Transcriptome-based identification and validation of reference genes for plant-bacteria interaction studies using <i>Nicotiana benthamiana</i>

RT-qPCR is a widely used technique for the analysis of gene expression. Accurate estimation of transcript abundance relies strongly on a normalization that requires the use of reference genes that are stably expressed in the conditions analyzed. Initially, they were adopted from those used in Northe...

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Detalles Bibliográficos
Autores principales: Pombo, Marina Alejandra, Ramos, Romina Nair, Zheng, Yi, Fei, Zhangjun, Martin, Gregory B., Rosli, Hernán Guillermo
Formato: Articulo
Lenguaje:Inglés
Publicado: 2019
Materias:
Ciencias Agrarias
Ciencias Naturales
Nicotiana benthamiana
Pseudomonas fuorescens
leaves
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/107857
http://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC6367355&blobtype=pdf
https://www.nature.com/articles/s41598-018-38247-2
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-107857
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Agrarias
Ciencias Naturales
Nicotiana benthamiana
Pseudomonas fuorescens
leaves
spellingShingle Ciencias Agrarias
Ciencias Naturales
Nicotiana benthamiana
Pseudomonas fuorescens
leaves
Pombo, Marina Alejandra
Ramos, Romina Nair
Zheng, Yi
Fei, Zhangjun
Martin, Gregory B.
Rosli, Hernán Guillermo
Transcriptome-based identification and validation of reference genes for plant-bacteria interaction studies using <i>Nicotiana benthamiana</i>
topic_facet Ciencias Agrarias
Ciencias Naturales
Nicotiana benthamiana
Pseudomonas fuorescens
leaves
description RT-qPCR is a widely used technique for the analysis of gene expression. Accurate estimation of transcript abundance relies strongly on a normalization that requires the use of reference genes that are stably expressed in the conditions analyzed. Initially, they were adopted from those used in Northern blot experiments, but an increasing number of publications highlight the need to fnd and validate alternative reference genes for the particular system under study. The development of high-throughput sequencing techniques has facilitated the identifcation of such stably expressed genes. <i>Nicotiana benthamiana</i> has been extensively used as a model in the plant research feld. In spite of this, there is scarce information regarding suitable RT-qPCR reference genes for this species. Employing RNA-seq data previously generated from tomato plants, combined with newly generated data from <i>N. benthamiana</i> leaves infltrated with <i>Pseudomonas fuorescens</i>, we identifed and tested a set of 9 candidate reference genes. Using three diferent algorithms, we found that <i>NbUbe35</i>, <i>NbNQO</i> and <i>NbErpA</i> exhibit less variable gene expression in our pathosystem than previously used genes. Furthermore, the combined use of the frst two is sufcient for robust gene expression analysis. We encourage employing these novel reference genes in future RT-qPCR experiments involving <i>N. benthamiana</i> and <i>Pseudomonas</i> spp.
format Articulo
Articulo
author Pombo, Marina Alejandra
Ramos, Romina Nair
Zheng, Yi
Fei, Zhangjun
Martin, Gregory B.
Rosli, Hernán Guillermo
author_facet Pombo, Marina Alejandra
Ramos, Romina Nair
Zheng, Yi
Fei, Zhangjun
Martin, Gregory B.
Rosli, Hernán Guillermo
author_sort Pombo, Marina Alejandra
title Transcriptome-based identification and validation of reference genes for plant-bacteria interaction studies using <i>Nicotiana benthamiana</i>
title_short Transcriptome-based identification and validation of reference genes for plant-bacteria interaction studies using <i>Nicotiana benthamiana</i>
title_full Transcriptome-based identification and validation of reference genes for plant-bacteria interaction studies using <i>Nicotiana benthamiana</i>
title_fullStr Transcriptome-based identification and validation of reference genes for plant-bacteria interaction studies using <i>Nicotiana benthamiana</i>
title_full_unstemmed Transcriptome-based identification and validation of reference genes for plant-bacteria interaction studies using <i>Nicotiana benthamiana</i>
title_sort transcriptome-based identification and validation of reference genes for plant-bacteria interaction studies using <i>nicotiana benthamiana</i>
publishDate 2019
url http://sedici.unlp.edu.ar/handle/10915/107857
http://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC6367355&blobtype=pdf
https://www.nature.com/articles/s41598-018-38247-2
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