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Fast detection by loop-mediated isothermal amplification (LAMP) of the three begomovirus species infecting tomato in Panama

Abstract

Potato yellow mosaic Panama virus (PYMPV), Tomato leaf curl Sinaloa virus (ToLCSiV) and Tomato yellow mottle virus (TYMoV) of genus Begomovirus (family Geminiviridae) are the only three begomovirus species detected infecting tomato (Solanum lycopersicum L.) in Panama. PYMPV, ToLCSiV and TYMoV induce symptoms of stunting, yellowing, curling, distortion of leaves and reduction of fruit size and cause important economic loses. A loop-mediated amplification under isothermal conditions (LAMP) assay was developed for the individual detection of these three begomovirus species by using a set of three primer pairs specific per each one of them. Amplification products were visualized by gel electrophoresis or direct Gel-Red staining of DNA into the reaction tube. PYMPV, ToLCSiV and TYMoV were detected in total DNA extracts obtained from different plant tissues such as leaves, stems, flowers, fruits and roots of infected tomato plants collected in different production regions of Panama. LAMP sensitivity was similar to that of conventional PCR but, the first procedure was faster and cheaper than the last one. Moreover, all three viruses were successfully detected by LAMP and not by conventional PCR from sap extracts obtained from leaf tissues of infected tomato plants which were embedded into 3MM Whatman paper and stored several days, facilitating the samples processing as well as the material movement among different laboratories. Therefore, LAMP is a specific, rapid and cheap procedure to detect all three begomoviruses infecting tomato in Panama and it is suitable for field surveys and sanitation programs.

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Acknowledgements

This work was supported by the project FID14-020 of the Secretaría Nacional de Ciencia, Tecnología e Innovación (SENACYT) of Panama. Dr. J.A. Herrera-Vázquez was recipient of a National Research Investigator award (SNI) from the SENACYT for a training stage in the Virology laboratory of IVIA under Dr. L. Galipienso’s supervision. A. Puchades was recipient of a specialization fellowship from IVIA and C. Carpino was recipient of a predoctoral fellowship from the University of Palermo, Sicily (Italy). We thank Dr. M. I. Font-San-Ambrosio (UPV, Valencia, Spain) for providing the TYLCV and ToLCNDV isolates used in this work.

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Correspondence to L. Galipienso.

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All authors in this manuscript have read and approved current version of the article. The part of the paper corresponding to detection of PYMPV, ToLCSiV and TYMoV by conventional PCR from tomato field samples which was previously published has been referenced in the manuscript.

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Herrera-Vásquez, J.A., Puchades, A.V., Elvira-González, L. et al. Fast detection by loop-mediated isothermal amplification (LAMP) of the three begomovirus species infecting tomato in Panama. Eur J Plant Pathol 151, 243–250 (2018). https://doi.org/10.1007/s10658-017-1358-7

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  • DOI: https://doi.org/10.1007/s10658-017-1358-7

Keywords

  • PYMPV
  • ToLCSiV
  • TYMoV
  • LAMP
  • Begomovirus
  • Solanum lycopersicum