Phloem

This second edition volume expands on the previous edition with updates on the latest innovative techniques and protocols used to study phloem. The chapters in this book cover the whole spectrum of scientific techniques from molecular biology to anatomical studies; imaging and tracer-based analyses; theoretical and modeling approaches; and the whole range of functional aspects of the phloem, from molecular-scale processes to its role in whole plant performance and responses to stress. 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.

Cutting-edge and thorough, Phloem: Methods and Protocols, Second Edition is a practical and detailed reference for scientists with different backgrounds. This volume provides enough context of methodological principles to help researchers select the most appropriate methods for their specific questions, and will inspire them to combine techniques from different subfields of phloem studies to learn more about this sensitive and complex tissue.

Preparation of Tissue Sections for Light-Microscopic Analysis of Phloem Anatomy. - Transmission Electron Microscopy of the Phloem. - Immunolabeling Sieve Element Cell Walls with the LM26 Antibody. - Quantification of Callose Deposition in Phloem of Woody Stems Using Supervised Machine Learning-Driven Automated Image Analysis with the
IlastiKlean
R Package. - Anatomical Features of Foliar Vasculature as Flux-Capacity Proxies. - EDTA-Facilitated Collection of Phloem Exudates for Chemical Analyses. - Using Aphids to Sample Phloem Sap. - Measurement of Inner Bark and Leaf Osmolality. - Infrared Microimaging of Sucrose Distribution in Plant Vascular Tissues. - Identification of Phloem Cell Transcriptome via Single Cell RNA-Sequencing. - Live-Cell Imaging of Fluorescently-Tagged Phloem Proteins with Confocal Microscopy. - Immunohistochemical Localization of Proteins in the Phloem: Problems and Solutions. - Analysis of Phloem Sterols in Plants using EDTA-Facilitated Exudation and GC-MS. - Visual Detection of Inorganic Orthophosphate in Phloem Cells. - Characterization of Viruses in Phloem by Correlative X-Ray Microtomography ( CT)-Volume Electron Microscopy (vEM) Imaging. - Go with the Phloem: A Review of the Methods to Collect Plant Phloem Tissue. - Detecting Rapid Changes in Carbon Transport and Partitioning with Carbon-11 (
¹¹
C). - Using
¹³
C to Quantify Phloem Transport on Tall Plants in the Field. - Non-Invasive Determination of Phloem Transport Speed with
¹⁴
C. - Measuring Phloem Transport Velocity on a Tissue-Level Using a Phloem-Mobile Dye. - Determination of the Phloem Mass Flow Velocity and Calculation of the Volumetric Flow Rate. - Measuring Esculin Export from Leaves as Proxy for Sucrose Loading Rates in the Phloem. - Relative Quantification of Phloem Loading with
¹³
CO
₂
in Grass Leaves. - Predicting Phloem Loading Types with Electron Microscopy. - Measurement of Subcellular Metabolite Concentrations in Relation to Phloem Loading. - Methods to Estimate Carbon Export from Leaves. - Neutron Imaging to Reveal Phloem Function in Trees. - A Dual-Reporter System for the Analysis of Phloem

Structural and Signaling Responses In Vivo. - Regulatory DNA Sequences Controlling Gene Expression in Leaf Phloem Cells. - Quantifying Fire-Induced Phloem Damage. - Methods for Assessing the Role of Phloem Transport in Plant Stress Responses. - Isotope Measurements of Phloem-Transported Organic Matter: Tools to Obtain Information on Gas Exchange Physiology and Transport Processes, and to Better Understand Tree Ring Archives. - High-Throughput Hypocotyl Grafting and Vascular Reconnection Assessment in Arabidopsis Seedlings. - Efficient Establishment of Interfamily Heterograft of
Nicotiana benthamiana
and
Arabidopsis thaliana. -
VISUAL-PH and VISUAL-CC: Modified Culture Systems for Phloem Differentiation in Arabidopsis. - Measurement of Phloem-Based Electrical Signals in Plants. - Modeling the Hydraulic Conductivity of Phloem Sieve Elements. - Phloem Catastrophe: Identifying Plant Tipping Points through Bifurcation Analysis. - Predicting Phloem Transport Rate in Crops.