Abstract
Aspergillus flavus and its aflatoxins pose serious threats to human and animal health, negatively affecting agricultural productivity and the global economy. Although chemical preservatives are widely used, their effectiveness remains limited by increased fungal resistance and environmental concerns, highlighting the need for sustainable alternatives. Microbial volatile organic compounds (mVOCs) represent a promising biocontrol strategy. Here, we investigate how glutamine regulates mVOC biosynthesis in Streptomyces alboflavus TD-1 and enhances its antifungal activity against A. flavus. Antifungal assays showed that supplementation with 40 mM glutamine significantly enhanced inhibitory activity, leading to 69.0% inhibition of conidial germination and 64.5% inhibition of mycelial biomass. Transcriptome profiling identified 283 differentially expressed genes, including the two-component system regulator gluR, which was strongly upregulated. CRISPR/Cas9-mediated disruption of gluR confirmed its regulatory role. Specifically, the mutant strain produced reduced levels of antifungal mVOCs, such as dimethyl trisulfide and o-anisidine, and exhibited diminished inhibition of A. flavus. Collectively, these findings demonstrate that exogenous glutamine enhances the mVOC-mediated suppression of A. flavus by S. alboflavus TD-1 through nutrient-sensing and transcriptional regulation of volatile biosynthesis. Although aflatoxin levels were not quantified in this study, the enhanced growth inhibition and the identified mVOC shifts provide a mechanistic basis for future studies that directly quantify aflatoxin production under storage-relevant conditions.