Abstract: In order to determine the resistance of Fusarium fujikuroi to phenamacril in Zhejiang Province and the alternative agent for its control, this study analyzed the resistance frequency, resistance level and resistance mechanism of 196 F. fujikuroi isolates collected from Hangzhou City and Jinhua City in Zhejiang Province, and also evaluated the feasibility of mefentrifluconazole against phenamacril-resistant F. fujikuroi strains via mycelium growth rate method, pot method and field efficacy test. The results showed that in 2021, 10 out of 86 isolates of F. fujikuroi isolated from Hangzhou City and Jinhua City were phenamacril-resistant isolates, with a resistance frequency was 11.63%; in 2022, 22 out of 110 isolates of F. fujikuroi isolated from Hangzhou City and Jinhua City were phenamacril-resistant isolates, and the resistance frequency was 20.00%. The resistance levels of the resistant F. fujikuroi isolates ranged from 145.963-235.256, and the average resistance level was 185.379, indicating a high-level resistance. The corresponding resistance mechanism was caused by K218T mutation of Myosin-5. The EC₅₀ values of the phenamacril -resistant and -sensitive strains to mefentrifluconazole ranged from 0.108 to 0.342 μg/mL, and the average EC₅₀ value were 0.221 μg/mL. In the pot experiments, the application of mefentrifluconazole 10 µg/mL can significantly inhibit the excessive growth of rice caused by phenamacril-resistant strains. Additionally, in the field assay, mefentrifluconazole 400 g/L SC 5000-fold dilution also showed an excellent control efficacy on rice bakanae disease. The results of this study indicated that mefentrifluconazole could be used as an alternative agent for the prevention and control of rice bakanae disease and delay the resistance of F. fujikuroi to phenamacril.