Chloroplast Research in Arabidopsis

Research into chloroplast biology has been hampered by heterogeneous methodologies and subject species. Now Arabidopsis has been adopted as the model research organism, and this second volume covers numerous aspects of it including multi-protein complexes.

Chloroplasts are essential for the survival and flourishing of life on Earth. Over the years, chloroplast biology has been studied in a variety of different organisms, leading to the significant disadvantage that findings which were made by using different experimental systems or species were not always directly cross-comparable. The relatively recent adoption of Arabidopsis thaliana as the model organism of choice for plant science research, across the globe, has led to its emergence as a pre-eminent system for research on chloroplasts and other types of plastid. In Chloroplast Research in Arabidopsis: Methods and Protocols , expert researchers bring together some of the most important, modern techniques and approaches for chloroplast research, with the unifying theme of Arabidopsis as the model system. Volume II explores topics such as multiprotein complexes, protein-protein interactions, omics and large-scale analyses, proteomics and suborganellar fractionation, as well as photosynthesis and biochemical analysis. 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.

Authoritative and convenient, Chloroplast Research in Arabidopsis: Methods and Protocols serves as an ideal reference for all researchers with a general interest in chloroplasts, plastids, or related processes.

Brings together the most important and current techniques in chloroplast research through the unifying lens of the Arabidopsis model system Features subjects like protein aspects, omics studies, and photosynthesis and biochemical analysis Includes expert tips and implementation advice to guarantee ideal experimental results

Inhalt

One- and Two-Dimensional Blue Native (BN) PAGE and Immunodetection of Low-Abundance Chloroplast Membrane Protein Complexes.- Analysis of Thylakoid Protein Complexes by Two Dimensional Electrophoretic Systems.- Preparation of Multiprotein Complexes from Arabidopsis Chloroplasts Using Tandem Affinity Purification.- Studying Interactions between Chloroplast Proteins in Intact Plant Cells Using Bimolecular Fluorescence Complementation (BiFC) and Förster Resonance Energy Transfer (FRET).- Studying Chloroplast Protein Interactions In Vitro: An Overview of the Available Methods.- Proteome Databases and Other Online Resources for Chloroplast Research in Arabidopsis.- Use of Transcriptomics to Analyse Chloroplast Processes in Arabidopsis.- Use of Non-Aqueous Fractionation and Metabolomics to Study Chloroplast Function in Arabidopsis.- Chloroplast Phenomics: Systematic Phenotypic Screening of Chloroplast Protein Mutants in Arabidopsis.- Preparation of Envelope Membrane Fractions from Arabidopsis Chloroplasts for Proteomic Analysis and Other Studies.- Preparation of Stroma, Thylakoid Membrane, and Lumen Fractions from Arabidopsis thaliana Chloroplasts for Proteomic Analysis.- Preparation of Plastoglobules from Arabidopsis Plastids for Proteomic Analysis and Other Studies.- Preparation and Proteomic Analysis of Chloroplast Ribosomes.- The Workflow for Quantitative Proteome Analysis of Chloroplast Development and Differentiation, Chloroplast Mutants, and Protein Interactions by Spectral Counting.- Use of Phosphoproteomics to Study Post-Translational Protein Modifications in Arabidopsis Chloroplasts.- Use of a Pulse-Amplitude Modulated (PAM) Chlorophyll Fluorometer to Study the Efficiency of Photosynthesis inArabidopsis Plants.- Gas Exchange Measurements for the Determination of Photosynthetic Efficiency in Arabidopsis Leaves.- Measurement of the pH and Electric Field Developed Across Arabidopsis Thylakoids in the Light.- Measurement of Chloroplast ATP Synthesis Activity in Arabidopsis.- Methods for Analysis of Photosynthetic Pigments and Steady-State Levels of Intermediates of Tetrapyrrole Biosynthesis.- Analysis of Starch Metabolism in Chloroplasts.- The Analysis of Lipid Content and Quality in Arabidopsis Plastids.