A loop-mediated isothermal DNA amplification (LAMP) assay for detection of the clubroot pathogen Plasmodiophora brassicae

Abstract

Clubroot caused by Plasmodiophora brassicae is a serious threat to cruciferous crops around the world. The resting spores of P. brassicae are primary source of infection and can survive in soil for many years. Detection of resting spores in soil is essential for forecasting clubroot prevalence. Detection of P. brassicae has been relying on plant bioassays or polymerase chain reaction (PCR)-based methods. The loop-mediated isothermal DNA amplification (LAMP) is a promising approach for microorganism detection with the advantage of high sensitivity, being accurate and convenient to visualize. In this study, we developed a LAMP assay for detection of P. brassicae in soil, roots and seeds. This method can detect P. brassicae at a minimal amount of 1 fg plasmid DNA or 10 resting spores in the soil. Compared to conventional PCR, the LAMP was more sensitive in detection P. brassicae at the lower levels in soil samples. In conclusion, we elaborated a sensitive, accurate and easy-to-use LAMP assay to detect P. brassicae, which will facilitate to plan sustainable clubroot management.