Direct Detection of Cylindrocarpon destructans, Root Rot Pathogen of Ginseng by Nested PCR from Soil Samples

• Published in: Mycobiology Vol. 38; no. 1; pp. 33 - 38
• Main Authors: Jang, C.S., Chungnam National University, Daejeon, Republic of Korea, Lim, J.H., Chungnam National University, Daejeon, Republic of Korea, Seo, M.W., Chungnam National University, Daejeon, Republic of Korea, Song, J.Y., Chungnam National University, Daejeon, Republic of Korea, Kim, H.G., Chungnam National University, Daejeon, Republic of Korea
• Format: Journal Article
• Language: English
• Published: Korea (South) The Korean Society of Mycology 01-03-2010; 한국균학회
• Subjects: DIAGNOSIS; DIAGNOSTIC; DIAGNOSTICO; NECTRIA RADICICOLA; Nested PCR; PANAX PSEUDOGINSENG; Root rot; 생물학; Cylindrocarpon destructans; Root rot; Nested PCR; Diagnosis; Ginseng
• ISSN: 1229-8093; 2092-9323
• DOI: 10.4489/MYCO.2010.38.1.033

We have successfully applied the nested PCR to detect Cylindrocarpon destructans, a major pathogen causing root rot disease from ginseng seedlings in our former study. The PCR assay, in this study, was used to detect the pathogen from soils. The nested PCR using internal transcribed spacer (ITS) 1, 4 primer set and Dest 1, 4 primer set maintained the specificity in soils containing various microorganisms. For a soil DNA extraction method targeting chlamydospores, when several cell wall disrupting methods were tested, the combination of lyophilization and grinding with glass beads, which broke almost all the chlamydospores, was the strongest. The DNA extraction method which was completed based on the above was simple and time-saving because of exclusion of unnecessary stages, and efficient to apply in soils. As three ginseng fields whose histories were known were analyzed, the PCR assay resulted as our expectation derived from the field information. The direct PCR method will be utilized as a reliable and rapid tool for detecting and monitoring C. destructans in ginseng fields.