Through proteomic analysis using mass spectrometry, a total of 1,931 proteins enriched in Fo-EVs were identified. Among these proteins, 350 were found to contain signal peptides and 375 possessed transmembrane domains. GO annotation and enrichment analysis, using the whole-genome proteins as background, revealed that these Fo-EVs proteins were primarily involved in cell wall synthesis, vesicle transport, and pathogenicity-related metabolic pathways. Furthermore, nine potential fungal EVs markers were screened, including Hsp70, Rho GTPase family proteins, and SNARE proteins. This study established a comprehensive Fo-EVs protein library and marker database, providing valuable proteomic resources for understanding the biological functions of fungal extracellular vesicles.