Recruiting grape-isolated antagonistic yeasts for the sustainable bio-management of Botrytis cinerea on grapes

Botrytis cinerea is the causative agent of grey mould disease in grapes, which was linked to significant postharvest losses. This study examined three grape-isolated yeasts (Metschnikowia aff. fructicola, Metschnikowia pulcherrima, and Hansenispora uvarum) through in vitro and in vivo tests on detached grape berries against grey mould, as well as the elucidation of their possible mechanisms of action. The antifungal mechanism of action of yeasts was determined by the lytic enzyme activity, inhibition of spore germination, biofilm activity, iron depletion, diffusible metabolites, wound-site colonisation, mycocin, and volatile organic compounds (VOCs) production. The highest in vitro efficacy (83.13%) was observed on M. aff. fructicola, followed by M. pulcherrima (82.10%) and H. uvarum (71.66%). Metschnikowia yeasts exhibited comparable enzyme activities, including protease, β-1,3 glucanase, gelatinase chitinase, and cellulase, while H. uvarum had a poor enzymatic activity with chitinase and gelatinase. M. aff. fructicola showed relatively higher iron depletion activity than M. pulcherrima, while M. pulcherrima outperformed via diffusible metabolites. All yeast cultures significantly reduced spore germination by at least 86%. Overall, M. aff. fructicola exhibited the highest biocontrol activity with its iron depletion, inhibition of conidial germination, biofilm formation, VOCs, and well colonisation on grape berries. M. aff. fructicola 1-UDM outperformed all other yeasts by significantly reducing disease incidence and lesion diameter values (93.4% and 94.3%, respectively). Remarkably, H. uvarum VOCs demonstrated potential as a biofumigant for suppressing grey mould. All yeasts are well adapted to their ecological niche to bio-protect grapes from grey mould disease.