Fungal extracellular vesicles for double-stranded RNA delivery in crop protection
A novel approach using yeast-derived extracellular vesicles to encapsulate and deliver double-stranded RNA for silencing virulence genes in agricultural fungal pathogens, offering improved uptake and gene silencing efficiency over existing delivery methods.
Overview
This research explores the use of extracellular vesicles (EVs) derived from non-pathogenic yeast (Saccharomyces cerevisiae) as natural delivery vehicles for double-stranded RNA (dsRNA) targeting agricultural fungal pathogens. By exploiting the natural endocytosis processes of fungal cells, this approach aims to improve the uptake and cytoplasmic delivery of dsRNA for gene silencing, enabling more effective crop protection with reduced off-target effects.
The technology addresses a critical gap in agricultural biotechnology: the lack of efficient, biocompatible, and cost-effective dsRNA delivery systems. Existing methods such as viral particles and liposomes face limitations including low dsRNA protection, poor biocompatibility, and high production costs. Fungal EVs offer a scalable, natural alternative that can be produced through traditional fermentation.
Technical specifications
Key features:
- EVs from S. cerevisiae produced in liquid culture and purified using size-exclusion chromatography and tangential-flow filtration
- dsRNA encapsulation achieved through extrusion, targeting genes involved in ergosterol production and DICER proteins in pathogens such as Botrytis cinerea
- EV quality monitored by nanoparticle-tracking analysis and electron microscopy
- Encapsulation efficiency assessed using agarose gel electrophoresis
- Gene silencing capacity validated through in vitro assays on strawberry tissue
- Scalable production leveraging traditional fermentation methods
Potential applications:
- Targeted crop protection against fungal pathogens including B. cinerea (grey mould)
- Reduced reliance on conventional fungicides
- Foundation for broader RNA-based agricultural biocontrol strategies
Technology readiness level
This technology is currently at an early stage of development (TRL 2–3). Initial in vitro assays using dsRNA constructs targeting B. cinerea genes have been performed on strawberry tissue, though results remain inconclusive. The encapsulation of dsRNA in fungal EVs has not been previously demonstrated, though proof-of-concept evidence from mammalian EV RNA loading studies supports feasibility. The proposed 12-month validation plan includes EV production, purification, dsRNA encapsulation optimization, and gene silencing validation in existing strawberry assay systems.
About La Trobe University
La Trobe University is a comprehensive public research university based in Melbourne, serving metropolitan and regional Australia through a multi‑campus footprint. Its Research and Innovation Precinct at the Bundoora campus enables industry co‑location, shared facilities, and streamlined access to talent. Collaborations with state agencies and Melbourne’s hospital network, plus a digital innovation hub and accelerator programs, provide pathways for prototyping, validation, and co‑development. Research is supported by competitive national funding, including the Australian Research Council and National Health and Medical Research Council, alongside industry partnerships and Cooperative Research Centre programs. A dedicated technology transfer office manages IP, licensing, and startup formation.
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