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PLANTS INTERACTING WITH OTHER ORGANISMS

Diverse Subcellular Locations of Cryptogein-Induced Reactive Oxygen Species Production in Tobacco Bright Yellow-2 Cells^1,[W],[OA]

Department of Plant Science, Weizmann Institute of Science, Rehovot 76100, Israel (C.A., V.K., O.D., R.F.); and Jacob Blaustein Institute for Desert Research, Albert Katz Department of Dryland Biotechnologies, Ben-Gurion University, Beer Sheva 84105, Israel (M.S.)

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 (Nicotiana tabacum) Bright Yellow-2 suspension cells. Using Amplex Red and Amplex Ultra Red reagents, which 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'-dichlorofluorescein diacetate 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, 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.

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: Robert Fluhr (robert.fluhr{at}weizmann.ac.il).

^[W] The online version of this article contains Web-only data.

^[OA] Open Access articles can be viewed online without a subscription.

www.plantphysiol.org/cgi/doi/10.1104/pp.106.090902

^* Corresponding author; e-mail robert.fluhr{at}weizmann.ac.il; fax 972–8–9344181.

Received October 5, 2006; accepted January 27, 2007; published February 2, 2007.

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