4.7 Article Proceedings Paper

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

Journal

JOURNAL OF EXPERIMENTAL BOTANY
Volume 57, Issue 7, Pages 1537-1546

Publisher

OXFORD UNIV PRESS
DOI: 10.1093/jxb/erj129

Keywords

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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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 degrees C and 10 degrees 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 degrees 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 degrees C, 18 of these 22 spots still showed a 2-fold change; however, the alterations were, in general, more moderate than observed under 6 degrees 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.

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