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INTACT-based guard cell transcriptomes from a progressive drought time course reveal targets for modifying stomatal
Anna van Weringh1, Paul J Gamueda1, Hasna Khan1
1Department of Cell and Systems Biology/Centre for the Analysis of Genome Evolution and Function, University of Toronto, 25 Willcocks St., Toronto, ON, Canada M5S 3B2.
The Plant Cell
|September 19, 2025
Summary
Guard cells (GCs) uniquely respond to early drought stress by altering gene expression. This study reveals GCs tailor their responses to drought severity, identifying key genes for improving crop drought tolerance.
Area of Science:
- Plant Biology
- Environmental Stress Response
- Molecular Genetics
Background:
- Drought stress significantly limits global crop production.
- Guard cells (GCs) play a crucial role in regulating plant water loss.
- Understanding GC-specific responses to drought is vital for crop improvement.
Purpose of the Study:
- To investigate gene expression changes in guard cells during progressive drought stress.
- To compare GC-specific responses with those of bulk leaf tissue.
- To identify candidate genes involved in drought tolerance.
Main Methods:
- Utilized the INTACT (isolation of nuclei tagged in specific cell types) system for GC-specific RNA sequencing.
- Optimized the INTACT protocol for Arabidopsis thaliana leaf tissue with fixation.
- Generated RNA-seq data from GCs and bulk leaf tissue under varying drought conditions and re-watering.
- Performed thermal imaging screens on mutants of candidate drought-responsive genes.
Main Results:
- Guard cells exhibit unique gene expression changes at the earliest stages of drought stress.
- GCs demonstrate a tailored gene expression response to drought severity, differing from bulk leaf tissue.
- A significant percentage of moderate drought-responsive genes in GCs were not altered under severe drought.
- Ten candidate early drought-responsive genes were identified through thermal imaging, showing altered phenotypes under drought.
Conclusions:
- Guard cells possess a distinct and adaptable gene expression strategy for drought response.
- These findings provide novel insights into the molecular mechanisms of plant drought tolerance.
- Identified candidate genes offer potential targets for genetic engineering to enhance crop resilience to drought.
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