Multiplex real-time PCR for the detection of Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and pathogenic Xanthomonas species on tomato plants

A multiplex real-time PCR method based on fluorescent TaqMan® probes was developed for the simultaneous detection of the tomato pathogenic bacteria Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and bacterial spot-causing xanthomonads. The specificity of the multiple...

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Autores principales: Peňázová, Eliška, Dvořák, Miloň, Ragasová, Lucia, Kiss, Tomáš, Pečenka, Jakub, Čechová, Jana, Eichmeier, Aleš
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6946519/
https://www.ncbi.nlm.nih.gov/pubmed/31910230
http://dx.doi.org/10.1371/journal.pone.0227559
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author Peňázová, Eliška
Dvořák, Miloň
Ragasová, Lucia
Kiss, Tomáš
Pečenka, Jakub
Čechová, Jana
Eichmeier, Aleš
author_facet Peňázová, Eliška
Dvořák, Miloň
Ragasová, Lucia
Kiss, Tomáš
Pečenka, Jakub
Čechová, Jana
Eichmeier, Aleš
author_sort Peňázová, Eliška
collection PubMed
description A multiplex real-time PCR method based on fluorescent TaqMan® probes was developed for the simultaneous detection of the tomato pathogenic bacteria Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and bacterial spot-causing xanthomonads. The specificity of the multiplex assay was validated on 44 bacterial strains, including 32 target pathogen strains as well as closely related species and nontarget tomato pathogenic bacteria. The designed multiplex real-time PCR showed high sensitivity when positive amplification was observed for one pg of bacterial DNA in the cases of Clavibacter michiganensis subsp. michiganensis and Pseudomonas syringae pv. tomato bacteria and 100 pg for bacterial spot-causing xanthomonads. The reliability of the developed multiplex real-time PCR assay for in planta detection was verified by recognition of the target pathogens in 18 tomato plants artificially inoculated by each of the target bacteria and tomato samples from production greenhouses.
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spelling pubmed-69465192020-01-17 Multiplex real-time PCR for the detection of Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and pathogenic Xanthomonas species on tomato plants Peňázová, Eliška Dvořák, Miloň Ragasová, Lucia Kiss, Tomáš Pečenka, Jakub Čechová, Jana Eichmeier, Aleš PLoS One Research Article A multiplex real-time PCR method based on fluorescent TaqMan® probes was developed for the simultaneous detection of the tomato pathogenic bacteria Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and bacterial spot-causing xanthomonads. The specificity of the multiplex assay was validated on 44 bacterial strains, including 32 target pathogen strains as well as closely related species and nontarget tomato pathogenic bacteria. The designed multiplex real-time PCR showed high sensitivity when positive amplification was observed for one pg of bacterial DNA in the cases of Clavibacter michiganensis subsp. michiganensis and Pseudomonas syringae pv. tomato bacteria and 100 pg for bacterial spot-causing xanthomonads. The reliability of the developed multiplex real-time PCR assay for in planta detection was verified by recognition of the target pathogens in 18 tomato plants artificially inoculated by each of the target bacteria and tomato samples from production greenhouses. Public Library of Science 2020-01-07 /pmc/articles/PMC6946519/ /pubmed/31910230 http://dx.doi.org/10.1371/journal.pone.0227559 Text en © 2020 Peňázová et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Peňázová, Eliška
Dvořák, Miloň
Ragasová, Lucia
Kiss, Tomáš
Pečenka, Jakub
Čechová, Jana
Eichmeier, Aleš
Multiplex real-time PCR for the detection of Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and pathogenic Xanthomonas species on tomato plants
title Multiplex real-time PCR for the detection of Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and pathogenic Xanthomonas species on tomato plants
title_full Multiplex real-time PCR for the detection of Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and pathogenic Xanthomonas species on tomato plants
title_fullStr Multiplex real-time PCR for the detection of Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and pathogenic Xanthomonas species on tomato plants
title_full_unstemmed Multiplex real-time PCR for the detection of Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and pathogenic Xanthomonas species on tomato plants
title_short Multiplex real-time PCR for the detection of Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and pathogenic Xanthomonas species on tomato plants
title_sort multiplex real-time pcr for the detection of clavibacter michiganensis subsp. michiganensis, pseudomonas syringae pv. tomato and pathogenic xanthomonas species on tomato plants
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6946519/
https://www.ncbi.nlm.nih.gov/pubmed/31910230
http://dx.doi.org/10.1371/journal.pone.0227559
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