Signal transduction pathway(s) in guard cells after prolonged exposure to low vapour pressure deficit

<p>Keywords: Abscisic acid, Arabidopsis thaliana, calcium, CYP707As, desiccation, environmental factors, guard cells’ signalling pathway, hydrogen peroxide, natural variation, nitric oxide, photosystem II efficiency, RD29A, relative water content, secondary messengers, stomata, vapour pressure deficit, Vicia faba</p> <p>In short-term, guard cells close stomata in response to an increase in vapour pressure deficit (VPD) and they open the stomata after exposure to low VPDs. However, in long-term responses to low VPD, adaptation processes occur which make stomata less sensitive to stimuli which usually induce stomatal closure (stomatal malfunctioning). Cellular mechanism(s) leading to occurrence of stomatal malfunctioning is (are) still unknown. The aim of this project was to elucidate the processes that are involved in the malfunctioning of stomata after long-term exposure to low VPD. To elucidate whether the problem of stomatal malfunctioning is due to alterations in stomatal morphology and leaf anatomy or in the ABA signalling pathway, fava bean plants were grown at low or moderate VPDs and some plants that developed their leaves at moderate VPD were then transferred for four days to low VPD. Leaf anatomical and stomatal morphological alterations due to low VPD were not the main reason of stomatal malfunctioning in response to ABA and desiccation. Within one day exposure to low VPD, the level of foliar ABA decreased to the same level as in low VPD-grown plants, while the level of ABA-glucose ester was not affected. Spraying ABA during a 4-day exposure to low VPD maintained closing ability of the stomata after 4-day low VPD-exposure. Therefore, alteration in the signalling pathways due to low foliar ABA level was recognized as the main reason for stomatal malfunctioning after long-term low VPD-exposure. Coincidence in changes of Ca2+, ABA receptors, and positive and negative regulators of ABA signalling are proposed as early steps for stomatal malfunctioning induced by low VPD-exposure. Transcriptional activators, transcriptional repressors as well as E3 ligases are proposed for long-term adaptation of cellular processes which consequently cause decreased stomatal response to closing stimuli afterwards. In order to find the molecular mechanism(s) of stomatal malfunctioning, possible variation in stomatal response to closing stimuli was studied among <em>Arabidopsis thaliana </em>accessions after a 4-day low VPD-exposure. Accessions could be grouped to very sensitive, moderately sensitive and less sensitive to closing stimuli using principle component analysis. A positive correlation was found between foliar ABA level (before desiccation) and stomatal closure response to ABA (but not to desiccation) after exposure to different VPDs. Stomatal response to desiccation was positively correlated with the foliar ABA level after desiccation. In order to elucidate the molecular network underlying stomatal malfunctioning in response to ABA due to long-term low VPD-exposure, two groups of <em>Arabidopsis</em> accessions were used as accessions that maintained responsiveness to ABA after low VPD-exposure and accessions with low VPD induced non-ABA-responsive stomata. The foliar ABA content in all accessions correlated with the stomatal response to ABA: only when the ABA level was above a threshold value, stomata responded to ABA. After low VPD-exposure, mainly due to catabolism of ABA, the foliar ABA content decreased. This decrease in ABA level resulted in down regulation of <em>RD29A</em>, which caused decreased stomatal responsiveness to ABA.</p> <p>