Extracellular vesicle analysis platform for novel fungicide target discovery in Botrytis cinerea

Overview

This research program targets extracellular vesicles (EVs) produced by Botrytis cinerea, one of the world's most destructive crop pathogens. The project investigates whether EVs released by this fungus concentrate virulence factors such as effector proteins, RNAs, lipids, metabolites, and polysaccharides that facilitate plant infection. By systematically profiling EV cargo, the team aims to uncover previously unrecognized molecular targets that can be exploited for next-generation fungicide development. The work addresses a critical gap: while EVs have been documented in more than 20 fungal species, their presence and composition in B. cinerea remain uncharacterized. Outcomes include a curated pool of candidate virulence-associated compounds and a validated methodological pipeline for studying fungal EVs as a source of crop-protection targets.

Technical specifications

Research approach and capabilities:

• B. cinerea cultivation in liquid culture to produce sufficient biomass for EV isolation
• EV separation using size-exclusion chromatography combined with tangential-flow filtration
• EV quality assurance through nanoparticle tracking analysis and electron microscopy
• EV content characterization via proteomics and metabolomics using established analytical pipelines
• In vitro bioassays on strawberry plants to evaluate EV phytotoxic activity and validate candidate virulence factors
• Twelve-month project timeline covering isolation, characterization, and functional validation

Team expertise:

• Demonstrated capability to isolate EVs from previously unstudied pathogenic fungi, including Fusarium oxysporum f. sp. vasinfectum
• Prior evidence that EVs from pathogenic fungi trigger hypersensitive responses in host plant tissue, while EVs from non-pathogenic yeast do not, confirming a role in virulence
• Joint leadership combining EV biology, fungal pathology, and translational research mentorship

Technology readiness level

This is an early-stage discovery research program at Technology Readiness Level 2–3 (concept validation through laboratory demonstration). The methodology draws on validated EV isolation and characterization techniques already applied to other fungal pathogens, reducing technical risk. The 12-month plan progresses from establishing B. cinerea EV isolation conditions, through cargo profiling, to functional testing on strawberry plants. The team is actively seeking collaboration and funding support for equipment, consumables, instrument access, analytical costs, and mentorship on producing translatable research outcomes.