Plant Physiology Preview
Published on November 24, 2004; 10.1104/pp.104.046573
Received May 17, 2004
Returned for revision September 22, 2004
Accepted September 24, 2004
Inhibition of Blue Light-Dependent H+ Pumping by Abscisic Acid through Hydrogen Peroxide-Induced Dephosphorylation of the Plasma Membrane H+-ATPase in Guard Cell Protoplasts
Xiao Zhang , Hengbin Wang , Atsushi Takemiya , Chun-peng Song , Toshinori Kinoshita , and Ken-ichiro Shimazaki *
Department of Biology, Faculty of Science, Kyushu University, Ropponmatsu, Fukuoka, Japan 810-8560
* Corresponding author; email: kenrcb{at}mbox.nc.kyushu-u.ac.jp.
Blue light (BL)-dependent H+ pumping by guard cells, which drives stomatal opening, is inhibited by abscisic acid (ABA). We investigated this response with respect to the activity of plasma membrane H+-ATPase using Vicia guard cell protoplasts. ATP hydrolysis by the plasma membrane H+-ATPase, phosphorylation of the H+-ATPase, and the binding of 14-3-3 protein to the H+-ATPase stimulated by BL were inhibited by ABA at 10 µM. All of these responses were similarly inhibited by hydrogen peroxide (H2O2) at 1 mM. The ABA-induced inhibitions of BL-dependent H+ pumping and phosphorylation of the H+-ATPase were partially restored by ascorbate, an intracellular H2O2 scavenger. A single-cell analysis of the cytosolic H2O2 using 2',7'-dichlorofluorescin revealed that H2O2 was generated by ABA in guard cell protoplasts. We also indicated that H+ pumping induced by fusicoccin and the binding of 14-3-3 protein to the H+-ATPase were inhibited slightly (approximately 20%) by both ABA and H2O2. By contrast, H2O2 at 1 mM did not affect H+ pumping by the H+-ATPase in microsomal membranes. From these results, we concluded that inhibition of BL-dependent H+ pumping by ABA was due to a decrease in the phosphorylation levels of H+-ATPase and that H2O2 might be involved in this response. Moreover, there are at least two inhibition sites by ABA in the BL signaling pathway of guard cells.
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