Disinfecting Aspergillus flavus represents a promising strategy to mitigate aflatoxin contamination in agricultural soils and crops. In this study, the efficient disinfection of Aspergillus flavus using a g-C3N4/alpha-Fe2O3 heterojunction under visible light irradiation, along with the roles and mechanisms of the main reactive oxygen species involved in the disinfection process were demonstrated. The g-C3N4/alpha-Fe2O3 exhibited a high photocatalytic disinfection efficiency of up to 91 %, with hydroxyl radicals (center dot OH) identified as the main active species. The production of chitin in the cell walls of Aspergillus flavus was mainly interfered with center dot OH, leading to the destruction of cellular components such as carbohydrates, proteins, and lipids during the disinfection process. The metabolic interference induced by center dot OH resulted in cell structural damage and the release of essential intracellular constituents, ultimately leading to the death of Aspergillus flavus. These findings provided valuable insights into Aspergillus flavus control that was beneficial for its future agricultural applications.