Trichothecium roseum is an important postharvest pathogen that causes core rot in apples and contaminates apple juice with spores. Contact glow discharge electrolysis (CGDE) is an innovative non-thermal technique for microbial control in fresh produce, but its efficiency against T. roseum spores in apple juice and the underlying mechanism remain unclear. This study evaluated the inactivation efficiency of T. roseum spores in apple juice under different CGDE treatments and investigated the underlying mechanisms. The results showed that spore inactivation efficiency increased with the prolongation of the treatment time, reaching a reduction of 3.97 log units after 30 min. Spore inactivation followed a log-linear model. Mechanistic analysis revealed that CGDE treatment reduced the activity of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and ascorbate peroxidase (APX), leading to the accumulation of reactive oxygen species (ROS). This oxidative stress caused lipid peroxidation, reducing the unsaturated fatty acid and ergosterol content of cell membranes and producing malondialdehyde. Damage to spore cell membranes resulted in leakage of cellular contents, structural changes and damage to surface morphology. In conclusion, CGDE effectively inactivated T. roseum spores in apple juice by disrupting cell membrane integrity through ROS generation, highlighting its potential as a non-thermal microbial control method for postharvest applications.