Abstract In this study, we researched the chemical composition and the antifungal and antibacterial activity of volatile oil from Mentha spicata. The Gas chromatography/mass spectrometry (GC/MS) analysis of M. spicata showed that the main component was carvone (56.94%), followed by limonene (11.63%), sabinene hydrate (7.04%) and caryophyllene (4.06%). The antifungal activity of the volatile oil from M. spicata L. was determined with respect to plant pathogenic fungi, such as Fusarium oxysporum f. sp. radicis-lycopersici (Sacc.) W.C. Synder & H.N. Hans (FORL), Rhizoctonia solani J.G. Kühn. (R. solani), Alternaria solani (A. solani), and Verticillium dahliae Kleb (V. dahliae). The volatile oil was shown to have strong antifungal activity against plant pathogenic fungi. The result of the study was that at a dose of 12 μL petri-1, the volatile oil inhibited 100% of mycelium growth in V. dahliae, A. solani FORL and R. solani. Volatile oil exhibited remarkable activity against the selected bacterial strains of Xanthomonas spp. (ZI365, ZI366, ZI368, ZI370, ZI373, ZI375, ZI376, ZI378).