Plant Immunity Methods and Protocols

A great deal of effort is being invested in understanding the molecular mechanisms through which plants interact with pathogenic microbes.  In  Plant Immunity: Methods and Protocols, expert researchers in the field describe emerging technologies that can be applied to the most significant outstanding questions faced by scientists studying immunity in plants. The technologies in this detailed volume include methods for examining protein localization, protein complex purification, protein-protein interactions, transient and inducible gene expression, chromatin immunoprecipitation, microaspiration, laser microdissection, purification of fungal haustoria, and genetic manipulation of bacterial and oomycete pathogens.  These techniques are applicable to a wide range of topics, including molecular functionality of NB-LRR proteins and other immune signaling components, and functional characterization of effector proteins and other pathogen components that sabotage host immunity.  Written in the highly successful  Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

1. Studying NB-LRR Immune Receptor Localization by Agroinfiltration Transient Expression Patrick Cournoyer and S. P. Dinesh-Kumar 2. Fragment Complementation and Co-Immunoprecipitation Assays for Understanding R Protein Structure and Function Peter Moffett 3. Purification of Resistance Protein Complexes Using a Biotinylated Affinity (HPB) Tag Yiping Qi and Fumiaki Katagiri 4. Biochemical Purification of Native Immune Protein Complexes James M. Elmore and Gitta Coaker 5. Chromatin Immunoprecipitation (ChIP) to Identify Global Targets of WRKY Transcription Factor Family Members Involved in Plant Immunity Mario Roccaro and Imre E. Somssich 6. Dose Response to and Systemic Movement of Dexamethasone in the GVG Inducible Transgene System in Arabidopsis Xueqing Geng and David Mackey 7. Quantifying Alternatively Spliced mRNA via Capillary Electrophoresis Xue-Cheng Zhang and Walter Gassmann 8. Constructing Haustorium-Specific cDNA Libraries from Rust Fungi Ann-Maree Catanzariti, Rohit Mago, Jeff Ellis, and Peter Dodds 9. Microaspiration of Esophageal Gland Cells and cDNA Library Construction for Identifying Parasitism Genes of Plant-Parasitic Nematodes Richard S. Hussey, Guozhong Huang, and Rex Allen 10. Construction of Pseudomonas syringae pv. tomato DC3000 Mutant and Polymutant Strains Brian H. Kvitko and Alan Collmer 11. A Straightforward Protocol for Electro-Transformation of Phytophthora capsici Zoospores Edgar Huitema, Matthew Smoker, and Sophien Kamoun 12. Propagation, Storage, and Assays with Hyaloperonospora arabidopsidis, a Model Oomycete Pathogen of Arabidopsis John M. McDowell, Troy Hoff, Ryan G. Anderson, and Daniel Deegan 13. Assaying Effector Function In planta Using Double-Barreled Particle Bombardment Shiv D. Kale and Brett M. Tyler 14. Assays for Effector-Mediated Suppression of Programmed Cell Death in Yeast Yuanchao Wang and Qian Huang 15. Purification of Effector-Target Protein Complexes via Transient Expression in N. benthamiana Joe Win, Sophien Kamoun, and Alexandra M.E. Jones 16. Imaging Fluorescently-Tagged Phytophthora Effector Proteins Inside Infected Plant Tissue Petra C. Boevink, Paul R.J. Birch, and Stephen C. Whisson 17. Immunolocalization of Pathogen Effectors Eric Kemen, Kurt Mendgen, and Ralf T. Voegele 18. Laser Capture Microdissection of Nematode Feeding Cells Nagabhushana Ithal and Melissa G. Mitchum 19. Laser Microdissection of Plant-Fungus Interaction Sites and Isolation of RNA for Downstream Expression Profiling Divya Chandran, Noriko Inada, and Mary C. Wildermuth 20. Global Expression Profiling of RNA from Laser Microdissected Cells at Fungal-Plant Interaction Sites Divya Chandran, Greg Hather, and Mary C. Wildermuth 21. Visualizing Cellular Dynamics in Plant-Microbe Interactions Using Fluorescent-Tagged Proteins William Underwood, Serry Koh, and Shauna C. Somerville