Plasmodesmata: Methods and Protocols

ISBN-13: 9781493915224 / Angielski / Twarda / 2014 / 346 str.

Manfred Heinlein

Opis

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.

Spis treści

Part I: Overview

 

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