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A DNA-based real-time PCR assay for robust growth quantification of the bacterial pathogen Pseudomonas syringae on Arabidopsis thaliana
BACKGROUND: The interaction of Pseudomonas syringae with Arabidopsis is one of the most commonly used systems to study various bacterial—host interrelationships. Currently, most studies are based on the growth quantification of the pathogen to characterize resistance or virulence targets. However, t...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5117497/ https://www.ncbi.nlm.nih.gov/pubmed/27895701 http://dx.doi.org/10.1186/s13007-016-0149-z |
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author | Ross, Annegret Somssich, Imre E. |
author_facet | Ross, Annegret Somssich, Imre E. |
author_sort | Ross, Annegret |
collection | PubMed |
description | BACKGROUND: The interaction of Pseudomonas syringae with Arabidopsis is one of the most commonly used systems to study various bacterial—host interrelationships. Currently, most studies are based on the growth quantification of the pathogen to characterize resistance or virulence targets. However, the standard available method for determining bacterial proliferation in planta is laborious and has several limitations. RESULTS: Here we present an alternative robust approach, which is based on the quantification of bacterial DNA by real-time PCR. We directly compared this assay with the routinely used plate counting method to access bacterial titers in a number of well described Arabidopsis mutants. CONCLUSIONS: These studies showed that the DNA-based technique is highly reliable and comparable. Moreover, the technique is easily applicable, robust, and ideal for routine experiments or for larger scale analyses. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13007-016-0149-z) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5117497 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-51174972016-11-28 A DNA-based real-time PCR assay for robust growth quantification of the bacterial pathogen Pseudomonas syringae on Arabidopsis thaliana Ross, Annegret Somssich, Imre E. Plant Methods Methodology BACKGROUND: The interaction of Pseudomonas syringae with Arabidopsis is one of the most commonly used systems to study various bacterial—host interrelationships. Currently, most studies are based on the growth quantification of the pathogen to characterize resistance or virulence targets. However, the standard available method for determining bacterial proliferation in planta is laborious and has several limitations. RESULTS: Here we present an alternative robust approach, which is based on the quantification of bacterial DNA by real-time PCR. We directly compared this assay with the routinely used plate counting method to access bacterial titers in a number of well described Arabidopsis mutants. CONCLUSIONS: These studies showed that the DNA-based technique is highly reliable and comparable. Moreover, the technique is easily applicable, robust, and ideal for routine experiments or for larger scale analyses. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13007-016-0149-z) contains supplementary material, which is available to authorized users. BioMed Central 2016-11-21 /pmc/articles/PMC5117497/ /pubmed/27895701 http://dx.doi.org/10.1186/s13007-016-0149-z Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Methodology Ross, Annegret Somssich, Imre E. A DNA-based real-time PCR assay for robust growth quantification of the bacterial pathogen Pseudomonas syringae on Arabidopsis thaliana |
title | A DNA-based real-time PCR assay for robust growth quantification of the bacterial pathogen Pseudomonas syringae on Arabidopsis thaliana |
title_full | A DNA-based real-time PCR assay for robust growth quantification of the bacterial pathogen Pseudomonas syringae on Arabidopsis thaliana |
title_fullStr | A DNA-based real-time PCR assay for robust growth quantification of the bacterial pathogen Pseudomonas syringae on Arabidopsis thaliana |
title_full_unstemmed | A DNA-based real-time PCR assay for robust growth quantification of the bacterial pathogen Pseudomonas syringae on Arabidopsis thaliana |
title_short | A DNA-based real-time PCR assay for robust growth quantification of the bacterial pathogen Pseudomonas syringae on Arabidopsis thaliana |
title_sort | dna-based real-time pcr assay for robust growth quantification of the bacterial pathogen pseudomonas syringae on arabidopsis thaliana |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5117497/ https://www.ncbi.nlm.nih.gov/pubmed/27895701 http://dx.doi.org/10.1186/s13007-016-0149-z |
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