Feng Yalan, Kong Beibei, Zhang Jun, Chen Xianni, Yuan Jiale, Tang Xingwang, Ma Chao
College of Agriculture, Henan University of Science and Technology, Luoyang 471023, China.
Protein Pept Lett. 2018;25(3):260-274. doi: 10.2174/0929866525666180118103039.
Vernalization is one of the pivotal ways for plants to flower. The twodimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-offlight/ time-of-flight mass spectrometry (MALDI-TOF/TOF MS) were applied to analyze the changes in protein expression profiles in responding to vernalization in leaves of wheat seedling before (0d) and after (30d) of vernalization.
The main objective of this study was to analyze the vernalization-responsive proteins in winter wheat after vernalization.
Winter wheat seedling leaf proteins were extracted by phenol extraction coupled with ammonium acetate in methanol. 2-DE was conducted according to procedures described in the manual given by the GE manufacture. The selected protein spots were identified by MALDITOF/ TOF MS. Gene ontology (GO) classification was applied to classify the functions of the differentially expressed proteins. Pathway enrichment analysis identified significantly enriched metabolic pathways or signal transduction pathways relative to the whole proteins background.
The results of 2-DE and MALDI-TOF/TOF MS showed that among the 65 differentially expressed proteins that were successfully identified under vernalization, 30 were up-regulated whereas 35 were down-regulated after vernalization, respectively. These vernalization-responsive proteins were found to play roles in carbohydrate metabolism, protein metabolism, photosynthesis, defense and stress-resistance and may therefore participate in many biological processes in responding to vernalization. The enhanced accumulation of proteins after vernalization, such as thiamine thiazole synthase, late embryogenesis abundant protein, and glutathione-S-transferase, probably play vital roles in the mechanisms underlying vernalization response in wheat.
Our results indicated these vernalization-responsive proteins were found to be involved in protein metabolism, carbohydrate metabolism, photosynthesis, and stress resistance/ defense. The responses of plants to low temperature were very complex, involving in a wide range of cellular pathways for signal transduction, gene regulation, protein modifications, and metabolic regulation. Studying on wheat proteomic profiles in response to vernalization can improve our understanding the molecular mechanisms underlying vernalization in cereals. The results obtained in this study have provided a novel insight into the mechanisms underlying vernalization in cereal crops.
春化作用是植物开花的关键途径之一。采用双向凝胶电泳（2-DE）和基质辅助激光解吸/电离飞行时间/飞行时间质谱（MALDI-TOF/TOF MS）技术，分析了冬小麦幼苗叶片在春化处理前（0天）和春化处理后（30天）响应春化过程中蛋白质表达谱的变化。
本研究的主要目的是分析春化处理后冬小麦中响应春化的蛋白质。
采用苯酚提取法结合甲醇中的醋酸铵提取冬小麦幼苗叶片蛋白质。按照通用电气公司手册中描述的程序进行双向凝胶电泳。通过MALDI-TOF/TOF MS鉴定所选蛋白质斑点。应用基因本体（GO）分类对差异表达蛋白质的功能进行分类。通路富集分析确定相对于整个蛋白质背景显著富集的代谢途径或信号转导途径。
双向凝胶电泳和MALDI-TOF/TOF MS结果表明，在春化处理下成功鉴定的65种差异表达蛋白质中，春化处理后30种上调，35种下调。这些响应春化的蛋白质在碳水化合物代谢、蛋白质代谢、光合作用、防御和抗逆性中发挥作用，因此可能参与响应春化的许多生物学过程。春化处理后蛋白质积累的增加，如硫胺噻唑合酶、晚期胚胎发生丰富蛋白和谷胱甘肽-S-转移酶，可能在小麦春化响应机制中起重要作用。
我们的结果表明，这些响应春化的蛋白质参与蛋白质代谢、碳水化合物代谢、光合作用和抗逆性/防御。植物对低温的反应非常复杂，涉及广泛的细胞信号转导、基因调控、蛋白质修饰和代谢调控途径。研究小麦响应春化的蛋白质组图谱可以增进我们对谷物春化分子机制的理解。本研究获得的结果为谷类作物春化机制提供了新的见解。
