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首页> 外文学位 >Differential proteome analysis of Pseudomonas syringae maculicola M6 in response to infection into Arabidopsis thaliana.
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Differential proteome analysis of Pseudomonas syringae maculicola M6 in response to infection into Arabidopsis thaliana.

机译：丁香假单胞菌M6对感染拟南芥感染的差异蛋白质组分析。

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摘要

This study aimed to understand the bacterial responses upon infection of the host plant. An incompatible plant-pathogen interaction model using Arabidopsis thaliana (A. thaliana or AT) and Pseudomonas syringae pv. maculicola (Psm) M6 had been successfully established. Furthermore, an efficient differential centrifugation method was developed for the separation of bacterial pathogens from the infected A. thaliana culture cells for proteomic analyses.;Two dimensional gel electrophoretic (2-DE) analyses of Psm M6 collected at 12th hr post-infection into the AT suspension cells were performed. Compared with the proteome of Psm M6 grown in MS medium, 24 proteins were found to be differentially expressed in the proteome of Psm M6 which responded to the plant host response. With the provision of the MALDI-TOF mass spectrometer, 23 proteins were identified. A majority of these differentially expressed proteins were up-regulated and had known functions that were related to transportation, pathogenesis, phosphate uptake as well as amino acid synthesis. In contrast, 4 proteins were down-regulated. Their transcriptional expression profiles were also measured by quantitative RT-PCR method. With these home-made antibodies, the temporal expression profiles of several bacterial proteins (OrpD; BadH; PstS; PhnD; Tat-sp and GlpQ) were measured in Psm M6 post-infection into AT suspension cultured cells. For in vivo validation purposes, protein expression level of BadH and PstS were measured after infection of A. thaliana planta. It reinforced the idea that the results obtained using cultured plant cells are also repeatable in planta .;Further, the bacterial responses in relation to a massive production of NO and ROS in plants were studied upon bacterial attack. The reaction of NO and free O2- yields ONOO- , which converts tyrosine residues into nitrotyrosine residues. This process is called protein nitration. Therefore, it would be interesting to detect the extent of bacterial protein nitration in vivo. With the use of anti-nitrotyrosine antibodies in 2DE-western blotting, 19 proteins were found to be nitrated upon infection of A. thaliana suspension culture cells. Some of these proteins are known to be related to apoptosis. These results suggested that NO and ROS produced by the host plants modulated bacterial protein nitration in vivo and this nitration process may participate in biochemical sequences leading to bacterial cell death. All these findings clearly demonstrate the complexity of the molecular response of bacteria to the host environment.
机译：这项研究旨在了解宿主植物感染后的细菌反应。使用拟南芥（拟南芥或AT）和丁香假单胞菌PV的不兼容的植物-病原体相互作用模型。 maculicola（Psm）M6已成功建立。此外，还开发了一种有效的差速离心方法，用于从感染的拟南芥培养细胞中分离细菌病原体，以进行蛋白质组学分析。;在感染后12小时收集的Psm M6的二维凝胶电泳（2-DE）分析。进行AT悬浮细胞。与在MS培养基中生长的Psm M6蛋白质组相比，发现24种蛋白质在Psm M6蛋白质组中差异表达，而Psm M6蛋白质组对植物宿主反应有反应。使用MALDI-TOF质谱仪，鉴定出23种蛋白质。这些差异表达的蛋白质中的大多数被上调，并且具有与运输，发病机制，磷酸盐吸收以及氨基酸合成相关的已知功能。相反，有4种蛋白质被下调。还通过定量RT-PCR方法测量了它们的转录表达谱。使用这些自制抗体，在感染了AT悬浮培养细胞的Psm M6中测量了几种细菌蛋白（OrpD，BadH，PstS，PhnD，Tat-sp和GlpQ）的时间表达谱。为了体内验证的目的，在植物拟南芥感染后测量了BadH和PstS的蛋白表达水平。它加强了这样的想法，即使用培养的植物细胞获得的结果在植物中也可重复。 NO和游离O2-的反应生成ONOO-，它将酪氨酸残基转化为硝基酪氨酸残基。此过程称为蛋白质硝化。因此，检测体内细菌蛋白硝化的程度将是有趣的。通过在2DE-western印迹中使用抗硝基酪氨酸抗体，发现感染拟南芥悬浮培养细胞后会将19种蛋白质硝化。已知其中一些蛋白质与细胞凋亡有关。这些结果表明，宿主植物产生的NO和ROS在体内调节细菌的硝化作用，而这种硝化过程可能参与导致细菌细胞死亡的生化序列。所有这些发现清楚地证明了细菌对宿主环境的分子反应的复杂性。

著录项

  • 作者

    Butt, Kwok Chu.;

  • 作者单位

    Hong Kong Polytechnic University (Hong Kong).;

  • 授予单位 Hong Kong Polytechnic University (Hong Kong).;
  • 学科 Molecular biology.
  • 学位 Ph.D.
  • 年度 2008
  • 页码 361 p.
  • 总页数 361
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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