Abscisic acid and CO2 signalling via calcium sensitivity priming in guard cells, new CDPK mutant phenotypes and a method for improved resolution of stomatal stimulus-response analyses.

Abstract

Stomatal guard cells are the regulators of gas exchange between plants and the atmosphere. Ca(2+)-dependent and Ca(2+)-independent mechanisms function in these responses. Key stomatal regulation mechanisms, including plasma membrane and vacuolar ion channels have been identified and are regulated by the free cytosolic Ca(2+) concentration ([Ca(2+)](cyt)). Here we show that CO(2)-induced stomatal closing is strongly impaired under conditions that prevent intracellular Ca(2+) elevations. Moreover, Ca(2+) oscillation-induced stomatal closing is partially impaired in knock-out mutations in several guard cell-expressed Ca(2+)-dependent protein kinases (CDPKs) here, including the cpk4cpk11 double and cpk10 mutants; however, abscisic acid-regulated stomatal movements remain relatively intact in the cpk4cpk11 and cpk10 mutants. We further discuss diverse studies of Ca(2+) signalling in guard cells, discuss apparent peculiarities, and pose novel open questions. The recently proposed Ca(2+) sensitivity priming model could account for many of the findings in the field. Recent research shows that the stomatal closing stimuli abscisic acid and CO(2) enhance the sensitivity of stomatal closing mechanisms to intracellular Ca(2+), which has been termed 'calcium sensitivity priming'. The genome of the reference plant Arabidopsis thaliana encodes for over 250 Ca(2+)-sensing proteins, giving rise to the question, how can specificity in Ca(2+) responses be achieved? Calcium sensitivity priming could provide a key mechanism contributing to specificity in eukaryotic Ca(2+) signal transduction, a topic of central interest in cell signalling research. In this article we further propose an individual stomatal tracking method for improved analyses of stimulus-regulated stomatal movements in Arabidopsis guard cells that reduces noise and increases fidelity in stimulus-regulated stomatal aperture responses ( Box 1). This method is recommended for stomatal response research, in parallel to previously adopted blind analyses, due to the relatively small and diverse sizes of stomatal apertures in the reference plant Arabidopsis thaliana.