Botrytis cinerea, the causal agent of gray mold, can result in considerable preharvest and postharvest losses in many economically valuable plant species. Fungicides were widely used to minimize such losses, but fungicide resistances were detected frequently. In the present study, we collected 164 isolates from nectarine and cherry in China and tested the sensitivity to six fungicides. Among the tested isolates, 71 (43.3%) were resistant to azoxystrobin, 14 (8.5%) to cyprodinil, 7 (4.3%) to boscalid, 4 (2.4%) to carbendazim, 1 (0.6%) to iprodione, and no isolates were found to be resistant to fludioxonil. The EC₅₀ value and resistance factor (RF) of resistant isolates were determined. Fitness analysis showed that there were no significant differences between sensitive and resistant isolates for osmotic stress and pathogenicity, while more conidia production was observed for some resistant isolates. Control efficacy of fungicides showed that the resistant isolates could not be controlled efficiently by using corresponding fungicides. The point mutation G143A was detected in the Cyt b gene of the isolates resistant to azoxystrobin, while the point mutation H272R of SdhB gene was confirmed in boscalid-resistant isolates, and mutations E198V/A of TUB2 gene and mutation I365S of BcOs1 occurred in carbendazim-resistant and iprodione-resistant isolates, respectively. These results indicate that the occurrence of fungicide resistance greatly threatens the management of gray mold on stone fruits nectarine and cherry.

Gray mold, caused by Botrytis cinerea, is one of the most destructive diseases of strawberry in China. For resistance monitoring, 198 B. cinerea isolates were collected from strawberry greenhouses at 10 locations in Hubei Province. The isolates were screened for resistance to fungicides procymidone and zoxamide. In mycelium growth assays for procymidone, the mean values of effective concentration at which mycelium growth is inhibited by 50% (EC50) for sensitive (ProS) and resistant (ProR) isolates were 0.25 μg/mL and 2.21 μg/mL, respectively. The frequency of ProR isolates was 14%, and the highest frequency (48%) was observed in Yichang. Positive cross-resistance was detected for ProR isolates to other dicarboximide fungicides, but not to phenylpyrroles. Comparative analysis of fitness parameters revealed increased osmotic sensitivity of ProR isolates compared to ProS ones. Sequence analysis of the dicarboximide target gene BcOs1 revealed that ProR isolates carried either a single point mutation at codon 365 (I365S) or a pair of point mutations (Q369P and N373S). The mean EC50 values for zoxamide sensitive (ZoxS) and resistant (ZoxR) isolates were 0.31 μg/mL and 7.76 μg/mL, respectively. Only six (3%) isolates from three locations were found to be resistant to zoxamide. All ZoxR isolates were found resistant to carbendazim. Fitness parameters did not show significant difference between ZoxR and ZoxS isolates. Sequence analysis of the beta-tubulin gene in resistant isolates revealed four previously reported point mutations (E198A, E198K, F200Y and T351I). The mutation T351I was detected only in the isolates possessing E198K point mutation. Mutation F200Y was detected in a highly resistant isolate. Results of this study will be helpful for the management of fungicide resistance in B. cinerea.

Altogether, 192 Botrytis cinerea isolates collected from tomato greenhouses at different locations in Hubei Province were evaluated for their sensitivity to fungicides procymidone and zoxamide. The mean effective concentration to cause 50% growth inhibition (EC₅₀) values of procymidone for sensitive and resistant isolates were 0.25 and 3.60 μg/ml, respectively. The frequency of procymidone-resistant (Pro^R) isolates was 18%, and the highest frequency was recorded in Jingmen. Positive cross-resistance was observed for Pro^R isolates to other dicarboximide fungicides but not to phenylpyrroles. Significant differences were observed for fitness parameters (i.e., mycelial growth, osmotic sensitivity, and virulence between sensitive and resistant isolates). Amino acid sequence of the Bos1 gene revealed that Pro^R isolates carried either point mutations at codon 365 (I365S) or a pair of point mutations at codons 369 (Q369P) and 373 (N373S). For zoxamide, the mean EC₅₀ values for sensitive and resistant isolates were 0.22 and 5.32 μg/ml, respectively. Approximately 14% of the isolates were found to be resistant to zoxamide, and the highest frequency of resistance was also observed in Jingmen. There was positive cross-resistance for zoxamide-resistant (Zox^R) isolates to carbendazim. No significant differences were observed for fitness parameters between zoxamide-sensitive and Zox^R isolates. Sequence analysis of the β-tubulin gene of Botrytis cinerea revealed two previously reported point mutations (E198A and E198K) and one new point mutation (T351I). This new mutation was detected in only those isolates which possessed the E198K but not E198A substitution. This study allows for a better understanding of the resistance development profile in Hubei Province. Results will be useful for the improvement of fungicide resistance management strategies.