Ochratoxin A (OTA) is a toxic secondary metabolite produced by the Aspergillus species which can contaminate various food products. This study analysed the transcriptome of the Aspergillus westerdijkiae fc-1 strain under NaCl concentrations of 0, 20, and 100 g/L using RNA-Seq technology to examine gene transcriptional changes linked to osmotic stress and OTA production. Significant changes were observed in metabolic-pathways associated with carbohydrates, cellular communication, and hydrolase activity under 20 g/L NaCl. The HOG1 gene, associated with osmotic pressure regulation was down-regulated by 78.06%. In contrast, OTA biosynthesis genes otaA, otaB, and otaC were up-regulated by 3.26 fold, 1.99 fold, and 2.06 fold, respectively. Conversely, the otaD gene was down-regulated by 43.50%. At 100 g/L NaCl, pathways related to ion transport, peptide metabolism, ribosomal function, and transmembrane transporter protein activities were significantly up-regulated. The HOG1 gene was up-regulated by 28.32% and OTA biosynthesis genes otaA, otaB, otaC, and otaD showed up-regulation of 27.06%, 36.80%, 19.59%, and 5.72 fold, respectively. The study highlights the role of metabolic pathways in osmotic stress regulation and the correlations between HOG1 expression and OTA biosynthesis genes, providing insights for developing strategies to prevent OTA contamination in food.

Aspergillus westerdijkiae is a major producer of ochratoxin A (OTA), a highly toxic and carcinogenic mycotoxin found in various food and feed products. A. westerdijkiae produces excessive amount of OTA under various water activity (a_w) conditions that occur during food and feed storage. The biosynthetic gene clusters associated with OTA production include OTAbZIP, which plays a key role in controlling mycotoxin production in response to environmental conditions. This study explored the regulation of OTA biosynthesis in A. westerdijkiae fc-1, focusing on the OTAbZIP gene’s influence under a_w stress. The mycelium growth of A. westerdijkiae fc-1 wild type and OTAbZIP mutant strains increased by 40.7% and 50.5% under high water activity (0.96 a_w) respectively, at 6 days post-inoculation (dpi), indicating a stress on A. westerdijkiae fc-1. While OTAbZIP mutant did not produce OTA under both high and moderate a_W conditions. The wild type produced OTA and OTA biosynthetic gene expression levels were downregulated under high (0.96 a_w) and moderate (0.91 a_w) water activity. The expression level of hog1 gene in OTAbZIP mutant was significantly lower than in the wild type. Pathogenicity tests revealed that deletion of OTAbZIP did not significantly affect disease infection. This study shows that deleting OTAbZIP gene greatly reduces OTA production, affecting the strain’s adaptability to water activity stress.