Fusarium fujikuroi is the pathogen of rice bakanae disease, and is subclassified into gibberellin and fumonisin groups (G- and F-groups). Thiophanate-methyl, a benzimidazole fungicide, has been used extensively to control F. fujikuroi. Previous investigation showed that F-group strains are thiophanate-methyl sensitive (TMS), while most G-group strains are thiophanate-methyl resistant (TMR) in Japan. The minimum inhibitory concentration (MIC) in TMS strains was 1–10 μg mL-1, while that in TMR strains was higher than 100 μg mL-1. E198K and F200Y mutations in β2-tubulin were detected in TMR strains. A loop-mediated isothermal amplification-fluorescent loop primer (LAMP-FLP) method was developed for diagnosis of these mutations, and was applied to 37 TMR strains and 56 TMS strains. The result indicated that 100% of TMR strains were identified as having either the E198K mutation (41%) or the F200Y mutation (59%), while none of the TMS strains tested showed either mutation. We found one remarkable TMR strain in the F-group which had a F200Y mutation. These results suggest that E198K and F200Y mutations in β2-tubulin contribute to thiophanate-methyl resistance in F. fujikuroi.
Progress of loop-mediated isothermal amplification technique in molecular diagnosis of plant diseases
