Antifungal activity of total saponins originated from roots of <i>Medicago hybrida</i> (Pourret) Trautv. were evaluated <i>in vitro</i> against six pathogenic fungi and eight individual major saponin glycosides were tested against one of the most susceptible fungi. The total saponins showed fungitoxic effect at all investigated concentrations (0.01%, 0.05% and 0.1%) but their potency was different for individual fungi. The highest saponin concentration (0.1%) was the most effective and the inhibition of <i>Fusarium oxysporum</i> f. sp. <i>callistephi</i>, <i>Botrytis cinerea</i>, <i>Botrytis tulipae</i>, <i>Phoma narcissi</i>, <i>Fusarium oxysporum</i> f. sp. <i>narcissi</i> was 84.4%, 69.9%, 68.6%, 57.2%, 55.0%, respectively. While <i>Fusarium oxysporum</i> Schlecht., a pathogen of <i>Muscari armeniacum</i>, was inhibited by 9.5% only. Eight major saponin glycosides isolated from the total saponins of <i>M. hybrida</i> roots were tested against the mycelium growth of <i>Botrytis tulipae</i>. The mycelium growth of the pathogen was greatly inhibited by hederagenin 3-O-<i>β</i>-D-glucopyranoside and medicagenic acid 3-O-<i>β</i>-D-glucopyranoside. Medicagenic acid 3-O-<i>β</i>-D-glucuronopyranosyl-28-O-<i>β</i>-D-glucopyranoside and oleanolic acid 3-O-[<i>β</i>-D-glucuronopyranosyl(1→2)-<i>α</i>-L-galactopyranosyl]-28-O-<i>β</i>-D-glucopyranoside showed low fungitoxic activity. Medicagenic acid 3-O-a-D-glucopyranosyl- 28-O-β-D-glucopyranoside, hederagenin 3-O-[α-L- hamnopyranosyl(1→2)-β-D-glucopyranosyl(1→2)-β-D-glucopyranosyl]- 28-O-α-D-glucopyranoside and hederagenin 3-O-<i>β</i>-D-glucuronopyranosyl-28-O-<i>β</i>-D- lucopyranoside did not limit or only slightly inhibited growth of the tested pathogen. While 2<i>β</i>, 3<i>β</i>-dihydroxyolean-12 ene-23-al-28-oic acid 3-O-<i>β</i>-D-glucuronopyranosyl-28-O-<i>β</i>-D-glucopyranoside slightly stimulated mycelium growth of <i>B. tulipae</i>.