烟草叶片接种野火病菌后蛋白质表达差异

Protein differential expression of tobacco leaves induced with Pseudomonas syringae pv. tabaci

为了解烟草受野火病菌侵染后蛋白质表达变化情况,探讨烟草品种的抗病机制,本研究对烟草抗病品种‘K346’叶片进行人工接种,在接种后0～72h内分9次在接种点周围取无病原菌叶片组织提取叶片总蛋白,利用双向电泳技术分析接种处理叶片与未处理叶片的蛋白质表达差异,并选取6个减量表达蛋白点和4个增量表达蛋白点进行MALDI-TOF-MS质谱分析及数据库搜索鉴定,结果表明,6个减量表达蛋白包括参与植物呼吸代谢途径的ATP合成酶CF1α亚基、ATP合成酶CF1α链以及参与植物能量代谢循环途径的1,5-二磷酸核酮糖羧化酶/加氧酶相关蛋白;4个增量表达蛋白包括参与植物氨基酸代谢途径的谷氨酰胺合成酶、参与糖类和碳循环相关途径的高等植物光系统II氧复合物中Psbp A链蛋白、叶绿体景天庚酮糖-1,7-二磷酸酯酶和光捕获叶绿素a/b结合蛋白。经过蛋白质功能分析,推测烟草是通过加速植株生长、促进抗病相关糖和蛋白的表达、增强氨基酸代谢以及植株自身蛋白的修复等方式来提高抗病性。

In order to understand the changes of protein and investigate the resistance mechanism of tobacco after infected with Pseudomonas syringae pv. tabaci, leaves of resistant tobacco variety ＇ K346＇ were inoculated with the pathogen of wildfire disease, and leaf tissues around inoculation sites were sampled at 0-72 h after inoculation, and total proteins were extracted and differentially expressed proteins between treatment group and control （0 h sample） were analyzed using two-dimensional electrophoresis （2-DE）. Ten differentially expressed protein spots, including six down-regulated and four up-regulated, were selected for MALDI-TOF MS analysis and database searching. Six down-regulated proteins contained ATP synthase CFla subunit and ATP synthase CFla chain which participate in respiration metabolism pathways of plant, and ribulose-1, 5-biphosphate arboxylase/oxygenase-related proteins, which participate in the pathways of energy metabolism. Four up-regulated proteins contained glutamine synthetase, which participates in amino acid metabolism pathways of plant, photosystem II oxygen complexes Psbp A chain, chloroplast sedoheptulose-1,7-biphosphatase and light-harvesting chlorophyll a/b binding protein, which participate in sugar and carbon circulation relevant pathways. It was shown that tobacco increases resistance to wildfire disease through accelerating plant growth, promoting synthesis of resistance-associated sugar and proteins, enhancing amino acid metabolism and protein reparation.