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

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 Hsp17.6pGFP 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 Hsp17.6pGFP response dynamics. Hsp17.6pGFP 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 Hsp17.6pGFP 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.