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

园艺学报 ›› 2009, Vol. 36 ›› Issue (4): 507-512.

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

李玉红¹; 陈　鹏^2*;程智慧¹;张　静¹; 董　剑³

(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)

收稿日期:2008-10-17 修回日期:2009-03-17 出版日期:2009-04-25 发布日期:2009-04-25
通讯作者: 陈　鹏

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

LI Yu-hong¹,CHEN Peng^2*,CHENG Zhi-hui¹, ZHANG Jing¹, and DONG Jian³

(¹College of Horticulture, Northwest A & F University, Yangling, Shaanxi 712100, China; ²College of Life Sciences, Northwest A & F University, Yangling, Shaanxi 712100, China; ³College of Agronomy, Northwest A & F University, Yangling, Shaanxi 712100, China)

Received:2008-10-17 Revised:2009-03-17 Online:2009-04-25 Published:2009-04-25
Contact: CHEN Peng

摘要/Abstract

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。

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

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.

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

中图分类号:

李玉红;陈　鹏;程智慧;张　静;董　剑. 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[J]. 园艺学报, 2009, 36(4): 507-512.

LI Yu-hong;CHEN Peng;CHENG Zhi-hui;ZHANG Jing;and DONG Jian. Purification and Identification of a Pathogenesis-related Protein in Intercellular Fluids of Cucumber Leaves Induced by BTH[J]. ACTA HORTICULTURAE SINICA, 2009, 36(4): 507-512.

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