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作 者 ：李玉红;陈　鹏;程智慧;张　静;董　剑

期 刊 ：园艺学报 2009年 36卷 04期 页码：507-512

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；

Keywords:cucumber, Pseudoperonospora cubensis, BTH, pathogenesis-related proteins, MALDI-TOF MS, Western blotting,

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27 kD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-TOFMS) 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。

Abstract:

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 exp ression of a 27 kD p rotein in intercellular fluids of cucumber leaves. This protein was purified by electroelution combined with SDS - PAGE, and analyzed by MALDI-TOF MS. The obtained pep tide mass fingerp rinting ( 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 andWestern blotting.

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