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The RootScope: a simple high-throughput screening system for quantitating gene expression dynamics in plant roots

BACKGROUND: High temperature stress responses are vital for plant survival. The mechanisms that plants use to sense high temperatures are only partially understood and involve multiple sensing and signaling pathways. Here we describe the development of the RootScope, an automated microscopy system f...

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Autores principales: Kast, Erin J, Nguyen, Minh-Duyen T, Lawrence, Rosalie E, Rabeler, Christina, Kaplinsky, Nicholas J
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3852858/
https://www.ncbi.nlm.nih.gov/pubmed/24119322
http://dx.doi.org/10.1186/1471-2229-13-158
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author Kast, Erin J
Nguyen, Minh-Duyen T
Lawrence, Rosalie E
Rabeler, Christina
Kaplinsky, Nicholas J
author_facet Kast, Erin J
Nguyen, Minh-Duyen T
Lawrence, Rosalie E
Rabeler, Christina
Kaplinsky, Nicholas J
author_sort Kast, Erin J
collection PubMed
description BACKGROUND: High temperature stress responses are vital for plant survival. The mechanisms that plants use to sense high temperatures are only partially understood and involve multiple sensing and signaling pathways. Here we describe the development of the RootScope, an automated microscopy system for quantitating heat shock responses in plant roots. RESULTS: The promoter of Hsp17.6 was used to build a Hsp17.6(p):GFP transcriptional reporter that is induced by heat shock in Arabidopsis. An automated fluorescence microscopy system which enables multiple roots to be imaged in rapid succession was used to quantitate Hsp17.6(p):GFP response dynamics. Hsp17.6(p):GFP signal increased with temperature increases from 28°C to 37°C. At 40°C the kinetics and localization of the response are markedly different from those at 37°C. This suggests that different mechanisms mediate heat shock responses above and below 37°C. Finally, we demonstrate that Hsp17.6(p):GFP expression exhibits wave like dynamics in growing roots. CONCLUSIONS: The RootScope system is a simple and powerful platform for investigating the heat shock response in plants.
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spelling pubmed-38528582013-12-07 The RootScope: a simple high-throughput screening system for quantitating gene expression dynamics in plant roots Kast, Erin J Nguyen, Minh-Duyen T Lawrence, Rosalie E Rabeler, Christina Kaplinsky, Nicholas J BMC Plant Biol Methodology Article BACKGROUND: High temperature stress responses are vital for plant survival. The mechanisms that plants use to sense high temperatures are only partially understood and involve multiple sensing and signaling pathways. Here we describe the development of the RootScope, an automated microscopy system for quantitating heat shock responses in plant roots. RESULTS: The promoter of Hsp17.6 was used to build a Hsp17.6(p):GFP transcriptional reporter that is induced by heat shock in Arabidopsis. An automated fluorescence microscopy system which enables multiple roots to be imaged in rapid succession was used to quantitate Hsp17.6(p):GFP response dynamics. Hsp17.6(p):GFP signal increased with temperature increases from 28°C to 37°C. At 40°C the kinetics and localization of the response are markedly different from those at 37°C. This suggests that different mechanisms mediate heat shock responses above and below 37°C. Finally, we demonstrate that Hsp17.6(p):GFP expression exhibits wave like dynamics in growing roots. CONCLUSIONS: The RootScope system is a simple and powerful platform for investigating the heat shock response in plants. BioMed Central 2013-10-12 /pmc/articles/PMC3852858/ /pubmed/24119322 http://dx.doi.org/10.1186/1471-2229-13-158 Text en Copyright © 2013 Kast et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology Article
Kast, Erin J
Nguyen, Minh-Duyen T
Lawrence, Rosalie E
Rabeler, Christina
Kaplinsky, Nicholas J
The RootScope: a simple high-throughput screening system for quantitating gene expression dynamics in plant roots
title The RootScope: a simple high-throughput screening system for quantitating gene expression dynamics in plant roots
title_full The RootScope: a simple high-throughput screening system for quantitating gene expression dynamics in plant roots
title_fullStr The RootScope: a simple high-throughput screening system for quantitating gene expression dynamics in plant roots
title_full_unstemmed The RootScope: a simple high-throughput screening system for quantitating gene expression dynamics in plant roots
title_short The RootScope: a simple high-throughput screening system for quantitating gene expression dynamics in plant roots
title_sort rootscope: a simple high-throughput screening system for quantitating gene expression dynamics in plant roots
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3852858/
https://www.ncbi.nlm.nih.gov/pubmed/24119322
http://dx.doi.org/10.1186/1471-2229-13-158
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