Mycobiology

The Korean Society of Mycology (한국균학회)
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Multiplex TaqMan qPCR Assay for Detection, Identification, and Quantification of Three Sclerotinia Species

  • Dong Jae Lee (Department of Biological Science, Kunsan National University) ;
  • Jin A Lee (Department of Biological Science, Kunsan National University) ;
  • Dae-Han Chae (Moghu Research Center Ltd) ;
  • Hwi-Seo Jang (Moghu Research Center Ltd) ;
  • Young-Joon Choi (Department of Biological Science, Kunsan National University) ;
  • Dalsoo Kim (Moghu Research Center Ltd)
  • Received : 2022.08.25
  • Accepted : 2022.09.29
  • Published : 2022.10.31
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Abstract

White mold (or Sclerotinia stem rot), caused by Sclerotinia species, is a major air, soil, or seed-transmitted disease affecting numerous crops and wild plants. Microscopic or culture-based methods currently available for their detection and identification are time-consuming, laborious, and often erroneous. Therefore, we developed a multiplex quantitative PCR (qPCR) assay for the discrimination, detection, and quantification of DNA collected from each of the three economically relevant Sclerotinia species, namely, S. sclerotiorum, S. minor, and S. nivalis. TaqMan primer/probe combinations specific for each Sclerotinia species were designed based on the gene sequences encoding aspartyl protease. High specificity and sensitivity of each probe were confirmed for sclerotium and soil samples, as well as pure cultures, using simplex and multiplex qPCRs. This multiplex assay could be helpful in detecting and quantifying specific species of Sclerotinia, and therefore, may be valuable for disease diagnosis, forecasting, and management.

Keywords

Acknowledgement

This work was supported by Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry (IPET) through Agricultural Machinery/Equipment Localization Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) [grant number 321056-05].

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