The ochratoxin A story in grapes and wine: Ecology, genomic and risk management.

Ochratoxin A (OTA) is a potent pentaketide nephrotoxin diffusely distributed in food and feed products (grains, legumes, coffee, dried fruits, meats, beer and wine); it is also carcinogenic, neurotoxic, teratogenic and immunotoxic. This mycotoxin is produced by species of genus Aspergillus and Penicillium. OTA is the primarily mycotoxin risk in wine and dried vine fruits. Several studies focused on Aspergillus section Nigri, due to their role as causative agents of black rot of grapes, and subsequently as cause of ochratoxin A contamination. Nine different black Aspergillus species have been identified on grapes with different secondary metabolites profiles. These species are often difficult to be identified by means of classical methods. The polyphasic approach used in our studies led to characterization of three new non toxigenic species occurring on grapes: A. brasiliensis, A. ibericus and A. uvarum. However, the main source of OTA contamination in grapes is A. carbonarius, followed by A. niger and A. welwitschiae. This contamination is strongly related to climatic conditions, geographical regions (South Mediterranean climate is highly conducive), grape varieties, damage by insects, growing season (high susceptibility from early veraison to harvest, with a peak at ripening), and great variations may occur from one year to another. Differently from other mycotoxins, the genes and the enzymatic stages involved in OTA biosynthesis pathway have remained unknown for long. In last years, genomics, transcriptomics and proteomics studies have provided new information to better define the molecular key steps of OTA biosynthesis. Genome sequencing of A. carbonarius led us to predict OTA cluster and to elucidate the key role of three genes (AcOTApks, AcOTAnrps and AcOTAhal) and the order of the enzymatic steps of the biosynthesis pathway. Other predicted genes in the cluster have been identified and analysed, such as a p450 monooxygenase and a transcription factor gene, likely involved in the structural and regulatory mechanisms of OTA production. Furthermore, transcriptomic analyses are in progress to study and clarify the complex genetic picture of the fungus during OTA biosynthesis at a deeper level. Interestingly, recent studies on climate change effects evidenced the influence of raising temperature and CO2 levels on OTA production increase. Managing OTA contamination to reduce risks in grapes implies several strategies, such as implementations of good agricultural practices and risk maps, in association with the use of insecticides and fungicides when favourable climatic conditions occur. In addition, corrective actions can be adopted in wineries.