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http://dx.doi.org/10.1080/12298093.2022.2131999

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)
Publication Information
Mycobiology / v.50, no.5, 2022 , pp. 382-388 More about this Journal
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
asp gene; phylogeny; TaqMan probe; sclerotia; white mold;
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