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

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1531; p. 141
• Main Authors: Aung, Kyaw, Xin, Xiufang, Mecey, Christy, He, Sheng Yang
• Format: Journal Article
• Language: English
• Published: United States 01-01-2017
• Subjects: Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Gene Expression; Genes, Reporter; Host-Pathogen Interactions - immunology; Intracellular Space - metabolism; Microscopy, Confocal; Nicotiana - microbiology; Plant Diseases - immunology; Plant Diseases - microbiology; Plants, Genetically Modified; Protein Transport; Pseudomonas syringae - genetics; Pseudomonas syringae - metabolism; Solanum lycopersicum - immunology; Solanum lycopersicum - microbiology; Transformation, Genetic; Type III Secretion Systems - genetics; Type III Secretion Systems - metabolism; Arabidopsis thaliana; Bacterial pathogenesis; Tobacco; Agrobacterium; Type III secretion; Plant pathogen; Plant immunity; Confocal microscopy
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-6649-3_12

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.