苹果叶片应答轮纹病菌胁迫的叶绿体蛋白质组学分析  

Proteomic analysis of pathogen-responsive proteins from the chloroplast of apple leaves induced by Botryosphaeria dothidea

作　　者：肖龙[1,2] 张彩霞[1,2] 宗泽冉 田义[1,2] 张利义[1,2] 丛佩华[1,2] 

机构地区：[1]农业部园艺作物种质资源利用重点实验室,辽宁兴城125100 [2]中国农业科学院果树研究所,辽宁兴城125100

出　　处：《果树学报》2016年第11期1357-1366,共10页Journal of Fruit Science

基　　金：农业部现代农业产业技术体系建设专项资金(CARS-28);国家科技支撑计划项目(2013BAD02B01)

摘　　要：【目的】开展苹果叶片应答轮纹病菌胁迫的叶绿体蛋白质组学研究,筛选苹果叶片应答轮纹病胁迫后差异表达的叶绿体蛋白。【方法】以未受轮纹病菌胁迫和受轮纹病胁迫的苹果叶片为试材,分别提取叶片叶绿体及其蛋白,利用双向电泳技术(2-DE)结合质谱技术(MALDI-TOF-TOF/MS)检测并分析差异表达蛋白点。【结果】叶绿体蛋白样品经双向电泳分离后,获得了背景清晰的双向电泳凝胶图谱。经质谱检测及数据库检索,成功鉴定得到21个差异表达蛋白点,按照功能划分为6类,分别为光合作用相关、能量代谢相关、蛋白代谢相关、氨基酸合成相关、抗氧化相关及未知功能蛋白。【结论】苹果叶片受轮纹病菌侵染后,叶绿体光合作用、能量代谢等多个代谢过程中的相关蛋白差异表达,应答轮纹病菌的胁迫。[ Objective] At present, apple ring rot is one of the three most serious apple diseases in the major apple producing areas in China. The apple ring rot seriously affects yield and quality. Proteome is the direct embodiment of life activities in plants, and proteomic study is a powerful tool to reveal the mecha- nisms of life activities. It is an important research approach to reveal the mechanisms of plant-disease in- teractions by examining the proteomic changes after the host plants are infected by diseases. In this study, a series of experiments were set up with excised apple leaves subject to the infection by the pathogenic fungus causing apple ring rot. Chloroplasts and chloroplast proteins were extracted from the leaves. With two-dimensional electrophoresis （2-DE） and mass spectrometry （MS）, differentially expressed chloroplast proteins in apple leaves after infection by apple ring rot were screened out so as to reveal proteomic response in chloroplasts to the biotic stress of apple ring rot. [Methods] The experimental materials were the apple leaves infected by apple ring rot pathogenic fungus and the uninfected healthy leaves. The leaf samples were collected from the apple breeding nursery of Institute of Pomology of Chinese Academy of Agricultural Sciences （CAAS） in Xingcheng, Liaoning province, China. The activated pathogenic fungus of apple ring rot were inoculated in PDA medium and cultured in an incubator to produce spores at 75% humidity, 25 ℃ and 24 h light. The spore suspension was prepared with the conidiospore collected from the surface of the PDA medium. Healthy young apple leaves were collected and washed with sterilized wa- ter. After 2 days of incubation in aseptic conditions at 75% humidity, 16℃ and 12 h light, the leaves showing diseases were discarded, and those with no diseases were chosen as the experimental materials for the infection experiments below. Some of the leaf materials were inoculated with the spore suspension using sterilized cotton （as shown in Fig. 1）,

关 键 词：苹果叶片 叶绿体蛋白 轮纹病菌 胁迫 差异蛋白质组 双向电泳 

分 类 号：S661.1[农业科学—果树学]