European Journal of Plant Pathology

, Volume 143, Issue 4, pp 779–787 | Cite as

Simultaneous detection of Clavibacter michiganensis subsp. michiganensis, Pepino mosaic virus and Mexican papita viroid by non-radioactive molecular hybridization using a unique polyprobe

  • Erika Janet Zamora-Macorra
  • Daniel Leobardo Ochoa-Martínez
  • Guadalupe Valdovinos-Ponce
  • Reyna Rojas-Martínez
  • Sergio Ramírez-Rojas
  • Jesús Ángel Sánchez-Navarro
  • Vicente Pallás
  • Frederic Aparicio
Article
  • 273 Downloads

Abstract

Clavibacter michiganensis subsp. michiganensis, Pepino mosaic virus, and Mexican papita viroid are three economically important pathogens that infect tomato crops. In this work, a polyprobe (poly-3) was developed and evaluated for the simultaneous detection of these pathogens in tomato plants by non-isotopic molecular hybridization. The endpoint detection limit of the poly-3 with C. michiganensis subsp. michiganensis cell cultures was 106 CFU/ml. No differences were found in terms of the sensitivity and specificity when the individual riboprobes were compared to the poly-3 for the detection of the three pathogens. The analysis of 80 tomato field samples by the poly-3 and RT-PCR techniques rendered the same number of positive samples for each pathogen. As far as we know this is the first time that three pathogens with very different life cycle styles (bacteria, virus and viroid) are simultaneously detected in a single assay. The possibility of using this poly-3 technology for the routine diagnosis of field tomato samples is discussed.

Keywords

Tomato Clavibacter michiganensis subsp. michiganensis Pepino mosaic virus Mexican papita viroid Polyriboprobe Non-isotopic molecular hybridization 

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Copyright information

© Koninklijke Nederlandse Planteziektenkundige Vereniging 2015

Authors and Affiliations

  • Erika Janet Zamora-Macorra
    • 1
  • Daniel Leobardo Ochoa-Martínez
    • 1
  • Guadalupe Valdovinos-Ponce
    • 1
  • Reyna Rojas-Martínez
    • 1
  • Sergio Ramírez-Rojas
    • 2
  • Jesús Ángel Sánchez-Navarro
    • 3
  • Vicente Pallás
    • 3
  • Frederic Aparicio
    • 3
  1. 1.Colegio de PostgraduadosTexcocoMéxico
  2. 2.Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias- ZacatepecMorelosMéxico
  3. 3.Instituto de Biología Molecular y Celular de PlantasUniversidad Politécnica de Valencia (UPV-CSIC)ValenciaSpain

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