Location: Emerging Pests and Pathogens Research
Project Number: 8062-22410-007-054-I
Project Type: Interagency Reimbursable Agreement
Start Date: Aug 1, 2024
End Date: Jul 15, 2025
Objective:
Overall Goal: Engineer an entomopathogenic fungus to express dsRNA for delivery via a novel autodissemination trap for efficient RNAi-based control of wood-boring insects.
Specific Objective 1: Develop and test dsRNA targets against EAB and quantify LC50 and LD50 for optimal dosage. This objective will screen 5-10 dsRNA candidate genes in EAB for lethality to select the best 1-2 candidates. For these, dose response curves will identify the effective dose for application.
Specific Objective 2: Engineer an entomopathogenic fungal strain expressing effective RNAi targets in EAB and test efficacy in topical applications. This objective will genetically engineer the entomopathogenic fungus Beauveria bassiana GHA to express dsRNA from the top 1-2 candidate loci and screen engineering fungal isolates for efficacy in lethality towards EAB.
Specific Objective 3: Use novel oviposition traps for delivery of fungi and dsRNA. This objective will test deployment of the fungal strain expressing dsRNA in a novel oviposition trap in the greenhouse for proof of concept as an application method.
Approach:
Specific Objective 1: Develop and test dsRNA targets against EAB. ARS will i) synthesize dsRNA targeting EAB essential genes, ii) test the efficacy of these dsRNA in a highly efficient larvae and adult oral ingestion assays of dsRNA, and also in topical applications to eggs.
Specific Ojbective 2: Engineer an entomopathogenic fungal strain expressing effective RNAi targets in EAB and test efficacy in topical applications. ARS will i) introduce an integrate a vector expressing the top 1-2 dsRNAs into the commercial insecticide fungus Beauveria bassiana GHA using fungal transformation, ii) test efficacy of the fungus in delivering dsRNA to the insect using topical applications to eab adults and eggs.
Specific Objective 3 Novel oviposition traps for delivery of fungi and dsRNA. The engineered fungus will be tested in growth chambers or the greenhouse in oviposition traps as a method to lure and deliver the fungus to adult eab females.
