苹果枝条表皮应答轮纹病菌侵染的蛋白质组学分析

Proteomic analysis of pathogen-responsive proteins from the bark of apple branches induced by Botryosphaeria dothidea

蛋白质提取方法是双向电泳分析的关键。本研究中,我们通过优化提取条件,建立了适于苹果枝条表皮总蛋白提取的技术体系。在此基础上,开展了苹果枝条表皮应答轮纹病菌侵染的蛋白质组学研究,以期明确轮纹病菌胁迫下苹果枝条表皮细胞参与抗病反应的关键蛋白。具体方法是以未接菌及接菌后的苹果枝条表皮为材料,分别提取总蛋白,利用双向凝胶电泳技术结合质谱(MALDI-TOF-TOF/MS)检测分析差异表达蛋白。经质谱检测及数据库检索,共有22个蛋白点得到成功鉴定,按照功能划分为光合作用相关、糖类及能量代谢相关、防御反应相关、蛋白质合成相关及未知功能蛋白等5类。其中参与防御反应的病程相关蛋白(APX、β-1,3-葡聚糖酶、Mal d1、PR-1)可能是苹果枝条表皮细胞应答轮纹病菌胁迫过程中参与抗病反应的关键蛋白。

The optimal protein extraction method is the key for proteomics research by tw o-dimensional electrophoresis（ 2-DE）. In this study,in order to establish a suitable method for extraction of total protein from the bark of apple branches,the extraction condition for the 2-DE w as optimized. To elucidate the key proteins related to disease-resistance of apple branches induced by Botryosphaeria dothidea,total proteins w ere extracted from control and infected branches, and separated by tw o-dimensional electrophoresis. The differentially expressed proteins w ere then identified by M ALDI-TOF-TOF / M S. In total,25 differentially expressed proteins w ere detected by Image M aster Softw are. After tryptic digestion,M ALDI-TOF-TOF / M S analysis and database searching,22 protein spots w ere finally identified. The predicted function of 22 proteins w ere related to photosynthesis,glucose / energy metabolism,defense responses,protein synthesis and other unknow n functions.The pathogenesis-related proteins（ APX,β-1,3-glucanase,M al d1 and PR-1） involved in defense responses and differentially expressed in apple branches may play a key role in resistance to B. dothidea.