The present study investigated the antifungal potential of hydroalcoholic extracts of propolis against the causal agent of grey mould, Botrytis cinerea, by in vitro and in vivo assays. Five different propolis from different Italian regions were subjected to hydroalcoholic extraction using different ethanol concentrations and extraction methods. The preliminary bio-assay showed significant inhibitory effects on B. cinerea mycelial growth of propolis extracts obtained using 90% ethanol and subjected to sonication. The calculation of EC50 values, based on the demonstrated efficacy of non-volatile and volatile metabolites of propolis extracts, was useful to understand the main fraction involved in the antifungal activity of the samples and to perform the in vivo assay on grape and blueberry fruits. Three of the propolis extracts showed a high amount of genistein. Conversely, the other two propolis showed a fair amount of apigenin, caffeic acid, chrysin, ferulic acid, kaempferol, luteolin, p-coumaric acid and quercetin. From the volatile analysis of propolis, the main compounds detected were α-cadinol, α-eudesmol, calamenene, cadinol, benzyl acetate, benzyl benzoate and benzyl alcohol. The results suggest that propolis extracts have potential as an effective postharvest antifungal treatment, with varying degrees of efficacy depending on the extraction method and the type of propolis metabolites.

Pleurostoma richardsiae is associated with host trunk diseases, known to cause dieback, cankers and wilting of woody trees, and human infections. This fungus was isolated from wood tissues of declining olive trees and grapevines showing esca disease symptoms, in the Apulia region of Italy. Fungus detection has been based on morphological and molecular features, which are time-consuming to identify and require well-trained personnel. Improvement of Pl. richardsiae detection in olive was achieved through development of real time loop-mediated isothermal amplification targeting the intergenic spacer (IGS) region of the fungus. Specificity of the assay was confirmed using ten Pl. richardsiae strains and 36 other fungus strains of species usually isolated from declining olive trees. The achieved limit of detection was 7.5 × 10-2 ng μL-1 of Pl. richardsiae genomic DNA. A preliminary validation of RealAmp was also performed using material from infected olive plants artificially inoculated in a greenhouse.