外源茉莉酸甲酯诱导的水稻叶片蛋白质差异表达分析  

Proteomic analysis of differentially expressed proteins induced by methyl jasmonate in rice leaves

作　　者：张智慧[1,2] 聂燕芳[3] 何磊[1,2] 高元媛[1,2] 李云锋[1,2] 王振中[1,2] 

机构地区：[1]华南农业大学植物病理生理学研究室,广州510642 [2]教育部生物防治工程研究中心,广州510642 [3]华南农业大学制药工程系,广州510642

出　　处：《植物病理学报》2012年第2期154-163,共10页Acta Phytopathologica Sinica

基　　金：国家自然科学基金资助项目(30971887;30600399);广东省自然科学基金资助项目(10151064201000008;07006683)

摘　　要：以抗稻瘟病水稻近等基因系C101LAC及CO39为材料,应用差异蛋白组学技术对外源茉莉酸甲酯(MeJA)诱导8 h和12 h后的水稻叶片蛋白质差异表达变化进行了分析。采用PEG-4000预沉淀法提取水稻叶片总蛋白,经双向电泳、凝胶染色和图像分析,结果表明:与对照相比MeJA处理后的2个水稻品系中共有21个蛋白质表现为2倍以上的表达差异。其中,MeJA处理8 h后,CO39中的差异表达蛋白质点有5个,C101LAC中有8个,U5为2个品系中均新增的蛋白质点。Me-JA处理12 h后,CO39中有6个差异表达蛋白质点,C101LAC中则有5个。经胶内酶解、MALDI-TOF/TOF质谱分析及数据库检索,成功对21个差异表达蛋白质进行了鉴定,其中新增的蛋白质点U5被鉴定为内切β-1,3-1,4-葡聚糖酶。依据GO数据库和UniProtKB数据库提供的蛋白质注释信息,按照功能可将21个差异表达蛋白质分为8类,分别在植物抗性、氨基酸代谢、信号转导、光合作用、光呼吸、糖代谢、蛋白质合成与分解及细胞代谢等方面发挥作用。Proteomic approaches using two-dimensional electrophoresis （2-DE） were conducted to identify the differentially expressed proteins in rice leaves in response to methyl jasmonate （MeJA）. Proteins were ex- tracted from rice leaves at 8 h and 12 h after treatment with 0.1 mmol/L MeJA and then fractionated using PEG 4000-mediated prefractionation technique before separation by 2-DE. In total, twenty-one proteins resolved in the 2-DE gels were up-regulated or down-regulated in the inoculated rice leaves compared with the controls. At 8 h after treatment with MeJA, 5 and 8 differentially expressed proteins in CO39 and C101LAC were identified, respectively. Among them, U5 protein spot was specifically induced in both two rice cultivars. At 12 h, changes of 6 proteins and 5 proteins were detected in CO39 and in C101LAC, respectively. These differentially displayed proteins were successfully identified by in-gel digestion, MALDI-TOF/TOF analysis and database searching. The protein spot U5 was identified as endo-1,3-1,4-beta-D-glucanase. According to protein annotations from GO database and UniProtKB database, the 21 MeJA-responsive proteins were classified into eight categories based on their putative function reported： 1 ） plant defense response, 2 ） amino-acid biosynthesis, 3 ） signal transduction, 4 ） photosynthesis, 5 ） photorespiratory, 6 ） carbohydrate metabolism, 7） protein biosynthesis and proteolysis and 8） metabolic process.

关 键 词：水稻 茉莉酸甲酯 蛋白组学 

分 类 号：S435.111.41[农业科学—农业昆虫与害虫防治]