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http://dx.doi.org/10.5423/RPD.2004.10.1.053

Detection of Cucumber green mottle mosaic virus in Bottle Gourd Seeds by RT-PCR  

Lee, Sook-Kyung (전북대학교 농생물학과)
Song, Wan-Yeob (전북대학교 농업과학기술연구소)
Kim, Hyung-Moo (전북대학교 농생물학과)
Publication Information
Research in Plant Disease / v.10, no.1, 2004 , pp. 53-57 More about this Journal
Abstract
Cucumber green mottle mosaic virus (CGMMV) was a major pathogen of watermelon and had affected seriously to watermelon production in Korea. Rapid and sensitive detection method of CGMMV associated with bottle gourd (Lagenafia siceraria) seeds was developed by using RT-PCR in this study. A pair of primeri Wmfl and Wmrl, specific for CGMMV was designed from coat protein gene sequences of CGMMV-W and used for amplifying 420 bp product in RT-PCR. To simplify the virus extraction procedure and reduce an inhibitor from the extract for the RT-PCR, some methods using ethanol precipitation, double filtration, polyethylene glycol precipitation and phenol/chloroform/isoamyl alcohol extraction procedure were compared and the phenol/chloroform/isoamyl alcohol extraction procedure was selected by its enhanced sensitivity. This detection method using the selected extraction step and the primers for RT-PCR could reliably detect an infected level of one CGMMV-infested seed in 1,000 seeds. This rapid and sensitive RT-PCR assay provides auseful tool for the specific detection of CGMMV in bottle gourd seed samples containing high levels of back-ground inhibitors.
Keywords
Bottle gourd seed; CGMMV; phenol/chloroform/isoamyl alcohol extraction procedure; RT-PCR;
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