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Resistance of benzimidazole fungicides is related to the point mutation of the  β -tubulin gene in  Sclerotinia sclerotiorum . The point mutation at codon 198 (GAG → GCG, E198A) occurs in more than 90% of field resistant populations in China. Traditional detection methods of benzimidazole-resistant mutants of  S. sclerotiorum  are time-consuming, tedious and inefficient. To establish a suitable and rapid detection of benzimidazole-resistant mutants of  S. sclerotiorum , an efficient and simple method with high specificity was developed based on loop-mediated isothermal amplification (LAMP). Eight sets of LAMP primers were designed and four sets were optimized to specially distinguish benzimidazole-resistant mutants of  S. sclerotiorum . With the optimal LAMP primers, the concentration of LAMP components was optimized and the reaction conditions were set as 60–64 °C for 60 min. This method had a good specificity, sensitivity, stability and repeatability. In the 1491 sclerotia, 614 (41.18%) were positive by LAMP, and 629 (42.19%) positive by MIC. Therefore, the LAMP assay is more feasible to detect benzimidazole-resistant mutants of  S. sclerotiorum  than traditional detection methods.

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Development and application of loop-mediated isothermal amplification for detecting the highly benzimidazole-resistant isolates in Sclerotinia sclerotiorum