mRNA from selected genes is useful for specific detection and quantification of viable Xanthomonas citri subsp citri

The purpose of this study was to assess the stability of mRNA and rRNA for evaluation of viability for Xanthomonas citri subsp. citri (Xcc). Total RNA from Xcc suspensions subjected to different stress treatments (high temperature or chemical treatment with sodium orthophenylphenate at different con...

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Autores principales: Golmohammadi, Morteza, Llop, Pablo, Scuderi, Giuseppe, Gell, I., Graham, James H., Cubero, Jaime
Formato: article
Lenguaje:Inglés
Publicado: 2017
Acceso en línea:http://hdl.handle.net/20.500.11939/5289
https://bsppjournals.onlinelibrary.wiley.com/doi/epdf/10.1111/j.1365-3059.2011.02526.x
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author Golmohammadi, Morteza
Llop, Pablo
Scuderi, Giuseppe
Gell, I.
Graham, James H.
Cubero, Jaime
author_browse Cubero, Jaime
Gell, I.
Golmohammadi, Morteza
Graham, James H.
Llop, Pablo
Scuderi, Giuseppe
author_facet Golmohammadi, Morteza
Llop, Pablo
Scuderi, Giuseppe
Gell, I.
Graham, James H.
Cubero, Jaime
author_sort Golmohammadi, Morteza
collection ReDivia
description The purpose of this study was to assess the stability of mRNA and rRNA for evaluation of viability for Xanthomonas citri subsp. citri (Xcc). Total RNA from Xcc suspensions subjected to different stress treatments (high temperature or chemical treatment with sodium orthophenylphenate at different concentrations) was extracted at different time periods post-treatment (0, 3, 24 and 48 h) and analysed by quantitative real-time reverse transcription PCR (Q-RT-PCR). Primers were designed from selected fragments of rRNA and mRNA from genes involved in bacterial fitness, virulence or general metabolic mechanisms (gumD, rpfB, avrBs2 and gyrB). After stress treatment, only a 445-bp fragment from the gumD mRNA was detected in live Xcc cells specifically, whereas other RNA fragments, as well as DNA targets, were detected in both viable and nonviable cells. Statistical analyses demonstrated that the amount of some transcripts from genes involved in xanthan synthesis, pathogenicity factor regulation and DNA processing was significantly reduced after lethal treatments. The amplification of the 445-bp product from gumD mRNA was demonstrated to be useful for the detection of viable Xcc; the product was detected specifically from viable bacteria on leaf and citrus fruit surfaces and in citrus canker lesions. Instability of long RNA fragments can be used as a practical tool for the study of survival of citrus canker bacteria or for diagnostic purposes when the presence of viable bacteria needs to be confirmed.
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institution Instituto Valenciano de Investigaciones Agrarias (IVIA)
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spelling ReDivia52892025-04-25T14:41:54Z mRNA from selected genes is useful for specific detection and quantification of viable Xanthomonas citri subsp citri Golmohammadi, Morteza Llop, Pablo Scuderi, Giuseppe Gell, I. Graham, James H. Cubero, Jaime The purpose of this study was to assess the stability of mRNA and rRNA for evaluation of viability for Xanthomonas citri subsp. citri (Xcc). Total RNA from Xcc suspensions subjected to different stress treatments (high temperature or chemical treatment with sodium orthophenylphenate at different concentrations) was extracted at different time periods post-treatment (0, 3, 24 and 48 h) and analysed by quantitative real-time reverse transcription PCR (Q-RT-PCR). Primers were designed from selected fragments of rRNA and mRNA from genes involved in bacterial fitness, virulence or general metabolic mechanisms (gumD, rpfB, avrBs2 and gyrB). After stress treatment, only a 445-bp fragment from the gumD mRNA was detected in live Xcc cells specifically, whereas other RNA fragments, as well as DNA targets, were detected in both viable and nonviable cells. Statistical analyses demonstrated that the amount of some transcripts from genes involved in xanthan synthesis, pathogenicity factor regulation and DNA processing was significantly reduced after lethal treatments. The amplification of the 445-bp product from gumD mRNA was demonstrated to be useful for the detection of viable Xcc; the product was detected specifically from viable bacteria on leaf and citrus fruit surfaces and in citrus canker lesions. Instability of long RNA fragments can be used as a practical tool for the study of survival of citrus canker bacteria or for diagnostic purposes when the presence of viable bacteria needs to be confirmed. 2017-06-01T10:12:04Z 2017-06-01T10:12:04Z 2012 JUN 2012 article publishedVersion Golmohammadi, M., Llop, P., Scuderi, G., Gell, I., Graham, J. H. & Cubero, J. (2012). mRNA from selected genes is useful for specific detection and quantification of viable Xanthomonas citri subsp citri. Plant Pathology, 61(3), 479-488. 0032-0862 http://hdl.handle.net/20.500.11939/5289 10.1111/j.1365-3059.2011.02526.x https://bsppjournals.onlinelibrary.wiley.com/doi/epdf/10.1111/j.1365-3059.2011.02526.x en openAccess Impreso
spellingShingle Golmohammadi, Morteza
Llop, Pablo
Scuderi, Giuseppe
Gell, I.
Graham, James H.
Cubero, Jaime
mRNA from selected genes is useful for specific detection and quantification of viable Xanthomonas citri subsp citri
title mRNA from selected genes is useful for specific detection and quantification of viable Xanthomonas citri subsp citri
title_full mRNA from selected genes is useful for specific detection and quantification of viable Xanthomonas citri subsp citri
title_fullStr mRNA from selected genes is useful for specific detection and quantification of viable Xanthomonas citri subsp citri
title_full_unstemmed mRNA from selected genes is useful for specific detection and quantification of viable Xanthomonas citri subsp citri
title_short mRNA from selected genes is useful for specific detection and quantification of viable Xanthomonas citri subsp citri
title_sort mrna from selected genes is useful for specific detection and quantification of viable xanthomonas citri subsp citri
url http://hdl.handle.net/20.500.11939/5289
https://bsppjournals.onlinelibrary.wiley.com/doi/epdf/10.1111/j.1365-3059.2011.02526.x
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