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The use of quantitative imaging to investigate regulators of membrane trafficking in Arabidopsis stomatal closure
Author(s) -
Bourdais Gildas,
McLachlan Deirdre H.,
Rickett Lydia M.,
Zhou Ji,
Siwoszek Agnieszka,
Häweker Heidrun,
Hartley Matthew,
Kuhn Hannah,
Morris Richard J.,
MacLean Dan,
Robatzek Silke
Publication year - 2019
Publication title -
traffic
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.677
H-Index - 130
eISSN - 1600-0854
pISSN - 1398-9219
DOI - 10.1111/tra.12625
Subject(s) - biology , arabidopsis , guard cell , microbiology and biotechnology , gtpase , mutant , endomembrane system , genetic screen , small gtpase , computational biology , gene , live cell imaging , genetics , signal transduction , cell , endoplasmic reticulum , golgi apparatus
Expansion of gene families facilitates robustness and evolvability of biological processes but impedes functional genetic dissection of signalling pathways. To address this, quantitative analysis of single cell responses can help characterize the redundancy within gene families. We developed high‐throughput quantitative imaging of stomatal closure, a response of plant guard cells, and performed a reverse genetic screen in a group of Arabidopsis mutants to five stimuli. Focussing on the intersection between guard cell signalling and the endomembrane system, we identified eight clusters based on the mutant stomatal responses. Mutants generally affected in stomatal closure were mostly in genes encoding SNARE and SCAMP membrane regulators. By contrast, mutants in RAB5 GTPase genes played specific roles in stomatal closure to microbial but not drought stress. Together with timed quantitative imaging of endosomes revealing sequential patterns in FLS2 trafficking, our imaging pipeline can resolve non‐redundant functions of the RAB5 GTPase gene family. Finally, we provide a valuable image‐based tool to dissect guard cell responses and outline a genetic framework of stomatal closure.