Plant Physiology Preview
Published on February 2, 2007; 10.1104/pp.106.090902
OPEN ACCESS ARTICLE
Received October 5, 2006
Accepted January 27, 2007
Diverse Subcellular Locations of Cryptogein-induced ROS Production in Tobacco BY-2 Cells
Cher Ashtamker , Vladimir Kiss , Moshe Sagi , Olga Davydov , and Robert Fluhr *
Department of Plant Science, Weizmann Institute of Science, P.O. Box 26, Rehovot 76100, Israel; The Jacob Blaustein Institute for Desert Research, The Albert Katz Department of Dryland Biotechnologies, Ben-Gurion University, P.O. Box 653, Beer Sheva 84105, Israel
* Corresponding author; email: robert.fluhr{at}weizmann.ac.il.
Reactive oxygen species (ROS) play a crucial role in many cellular responses and signaling pathways including the oxidative burst defense response to pathogens. We have examined very early events in cryptogein-induced ROS production in tobacco BY-2 suspension cells. Using Amplex red and Amplex ultra red reagents that report real-time H2O2 accumulation in cell populations, we show that the internal signal for H2O2 develops more rapidly than the external apoplastic signal. Subcellular accumulation of H2O2 was also followed in individual cells using the 2', 7'-dichlorofluorescin diacetate (DCF) fluorescent probe. Major accumulation was detected in endomembrane, cytoplasmic and nuclear compartments. When cryptogein was added, the signal developed first in the nuclear region and only after a short delay in the cell periphery. Interestingly, isolated nuclei were capable of producing H2O2 in a calcium dependent manner, implying that nuclei can serve as a potential active source of ROS production. These results show complex spatial compartmentalization for ROS accumulation and an unexpected temporal sequence of events that occurs after cryptogein application suggesting novel intricacy in ROS signaling cascades.
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