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First published online November 30, 2007; 10.1104/pp.107.106450

Plant Physiology 146:505-514 (2008)
© 2008 American Society of Plant Biologists

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CELL BIOLOGY AND SIGNAL TRANSDUCTION

Cytoplasmic Calcium Increases in Response to Changes in the Gravity Vector in Hypocotyls and Petioles of Arabidopsis Seedlings1

Masatsugu Toyota, Takuya Furuichi, Hitoshi Tatsumi and Masahiro Sokabe*

Department of Physiology, Nagoya University Graduate School of Medicine, Showa-ku, Nagoya, Aichi 466–8550, Japan (M.T., T.F., H.T., M.S.); Molecular Plant Physiology, University of Erlangen, Erlangen D–91058, Germany (T.F.); International Cooperative Research Project/Solution Oriented Research for Science and Technology, Cell Mechanosensing Project, Japan Science and Technology Agency, Showa-ku, Nagoya, Aichi 466–8550, Japan (M.S.); and Department of Molecular Physiology, National Institute for Physiological Sciences, Okazaki, Aichi 444–8585, Japan (M.S.)

Plants respond to a large variety of environmental signals, including changes in the gravity vector (gravistimulation). In Arabidopsis (Arabidopsis thaliana) seedlings, gravistimulation is known to increase the cytoplasmic free calcium concentration ([Ca2+]c). However, organs responsible for the [Ca2+]c increase and the underlying cellular/molecular mechanisms remain to be solved. In this study, using Arabidopsis seedlings expressing apoaequorin, a Ca2+-sensitive luminescent protein in combination with an ultrasensitive photon counting camera, we clarified the organs where [Ca2+]c increases in response to gravistimulation and characterized the physiological and pharmacological properties of the [Ca2+]c increase. When the seedlings were gravistimulated by turning 180°, they showed a transient biphasic [Ca2+]c increase in their hypocotyls and petioles. The second peak of the [Ca2+]c increase depended on the angle but not the speed of rotation, whereas the initial peak showed diametrically opposite characters. This suggests that the second [Ca2+]c increase is specific for changes in the gravity vector. The potential mechanosensitive Ca2+-permeable channel (MSCC) inhibitors Gd3+ and La3+, the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA), and the endomembrane Ca2+-permeable channel inhibitor ruthenium red suppressed the second [Ca2+]c increase, suggesting that it arises from Ca2+ influx via putative MSCCs in the plasma membrane and Ca2+ release from intracellular Ca2+ stores. Moreover, the second [Ca2+]c increase was attenuated by actin-disrupting drugs cytochalasin B and latrunculin B but not by microtubule-disrupting drugs oryzalin and nocodazole, implying that actin filaments are partially involved in the hypothetical activation of Ca2+-permeable channels. These results suggest that the second [Ca2+]c increase via MSCCs is a gravity response in the hypocotyl and petiole of Arabidopsis seedlings.


1 This work was supported by the Japan Society for the Promotion of Science for Young Scientists (research fellowships to M.T.), by International Cooperative Research Project/Solution Oriented Research for Science and Technology (Japan Science and Technology Agency; grant to M.S.), by the Ministry of Education, Science, Sports and Culture (grant-in-aid for General Scientific Research nos. 13480216 to M.S. and 14580769 to H.T., grant-in-aid for Scientific Research on Priority Areas no. 15086270 to M.S., and grant-in-aid for Creative Research no. 16GS0308 to M.S.), and by the Japan Space Forum (to H.T. and M.S.).

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Masahiro Sokabe (msokabe{at}med.nagoya-u.ac.jp).

www.plantphysiol.org/cgi/doi/10.1104/pp.107.106450

* Corresponding author; e-mail msokabe{at}med.nagoya-u.ac.jp.

Received July 31, 2007; accepted November 16, 2007; published November 30, 2007.







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