Amme Steffen, Matros Andrea, Schlesier Bernhard, Mock Hans-Peter
Leibniz Institute of Plant Genetics and Crop Plant Research, Corrensstrasse 3, D-06466 Gatersleben, Germany.
J Exp Bot. 2006;57(7):1537-46. doi: 10.1093/jxb/erj129. Epub 2006 Mar 30.
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.
为了分析拟南芥植物的冷胁迫响应,进行了一项基于二维凝胶电泳的蛋白质组研究。重点是监测长时间暴露后蛋白质组的整体变化,而非短期响应。使用了两种不同的温度条件(6摄氏度和10摄氏度),并将植物暴露于冷胁迫1周。在将植物重新转移到对照条件下再过1周后,也监测了蛋白质模式。为了监测对两种冷胁迫条件响应的逐渐变化,采用DIGE技术并加入内标进行分析。在使用6摄氏度的实验中,发现22个斑点的表达至少有2倍变化;其中18个增加,4个减少。当植物暴露于10摄氏度时,这22个斑点中有18个仍显示2倍变化;然而,总体而言,这些变化比在6摄氏度下观察到的更为温和。通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)和电喷雾串联质谱(ESI-MS/MS)进行斑点鉴定。许多鉴定出的蛋白质此前已在冷胁迫响应的背景下被描述过,这表明这种蛋白质组方法对于进一步深入研究的有效性。