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

Abstract

BackgroundHigh 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.ResultsThe 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.ConclusionsThe RootScope system is a simple and powerful platform for investigating the heat shock response in plants.

Breeding Cultivars for Heat Stress Tolerance in Staple Food Crops

“…Recently, high-resolution thermal imaging system was used to precisely measure the leaf temperature [194]. The phenotyping platforms established to screen for HT include "HTpheno" for image analysis [195] and "Rootscope" used to quantify heat-shock responses in plants [196]. In the near future, the new-generation phenomics platforms would allow cost-effective and user-friendly screening for HT in crop plants.…”

Integrated “omics” approaches to sustain global productivity of major grain legumes under heat stress

“…Similarly, to measure leaf temperature, high‐resolution thermal imaging system was used (Benavente, García‐Toledano, Carrillo, & Quemada, ; Fahlgren et al., ; Prashar & Jones, ) (for details, see Humplík, Lazár, Husičková, & Spíchal, ). To have better insight into heat shock response in plant, “root scope” phenotyping system was developed (Kast, Nguyen, Lawrence, Rabeler, & Kaplinsky, ). The chlorophyll fluorescence ( Fv/Fm ) under HS has been implicated in wheat as an important parameter to assess HS tolerance (Haque, Kjaer, Rosenqvist, Sharma, & Ottosen, ; Sharma, Andersen, Ottosen, & Rosenqvist, , ; Shirdelmoghanloo, Cozzolino, Lohraseb, & Collins, ).…”

“…To avoid laboratory-based testing of heat tolerance, "heat tolerance testing system" (HTTS) has been introduced to measure the impact of HS in situ under "natural solar irradiation" condition (Buchner, Karadar, Bauer, & Neuner, 2013). Despite remarkable progress in the establishment of HTPP in cereal and other crops for HS (Kast et al, 2013;Munns, James, Sirault, Furbank, & Jones, 2010), the immense potential of these systems is yet to be elucidated in to specific stress including HS (Arbona et al, 2013;Rodziewicz et al, 2014). Ultimately, outcome of these complex networks enables in characterizing phenotype of plant under stress, viz.…”

Heat stress in crop plants: its nature, impacts and integrated breeding strategies to improve heat tolerance

“…Rootscope represents another phenotyping system that is recently used to quantify heat‐shock responses in plants (Kast et al. ). Excellent reviews were published in recent years describing applications, benefits and challenges of the new‐generation phenotyping systems (Sozzani and Benfey , Cobb et al.…”

“…In recent years, various platforms were established for phenotyping, including 'HTpheno' for image analysis (Hartmann et al 2011), 'PHENOPSIS DB' to perform image analyses under different environmental conditions in Arabidopsis (Fabre et al 2011), 'Gia Roots' for root architecture analysis (Galkovskyi et al 2012) and 'Phenoscope' for spatial homogeneity (Tisne et al 2013). Rootscope represents another phenotyping system that is recently used to quantify heat-shock responses in plants (Kast et al 2013). Excellent reviews were published in recent years describing applications, benefits and challenges of the new-generation phenotyping systems (Sozzani andBenfey 2011, Cobb et al 2013).…”