Plant Mitochondria Methods and Protocols

This detailed volume presents a wide range of techniques for plant mitochondrial analysis, ranging from tried-and-tested work horse techniques to the latest innovations. Within its pages, it explores subjects such as affinity-based isolation of mitochondria with magnetic beads, mitochondrial quality assessment protocols, measurement of uptake and release of specific metabolites, mitochondrial protein identification and visualization, as well as gene splicing and editing, and much more. 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. 

Purification of Leaf Mitochondria from Arabidopsis thaliana Using Percoll Density Gradients.- Cell Type-Specific Isolation of Mitochondria in Arabidopsis.- Isolation of Plant Mitochondria Using Affinity Purification.- Isolation of Highly Purified, Intact, and Functional Mitochondria from Potato Tubers Using a Two-in-One Percoll Density Gradient.- Integrity Assessment of Isolated Plant Mitochondria.- High-Throughput Oxygen Consumption Measurements in Leaf Tissue Using Oxygen Sensitive Fluorophores.- Assessment of Respiratory Enzymes in Intact Cells by Permeabilization with Alamethicin.- Assessing the Kinetics of Metabolite Uptake and Utilization by Isolated Mitochondria Using Selective Reaction Monitoring-Mass Spectrometry (SRM-MS).- Complexome Profiling of Plant Mitochondrial Fractions.- High Throughput BN-PAGE for Mitochondrial Respiratory Complexes.- Assessment of Mitochondrial Protein Composition and Purity by Mass Spectroscopy.- Mitochondrial GFP-Tagged Protein Localization Using Transient Transformations in Arabidopsis thaliana.- Assessment of Mitochondrial Protein Topology and Membrane Insertion.- Assessment of Protein Synthesis in Mitochondria Isolated from Rosette Leaves and Liquid Culture Seedlings of Arabidopsis.- Markers for Mitochondrial ROS Status.- Mass Spectrometry-Based Quantitative Cysteine Redox Proteome Profiling of Isolated Mitochondria Using Differential iodoTMT Labeling.- Analysis of the Plant Mitochondrial Transcriptome.- Quantification of Mitochondrial RNA Editing Efficiency Using Sanger Sequencing Data.- Nascent Transcript Sequencing for the Mapping of Promoters in Arabidopsis thaliana Mitochondria.- Assessment of Mitochondrial DNA Copy Number, Stability, and Repair in Arabidopsis.- Measuring the Activity of DNA Repair Enzymes in Isolated Mitochondria.- MitoTALENs: A Method for Targeted Gene Disruption in Plant Mitochondrial Genomes.