The Plant Endoplasmic Reticulum (2^nd Ed., 2nd ed. 2024) Methods and Protocols Methods in Molecular Biology Series, Vol. 2772

This second edition provides new and updated methods detailing techniques and state-of-the-art approaches on the structure and function of plant endoplasmic reticulum (ER). Chapters guide readers through modern microscopy techniques, software protocols, purification, and analysis of ER membrane structure. 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 key tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, The Plant Endoplasmic Reticulum: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.

 Make it shine: Labelling the ER for light and fluorescence microscopy.- Electron microscopy techniques for 3D plant ER imaging.- Studying plant ER-PM contact site localised proteins using microscopy.- Preparation and imaging of specialised ER using super-resolution and TEM techniques.- Quantitation of ER morphology and dynamics.- Long-term Imaging of Endoplasmic Reticulum Morphology in Embryos During Seed Germination.- Dancing with the stars: Using image analysis to study the choreography of the endoplasmic reticulum and its partners and of movement within its tubules.- Preparation of highly enriched ER membranes using free-flow electrophoresis.-Preparation of ER microsomes from Arabidopsis thaliana.- ER membrane lipid composition and metabolism: lipidomic analysis.- 2in1 vectors improve in planta BiFC and FRET analysis.- Immunoprecipitation and FRET-FLIM to determine metabolons on the plant ER.- Using optical tweezers combined with total internal reflection microscopy to study interactions between the ER and Golgi in plant cells.- Protein biosynthesis and maturation in the ER.- ER membrane Protein Interactions using the Split-Ubiquitin System (SUS).- Analysis of protein glycosylation in the ER.- Unfolded Protein Response in Arabidopsis.- Imaging the ER and endomembrane system in cereal endosperm.- Multi-omics resources for understanding gene regulation in response to ER stress in plants.- Variable angle epifluorescence microscopy for single particle tracking in the plant ER.- Use of super resolution live cell imaging to distinguish endoplasmic reticulum - nuclear envelope subcellular localisation.- Using ER-targeted photoconvertible fluorescent proteins in living plant cells.- In vivo analysis of ER-associated protein degradation and ubiquitination in Arabidopsis thaliana.- Overexpression of endoplasmic reticulum proteins from Arabidopsis thaliana in baculovirus.- Observing ER dynamics over long timescales using light sheet fluorescence microscopy.- Gene stacking and stoichiometric expression of ER-targeted constructs using “2A” self-cleaving peptides.- Automated Segmentation and 3D Reconstruction of Different Membranes from Confocal Z-stacks.- Visualizing orientation and topology of ER membrane proteins in planta.- Organ specific microsomes from dark grown hypocotyls of Arabidopsis thaliana.- Studying secretory protein synthesis in Nicotiana benthamiana protoplasts.- Analyzing photoactivation with diffusion models to study transport in the Endoplasmic Reticulum network.