We performed a comparative study to determine the proteome of extracellular vesicles (EVs) from the cotton pathogen Fusarium oxysporum f. sp. vasinfectum (Fov), recovered from two growth conditions in vitro. Label-free quantitative protemics was used to find significant enrichment of proteins between EV samples, the secretome (secreted-soluble proteins) and the cell lysate. Our results show that some proteins were exclusive to EVs and were upregulated compared to the secretome or cell lysate.