Extracellular vesicles (EVs) are tiny messengers that convey bioactive molecules from donor to recipient cells, leading to changes in their physiology and function. We investigated the role of EVs in shaping growth and the biofilm biology of the tree pathogen Fusarium circinatum and its interaction with the susceptible host, Pinus patula. Vesicles were collected from fungal planktonic and biofilm cultures and from pine seedling needles and roots. The physical properties of these vesicles were analysed using nanoparticle tracking analysis and transmission electron microscopy, which revealed a diverse range of sizes and shapes, respectively. Furthermore, uptake of vesicles by conidia was conducted. The results demonstrated that F. circinatum EVs significantly but variably affected spore viability during the early phase (2-4 h) although they enhanced fungal biofilm integrity. In contrast, P. patula EVs greatly inhibited hyphal formation and biofilm biomass, but failed to inhibit matrix production in the fungal biofilm. Our results therefore show that conidial germination is essential for late fungal development including hyphal and biofilm formation while matrix production is a counter measure against harsh environmental conditions including the effects of plant-derived EVs.