ABSTRACTRoot rot is one of the main diseases affecting Aconitum carmichaelii Debx. during cultivation, seriously limiting yields of this herb. Currently, there is no effective control measure for aconite root rot. The antifungal activities of antagonistic strains against aconite root rot pathogens (Fusarium proliferatum, Fusarium solani, and Fusarium oxysporum) were investigated in this study. Three antagonistic strains, JKT7, JKT28 and JKT39, were screened and identified as Trichoderma asperellum, Trichoderma hamatum and Trichoderma virens, respectively. Dual culture tests showed that the inhibition rates of the three Trichoderma strains on the pathogens were all approximately 70%. The volatile metabolites had inhibitory effects on the mycelial growth of pathogens, while the nonvolatile metabolites in the culture filtrates did not show significant inhibitory effects. The volatile components analyzed by GC‒MS were mainly ketones, esters, and alcohols. These results indicate that these strains of Trichoderma and their secondary metabolites have antimicrobial activities against the pathogens of aconite root rot. This study could provide a scientific basis for the biocontrol of aconite root rot.