Location: Emerging Pests and Pathogens Research
Project Number: 8062-22410-007-059-A
Project Type: Cooperative Agreement
Start Date: Aug 1, 2024
End Date: Jun 15, 2025
Objective:
1. Develop and test dsRNA targets against EAB and quantify LC50 and LD50 for optimal dosage.
2. Engineer an entomopathogenic fungal strain expressing effective RNAi targets in EAB and test efficacy in topical applications.
3. Use novel oviposition traps for delivery of fungi and dsRNA.
Approach:
1. 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. We will Synthesize dsRNA for several genes previously shown to activate RNAi in EAB and screen 5-10 of these targets though larval and adult oral ingestion assays and topical application to eggs
2. Engineer an entomopathogenic fungal strain expressing effective RNAi targets in EAB and test efficacy in topical applications. We will engineer an entomopathogenic fungus (Beauveria bassiana GHA) to express dsRNA targeting the top 1-2 candidate genes from objective 1 using protoplast mediated transformation and/or CRISPR/CAS9 approaches. After screening for successful expression of dsRNAi, we will test application to the insect using topical application of fungal spores.
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.
