A Loop-Mediated Isothermal DNA Amplification (LAMP) Assay for Detection of the Clubroot Pathogen Plasmodiophora brassicae

Plant Dis. 2022 Jun;106(6):1730-1735. doi: 10.1094/PDIS-11-21-2430-RE. Epub 2022 May 5.

Abstract

Clubroot caused by Plasmodiophora brassicae is a serious threat to cruciferous crops around the world. The resting spores of P. brassicae are a 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 PCR-based methods. The loop-mediated isothermal DNA amplification (LAMP) is a promising approach for microorganism detection with the advantage of high sensitivity, accuracy, and convenience in viewing. 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 of plasmid DNA or 10 resting spores in the soil. Compared with conventional PCR, the LAMP was more sensitive in detection of 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 sustainable clubroot management and planning.

Keywords: LAMP; Plasmodiophora brassicae; clubroot; molecular detection.

MeSH terms

  • Biological Assay
  • DNA
  • Molecular Diagnostic Techniques
  • Nucleic Acid Amplification Techniques
  • Plant Diseases / genetics
  • Plasmodiophorida* / genetics
  • Soil
  • Spores, Protozoan

Substances

  • Soil
  • DNA

Supplementary concepts

  • LAMP assay