Source: American Phytopathological Society

SUMMARY

Identification of Soybean mosaic virus Strains by RT-PCR/RFLP Analysis of Cylindrical Inclusion Coding Region. 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-Yong Lee, National Institute of Crop Science. Plant Dis. D-2004-0322-02R, 2004 (online). Accepted for publication 21 January 2004.

Soybean mosaic virus (SMV) is the most important viral disease wherever soybean is grown. It causes serious damage and effects on plant growth. Therefore, detection and identification of SMV strains is very important both for soybean cultivation and breeding SMV-resistant cultivars. Nevertheless, the classification of SMV strains mostly depends on conventional methods, which are laborious and time consuming, and uses differential soybean cultivars. A rapid and simple method using reverse-transcriptase polymerase chain reaction/restriction fragment length polymorphism (RT-PCR/RFLP) for identification and differentiation of SMV strains has been developed. In this method, a primer pair amplifying a 1,385-bp fragment of the cylindrical inclusion (CI) coding region at position 4,176 to 5,560 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 eight seedborne SMV isolates from local soybean cultivars, could be differentiated by RT-PCR/RFLP analysis. The sensitivity of RT-PCR permitted detection of SMV from plants with necrotic symptoms in which the number of virus particles was too low to be detected by immunological techniques.

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