Tulbaghia violacea (Harv) Exerts its Antifungal Activity by Reducing Ergosterol Production in Aspergillus flavus

Opportunistic infections in immunosuppressed patients have led to an increase in fungal infections, with Aspergillus being one of the main causative agents. Medicinal plants exhibiting antifungal activity have the potential to be used as chemotherapeutic agents. However, often their mechanisms of action are not fully researched. Tulbaghia violacea exhibits antifungal activity towards Candida, Aspergillus flavus and Aspergillus parasiticus but its mode of action has only recently begun to be investigated. This study aimed to ascertain the effect of T. violacea rhizome extracts on ergosterol production in A. flavus and the mechanism of inhibition. The MIC of a T. violacea rhizome extract against A. flavus was first determined, using a broth dilution assay, to be 15 mg/ml. Thereafter, the culture was subjected to sub-inhibitory concentrations of the extract before sterol intermediates of the ergosterol biosynthetic pathway were isolated and analysed for dose-dependent accumulation. Analysis by reverse-phase HPLC displayed a decline in ergosterol production in a dose-dependent manner when exposed to increasing concentrations of T. violacea extract. Quantification of the sterol intermediates of the ergosterol pathway indicated a definite accumulation of 2,3-oxidosqualene. The results prove that the plant extract affected ergosterol synthesis by inhibiting oxidosqualene cyclase. This prevented the formation of downstream intermediates of the ergosterol pathway ultimately resulting in inhibition of ergosterol production.