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http://dx.doi.org/10.5423/RPD.2015.21.3.180

A Simple and Reliable Molecular Detection Method for Tomato yellow leaf curl virus in Solanum lycopersicum without DNA Extraction  

Yoon, Ju-Yeon (Virology Unit, Department of Horticultural Environment, National Institute of Horticultural and Herbal Science, Rural Development Administration)
Kim, Su (Vegetable Fruit Unit, Department of Vegetable Science, National Institute of Horticultural and Herbal Science, Rural Development Administration)
Choi, Gug-Seoun (Virology Unit, Department of Horticultural Environment, National Institute of Horticultural and Herbal Science, Rural Development Administration)
Choi, Seung-Kook (Virology Unit, Department of Horticultural Environment, National Institute of Horticultural and Herbal Science, Rural Development Administration)
Publication Information
Research in Plant Disease / v.21, no.3, 2015 , pp. 180-185 More about this Journal
Abstract
In the present work, a pair of primers specific to Tomato yellow leaf curl virus (TYLCV) was designed to allow specific amplification of DNA fragments from any TYLCV isolates using an extensive alignment of the complete genome sequences of TYLCV isolates deposited in the GenBank database. A pair of primers which allows the specific amplification of tomato ${\beta}$-tubulin gene was also analyzed as an internal PCR control. A duplex PCR method with the developed primer sets showed that TYLCV could be directly detected from the leaf crude sap of infected tomato plants. In addition, our developed duplex PCR method could determine PCR errors for TYLCV diagnosis, suggesting that this duplex PCR method with the primer sets is a good tool for specific and sensitive TYLCV diagnosis. The developed duplex PCR method was further verified from tomato samples collected from some farms in Korea, suggesting that this developed PCR method is a simple and reliable tool for rapid and large-scale TYLCV detections in tomato plants.
Keywords
Detection; Duplex PCR; Internal Control; Tomato; TYLCV;
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