Phloem : Methods and Protocols

This detailed book presents methods that enable the determination of parameters relevant to phloem research in the most efficient ways, putting plant scientists with access to adequate instrumentation in the position to answer any phloem-related question. The collection explores techniques that have been used for decades, such as tracing phloem transport with carbon isotopes, as well as recent developments, such as esculin-based assays of phloem transport and super-resolution microscopy of phloem proteins. As such, the book presents the state-of-the-art in phloem research and, at the same time, a starting point for the development of new methods that will fill the remaining gaps in the phloem researcher’s toolbox in the future. 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 protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and comprehensive, __Phloem: Methods and Protocols__ is an ideal guide for all researchers exploring this challenging area of plant biology. Front Matter ....Pages i-xiv Front Matter ....Pages 1-1 Optimal Preparation of Tissue Sections for Light-Microscopic Analysis of Phloem Anatomy (Marcelo Rodrigo Pace)....Pages 3-16 Transmission Electron Microscopy of the Phloem with Minimal Artifacts (Pascal Hunziker, Alexander Schulz)....Pages 17-27 Scanning Electron Microscopy of the Phloem (Daniel L. Mullendore)....Pages 29-35 Noninvasive Investigation of Phloem Structure by 3D Synchrotron X-Ray Microtomography (Jussi-Petteri Suuronen, Tuula Jyske)....Pages 37-54 Quantification of Leaf Phloem Anatomical Features with Microscopy (Jared J. Stewart, Onno Muller, Christopher M. Cohu, Barbara Demmig-Adams, William W. Adams III)....Pages 55-72 Methods of Phloem Visualization: A Clear Future in Sight? (Elisabeth Truernit)....Pages 73-79 Front Matter ....Pages 81-81 Super-Resolution Microscopy of Phloem Proteins (Ryan C. Stanfield, Alexander Schulz)....Pages 83-94 Live-Cell Imaging of Fluorescently Tagged Phloem Proteins with Confocal Microscopy (Thibaud Cayla, Rozenn Le Hir, Sylvie Dinant)....Pages 95-108 Immunohistochemical Localization of Proteins in the Phloem: Problems and Solutions (Christina Kühn)....Pages 109-122 Front Matter ....Pages 123-123 Studying Phloem Loading with EDTA-Facilitated Phloem Exudate Collection and Analysis (Qiyu Xu, Yunjuan Ren, Johannes Liesche)....Pages 125-133 Measurement of Inner Bark and Leaf Osmolality (Teemu Paljakka, Anna Lintunen, Yann Salmon, Teemu Hölttä)....Pages 135-142 Front Matter ....Pages 143-143 Using 13C to Quantify Phloem Transport on Tall Plants in the Field (Daniel Epron, Masako Dannoura, Caroline Plain)....Pages 145-151 Noninvasive Determination of Phloem Transport Speed with Carbon-14 (14C) (Christopher Vincent, Peter E. H. Minchin, Johannes Liesche)....Pages 153-162 Detecting Rapid Changes in Carbon Transport and Partitioning with Carbon-11 (11C) (Benjamin A. Babst, Richard Ferrieri, Michael Schueller)....Pages 163-176 In Vivo Veritas: Radiotracers in Studies of Phloem Transport of Carbohydrate (Michael R. Thorpe, Peter E. H. Minchin)....Pages 177-194 Measuring Phloem Transport Velocity in Arabidopsis Seedlings Using the Fluorescent Coumarin Glucoside, Esculin (Kirsten Knox)....Pages 195-201 Measuring Phloem Transport Velocity on a Tissue Level Using a Phloem-Mobile Dye (Jessica A. Savage, Maciej A. Zwieniecki)....Pages 203-211 Front Matter ....Pages 213-213 Quantification of Symplasmic Phloem Loading Capacity with Live-Cell Microscopy (Helle Juel Martens, Chen Gao, Johannes Liesche)....Pages 215-221 Assessing Long-Distance Carbon Partitioning from Photosynthetic Source Leaves to Heterotrophic Sink Organs with Photoassimilated [14C]CO2 (Umesh P. Yadav, Mearaj A. Shaikh, John Evers, Kamesh C. Regmi, Roberto A. Gaxiola, Brian G. Ayre)....Pages 223-233 Measurement of Subcellular Metabolite Concentrations in Relation to Phloem Loading (Gertrud Lohaus)....Pages 235-251 Measuring Sucrose Transporter Activities Using a Protoplast-Esculin Assay (Theresa Rottmann, Ruth Stadler)....Pages 253-266 The AtSUC2 Promoter: A Powerful Tool to Study Phloem Physiology and Development (Ruth Stadler, Norbert Sauer)....Pages 267-287 Front Matter ....Pages 289-289 Using Aphids to Measure Turgor Pressure Inside Sieve Elements (Nick Gould, Peter E. H. Minchin, Michael R. Thorpe)....Pages 291-299 Stem Compression: A Means to Reversibly Reduce Phloem Transport in Tree Stems (Nils Henriksson, Tim T. Rademacher)....Pages 301-310 Methods for Assessing the Role of Phloem Transport in Plant Stress Responses (Sanna Sevanto)....Pages 311-336 Front Matter ....Pages 337-337 Modeling the Hydraulic Conductivity of Phloem Sieve Elements (Kaare H. Jensen)....Pages 339-344 Using a Multi-compartmental Metabolic Model to Predict Carbon Allocation in Arabidopsis thaliana (Maksim Zakhartsev)....Pages 345-369 A Mechanistic Model to Predict Distribution of Carbon Among Multiple Sinks (André Lacointe, Peter E. H. Minchin)....Pages 371-386 Using a Mathematical Model of Phloem Transport to Optimize Strategies for Crop Improvement (Motohide Seki)....Pages 387-395 Making Microfluidic Devices that Simulate Phloem Transport (Jean Comtet)....Pages 397-408 Front Matter ....Pages 409-409 Efficient Establishment of Interfamily Heterograft of Nicotiana benthamiana and Arabidopsis thaliana (Koji Okayasu, Michitaka Notaguchi)....Pages 411-420 Identification of Phloem Mobile mRNAs Using the Solanaceae Heterograft System (Chao Xia, Yi Zheng, Jing Huang, Zhangjun Fei, Cankui Zhang)....Pages 421-431 Analyzing and Predicting Phloem Mobility of Macromolecules with an Online Database (Daogang Guan, Yiji Xia, Shoudong Zhang)....Pages 433-438 Measurement of Electropotential Waves in Intact Phloem Sieve Elements Using Microelectrodes (Matthias R. Zimmermann, Jens B. Hafke, Maria K. Paulmann, Alexandra C. U. Furch)....Pages 439-447 Measurement of Electropotential Waves in Intact Sieve Elements Using Aphids as Bioelectrodes (Alexandra C. U. Furch, Matthias R. Zimmermann, Torsten Will)....Pages 449-457 What Can Cell Culture Systems Reveal About Sieve Element Differentiation? (Masato Saito, Yuki Kondo)....Pages 459-466 Back Matter ....Pages 467-473 "This detailed book presents methods that enable the determination of parameters relevant to phloem research in the most efficient ways, putting plant scientists with access to adequate instrumentation in the position to answer any phloem-related question. The collection explores techniques that have been used for decades, such as tracing phloem transport with carbon isotopes, as well as recent developments, such as esculin-based assays of phloem transport and super-resolution microscopy of phloem proteins. As such, the book presents the state-of-the-art in phloem research and, at the same time, a starting point for the development of new methods that will fill the remaining gaps in the phloem researcher's toolbox in the future. 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 protocols, and tips on troubleshooting and avoiding known pitfalls." -- Prové de l'editor