BTH诱导的一种黄瓜叶片胞间隙病程相关蛋白的纯化及鉴定  

Purification and Identification of a Pathogenesis-related Protein in Intercellular Fluids of Cucumber Leaves Induced by BTH

作　　者：李玉红[1] 陈鹏[2] 程智慧[1] 张静[1] 董剑[3] 

机构地区：[1]西北农林科技大学园艺学院,陕西杨凌712100 [2]西北农林科技大学生命科学学院,陕西杨凌712100 [3]西北农林科技大学农学院,陕西杨凌712100

出　　处：《园艺学报》2009年第4期507-512,共6页Acta Horticulturae Sinica

基　　金：国家‘十一五’科技支撑计划项目(2006BAD07B01);公益性行业(农业)科研专项(nyhyzx07-007);农业部蔬菜遗传与生理重点开放实验室资助项目

摘　　要：对苯并噻二唑(BTH)处理和接种霜霉病菌后的黄瓜幼苗叶片胞间隙液中病程相关蛋白的诱导积累及纯化鉴定进行了研究。结果表明,BTH处理和接种霜霉病菌均可诱导黄瓜叶片胞间隙液产生分子量为27kD的新蛋白,利用SDS-PAGE结合电洗脱的方法纯化了该蛋白。通过基体辅助激光解析电离飞行时间质谱(MALDI-TOF MS)分析,将所得的肽指纹图谱(PMF)在NCBInr蛋白质数据库中比对,发现该蛋白是一种酸性几丁质酶。酶活性测定及Western blotting分析进一步证实了上述结果。This article reported the induction, purification and identification of a pathogenesis-related protein from the intercellular fluids of cucumber seedlings, which were treated with benzothiadiazole （BTH） or Pseudoperonospora cubensis. Our results demonstrated that both BTH treatment and Pseudoperonospora cubensis inoculation could induce the expression of a 27 kD protein in intercellular fluids of cucumber leaves. This pro- tein was purified by electroelution combined with SDS - PAGE, and analyzed by MALDI-TOF MS. The obtained peptide mass fingerprinting （PMF） was searched in the protein data bank, indicating that it was an acidic chitinase. This was confirmed further by the assay of chitinase activity and Western blotting.

关 键 词：黄瓜 霜霉病 苯并噻二唑 病程相关蛋白 质谱分析 Western BLOTTING 

分 类 号：S642.2[农业科学—蔬菜学]