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The vacuolar Ca2+-activated channel TPC1 regulates germination and stomatal movement

Abstract

Cytosolic free calcium ([Ca2+]cyt) is a ubiquitous signalling component in plant cells1. Numerous stimuli trigger sustained or transient elevations of [Ca2+]cyt that evoke downstream stimulus-specific responses. Generation of [Ca2+]cyt signals is effected through stimulus-induced opening of Ca2+-permeable ion channels that catalyse a flux of Ca2+ into the cytosol from extracellular or intracellular stores. Many classes of Ca2+ current have been characterized electrophysiologically in plant membranes2. However, the identity of the ion channels that underlie these currents has until now remained obscure. Here we show that the TPC1 (‘two-pore channel 1’) gene of Arabidopsis thaliana encodes a class of Ca2+-dependent Ca2+-release channel that is known from numerous electrophysiological studies as the slow vacuolar channel3,4,5. Slow vacuolar channels are ubiquitous in plant vacuoles, where they form the dominant conductance at micromolar [Ca2+]cyt. We show that a tpc1 knockout mutant lacks functional slow vacuolar channel activity and is defective in both abscisic acid-induced repression of germination and in the response of stomata to extracellular calcium. These studies unequivocally demonstrate a critical role of intracellular Ca2+-release channels in the physiological processes of plants.

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Competing interests

The authors declare that they have no competing financial interests.

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Acknowledgements

We thank M. Fischer and R. V. Jolley for initial experiments, L. Skiera and S. Kilmartin for technical assistance, P. O'Toole for advice on confocal microscopy, G. Park for preparing the pART7GFP construct, M. G. Palmgren for providing AHA2 antibodies, and J. I. Schroeder for discussions. This work was funded by the Biotechnology and Biological Sciences Research Council and an award by the Koerber Foundation to D.S.

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Competing interests

The authors declare that they have no competing financial interests.

Correspondence to Dale Sanders.

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Further reading

Figure 1: Topology and vacuolar localization of the Arabidopsis TPC1 protein.
Figure 2: Characterization of TPC1-misexpressing Arabidopsis thaliana lines.
Figure 3: Electrophysiological characterisation of Arabidopsis mesophyll vacuolar membranes.
Figure 4: Phenotypic characterization of tpc1-2 mutants and TPC1 overexpressing lines.

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