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<i>Botryosphaeria dothidea</i>, a notorious plant pathogen, is responsible for causing chestnut rot during postharvest storage. This research aimed to assess the antifungal properties of perilla essential oil (PEO) and perillaldehyde (PAE) against <i>B. dothidea</i>. PEO’s and PAE’s inhibitory effects on <i>B. dothidea</i> were investigated using an agar dilution method, a fumigation method, and an in vivo assay in chestnuts and shell buckets. Based on the results of gas chromatography-mass spectrometry, it was confirmed that the main component of PEO was elemicin. The antifungal mechanism of PEO and PAE against <i>B. dothidea</i> was investigated by conducting staining experiments of the fungal cell wall and cell membrane. PEO and PAE strongly inhibit the mycelial growth of <i>B. dothidea</i> in a dose-dependent manner. The inhibitory mechanism is mainly related to the destruction of the integrity of the fungal cell wall and plasma membrane. Notably, PEO retains its antifungal efficacy against <i>B. dothidea</i> in chestnuts, effectively prolonging their storage life. These findings indicate that PEO and PAE are nontoxic, eco-friendly botanical fungicides, holding promise for controlling postharvest chestnut rot.