Collaborative Research: Arabidopsis 2010: Dissecting Cortical Actin Function during Arabidopsis-Pseudomonas Interactions

植物病原细菌与它们侵入的宿主细胞和组织之间存在着微妙而复杂的相互作用。 新的信息表明，一个动态的细丝网络，细胞骨架，是必不可少的抵抗和敏感性植物病原体。 然而，究竟是哪些来自细菌的分子刺激细胞骨架反应，以及哪些植物蛋白质是这些反应所必需的仍然知之甚少。 在这个项目中，研究人员将剖析细菌效应蛋白和植物宿主细胞骨架之间串扰的遗传基础。 利用分子遗传学工具和先进的成像技术，研究人员将测试特定病原体效应蛋白干扰植物肌动蛋白结合蛋白（ABPs）的假设，这反过来又导致防御信号传导过程中植物细胞骨架的改变。 将开发肌动蛋白和肌动蛋白结合蛋白突变体的工具箱，并开发用于将细菌效应物引入宿主细胞的递送系统。 通过这项工作，PI旨在鉴定细菌效应蛋白和39种植物基因/蛋白的功能，以揭示信号网络。 首席PI Staiger将协调各组之间的活动，并利用最先进的成像模式来评估ABP收集中肌动蛋白动力学的缺陷和对效应蛋白的反应。 Day和Chang实验室将使用遗传学方法来剖析串扰是否涉及病原体相关的分子模式识别反应（PTI）或效应器触发的免疫（ETI），或两者兼而有之。这两个实验室还将使用一种新的细菌传递系统，将肌动蛋白动力学的变化与相应的细菌效应蛋白联系起来。这两个实验室还将为蛋白质-蛋白质相互作用测定和亚细胞定位制备构建体。在这项工作中创建的所有项目数据和资源将在一个联合网站上及时传播：http://actinpathogen-network.msu.edu。 更广泛的影响，这项工作将有助于深入了解主机？病原体信号，并将使作物的修改，以防止疾病，破坏农业生产力。 作为一个独特的外展组成部分，我们将建立一个夏季研讨会，培训所有项目科学家和其他感兴趣的各方，包括当地高中生，在先进的方法成像细胞动力学。

An exquisite and intricate crosstalk exists between bacterial phytopathogens and the host cells and tissues they invade. New information suggests that a dynamic network of filaments, the cytoskeleton, is essential for both resistance and susceptibility to plant pathogens. However, exactly which molecules from the bacterium stimulate cytoskeletal responses, and which plant proteins are necessary for these responses remains poorly understood. In this project, the investigators will dissect the genetic basis for crosstalk between bacterial effector proteins and the plant host cytoskeleton. Using both molecular genetics tools and advanced imaging technologies, the investigators will test the hypothesis that specific pathogen effector proteins perturb plant actin-binding proteins (ABPs), which in turn leads to alterations in the plant cytoskeleton during defense signaling. A toolbox of actin and actin-binding protein mutants will be developed, and a delivery system for introducing bacterial effectors into host cells will be exploited. Through this work, the PIs aim to identify the function of bacterial effector proteins and 39 plant genes/proteins in order to uncover the signaling network. Lead PI Staiger will coordinate activities between the groups and exploit state-of-the-art imaging modalities to evaluate the defects in actin dynamics in the ABP collection and in response to effector proteins. The Day and Chang laboratories will use genetic approaches to dissect whether crosstalk involves pathogen-associated molecular pattern recognition responses (PTI) or effector-triggered immunity (ETI), or both. These two labs will also use a novel bacterial delivery system to associate changes in actin dynamics to a corresponding bacterial effector protein. The two labs will also prepare constructs for protein-protein interaction assays and subcellular localization. All project data and resources created in this work will be disseminated in a timely fashion at a joint website: http://actinpathogen-network.msu.edu. The broader impact of this work will contribute to a deep understanding of host?pathogen signaling and will enable crop modifications to protect against diseases that devastate agricultural productivity. As a unique outreach component, we will establish a summer workshop to train all project scientists and other interested parties, including local high school students, in advanced methods for imaging cellular dynamics.