Aspergillus flavus is a widespread pathogen affecting crops like peanuts, contributing significantly to mycotoxin contamination and subsequent crop losses. Discriminating between toxigenic and non-toxigenic strains is crucial, yet conventional methods are often cumbersome and time-consuming. In this study, we developed rapid molecular tools to differentiate between these strains. Using morphological characteristics and species-specific PCR-sequencing, we identified isolates collected from peanut seeds in southern Georgia. Through primer optimization and qPCR targeting aflatoxin regulatory genes, we successfully distinguished aflatoxin-producing and non-producing isolates. Additional genes involved in aflatoxin biosynthesis were also analyzed, showing clear expression distinctions. Our findings demonstrate the specificity and efficiency of these primer sets, providing a valuable tool for managing A. flavus contamination in peanut seed lots. Additionally, research on the seed microbiome's impact on mycotoxin production remains limited. In this study, we assessed microbial communities in peanut seeds collected over various years using ITS gene sequencing. Our results revealed a diverse microbial population, including A. flavus and other fungal pathogens, highlighting the complexity of seed microbiota. This approach offers novel insights into peanut seed-associated microbiomes and aflatoxin contamination, shedding light on the correlation between microbial communities and aflatoxin pollution.
