1. Plasmodesmata: Channels for Intercellular Signaling During Plant Growth and Development
Iris Sevilem, Shri Ram Yadav, and Ykä Helariutta
2. Plasmodesmata: Channels for Viruses on the Move
Manfred Heinlein
Part II: Imaging of Plasmodesmata
3. Imaging Plasmodesmata with High Resolution Scanning Electron Microscopy
Deborah A. Barton and Robyn L. Overall
4. Preparative Methods for Imaging Plasmodesmata at Super-Resolution
Karen Bell and Karl Oparka
Part III: Structural Analysis of Plasmodesmata
5. Isolation of Plasmodesmata from Arabidopsis Suspension Culture Cells
Magali S. Grison, Lourdes Fernandez-Calvino, Sébastien Mongrand, and Emmanuelle M.F. Bayer
6. Immunofluorescence Detection of Callose Deposition around Plasmodesmata Sites
Ali Pendle and Yoselin Benitez-Alfonso
7. Imaging Callose at Plasmodesmata Using Aniline Blue: Quantitative Confocal Microscopy
Raul Zavaliev and Bernard L. Epel
8. Localization of Fluorescently-Tagged Protein to Plasmodesmata by Correlative Light and Electron Microscopy
Shannon Modla, Jeffrey L. Caplan, Kirk J. Czymmek, and Jung-Youn Lee
Part IV: Analysis of Plasmodesmata Conductivity and Regulation
9. Quantification of Plant Cell Coupling with Live-Cell Microscopy
Johannes Liesche and Alexander Schulz
10. Drop-ANd-See: A Simple, Real-Time, and Non-Invasive Technique for Assaying Plasmodesmal Permeability
Weier Cui, Xu Wang, and Jung-Youn Lee
11. Mapping Symplasmic Fields at the Shoot Apical Meristem Using Iontophoresis and Membrane Potential Measurements
Christiaan van der Schoot and Päivi L.H. Rinne
12. Analysis of the Conductivity of Plasmodesmata by Microinjection
Friedrich Kragler
13. Investigating Plasmodesmata Genetics with Virus-Induced Gene Silencing and an Agrobacterium-Mediated GFP Movement Assay
Jacob O. Brunkard, Tessa M. Burch-Smith, Anne M. Runkel, and Patricia Zambryski
14. Probing Plasmodesmata Function with Biochemical Inhibitors
Rosemary G. White
Part V: Studying Macromolecular Trafficking
15. GAL4 Transactivation-Based Assay for the Detection of Selective Intercellular Protein Movement
Dhinesh Kumar, Huan Chen, Yeonggil Rim, and Jae-Yean Kim
16. Techniques for Assessing the Effects of Pharmacological Inhibitors on Intercellular Protein Movement
Shuang Wu and Kimberly L. Gallagher
17. Probing Protein Targeting to Plasmodesmata Using Fluorescence Recovery after Photo-Bleaching
Kathryn M. Wright and Katrin M. MacKenzie
18. The Tracking of Intercellular Small RNA Movement
Christophe Himber and Patrice Dunoyer
19. Analysis of the Role of Myosins in Targeting Proteins to Plasmodesmata
Martin Di Donato and Khalid Amari
20. Pumilio-Based RNA In Vivo Imaging
Jens Tilsner
21. In Vivo RNA Labeling Using MS2
Eduardo Peña, Manfred Heinlein, and Adrian Sambade
Plasmodesmata: Methods and Protocols provides a unique compendium of currently applied methods to explore plasmodesmata (PD). Opening with overviews of PD structure and their function during development as well as the role of viruses as models for addressing cellular mechanisms in macromolecular trafficking and movement through PD, the book continues with detailed experimental protocols for imaging PD, the isolation and structural analysis of PD, the analysis of PD conductivity and regulation, and for studying cellular mechanisms by which macromolecules are targeted to PD and transported through the channel. 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 tips on troubleshooting and avoiding known pitfalls.
Practical and easy to use, Plasmodesmata: Methods and Protocols delivers detailed information about specific practical procedures in order to enable their reproduction and application in PD-related research in laboratories around the world.