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Open Access Highly Accessed Methodology article

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

Erin J Kast, Minh-Duyen T Nguyen, Rosalie E Lawrence, Christina Rabeler and Nicholas J Kaplinsky*

Author Affiliations

Department of Biology, Swarthmore College, Swarthmore, PA, 19081, USA

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BMC Plant Biology 2013, 13:158  doi:10.1186/1471-2229-13-158

Published: 12 October 2013

Abstract

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 Hsp17.6p: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 Hsp17.6p:GFP response dynamics. Hsp17.6p: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 Hsp17.6p: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.

Keywords:
Heat shock; Thermometer; Thermostat; Automated imaging; Quantitative imaging; Temperature; Arabidopsis