Source: UNIV OF WISCONSIN submitted to NRP
BIOPESTICIDES - A VITAL PART OF SUSTAINABLE CROP PRODUCTION SYSTEMS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1031456
Grant No.
2021-67021-41088
Cumulative Award Amt.
$151,420.80
Proposal No.
2023-03558
Multistate No.
(N/A)
Project Start Date
Sep 1, 2023
Project End Date
Aug 31, 2025
Grant Year
2024
Program Code
[A1521]- Agricultural Engineering
Recipient Organization
UNIV OF WISCONSIN
21 N PARK ST STE 6401
MADISON,WI 53715-1218
Performing Department
(N/A)
Non Technical Summary
Modern agriculture relies heavily on the use of pesticides to protect crops. In response to society's need for safer and more environmentally friendly alternatives to synthetic pesticides, we have designed the pilot plant for production of biopesticidal extracts from mustard seed from which the oil has been removed as a feedstock for biofuel. Because the success in the form of grower adoption requires consistent and economically viable product, the overall goal of the project is to provide a biopesticides formulation that can be grower-implemented. To increase the extract value and the quality of biopesticides, we propose to design and optimize an ultrafiltration system that would allow us to separate multiple biologically active products such as enzymes from mustard extracts. We will 1) produce water-based pesticidal extracts from yellow and oriental mustards; 2) design ultrafiltration system for myrosinase enzyme separation from mustard extract; 3) optimize drying technique for shelf-stabilization of enzyme; and 4) assess the efficiency of the produced biopesticide formulations. By producing multiple high value products from mustard extract we will provide insensitive for growing mustard, a great rotational and cover crop that can be used toimprove soil health and reduce pest pressure.
Animal Health Component
40%
Research Effort Categories
Basic
20%
Applied
40%
Developmental
40%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
4025220202035%
5111899100035%
5111899200030%
Knowledge Area
511 - New and Improved Non-Food Products and Processes; 402 - Engineering Systems and Equipment;

Subject Of Investigation
1899 - Oilseed and oil crops, general/other; 5220 - Pesticides;

Field Of Science
2020 - Engineering; 1000 - Biochemistry and biophysics; 2000 - Chemistry;
Goals / Objectives
The overall goal of the project is to provide an efficient biopesticide formulation for controlling a wide range of pests that can be grower-implemented and provide insensitive for mustard production. In addition to conventional agriculture, biopesticides are vital for organic and high value agricultural and horticultural crops for which the few alternative pest control products availableare expensive, ineffective, or harmful to humans or the environment. Organic cropland has increased from 1.3 million acres in 2000 to over 3 million acres in 2014. Organic food revenues in the U.S. have grown to $31 billion a year. Organic vegetable and fruit acres, crops most relevant to the proposed products, have increased 300% during that same time period. California continues to lead in certified organic cropland, with nearly half of its total 405,000 acres used for fruit/vegetable production. These 200,000 acres are the initial target for our mustard meal-based pest control products.The specific objectives for the projects are:Objective 1. Produce water-based extracts from mustard meals.Objective 2. Design an ultrafiltration system for enzyme separation from mustard meal extractObjective 3. Optimize a drying technique for shelf-stabilization of the enzyme.Objective 4. Asses the efficacy of the produced biopesticide formulations.
Project Methods
To test our hypothesis, we willdesign and build the ultrafiltration system that will be interfaced with our extraction pilot plant to separate multiple biologically active compounds from mustard meal extracts.Objective 1: Produce water-based extracts from mustard meals. Glucosinolate containingextracts produced from mustard seed meals will be formulated into a shelf-stable powder. The concentration of glucosinolates and their hydrolysisproducts will be determined by the appropriate analytical technique.Objective 2: Design an ultrafiltration system for enzyme separation from mustard meal extract.The ultrafiltration system will be designed and used to separate enzyme present in the mustard seed meal extracts. Myrosinase enzyme separation will be performed using aqueous solution. Mustard extract will be separated using hollow-fiber membrane surface with a positive pressure relative to the permeate. The feed solution will be recirculated through the TFF system until the desired enzyme concentration is reached.Objective 3: Optimize drying technique for shelf-stabilization of the enzyme.The concentrated myrosinase extract will be collected and freeze dried. Spray-drying conditions for myrosinase extract will be optimized using a bench scale spray drier. Myrosinase activity will be assessed for each evaporation configuration, and conditions to assure stability of the enzyme and the preservation of catalytic activity will be determined. Activity will be measured by a spectrophotometric assay with sinigrin as substrate by measuring its decomposition as indicated by the decrease in absorbance at 227 nm.Objective 4: Assess the efficacy of the produced biopesticide formulations.Efficacy of B. juncea and S. alba extracts formulated using concentrated myrosinase will be tested to control pest weeds in ornamental production and perennial field crops. Trials will be conducted in the University of Idaho greenhouse. The first trial will test the extract's effect on 1) cash crop ornamentals (such as cyclamen or chrysanthemums) to determine if the product will control pest weeds in commercial ornamental nurseries and 2) agricultural crops (such as wheat and strawberries). Both pre- and post- emergence application of the extract will be conducted.

Progress 09/01/23 to 08/31/24

Outputs
Target Audience:Target audiences include members of the research community, organic farmers who will potentially utilize the proposed biopesticide and industry partners. Organic producers are a specific target audience for some of our potential commercial products since we produce natural pesticides that can be registered with EPA as biopesticides. Industries associated with potato storage and biopesticide production and sales are also targets. Changes/Problems:Due to the demonstrated superiority of liquid extract form, further research effort will be focused on optimizing liquid rather than dry extract formulation as was originally planned (Objective 3). The optimization will include ionic thiocyanate optimizations, liquid stability tests, and addition ofadjuvants. What opportunities for training and professional development has the project provided?Training and professional development were provided for a graduate student involved in the project to assess the developed biopesticide's effect on weed density and emergence under the field conditions. How have the results been disseminated to communities of interest?The results were shared with the industrial partner and research community through personal communications. What do you plan to do during the next reporting period to accomplish the goals?During the next funding period, we will assess the dynamicsof liquid mustard extract application to soil. The obtained knowledgewill allow for better weed control. It will also provide the information for any additional modifications of the mustard seed extraction procedure such as relative ratios of seed meal to water and the addition of chemicals that facilitate the production and purification of ionic thiocyanate.

Impacts
What was accomplished under these goals? During the reporting period, water-based extracts from mustard meals were produced and concentrated using the designedultrafiltration system for enzyme separation from mustard meal extract. The extract was concentrated and tested under field conditions. A pre- and post-plant trials were conducted at theSoil Stewards Organic farm (Moscow, ID, USA), a certified organic farm in Northern Idaho. S. alba extract was formulated in a liquid and dry form to compare the application logistics and efficacy. Extracts were applied before and after planting potato seeds. The application rate of ionic thiocyanate in the extract was equivalent to 0.9 kg ionic thiocyanate per square meter of soil. Based on the weed density, application of liquid extract form produced more consistent results and provided better weed control.For pre-planting treatment, i) potato yield was significantly higher by 70%, ii) the emergence of weeds before planting potatoes wassignificantly reduced by 92%, and iii) weed dry weight was significantly reduced by 74% at theend of potato season compared to the non-treated control. For post-planting treatment, there wasa significant reduction in weed dry weight by 57% but no significant impact on potato tuber yieldcompared to the non-treated control. For monitoring of ionic thiocyanate fate in soil, a methodology based on the use of an ion-selective electrode was developed. The developed methodology provides an economic and accessible way for laboratory and field analysis of ionic thiocyanate concentrations in soil amended with the mustard extract. The method allows for detection of ionic thiocyanate concentrations as low as 0.15 ppm. Recovery of ionic thiocyanate from soil (sandy soil, silt loam soil) varied from 95.2 to 103.0% in 20 - 200 ppm range of spiked concentrations.

Publications

  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Popova, I.; Shrestha, D.; Morra, M. Pilot-Scale Extraction of Multiple Biopesticidal Compounds from Mustard Biofuel by-Products. Ind. Crops Prod. 2023, 199, 116775. https://doi.org/https://doi.org/10.1016/j.indcrop.2023.116775.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Chhetri, P., Tuladhar, A., Popova, I. 2023 Effect of Sinapis Alba Seed Meal Extract as a Pre- or Post-Planting Bioherbicide in Organic Potato Production ASA, CSSA, SSSA International Annual Meeting, St. Louis, MO, November 2023.