Crocus sativus is an important crop for the production of saffron and bioactive compounds. Plant endophytic fungi are a source of secondary metabolites additional to those produced by the plant itself. We analysed the biodiversity of endophytic fungi present in corms, stems, leaves, tepals, and stigmas of C. sativus from ten Italian sites; furthermore, we isolated putative pathogenic fungi from rotten plants. We used an in vitro isolation approach followed by molecular analysis of the internal transcribed spacer (ITS rDNA) region. We obtained 165 strains belonging to 39 OTUs, spreading over 26 genera and 29 species. Dark septate endophytes of the genus Cadophora and the species Talaromyces pinophilus dominated in corms, while Alternaria alternata, Epicoccum spp., T. pinophilus, Mucor fragilis, and Stemphylium vesicarium dominated in other tissues. The most frequently isolated pathogens were Fusarium oxysporum and Rhizopus oryzae. Endophytic communities significantly differed among tissues and life stages, whereas differences among cultivation sites were not statistically supported. Several endophytes were hypothesized to have changing trophic modes and/or to be latent pathogens in C. sativus. All strains were conserved ex-situ for future bioactivity tests and production of metabolites.

The ectomycorrhizal fungus Tuber magnatum produces the white truffle appreciated worldwide for its unique aroma. With respect to other Tuber spp. of economic interest, T. magnatum presents a narrower geographical range. This species has, in fact, long been considered endemic to Italy. However, over the last few decades several reports have documented the presence of white truffles in different Mediterranean countries and in particular in various areas of south-east Europe. In this study, samples from several Pannonian and Balkan countries such as Hungary, Serbia, Romania, Bulgaria and Greece have been collected and genotyped with microsatellite markers and the data merged with those available for Italian populations. Our objectives were to test whether Italian and south-east European populations are differentiated and to evaluate the genetic diversity of T. magnatum all over its distributional range. We show the genetic structure of T. magnatum populations with the differentiation of four main groups: northern Italy, central-northern Italy, southern Italy and the Balkan/Pannonian region. The present study allowed us to refine the evolutionary history of T. magnatum and track the possible post-glacial expansion route of this species. The assessment of T. magnatum’s genetic structure is not only of scientific relevance, but it is also important for the conservation and market traceability of this prestigious fungus.

Ectomycorrhizal ascomycetes belonging to the genus Tuber produce edible fruiting bodies known as truffles. Tuber aestivum, in particular, is a fungus appreciated worldwide and has a natural distribution throughout Europe. Most of the molecular studies conducted on this species have been focused on the question as to whether or not T. aestivum and the morphologically similar T. uncinatum are conspecific. Conversely, only a handful of studies have assessed the level and distribution of genetic diversity and occurrence of phylogeographic patterns in this species. Here, we analyzed the genetic diversity of T. aestivum over a wide geographic range, performing an extensive sampling of specimens from Turkey, which is novel, to the best of our knowledge. We compared the internal transcribed spacer (ITS) profiles of 45 samples collected in different Turkish areas with those of 144 samples from all over Europe. We identified 63 haplotypes, 32 of which were exclusively present in Turkey. The majority of these haplotyes were also population specific. Haplotype network analysis and statistical tests highlighted the presence of a genetic structure and phylogeographic pattern, with three spatially distinct genetic clusters (northeastern Europe, southern Europe, and Turkey), with Turkey representing a diversity hotspot. Based on these results, we hypothesize the presence of glacial refugia for T. aestivum in Turkey, whereas European populations likely experienced a population bottleneck. The possible occurrence of cryptic species among Turkish T. aestivum samples also emerged. Our results are of practical relevance for the marketing of T. aestivum truffles and mycorrhizal seedlings and the preservation of the biodiversity of this species.

Humulus lupulus L. (hop) is a crucial crop within the brewing industry and a rich source of bioactive compounds. Traditionally concentrated in northeast regions of Europe, hop cultivation has expanded towards southern territories such as Italy over recent decades. Managing phytosanitary threats in Mediterranean climates poses challenges due to limited knowledge and registered agrochemicals. In pursuit of eco-friendly alternatives for disease management, we isolated 262 endophytic fungal strains from wild hop roots, stems, leaves, and flowers. Through phylogenetic analyses, we identified 51 operational taxonomic units. Dominant species such as Ilyonectria macrodidyma, Penicillium sp., Diaporthe columnaris, Plectosphaerella cucumerina, and Fusarium oxysporum were exclusive to roots. In contrast, Alternaria spp. and Epicoccum spp. were prevalent in other tissues, and Botrytis cinerea was exclusively detected in female flowers. We tested seven isolates—Epicoccum sp., Aureobasidium pullulans, Plectosphaerella cucumerina, Stemphylium vesicarium, Periconia byssoides, Talaromyces wortmannii, and Nigrospora sphaerica—against the four phytopathogenic fungi Alternaria sp., Fusarium oxysporum, Botrytis cinerea, and Sclerotinia sclerotiorum. All endophytes exhibited antagonistic effects against at least one pathogen, with Plectosphaerella cucumerina showing the strongest inhibition against Alternaria sp. This study marks the first exploration of endophytic fungi from various hop tissues. All isolated strains were ex situ conserved for future bioactivity assessments and biotechnological applications. Original data with a key relevance for the environmentally friendly management of plant diseases are provided.

Bacterial growing resistance to antibiotics poses a critical threat to global health. This study investigates, for the first time, the antibiofilm properties of Vicia ervilia agglutinin (VEA) from six different V. ervilia accessions against pathogenic bacteria, and the yeast Candida albicans. In the absence of antimicrobial properties, purified VEA significantly inhibited biofilm formation, both in Gram-positive and Gram-negative bacteria, but not in C. albicans. With an inhibitory concentration ranging from 100 to 500 µg/ml, the VEA antibiofilm activity was more relevant against the Gram-positive bacteria Streptococcus aureus and Staphylococcus epidermidis, whose biofilm was reduced up to 50% by VEA purified from accessions #5 and #36. VEA antibiofilm variability between accessions was observed, likely due to co-purified small molecules rather than differences in VEA protein sequences. In conclusion, VEA seed extracts from the accessions with the highest antibiofilm activity could represent a valid approach for the development of an effective antibiofilm agent.