Multiplex TaqMan qPCR Assay for Detection, Identification, and Quantification of Three Sclerotinia Species
10.1080/12298093.2022.2131999
- Author:
Dong Jae LEE
1
;
Jin A LEE
;
Dae-Han CHAE
;
Hwi-Seo JANG
;
Young-Joon CHOI
;
Dalsoo KIM
Author Information
1. Department of Biological Science, Kunsan National University, Gunsan, Korea
- Publication Type:RESEARCH ARTICLE
- From:Mycobiology
2022;50(5):382-388
- CountryRepublic of Korea
- Language:English
-
Abstract:
White mold (orSclerotinia stem rot), caused bySclerotinia species, is a major air, soil, or seed-transmitted disease affecting numerous crops and wild plants. Microscopic or culturebased 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 Sclerotiniaspecies 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.
