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Identification of Soybean mosaic virus Strains by RT-PCR/RFLP Analysis of Cylindrical Inclusion Coding Region

June 2004 , Volume 88 , Number  6
Pages  641 - 644

Yul-Ho Kim , National Institute of Crop Science, RDA, Suwon 441-857 Korea ; Ok-Sun Kim , National Seed Management Office, MAF, Suwon 442-400, Korea ; Jae-Hwan Roh , Jung-Kyung Moon , and Soo-In Sohn , National Institute of Crop Science ; Sang-Chul Lee , Department of Agronomy, Kyungpook National University, Taegu 702-701, Korea ; and Jang-Young Lee , National Institute of Crop Science

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Accepted for publication 21 January 2004.

A reverse-transcriptase polymerase chain reaction/restriction fragment length polymorphism (RT-PCR/RFLP) was employed successfully for detection and identification of Soybean mosaic virus (SMV) strains. A primer pair amplifying a 1,385-bp fragment of the cylindrical inclusion (CI) coding region was used for RT-PCR and the RFLP profiles of the RT-PCR products were compared after restriction digestion with RsaI, EcoRI, or AccI restriction endonucleases. These enzymes were chosen based on the nucleotide sequences of SMV strains G2, G5, G5H, G7, and G7H in the CI coding region. These five strains, as well as seedborne SMV isolates from local soybean cultivars, could be differentiated by RT-PCR/RFLP analysis. The results correlated well with strain identification by symptom phenotypes produced on differential cultivars inoculated with strains and isolates. The sensitivity of RT-PCR enabled detection of SMV from plants with necrotic symptoms in which the number of virus particles was too low to be detected by enzyme-linked immunosorbent assay.

© 2004 The American Phytopathological Society