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This second edition volume expands on the previous edition with new and updated chapters that explore our current view on plastid evolution, structure, and function. The chapters in this book are organized into three parts and cover topics such as plastid evolution, plasticity, and functional and structural diversity; techniques used to visualize, fractionate, purify, and study primary plastids from plant materials, and secondary plastids; and methods to analyze plastids by integrated biology strategies based on genetics, genomics, proteomics, and lipidomics. 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 authoritative, Plastids: Methods and Protocols, Second Edition is a valuable resource that willhelp students, engineers, and researchers further explore and understand this fascinating organelle.
List of contents
How Did Thylakoids Emerge in Cyanobacteria and How Were the Primary Chloroplast and Chromatophore Acquired?.- Complex Endosymbioses I: From Primary to Complex Plastids, Serial Endosymbiotic Events.- Complex Endosymbiosis II: The Non-Photosynthetic Plastid of Apicomplexa Parasites (The Apicoplast) and Its Integrated Metabolism.- Diversification of Plastid Structure and Function in Land Plants.- The Main Functions of Plastids.- Plastid Transient and Stable Interactions with Other Cell Compartments.- Purification of Chloroplast Envelope, Thylakoids, and Stroma from Angiosperm Leaves.- Isolation of the Inner and Outer Membranes of the Chloroplast Envelope from Angiosperms.- Analysis of a Super-Complex at Contact Sites between Mitochondria and Plastids.- Isolation of High-Quality Plastids from the Diatom Phaeodactylum tricornutum.- Determining the Subcellular Localization of Proteins in the Different Membranes of Diatom Secondary Plastid.- Monitoring of Lipid Fluxes between Host and Plastid-Bearing Apicomplexan Parasites.- Extraction and Quantification of Lipids from Plant or Algae.- Quantitative Assessment of the Chloroplast Lipidome.- The Use of Nanopore Sequencing to Analyze the Chloroplast Transcriptome Part I: Library Preparation.- The Use of Nanopore Sequencing to Analyze the Chloroplast Transcriptome Part II: Bioinformatic Analyzes and Virtual RNA Blots.- Targeted Gene Editing of Nuclear-Encoded Plastid Proteins in Phaeodactylum tricornutum via CRISPR/Cas9.- Isolation of Cytosolic Ribosomes Associated with Plant Mitochondria and Chloroplasts.- A Practical Guide to the Representation of Protein Regulation in the Web Application ChloroKB.
