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This study aimed to focus on the evaluation of the antifungal activity of extract prepared from Zizyphus lotus leaves harvested from the mountains of the Medea region in the north of Algeria. The extracts were prepared by maceration of leaves in sterile distilled water, methanol, chloroform and ethyl acetate. In vitro antifungal activity of the extracts was evaluated at concentrations of 5, 2.5, 1.25 and 0.625 mg mL-1 by the direct contact method on the mycelial growth of toxigenic molds of hydroponic barley namely Fusarium aveaceanum, Fusarium culmorum, Alternaria sp. and Aspergillus niger. Chloroform and ethyl acetate extracts exhibited pronounced antifungal potential. The inhibition potential varied proportionally with the concentrations. The fungal growth inhibition potentials varied significantly from 5.9–97%. Thus, Z. lotus ethyl acetate extract could be used in the food industry for better phytoprotection of barley hydroponic culture and as a natural conservative of its grains in places of storage.