Resistance Development to Procymidone and Boscalid in Causing Black Spot Disease on .

is a valuable medicinal plant threatened by black spot disease caused by for which application of fungicides is the primary strategy. However, the detection and characterization of fungicide resistance status in has not yet been reported. A total of 106 and 128 single-spore isolates of were recovered in 2015 and 2021, respectively, from 52 and 56 fields located in Jinhua, Wenzhou, Hangzhou, and Ningbo, where 90% of is produced. In 2015, moderate resistance to procymidone (Pro), a dicarboximide fungicide that has been applied for more than 10 years, was detected at a rate of 23.6%, and the resistance frequency increased to as high as 44.5% in 2021. For the SDHI boscalid, the EC values ranged from 0.013 to 0.976 μg/ml, with the baseline EC value of 0.57 ± 0.29 μg/ml in 2015, while the sensitivity profile showed prominent characteristics of a disruptive distribution, with a resistance frequency of 22.6% in 2022. Positive cross-resistance was observed between procymidone and iprodione (ρ = 0.897, = 0.000) and between boscalid and pydiflumetofen ( = 0.937, = 0.000). No fitness penalty in growth, sporulation, germination, or pathogenicity was found for the Pro or the boscalid-resistant (Bos) isolates. P894L of s was observed in all the tested Pro isolates, and the double point mutation (P894L + S1277L) was detected in 33.3% of the Pro isolates. Four types of mutations in (G14W in B, V181G in D, A47T in D, and G14W in B + A47T in D) were found in the Bos isolates.