Antifungal potency and modes of action of a novel olive tree defensin against closely related ascomycete fungal pathogens

Abstract

Antimicrobial peptides play a pivotal role in the innate immunity of plants. Defensins are cysteine-rich antifungal peptides with multiple mechanisms of action (MOA). A novel Oleaceae-specific defensin gene family was discovered in the genome sequences of the wild and cultivated species of a perennial olive tree, Olea europaea. Antifungal properties of an olive tree defensin OefDef1.1 were investigated against a necrotrophic ascomycete fungal pathogen Botrytis cinerea in vitro and in planta. OefDef1.1 displayed potent antifungal activity against this pathogen by rapidly permeabilizing the plasma membrane of the conidial and germling cells. Interestingly, it was translocated to the cytoplasm and induced reactive oxygen species in the germlings, but not in the conidia. In medium containing high concentrations of Na1+, antifungal activity of OefDef1.1 against B. cinerea was significantly reduced. In contrast, OefDef1.1_V1 variant in which the γ-core motif of OefDef1.1 was replaced by that of a Medicago truncatula defensin MtDef4 displayed Na1+-tolerant antifungal activity and was more potent in reducing the virulence of B. cinerea in planta. OefDef1.1 also exhibited potent antifungal activity against three hemibiotrophic ascomycete pathogens Fusarium graminearum, F. oxysporum and F. virguliforme. Significant differences were observed among the four pathogens in their responses to OefDef1.1 in growth medium with or without the high concentrations of Na1+. The varied responses of closely related ascomycete pathogens to this defensin have implications for engineering disease resistance in plants.