P-317 - Exploring Cross-Kingdom RNA Exchange via Extracellular Vesicles in the Verticillium longisporum-Brassica napus Interaction

Verticillium longisporum is a vascular pathogen that causes severe yield losses in Brassica crops by colonizing the xylem and forming persistent microsclerotia. While its infection strategy is well characterized, the molecular dialogue between the fungus and its host remains poorly understood. Recent evidence suggests that extracellular vesicles (EVs) play a crucial role in cross-kingdom communication, potentially delivering RNA molecules that modulate the infection process. To better understand the role of EVs in host-pathogen interactions, we are developing a workflow for the isolation and characterization of EVs from Brassica napus roots infected with V. longisporum. Apoplastic wash fluids are collected to obtain EV-enriched fractions via ultracentrifugation. These vesicles, originating from both fungal and plant sources, are then analyzed for their RNA and protein content. Establishing this method will provide the foundation for future studies on the molecular mechanisms of EV-mediated cross-kingdom communication.