives:Control of soil-borne pests and diseases has traditionally been accomplished by soil fumigation. The most widely used compound has been volatile methyl bromide which is toxic and has been phased out as a class 1 ozone depleting substance under the Montreal Protocol (https://www.epa.gov/ods-phaseout/methyl-bromide). Thus, alternative approaches to manage pests are needed to maintain high crop production. Using natural compounds instead of synthetic compounds for pest control is much desirable because they are environmentally friendly. Biofumigation is such an approach which utilizes naturally occurring toxic isothiocyanates evolved as hydrolysis products from glucosinolate-containing plants applied to soil [9-11]. It has been demonstrated that biofumigation could provide good management of nematodes, weeds, fungi, and bacteria for potato cropping systems [12-14]. Since Idaho leads the United States in potato production and produces nearly 1/3 of all U.S.potatoes (Idaho State Department of Agriculture -https://agri.idaho.gov/main/about/about-idaho-agriculture/idaho-crops), harnessing toxicisothiocynates derived from glucosinolate for pest control is pertinent approach for Idaho's and Pacific Northwest's potato industry. However, up to date, there is no report of slow control release of glucosinolate-derived biopesticide. Nanotechnology may offer a way to control delivery of aforementioned biopesticide and to achieve greater effectiveness of much small dose in an extended releasing time frame withoutnegative environmental consequences.For this seed grant proposal, PD will initiate a proof-of-concept investigation using biodegradable nanoparticles to incorporate glucosinolate and myrosinase into separate domains and then trigger the formation and release of isothiocyanate. The successful outcome of this proposed study will lay the framework for PD to pursue future AFRI grants. To accomplish the goals set by this seed grant project, specific studies will be conducted with the following objectives.Objective 1:To design and synthesize glucosinolate-incorporated biotinylated chitosan nanoparticlesObjective 2:To construct expression vectors containing myrosinase-streptavidin fusion gene for bacterial transformation, protein expression and purificationObjective 3:To assemble multifunctional chitosan nanoparticles with glucosinolate andmyrosinase confined in separate compartmentsObjective 4:To trigger the formation and release of isothiocyanate from the multifunctional chitosan nanoparticles