TY - JOUR A1 - Dindas, Julian A1 - Dreyer, Ingo A1 - Huang, Shouguang A1 - Hedrich, Rainer A1 - Roelfsema, M. Rob G. T1 - A voltage-dependent Ca\(^{2+}\) homeostat operates in the plant vacuolar membrane JF - New Phytologist N2 - Cytosolic calcium signals are evoked by a large variety of biotic and abiotic stimuli and play an important role in cellular and long distance signalling in plants. While the function of the plasma membrane in cytosolic Ca\(^{2+}\) signalling has been intensively studied, the role of the vacuolar membrane remains elusive. A newly developed vacuolar voltage clamp technique was used in combination with live-cell imaging, to study the role of the vacuolar membrane in Ca\(^{2+}\) and pH homeostasis of bulging root hair cells of Arabidopsis. Depolarisation of the vacuolar membrane caused a rapid increase in the Ca\(^{2+}\) concentration and alkalised the cytosol, while hyperpolarisation led to the opposite responses. The relationship between the vacuolar membrane potential, the cytosolic pH and Ca2+ concentration suggests that a vacuolar H\(^{+}\)/Ca\(^{2+}\) exchange mechanism plays a central role in cytosolic Ca2+ homeostasis. Mathematical modelling further suggests that the voltage-dependent vacuolar Ca\(^{2+}\) homeostat could contribute to calcium signalling when coupled to a recently discovered K\(^{+}\) channel-dependent module for electrical excitability of the vacuolar membrane. KW - voltage clamp KW - Arabidopsis thaliana KW - calcium signalling KW - computational cell biology KW - cpYFP cytosolic pH reporter KW - R-GECO1 cytosolic Ca\(^{2+}\) reporter KW - TPC1 channel KW - vacuolar membrane Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-259627 VL - 230 IS - 4 ER - TY - JOUR A1 - Huang, Shouguang A1 - Ding, Meiqi A1 - Roelfsema, M. Rob G. A1 - Dreyer, Ingo A1 - Scherzer, Sönke A1 - Al-Rasheid, Khaled A. S A1 - Gao, Shiqiang A1 - Nagel, Georg A1 - Hedrich, Rainer A1 - Konrad, Kai R. T1 - Optogenetic control of the guard cell membrane potential and stomatal movement by the light-gated anion channel GtACR1 JF - Science Advances N2 - Guard cells control the aperture of plant stomata, which are crucial for global fluxes of CO\(_2\) and water. In turn, guard cell anion channels are seen as key players for stomatal closure, but is activation of these channels sufficient to limit plant water loss? To answer this open question, we used an optogenetic approach based on the light-gated anion channelrhodopsin 1 (GtACR1). In tobacco guard cells that express GtACR1, blue- and green-light pulses elicit Cl\(^-\) and NO\(_3\)\(^-\) currents of -1 to -2 nA. The anion currents depolarize the plasma membrane by 60 to 80 mV, which causes opening of voltage-gated K+ channels and the extrusion of K+. As a result, continuous stimulation with green light leads to loss of guard cell turgor and closure of stomata at conditions that provoke stomatal opening in wild type. GtACR1 optogenetics thus provides unequivocal evidence that opening of anion channels is sufficient to close stomata. KW - abscisic-acid activation KW - Arabidopsis thaliana KW - H+-atpase KW - signal transduction KW - potassium channel KW - intact plants KW - K+ channels KW - R-type KW - CO2 KW - SLAC1 Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-260925 VL - 7 IS - 28 ER -