Plant Physiol. Drug Metab Dispos
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Plant Physiology 141:397-403 (2006)
© 2006 American Society of Plant Biologists

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BREAKTHROUGH TECHNOLOGIES

Expression and Characterization of a Redox-Sensing Green Fluorescent Protein (Reduction-Oxidation-Sensitive Green Fluorescent Protein) in Arabidopsis

Keni Jiang, Christian Schwarzer, Elizabeth Lally, Shibo Zhang, Steven Ruzin, Terry Machen, S. James Remington and Lewis Feldman*

Department of Plant and Microbial Biology (K.J., E.L., S.Z., S.R., L.F.) and Department of Molecular and Cell Biology, Division of Cell and Developmental Biology (C.S., T.M.), University of California, Berkeley, California 94720; and Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403–1229 (J.R.)

Arabidopsis (Arabidopsis thaliana) was transformed with a redox-sensing green fluorescent protein (reduction-oxidation-sensitive green fluorescent protein [roGFP]), with expression targeted to either the cytoplasm or to the mitochondria. Both the mitochondrial and cytosolic forms are oxidation-reduction sensitive, as indicated by a change in the ratio of 510 nm light (green light) emitted following alternating illumination with 410 and 474 nm light. The 410/474 fluorescence ratio is related to the redox potential (in millivolts) of the organelle, cell, or tissue. Both forms of roGFP can be reduced with dithiothreitol and oxidized with hydrogen peroxide. The average resting redox potentials for roots are –318 mV for the cytoplasm and –362 mV for the mitochondria. The elongation zone of the Arabidopsis root has a more oxidized redox status than either the root cap or meristem. Mitochondria are much better than the cytoplasm, as a whole, at buffering changes in redox. The data show that roGFP is redox sensitive in plant cells and that this sensor makes it possible to monitor, in real time, dynamic changes in redox in vivo.

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: Lewis Feldman (feldman{at}nature.berkeley.edu).

www.plantphysiol.org/cgi/doi/10.1104/pp.106.078246.

* Corresponding author; e-mail feldman{at}nature.berkeley.edu; fax 510–642–4995.

Received January 31, 2006; returned for revision March 28, 2006; accepted March 31, 2006.




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