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Plants cannot avoid environmental challenges and are constantly threatened by diverse biotic and abiotic stresses. However, plants have developed a unique immune system to defend themselves against the invasion of various pathogens. Melatonin, N‐acetyl‐5‐methoxytryptamine has positive physiological effects in plants that are involved in disease resistance. The processes underlying melatonin‐induced pathogen resistance in plants are still unknown. The current study explores how melatonin regulates the plant‐disease interaction in maize. The results showed that 400 μM melatonin strongly reduced the disease lesion on maize stalks by 1.5 cm and corn by 4.0 cm caused by Fusarium graminearum PH‐1. Furthermore, after treatment with melatonin, the plant defense enzymes like SOD significantly increased, while POD and APX significantly decreased compared to the control. In addition, melatonin can also improve maize's innate immunity, which is mediated by melatonin treatments through the salicylic acid signaling pathway, and up‐regulate the defense‐associated expression of PR1, LOX1, OXR, serPIN, and WIPI genes in maize. Melatonin not only inhibits the disease in the maize stalks and corn, but also down‐regulates the deoxynivalenol (DON) production‐related expression of genes Tri1, Tri4, Tri5, and Tri6 in maize. Overall, this study sheds new light on the mechanisms by which melatonin regulates antioxidant enzymes and defense‐related genes involved in plant immunity to effectively suppress plant diseases. [ABSTRACT FROM AUTHOR]