\u3cem\u3eHSFA1d\u3c/em\u3e Regulates The Kinetics Of Heat-Induced \u3cem\u3eHSP17.6\u3c/em\u3e Expression In Arabidopsis

Arabidopsis contains four HSFA1 heat shock transcription factors (HSFA1a, HSFA1b, HSFA1d, and HSFA1e) that regulate the primary response to high temperature stress responses. These genes have overlapping functions and, while double and triple HSFA1 mutants have thermotolerance phenotypes, these genes have no reported single mutant thermotolerance phenotypes. We used an automated fluorescence microscopy system to quantitate the expression of a HSP17.6:GFP reporter with high temporal resolution to show that HSFA1d is required for normal heat-induced HSP17.6 expression. HSP17.6 expression is reduced and delayed in hsfa1d-1 mutants. This finding highlights the power of using gene expression kinetics as a quantitative phenotype for discovering the function of genes that exhibit functional redundancy

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 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 Hsp17.6p: GFP response dynamics. Hsp17.6(p):GFP signal increased with temperature increases from 28 degrees C to 37 degrees C. At 40 degrees C the kinetics and localization of the response are markedly different from those at 37 degrees C. This suggests that different mechanisms mediate heat shock responses above and below 37 degrees 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