Type 3 Secretion Systems pp 141-153

Part of the Methods in Molecular Biology book series (MIMB, volume 1531) | Cite as

Subcellular Localization of Pseudomonas syringae pv. tomato Effector Proteins in Plants

  • Kyaw Aung
  • Xiufang Xin
  • Christy Mecey
  • Sheng Yang He
Protocol

Abstract

Animal and plant pathogenic bacteria use type III secretion systems to translocate proteinaceous effectors to subvert innate immunity of their host organisms. Type III secretion/effector systems are a crucial pathogenicity factor in many bacterial pathogens of plants and animals. Pseudomonas syringae pv. tomato (Pst) DC3000 injects a total of 36 protein effectors that target a variety of host proteins. Studies of a subset of Pst DC3000 effectors demonstrated that bacterial effectors, once inside the host cell, are localized to different subcellular compartments, including plasma membrane, cytoplasm, mitochondria, chloroplast, and Trans-Golgi network, to carry out their virulence functions. Identifying the subcellular localization of bacterial effector proteins in host cells could provide substantial clues to understanding the molecular and cellular basis of the virulence activities of effector proteins. In this chapter, we present methods for transient or stable expression of bacterial effector proteins in tobacco and/or Arabidopsis thaliana for live cell imaging as well as confirming the subcellular localization in plants using fluorescent organelle markers or chemical treatment.

Key words

Plant pathogen Bacterial pathogenesis Type III secretion Plant immunity Tobacco Arabidopsis thaliana Agrobacterium Confocal microscopy 

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Kyaw Aung
    • 1
    • 3
  • Xiufang Xin
    • 1
  • Christy Mecey
    • 1
  • Sheng Yang He
    • 1
    • 2
    • 3
  1. 1.Department of Energy Plant Research LaboratoryMichigan State UniversityEast LansingUSA
  2. 2.Department of Plant BiologyMichigan State UniversityEast LansingUSA
  3. 3.Howard Hughes Medical InstituteMichigan State UniversityEast LansingUSA

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