Jones Alexandra M E, Thomas Vincent, Bennett Mark H, Mansfield John, Grant Murray
Department of Agricultural Science, Imperial College London, Wye TN25 5AH, United Kingdom.
Plant Physiol. 2006 Dec;142(4):1603-20. doi: 10.1104/pp.106.086231. Epub 2006 Oct 6.
Alterations in the proteome of Arabidopsis (Arabidopsis thaliana) leaves during responses to challenge by Pseudomonas syringae pv tomato DC3000 were analyzed using two-dimensional gel electrophoresis. Protein changes characteristic of the establishment of disease, basal resistance, and resistance-gene-mediated resistance were examined by comparing responses to DC3000, a hrp mutant, and DC3000 expressing avrRpm1, respectively. The abundance of each protein identified was compared with that of selected transcripts obtained from comparable GeneChip experiments. We report changes in three subcellular fractions: total soluble protein, chloroplast enriched, and mitochondria enriched over four time points (1.5-6 h after inoculation). In total, 73 differential spots representing 52 unique proteins were successfully identified. Many of the changes in protein spot density occurred before significant transcriptional reprogramming was evident between treatments. The high proportion of proteins represented by more than one spot indicated that many of the changes to the proteome can be attributed to posttranscriptional modifications. Proteins found to show significant change after bacterial challenge are representative of two main functional groups: defense-related antioxidants and metabolic enzymes. Significant changes to photosystem II and to components of the mitochondrial permeability transition were also identified. Rapid communication between organelles and regulation of primary metabolism through redox-mediated signaling are supported by our data.
利用二维凝胶电泳分析了拟南芥叶片在应对丁香假单胞菌番茄致病变种DC3000侵染时蛋白质组的变化。通过分别比较对DC3000、一个hrp突变体以及表达avrRpm1的DC3000的反应,研究了疾病建立、基础抗性和抗性基因介导的抗性过程中蛋白质变化的特征。将鉴定出的每种蛋白质的丰度与从可比的基因芯片实验中获得的选定转录本的丰度进行比较。我们报告了在四个时间点(接种后1.5 - 6小时)三个亚细胞组分中的变化:总可溶性蛋白、叶绿体富集组分和线粒体富集组分。总共成功鉴定出73个差异点,代表52种独特的蛋白质。在处理之间明显的显著转录重编程之前,蛋白质斑点密度就发生了许多变化。由多个斑点代表的蛋白质比例很高,这表明蛋白质组的许多变化可归因于转录后修饰。发现细菌侵染后显示出显著变化的蛋白质代表两个主要功能组:与防御相关的抗氧化剂和代谢酶。还鉴定出了光系统II和线粒体通透性转换组分的显著变化。我们的数据支持了细胞器之间的快速通讯以及通过氧化还原介导的信号传导对初级代谢的调节。