This fully updated book aims to facilitate the study of the nanochannels that connect plant cells, known as plasmodesmata, and to instigate new research that will further advance our knowledge of these structures. Beginning with the general structural composition and regulation of plasmodesmata as well as their role in plant development and disease, the volume continues with chapters exploring plasmodesmata architectures and distribution in cell interfaces, approaches to dissect plasmodesmata composition, protocols to quantify changes in plasmodesmata permeability using fluorescent tracers and mobile proteins, as well as a section with protocols that contribute to plasmodesmata research but fall outside the previous classifications. Written for the highly successful Methods in Molecular Biology series, 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.
Authoritative and up-to-date, Plasmodesmata: Methods and Protocols, Second Edition serves as a vital guide for all plant scientists, both novice and expert, especially those studying the intricacies of cell-to-cell communication pathways.

Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Inhalt

Plasmodesmata Structural Components and Their Role in Signaling and Plant Development.- Function of Plasmodesmata in the Interaction of Plants with Microbes and Viruses.- Plasmodesmata Ultrastructure Determination Using Electron Tomography.- Ultrastructural Analysis and Three-Dimensional Reconstruction of Plasmodesmata.- Serial Block Electron Microscopy to Study Plasmodesmata in the Vasculature of Arabidopsis thaliana Roots.- Focused Ion Beam-Scanning Electron Microscopy for Investigating Plasmodesmal Densities.- Measuring Plasmodesmata Density on Cell Interfaces of Monocot Leaves Using 3D Immunolocalization and Scanning Electron Microscopy.- Super-Resolution Imaging of Plasmodesmata Using 3D-Structured Illumination Microscopy.- In Vivo Aniline Blue Staining and Semi-Automated Quantification of Callose Deposition at Plasmodesmata.- Immunofluorescence Detection of Callose in Plant Tissue Sections.- Callose Detection and Quantification at Plasmodesmata in Bryophytes.-Isolation of Plasmodesmata Membranes for Lipidomic and Proteomic Analysis.- Methods for Detection of Protein Interactions with Plasmodesmata-Localized Reticulons.- Studying Protein-Protein Interactions at Plasmodesmata by Measuring Förster Resonance Energy Transfer.- Quantifying the Organization and Dynamics of the Plant Plasma-Membrane across Scales Using Light Microscopy.- Using Steady-State Fluorescence Anisotropy to Study Protein Clustering.- Quantification of Cell-to-Cell Connectivity Using Particle Bombardment.- Investigating Plasmodesmata Function in Arabidopsis thaliana Using a Low-Pressure Bombardment System and GFP Movement Assay.- Quantifying Plasmodesmatal Transport with an Improved GFP Movement Assay.- An Arabidopsis Callus Grafting Method to Test Cell-to-Cell Mobility of Proteins.- Quantification of Plasmodesmata Permeability in Arabidopsis Leaves by Tracing the Movement of GFP.- Tracking Intercellular Movement of Fluorescent Proteins in Bryophytes.- Virus Genome-Based Reporter for Analyzing Viral Movement Proteins and Plasmodesmata Permeability.- Analysis of the Distribution of Symplasmic Tracers during Zygotic and Somatic Embryogenesis.- Quantifying Intercellular Movement and Protein Stoichiometry for Computational Modeling.- Spatiotemporal Specific Blocking of Plasmodesmata by Callose Induction.- A Forward Genetic Approach to Identify Plasmodesmal Trafficking Regulators Based on Trichome Rescue.- In Vivo Visualization of Mobile mRNA Particles in Plants Using BglG.- Multi-Angle In Vivo Imaging of the Arabidopsis thaliana Shoot Apical Meristem (SAM).- More Insights from Ultrastructural and Functional Plasmodesmata Data Using PDinsight.- Measuring Intercellular Interface Area in Plant Tissues Using Quantitative 3D Image Analysis.