In this work, liposomes loaded with the fungicide, Fludioxonil (FLUD), for the containment of fungal diseases in agriculture were developed. Three types of vesicles with different compositions were compared: (I) plain vesicles, composed of soy phosphatidylcholine and cholesterol; (II) PEG-coated vesicles, with an additional polyethylene glycol coating; and (III) cationic vesicles, containing didodecyldimethylammonium bromide. Nanometric-sized vesicles were obtained both by the micelle-to-vesicle transition method and by the extrusion technique, and encapsulation efficiency, drug loading content, and Zeta potential were determined for all the samples. The extruded and PEGylated liposomes were the most stable over time and together with the cationic ones showed a significant prolonged FLUD release capacity. The liposomes’ biological activity was evaluated on conidial germination, germ tube elongation and colony radial growth of the ascomycete Botrytis cinerea, a phytopathogenic fungus affecting worldwide many important agricultural crops in the field as well as in the postharvest phase. The extruded and PEGylated liposomes showed greater effectiveness in inhibiting germ tube elongation and colony radial growth of the fungal pathogen, even at 0.01 µg·mL⁻¹, the lowest concentration assessed.

Ochratoxin A (OTA) is a mycotoxin, a common contaminant of grapes and their derivatives, such as wine, and classified as possible human carcinogen (group 2B) by the International Agency for Research on Cancer (IARC). Aspergillus carbonarius is the main producer of OTA in grapes. The stability of the molecule and the poor availability of detoxification systems makes the control of A. carbonarius in vineyards the main strategy used to reduce OTA contamination risk. Several molecular methods are available for A. carbonarius detection, but the correlation between the abundance of fungal population and OTA contamination needs to be improved. This study aimed at the development of innovative quantitative PCR (qPCR) and digital droplet PCR (ddPCR) tools to quantify the mycotoxigenic fractions of A. carbonarius strains on grapes, based on the key gene AcOTApks in the pathway of OTA biosynthesis. Different primers/probe sets were assessed, based on their specificity and sensitivity. This method allowed to quantify up to 100 fg∙µL⁻¹ [cycle of quantification (Cq) = 37] and 10 fg∙µL⁻¹ (0.38 copies∙µL⁻¹) of genomic DNA (gDNA) from A. carbonarius mycelium in qPCR and ddPCR, respectively. The sensitivity as to artificially contaminated must samples was up to 100 conidia (Cq = 38) and 1 conidium (0.13 copies∙µL⁻¹) with qPCR and ddPCR, respectively. Finally, the methods were validated on naturally infected must samples, and the quantification of the fungus was in both cases highly correlated (r = +0.8) with OTA concentrations in the samples. The results showed that both analytical methods can be suitable for improving the sustainable management of OTA contamination in grapes and their derivatives.

Grapevine trunk diseases are a threat to table- and grape-wine cultivation worldwide. Phaeomoniella chlamydospora (Pch) is a vascular fungus recognized as one of the most important pathogens associated with grapevine trunk diseases. The relationships between xylem vessel features and Pch susceptibility of 10 table- and 17 wine-grape genotypes, as well as 3 rootstocks, were investigated by image analysis of 50 µm cross-sections and artificial Pch inoculation on one-year-old vine cuttings. Vessels were grouped in the diameter classes 1–30, 31–60, 61–90, 91–120, and >120 µm. Among the table-grape varieties, ‘Sable’, ‘Timco’, and ‘Red Globe’ showed higher densities of large vessels (>120 μm) than ‘Italia’, ‘Sugar Crisp’, and ‘Sugraone’. Among the wine-grape varieties, ‘Minutolo’, ‘Montepulciano’, ‘Primitivo’ CDTa19, and ‘Verdeca’ showed higher densities of large vessels than ‘Aglianico’, ‘Nero di Troia’, ‘Sangiovese’, and ‘Susumaniello’. In the rootstocks, the vessel diameters were 50.8, 54.0, and 60.9 μm for ‘34 E.M.’, ‘140 Ruggieri’, and ‘1103 Paulsen’, in that order. For table-grape varieties, Pch was re-isolated from 13.3% for ‘Sugar Crisp’ and ‘Sugraone’ cuttings up to 93.3% for ‘Timco’. For wine-grape varieties, Pch re-isolation ranged from 51.1% (‘Bombino nero’, ‘Negroamaro’ D15, and ‘Sangiovese’) to 81.1% (‘Montepulciano’), while for the rootstocks, the values were from 33 to 51%. A principal component analysis (PCA) revealed a positive correlation between the frequencies of large vessels and Pch re-isolation. In addition, in wine grapes and rootstocks, higher Pch re-isolation frequencies in the lateral parts of cuttings were correlated (r = 0.79) to a higher frequency of large vessels. The results highlight relationships between grapevine xylem vessel sizes and susceptibility to P. chlamydospora that are worthy of further research.

Histone post-translational modifications (HPTMs) can affect gene expression by rearranging chromatin structure. Between these, histone methylation is one of the most studied in filamentous fungi, and different conserved domains coding for methyltransferase were found in Aspergillus spp. genomes. In this work, the role of the histone methyltransferases AcDot1 and AcRmtA in the mycotoxigenic fungus Aspergillus carbonarius was investigated, obtaining knockout or overexpression mutants through Agrobacterium tumefaciens-mediated transformation (ATMT). A. carbonarius is responsible for grape-bunch rot, representing the major source of ochratoxin A (OTA) contamination on grapes. In vivo conditions, the deletion of Acdot1 or AcrmtA resulted in upregulation of growth when the isolates were cultivated on a minimal medium. The influence of Acdot1 on the OTA biosynthesis was differently affected by culture conditions. On rich media, an increase in OTA accumulation was observed, while on minimal medium, lower OTA concentrations were reported. The deletion of AcrmtA always resulted in lower OTA accumulation. However, the expression of OTA biosynthesis genes was regulated by both histone methyltransferases. Of the six analyzed OTA genes, three of them showed altered expression in the knockout mutants, and otaB and otaR1 were common between both mutants. Furthermore, both AcDot1 and AcRmtA play a role in oxidative stress response, induced by 1 mM hydrogen peroxide, by modulating growth, conidiation and OTA biosynthesis. Neither the deletion nor the overexpression of the Acdot1 or AcrmtA affected virulence, while both the sporulation and OTA production were negatively affected in vivo by the deletion of AcrmtA.

Polystigma amygdalinum the causal agent of Red Leaf Blotch (RLB), is responsible for one of the most important foliar diseases affecting almond [Prunus dulcis (Miller) D.A. Webb] in the Mediterranean Basin and the Middle East. The study is aimed at improving knowledge on RLB epidemiology and the role of environmental conditions in disease development. Field monitoring was conducted from 2022 to 2025 in three almond orchards located in Apulia (southern Italy) and characterized by different microclimatic conditions. A total of 39 cultivars, including Apulian local germplasm and international cultivars (‘Belona’, ‘Genco’, ‘Guara’, ‘Ferragnès’, ‘Filippo Ceo’, ‘Lauranne^® Avijor’, ‘Soleta’, and ‘Supernova’), were evaluated. Symptoms occurred from late spring to summer, resulting particularly severe on ‘Guara’ and ‘Lauranne^® Avijor’, whereas ‘Belona’, ‘Ferragnès’, ‘Genco’, and ‘Supernova’ exhibited the highest tolerance. To our knowledge, this is also the first report of RLB tolerance by ‘Filippo Ceo’, ‘Ficarazza’, ‘Centopezze’, and ‘Rachele piccola’ representing potential genetic resources for breeding programs. Moreover, these findings reinforced previous observations proving that RLB was less severe on medium-late and late cultivars. Disease incidence varied significantly among sites and years and was strongly associated with increased rainfall, higher relative humidity, and mild temperatures recorded in November, influencing disease occurrence in the following growing season. P. amygdalinum was consistently detected by qPCR in all RLB-affected tissues and, in some cases, from mixed early RLB + Pseudomonas-like symptoms. From some leaves with early RLB symptoms, P. amygdalinum was also successfully isolated in pure culture. Overall, our results provide clear evidence that P. amygdalinum is the sole fungal pathogen consistently associated with typical RLB symptoms in Apulia (southern Italy) and highlight important cultivar-dependent differences. Its frequent molecular detection in leaves showing atypical or mixed symptoms suggests unresolved epidemiological aspects requiring further investigation.

Agrivoltaic (AV) systems are emerging as a promising strategy to reconcile agricultural management and productivity with renewable energy generation, particularly in climate-vulnerable Mediterranean regions. This study evaluated the effects of static photovoltaic shading on the vineyard microenvironment, as well as the vegetative, productive, and microbiological responses of Vitis vinifera cv. ‘Falanghina’ grown under semi-arid conditions in Southern Italy, comparing high and low shading configurations with full sun (FS) exposure. Although the study was conducted during a single growing season, the vines in the AV system had been growing and adapting to their respective shading regimes for approximately eight years. The AV system modified the vineyard microclimate by reducing solar radiation, air temperature, and wind speed, thereby lowering atmospheric and soil water demand. These morphological adjustments were associated with improved photochemical performance and enhanced vine water status, as well as more effective gas exchange. The physiological and structural responses translated into variations in grapevine productivity: moderate shading (AVL) significantly increased yield per vine by 97.7% compared to FS conditions and by 54.6% compared to high shading (AVS). This increased productivity was achieved while maintaining berry quality comparable to the control, with only an 18% to 15% increase in titratable acidity with respect to FS vines. Beyond vine responses, under the assessed conditions, AV environment also influenced the endophytic culturable microbial community of phyllosphere, inducing early shifts in the relative abundance of dominant morphotypes without detectable changes in richness or Shannon diversity at either stage, although replication was limited. Under the conditions evaluated, AVL showed a possible favorable combination of physiological and productive responses. These findings support the potential of established agrivoltaic systems to mitigate environmental stress and maintain grapevine performance under Mediterranean conditions subjected to drought and high temperatures, while further multi-year studies are needed to assess the consistency of these responses across different growing seasons.