Journal of Plant Protection Research (2013) 53, 289-294

Przemyslaw Wieczorek and Aleksandra Obrepalska-Steplowska (2013)
Multiplex RT-PCR reaction for simultaneous detection of Tomato torrado virus and Pepino mosaic virus co-infecting Solanum lycopersicum
Journal of Plant Protection Research 53 (3), 289-294
Abstract: Tomato (Solanum lycopersicum L.) is cultivated all over the world and is a vegetable of significant economic importance. However, an increased production of the vegetable is directly connected with an elevated occurrence of pathogens limiting the production efficiency of the vegetable. Both, Tomato torrado virus and Pepino mosaic virus have been found to be serious disease factors. When not controlled, these viruses can significantly decrease tomato cultivation. In this article, we report a multiplex reverse transcription-polymerase chain reaction (RT-PCR) protocol for simultaneous detection of both, Tomato torrado virus (ToTV) and Pepino mosaic virus (PepMV) in virus infected plants. The assay was designed to specifically amplify the conserved regions of genomic ribonucleic acid (RNA) of both viruses. Moreover, the glycerandehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control of amplification to exclude false-negative assay results. High-resolution melt analysis of generated RT-PCR products was additionally performed to increase sensitivity and double-check the specificity of the reaction without the need of subsequent complementary deoxyribonucleic acid (cDNA) sequencing.
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Database assignments for author(s): Aleksandra Obrepalska-Steplowska

Research topic(s) for pests/diseases/weeds:
identification/taxonomy
surveys/sampling/distribution

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Pepino mosaic virus	Tomato (Solanum lycopersicum)
	Torradovirus lycopersici	Tomato (Solanum lycopersicum)	Poland
	Torradovirus lycopersici	Tomato (Solanum lycopersicum)	Spain (continental)