Microscopic investigations were conducted into the interaction of Colletotrichum acutatum on white and red strawberry (Fragaria ·ananassa) fruit surfaces. The results showed that, whilst the early interaction events were similar in both white and red fruits, after 24 h fungal colonization dramatically varied: in white fruits C. acutatum became quiescent as melanized appressoria, but on red fruits it displayed subcuticular necrotrophic invasion. A microarray analysis of white and red strawberries after 24 h of interaction with C. acutatum was performed, in order to reveal differences in gene expression possibly related to the different susceptibility of unripe and ripe fruits. Epi ⁄ catechin-related genes and fatty acid metabolism genes, involved in the production of quiescence-related molecules such as flavan-3-ols, proanthocyanidins and antifungal dienes, were found to be regulated during strawberry ripening, supporting a role for these molecules as preformed defence mechanisms. Besides several genes commonly regulated upon pathogen interaction, different genes were specifically transcribed only in white or red challenged fruits; a number of these, such as those coding for lectin and polyphenol oxidase, possibly account for specific pathogen-induced responses. The putative biological role of these genes in the different susceptibility of fruits to C. acutatum is discussed.

PR-10 proteins are a family of pathogenesis-related (PR) allergenic proteins playing multifunctional roles. The peach (Prunus persica) major allergen, Pru p 1.01, and its isoform, Pru p 1.06D, were found highly expressed in the fruit skin at the pit hardening stage, when fruits transiently lose their susceptibility to the fungal pathogen Monilinia spp. To investigate the possible role of the two Pru p 1 isoforms in plant defense, the recombinant proteins were expressed in Escherichia coli and purified. Light scattering experiments and circular dichroism spectroscopy showed that both proteins are monomers in solution with secondary structures typical of PR-10 proteins. Even though the proteins do not display direct antimicrobial activity, they both act as RNases, a function possibly related to defense. The RNase activity is different for the two proteins, and only that of Pru p 1.01 is affected in the presence of the cytokinin zeatin, suggesting a physiological correlation between Pru p 1.01 ligand binding and enzymatic activity. The binding of zeatin to Pru p 1.01 was evaluated using isothermal titration calorimetry, which provided information on the stoichiometry and on the thermodynamic parameters of the interaction. The structural architecture of Pru p 1.01 and Pru p 1.06D was obtained by homology modeling, and the differences in the binding pockets, possibly accounting for the observed difference in binding activity, were evaluated.

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Nine plant-volatile compounds were tested for their activity against Monilinia laxa, the cause of brown rot in stone fruit. In vitro trials on conidial germination and mycelial growth showed a consistent fungicidal activity of trans-2-hexenal, carvacrol, and citral, whereas trans-cinnamaldehyde, hexanal, (-)-carvone, eugenol, 2-nonanone, and p-anisaldehyde exhibited a progressively lower inhibition. The best inhibitor of conidial germination was trans-2-hexenal (effective dose for 50 and 90% inhibition [ED50 and ED95] = 7.53 and 9.4 μl/liter, respectively; minimal inhibitory concentration [MIC] = 12.3 μl/liter], whereas carvacrol was the best inhibitor of mycelial growth (ED50 and ED95 = 2 and 3.4 μl/liter, respectively; MIC = 6.1 μl/liter). The three most active compounds in in vitro studies also were tested in vivo as postharvest biofumigants. The best control of brown rot was with trans-2-hexenal (efficacy ranging from 46.2 to 80.3%, depending on cultivar), whereas citral and carvacrol resulted in a lower efficacy of 40 and 32.9%, respectively. Fumigation with trans-2-hexenal at concentrations that stopped decay did not cause any visible disorders to plum, whereas it was phytotoxic to apricot, peach, and nectarine and produced off-odors or off-flavors in all species of stone fruit tested.