Hormetic effects of fungicides are highly relevant to fungicide applications and management of plant-pathogenic fungi. Preconditioning (i.e., early exposure to relatively low doses of a toxicant) is a special form of hormesis, and fungicide preconditioning of phytopathogenic fungi is inevitable in the field. The present study showed that spraying the demethylation inhibitor (DMI) fungicide flusilazole at 0.1 µg/ml had stimulatory effects on the virulence of Sclerotinia sclerotiorum inoculated at 1 and 24 h after spraying. Flusilazole sprayed at 10 µg/ml showed inhibitory effects on the virulence of S. sclerotiorum inoculated during the first 3 days after spraying. Inoculations on the 5th, 7th, and 10th day after spraying did not show any significant inhibitory or stimulatory effects on the virulence. After growing for 2 days on potato dextrose agar (PDA) amended with flusilazole at a dose range from 0.0005 to 0.25 µg/ml as preconditioning treatments, mycelia were transferred onto PDA without fungicide and subsequent mycelial growth was slower than the nonpreconditioned control. However, after the preconditioned colonies were transferred onto PDA supplemented with flusilazole at 0.2 µg/ml, percent stimulations of mycelia growth compared with the control had a parabolic shape across the preconditioning flusilazole concentration range. Similarly, the mycelial growth of the preconditioned mycelial plugs on PDA amended with other DMI fungicides (prochloraz or tebuconazole) also showed a typical hormetic response, whereas mycelial growth on PDA amended with carbendazim or dimethachlone was inhibited in a dose-dependent manner. Preconditioning S. sclerotiorum with flusilazole on rapeseed plants elicited virulence stimulations in a dose-dependent manner similar to those on mycelial growth on PDA. After disease lesions developed on rapeseed leaves sprayed with flusilazole as the preconditioning treatment were inoculated onto rapeseed plants, virulence was inhibited on leaves without fungicide or sprayed with carbendazim or dimethachlone compared with the nonpreconditioned control, whereas virulence was stimulated on leaves sprayed with flusilazole, prochloraz, or tebuconazole, and the maximum percent stimulation was 10.2%. These results will advance our understanding of hormetic effects of fungicides and of preconditioning hormesis in particular.