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BREAKTHROUGH TECHNOLOGIES

Self-Reporting Arabidopsis Expressing pH and [Ca²⁺] Indicators Unveil Ion Dynamics in the Cytoplasm and in the Apoplast under Abiotic Stress^1,[w]

Institut für Pflanzenernährung und Bodenkunde, Christian-Albrechts-Universität, D–24098 Kiel, Germany (D.G., B.S.); Department of Plant Sciences, University of Oxford, South Parks Road, Oxford OX1 3RB, United Kingdom (M.R.K.); Institute of Cell and Molecular Biology (Botany), Mayfield Road, Edinburgh EH9 3JH, United Kingdom (A.J.T.); and Zentrum für Biochemie und Molekularbiologie, Universität Kiel, Am Botanischen Garten 9, 24118 Kiel, Germany (C.P.)

For noninvasive in vivo measurements of intra- and extracellular ion concentrations, we produced transgenic Arabidopsis expressing pH and calcium indicators in the cytoplasm and in the apoplast. Ratiometric pH-sensitive derivatives of the green fluorescent protein (At-pHluorins) were used as pH indicators. For measurements of calcium ([Ca²⁺]), luminescent aequorin variants were expressed in fusion with pHluorins. An Arabidopsis chitinase signal sequence was used to deliver the indicator complex to the apoplast. Responses of pH and [Ca²⁺] in the apoplast and in the cytoplasm were studied under salt and "drought" (mannitol) stress. Results are discussed in the frame of ion flux, regulation, and signaling. They suggest that osmotic stress and salt stress are differently sensed, compiled, and processed in plant cells.

¹ This work was supported by the Deutsche Forschungsgemeinschaft (grant nos. SA 359/12–3 to B.S. and D.G. and PL253/1–1 to C.P.) and by the European Commission (grant no. BIO4–CT97–5080 to C.P.).

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

^* Corresponding author; e-mail cplieth{at}zbm.uni-kiel.de; fax 49–431–880–4368.

Received August 28, 2003; returned for revision October 2, 2003; accepted November 25, 2003.

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