Cargando…

Transcriptome and proteome responses in RNAlater preserved tissue of Arabidopsis thaliana

Tissue preservation is a minimal requirement for the success of plant RNA and protein expression studies. The standard of snap-freezing in liquid nitrogen is not always practical or possible. RNAlater, a concentrated solution of ammonium and cesium sulfates, has become a standard preservative in the...

Descripción completa

Detalles Bibliográficos
Autores principales: Kruse, Colin P. S., Basu, Proma, Luesse, Darron R., Wyatt, Sarah E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5397022/
https://www.ncbi.nlm.nih.gov/pubmed/28423006
http://dx.doi.org/10.1371/journal.pone.0175943
_version_ 1783230185411182592
author Kruse, Colin P. S.
Basu, Proma
Luesse, Darron R.
Wyatt, Sarah E.
author_facet Kruse, Colin P. S.
Basu, Proma
Luesse, Darron R.
Wyatt, Sarah E.
author_sort Kruse, Colin P. S.
collection PubMed
description Tissue preservation is a minimal requirement for the success of plant RNA and protein expression studies. The standard of snap-freezing in liquid nitrogen is not always practical or possible. RNAlater, a concentrated solution of ammonium and cesium sulfates, has become a standard preservative in the absence of liquid nitrogen. Here, we demonstrate the effectiveness of RNAlater in preserving both RNA and proteins in Arabidopsis thaliana tissues for use in RNAseq and LC-MS/MS analysis of proteins. While successful in preserving plant material, a transcriptomic and proteomic response is evident. Specifically, 5770 gene transcripts, 84 soluble proteins, and 120 membrane-bound proteins were found to be differentially expressed at a log-fold change of ±1 (P ≤ 0.05). This response is mirrored in the abundance of post-translational modifications, with 23 of the 108 (21.3%) phosphorylated proteins showing altered abundance at a log-fold change of ±1 (P ≤ 0.05). While RNAlater is effective in preserving biological information, our findings warrant caution in its use for transcriptomic and proteomic experiments.
format Online
Article
Text
id pubmed-5397022
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-53970222017-05-04 Transcriptome and proteome responses in RNAlater preserved tissue of Arabidopsis thaliana Kruse, Colin P. S. Basu, Proma Luesse, Darron R. Wyatt, Sarah E. PLoS One Research Article Tissue preservation is a minimal requirement for the success of plant RNA and protein expression studies. The standard of snap-freezing in liquid nitrogen is not always practical or possible. RNAlater, a concentrated solution of ammonium and cesium sulfates, has become a standard preservative in the absence of liquid nitrogen. Here, we demonstrate the effectiveness of RNAlater in preserving both RNA and proteins in Arabidopsis thaliana tissues for use in RNAseq and LC-MS/MS analysis of proteins. While successful in preserving plant material, a transcriptomic and proteomic response is evident. Specifically, 5770 gene transcripts, 84 soluble proteins, and 120 membrane-bound proteins were found to be differentially expressed at a log-fold change of ±1 (P ≤ 0.05). This response is mirrored in the abundance of post-translational modifications, with 23 of the 108 (21.3%) phosphorylated proteins showing altered abundance at a log-fold change of ±1 (P ≤ 0.05). While RNAlater is effective in preserving biological information, our findings warrant caution in its use for transcriptomic and proteomic experiments. Public Library of Science 2017-04-19 /pmc/articles/PMC5397022/ /pubmed/28423006 http://dx.doi.org/10.1371/journal.pone.0175943 Text en © 2017 Kruse 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
Kruse, Colin P. S.
Basu, Proma
Luesse, Darron R.
Wyatt, Sarah E.
Transcriptome and proteome responses in RNAlater preserved tissue of Arabidopsis thaliana
title Transcriptome and proteome responses in RNAlater preserved tissue of Arabidopsis thaliana
title_full Transcriptome and proteome responses in RNAlater preserved tissue of Arabidopsis thaliana
title_fullStr Transcriptome and proteome responses in RNAlater preserved tissue of Arabidopsis thaliana
title_full_unstemmed Transcriptome and proteome responses in RNAlater preserved tissue of Arabidopsis thaliana
title_short Transcriptome and proteome responses in RNAlater preserved tissue of Arabidopsis thaliana
title_sort transcriptome and proteome responses in rnalater preserved tissue of arabidopsis thaliana
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5397022/
https://www.ncbi.nlm.nih.gov/pubmed/28423006
http://dx.doi.org/10.1371/journal.pone.0175943
work_keys_str_mv AT krusecolinps transcriptomeandproteomeresponsesinrnalaterpreservedtissueofarabidopsisthaliana
AT basuproma transcriptomeandproteomeresponsesinrnalaterpreservedtissueofarabidopsisthaliana
AT luessedarronr transcriptomeandproteomeresponsesinrnalaterpreservedtissueofarabidopsisthaliana
AT wyattsarahe transcriptomeandproteomeresponsesinrnalaterpreservedtissueofarabidopsisthaliana