Zou Junjie, Song Lianfen, Zhang Wenzheng, Wang Yi, Ruan Songlin, Wu Wei-Hua
State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
J Integr Plant Biol. 2009 May;51(5):438-55. doi: 10.1111/j.1744-7909.2009.00823.x. Epub 2009 Mar 30.
Proteomic analysis was applied to generating the map of Arabidopsis mature pollen proteins and analyzing the differentially expressed proteins that are potentially involved in the regulation of Arabidopsis pollen germination. By applying 2-D electrophoresis and silver staining, we resolved 499 and 494 protein spots from protein samples extracted from pollen grains and pollen tubes, respectively. Using the matrix-assisted laser desorption ionization time-of-flight mass spectrometry method, we identified 189 distinct proteins from 213 protein spots expressed in mature pollen or pollen tubes, and 75 new identified proteins that had not been reported before in research into the Arabidopsis pollen proteome. Comparative analysis revealed that 40 protein spots exhibit reproducible significant changes between mature pollen and pollen tubes. And 21 proteins from 17 downregulated and six upregulated protein spots were identified. Functional category analysis indicated that these differentially expressed proteins mainly involved in signaling, cellular structure, transport, defense/stress responses, transcription, metabolism, and energy production. The patterns of changes at protein level suggested the important roles for energy metabolism-related proteins in pollen tube growth, accompanied by the activation of the stress response pathway and modifications to the cell wall.
蛋白质组学分析被用于生成拟南芥成熟花粉蛋白质图谱,并分析可能参与拟南芥花粉萌发调控的差异表达蛋白质。通过二维电泳和银染,我们分别从花粉粒和花粉管提取的蛋白质样品中分离出499个和494个蛋白质斑点。使用基质辅助激光解吸电离飞行时间质谱法,我们从成熟花粉或花粉管中表达的213个蛋白质斑点中鉴定出189种不同的蛋白质,以及75种在拟南芥花粉蛋白质组研究中以前未报道过的新鉴定蛋白质。比较分析表明,40个蛋白质斑点在成熟花粉和花粉管之间表现出可重复的显著变化。并且从17个下调和6个上调的蛋白质斑点中鉴定出21种蛋白质。功能类别分析表明,这些差异表达的蛋白质主要参与信号传导、细胞结构、运输、防御/应激反应、转录、代谢和能量产生。蛋白质水平的变化模式表明能量代谢相关蛋白质在花粉管生长中起重要作用,同时伴随着应激反应途径的激活和细胞壁的修饰。