Fungi can grow in foods and commodities worldwide, leading to the production of secondary metabolites (SM) in these substrates. Mycotoxins are toxic SM, and contamination in agricultural and food products poses a serious risk to human health, livestock and the environment. In the current study, we aim to investigate the associated mycobiota of traditional foods, nuts and dried fruits originating from the Kurdistan Province in Iran, and analyse the presence of important mycotoxins in these substrates and selected isolated strains. Sampling was conducted in nine cities during the summer and autumn of 2022, and the mycological analysis resulted in a large fungal strain collection. Morphological examination of the strains showed that the majority of isolated fungi belonged to Alternaria, Aspergillus, Penicillium and Rhizopus. A large selection of strains was identified to species level using a molecular-based identification approach, which involved sequencing various barcode genes (e.g., BenA, CaM, GPDH, ITS and RPB2). Mycotoxin analysis of a representative selection of substrates, and determination of metabolite profiles from the isolated fungi, were performed using LC-MS/MS. This study investigates the mycobiota and mycotoxins in diverse substrates from Iran, and discusses the obtained data in relation to their prevalence and potential health concerns.