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Geonwoo Kim,Doeun Son,Sungyu Choi,Haifeng Liu,상현규 한국식물병리학회 2023 Plant Pathology Journal Vol.39 No.6
Botrytis cinerea is a major fungal plant pathogen that causes gray mold disease in strawberries, leading to a decrease in strawberry yield. While benzimidazole is widely used as a fungicide for controlling this disease, the increasing prevalence of resistant populations to this fungicide undermines its effectiveness. To investigate benzimidazole resistant B. cinerea in South Korea, 78 strains were isolated from strawberries grown in 78 different farms in 2022, and their EC50 values for benzimidazole were examined. As a result, 64 strains exhibited resistance to benzimidazole, and experimental tests using detached strawberry leaves and the plants in a greenhouse confirmed the reduced efficacy of benzimidazole to control these strains. The benzimidazole resistant strains identified in this study possessed two types of mutations, E198A or E198V, in the TUB2 gene. To detect these mutations, TaqMan probes were designed, enabling rapid identification of benzimidazole resistant B. cinerea in strawberry and tomato farms. This study utilizes TaqMan real-time polymerase chain reaction analysis to swiftly identify benzimidazole resistant B. cinerea, thereby offering the possibility of effective disease management by identifying optimum locations and time of application.
Sungyu Choi,Doeun Son,Hyunkyu Sang 한국농약과학회 2021 한국농약과학회 학술발표대회 논문집 Vol.2021 No.11
Aspergillus flavus is a causal agent of various crops and causes the greatest economic impacts. The use of antifungal agents results in development of drug resistance in A. flavus. A total of 123 A. flavus strains were collected from diverse regions and sources, and antifungal susceptibility tests of the strains to a demethylation inhibitor (DMI) fungicide, metconazole, were conducted. The results showed 26 strains were resistant to metconazole at 10μg ml-1. Three paralogous sterol 14α-demethylases (Cyp51A, Cyp51B, and Cyp51C), DMI fungicide targets, sequences of metconazole resistant and sensitive strains were investigated to find potential mutation(s) in three paralogous cyp51 genes associated with DMI resistance. Several mutations in cyp51 genes were found in metconazole resistant strains and currently candidate mutations associated with drug resistance are validated by the genetic transformation method with the CRISPR system. In addition, to establish the genetic transformation system in A. flavus, protoplasts of the field strain of A. flavus were transformed with the plasmid containing a green fluorescence protein (GFP), and the transformants exhibiting GFP were observed under the fluorescence microscope. This study reports metconazole resistant strains of A. flavus containing mutations in cyp51 genes in Korea for the first time and the genetic transformation tool developed in this study can be utilized for studying mechanisms of antifungal drug resistance in A. flavus.