The contamination of mycotoxin zearalenone (ZEN) in foods has been reported worldwide, resulting in potential risks to food safety. However, the toxic mechanism of ZEN on neurodegenerative diseases has not been fully elucidated. Therefore, this study conducted in vivo ZEN neurotoxicity assessment on Parkinson’s disease (PD)-related dopaminergic neurodegeneration and mitochondrial dysfunction using Caenorhabditis elegans. The results demonstrated that dopaminergic neuron damage was induced by ZEN exposure (1.25, 10, and 50 μM), and dopaminergic neuron-related behaviors were adversely affected subsequently. Additionally, the mitochondrial fragmentation was significantly increased by ZEN exposure. Moreover, upregulated expression of mitochondrial fission and cell apoptosis-related genes (drp-1, egl-1, ced-4, and ced-3) revealed the crucial role of DRP-1 on ZEN-induced neurotoxicity, which was further confirmed by drp-1 mutant and RNAi assays. In conclusion, our study indicates ZEN-induced dopaminergic neurodegeneration via DRP-1-involved mitochondrial fragmentation and apoptosis, which might cause harmful effects on PD-related symptoms.