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中国生物防治学报 ›› 2015, Vol. 31 ›› Issue (6): 913-920.DOI: 10.16409/j.cnki.2095-039x.2015.06.014

• 研究论文 • 上一篇    下一篇

内生细菌TF28诱导番茄抗病分子机制研究

张淑梅1,2, 李晶1,2, 姜威1,2, 孟利强1,2, 陈静宇1, 胡基华1,2, 曹旭1,2, 尹徳亮1   

  1. 1. 黑龙江省科学院微生物研究所, 哈尔滨 150010;
    2. 黑龙江省科学院高技术研究院, 哈尔滨 150001
  • 收稿日期:2014-05-26 出版日期:2015-12-08 发布日期:2015-12-17
  • 通讯作者: 尹徳亮
  • 作者简介:张淑梅(1969-),研究员,博士,E-mail:shumeizhang@yahoo.com
  • 基金资助:
    国家自然科学基金(30301780);哈尔滨市创新人才专项(2012RFXYNO25)

Molecular Mechanisms of Induced Resistance to Tomato Pathogens by Endophytic Bacterium TF28

ZHANG Shumei1,2, LI Jing1,2, JIANG Wei1,2, MENG Liqiang1,2, CHEN Jingyu1, HU Jihua1,2, CAO Xu1,2, YIN Deliang1   

  1. 1. Institute of Microbiology, Heilongjiang Academy of Sciences, Harbin 150010, China;
    2. High Technique Council, Heilongjiang Academy of Sciences, Harbin 150009, China
  • Received:2014-05-26 Online:2015-12-08 Published:2015-12-17

摘要: 为了解内生解淀粉芽胞杆菌Bacillus amyloliquefaciens TF28诱导番茄抗病分子机制,本文采用相对定量RT-PCR方法研究内生细菌TF28对番茄抗病信号传递途径中促分裂原活化蛋白激酶基因MAPK和转录因子WRKY基因表达的影响,采用双向电泳、质谱检测和生物信息学等蛋白质组学技术研究接种菌株TF28后番茄叶片差异蛋白的表达情况。结果表明接种菌株TF28后,番茄叶片中的基因MAPKWRKY上调表达,基因WRKY表达增幅高于基因MAPK,2个基因均在接种后2 d表达量最高。从接种内生细菌2 d后的番茄叶片中分离到373个蛋白点,与对照相比,有23个蛋白点特异表达,18个蛋白点上调表达,53个蛋白点下调表达,280个蛋白表达量与对照相当,对14个特异表达蛋白进行质谱分析和蛋白数据库检索,9个与抗病相关,3个与品质相关,2个与生长调控相关。

关键词: 内生细菌, 番茄, 抗病信号, MAPKWRKY基因, 表达, 差异蛋白

Abstract: The objective of this study is to elucidate molecular mechanisms of induced resistance to tomato pathogens by endophytic bacterium TF28. In this study, relatively quantitative RT-PCR method was used to analyze the impact of endophytic bacterium TF28 on the expression of MAPK and WRKY gene. Proteomics technology, including 2-D electrophoresis, mass spectrometry and bioinformation analysis, were used to analyze different proteins in tomato leaves between control and TF28 treatment. Results indicated that the expression of MAPK and WRKY gene in tomato leaves were up-regulated after inoculation with TF28. Expression of WRKY gene was higher than that of MAPK. Both two genes reached the maximum expression in leaves at 48 h after inoculation. A total of 373 proteins were obtained using 2-D electrophoresis, of which 18 showed up-regulated, 53 down-regulated and 23 specific expression in tomato leaves after inoculation with strain TF28 two days later, but 280 showed identical to the control. Mass spectrometry analysis and functional annotations were performed for 14 specific expression proteins and the results revealed that 9 was related to disease defense, 3 correlated with quality and 2 reffered to growth regulation.

Key words: endophytic bacterium, tomato, defense signal, MAPK and WRKY gene, expression, different proteins

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