Genome diversity, population structure and MALDI-TOF MS profiling of Aspergillus oryzae/flavus strains from fermentation and wild environments

Various strains of Aspergillus oryzae, regarded as a domesticated variant of aflatoxigenic Aspergillus flavus, are utilized in the soybean fermentation industry of Korea. This study compared A. oryzae/flavus strains isolated from various environments in Korea including industrial settings, Meju (brick of dried fermented soybeans), and wild conditions with globally reported strains using genomic analysis to determine their taxonomic positions and risk of mycotoxicity. Using population genomics, five distinct groups (A to E) were identified, with all aflatoxigenic Korean strains in Group C and non-aflatoxigenic Korean strains in Groups A, B, and E. Korean strains from Meju and wild conditions are distributed across Groups A and B, and most of the Korean industrial strains form a sub-cluster with Japanese industrial strains in Group A. Comparing secondary metabolite gene cluster mutation pattern, three gene clusters (Aflatoxin, Cyclopiazonic acid and Ditryptophenaline) were revealed as group specific ones. In aflatoxin and cyclopiazonic acid clusters, most of the Group C strains had intact regions compared to strains in other groups. Since most of the Group C strains produce aflatoxin and have intact Aflatoxin and Cyclopiazonic acid gene clusters, we considered that this group represent A. flavus. Profiling using MALDI-TOF MS analysis also distinguished Group C from Groups A, B and E by specific three proteomic peaks. Among the three peaks, those around 12,700 to 12,900 m/z (Da) are expected to correspond to AflF (nor B), an enzyme involved in Aflatoxin metabolism. These results showed taxonomic positions of Korean strains of A. oryzae/flavus from various environments and also showed possibility to differentiate between A. oryzae and A. flavus with genome and MALDI-TOF MS analysis.