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JXB Advance Access originally published online on March 30, 2006
Journal of Experimental Botany 2006 57(7):1537-1546; doi:10.1093/jxb/erj129
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© The Author [2006]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

RESEARCH PAPER

Proteome analysis of cold stress response in Arabidopsis thaliana using DIGE-technology

Steffen Amme, Andrea Matros, Bernhard Schlesier and Hans-Peter Mock*

Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstrasse 3, D-06466 Gatersleben, Germany

*To whom correspondence should be addressed. E-mail:mock{at}ipk-gatersleben.de

A proteome study based on 2-D gel electrophoresis was performed in order to analyse the cold-stress response of Arabidopsis plants. The emphasis was to monitor the overall changes in the protein complement after prolonged exposure rather than short-term responses. Two different temperature regimes were used (6 °C and 10 °C) and plants were exposed to cold-stress exposure for 1 week. Protein patterns were also monitored after re-shifting plants to control conditions for a further week. To monitor gradual changes in the response to the two cold-stress conditions, the analysis was performed with DIGE technology with the inclusion of an internal standard. In the experiments using 6 °C, 22 spots with at least 2-fold altered expression were found; among them 18 were increased and four were decreased. When plants were exposed to 10 °C, 18 of these 22 spots still showed a 2-fold change; however, the alterations were, in general, more moderate than observed under 6 °C. Spot identification was performed by MALDI-TOF and ESI-MS/MS. Many of the proteins identified have previously been described in the context of cold-stress responses, indicating the validity of this proteome approach for further in-depth studies.

Key words: 2-D DIGE, Arabidopsis, cold stress, differential in- gel electrophoresis, ESI-MS/MS, fluorescent dyes, MALDI-TOF, mass spectrometry, proteome analysis, two-dimensional gel electrophoresis


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