JXB Advance Access originally published online on November 26, 2007
Journal of Experimental Botany 2007 58(15-16):4183-4194; doi:10.1093/jxb/erm275
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© 2007 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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RESEARCH PAPER |
Inhibition of catalase activity as an early response of Arabidopsis thaliana cultured cells to the phytotoxin fusicoccin
Istituto di Biofisica del CNR-Sezione di Milano, Dipartimento di Biologia, Università degli Studi di Milano, Via Celoria 26, 20133 Milano, Italy
* To whom correspondence should be addressed. E-mail: nicoletta.beffagna{at}unimi.it
In Arabidopsis thaliana cells, fusicoccin (FC) treatment induced an early and marked increase in the extracellular H2O2 level. It also increased the huge hypo-osmotic stress-induced oxidative wave and, in addition, prevented the H2O2 peak drop. These effects were apparently not linked to changes in either cytoplasmic pH or cytoplasmic free calcium concentration, since they occurred independently of the activity state of the plasma membrane (PM) H+-ATPase and neither influx nor efflux of 45Ca2+ was modified by FC. In the presence of diphenylene iodonium (DPI), inhibiting the PM NADPH oxidase presumably responsible for reactive oxygen species (ROS) production, no apoplastic H2O2 development was detected either with or without FC. However, no increase in DPI-sensitive ferricyanide reduction, but rather a gradual decrease, occurred with FC. These results suggested that the H2O2 increase observed with FC was not due to a overproduction of ROS but, more probably, to a reduced capability of FC-treated cells to degrade the H2O2 formed. This view, at first supported by the finding that FC-treated cells failed to break down exogenously supplied H2O2, was clearly confirmed by a series of measurements on exogenous catalase activity, tested in cell-free media of FC-treated samples. This assay, in fact, allowed ascertainment and partial characterization of an as yet unidentified factor increasingly accumulating in the incubation medium of FC-treated cells, behaving as a non-competitive catalase inhibitor and able to reduce markedly the cell's capability for H2O2 scavenging.
Key words: Arabidopsis thaliana cells, catalase activity, cytoplasmic pH, fusicoccin, H2O2 scavenging, reactive oxygen species, stress
Received 24 July 2007; Revised 11 October 2007 Accepted 15 October 2007