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This detailed volume collects protocols from scientists who are actively engaged in developing or using protoplast technology. The book begins with chapters that focus on basic protoplast techniques and their utilities, such as protocols on protoplast isolation, transfection, and regeneration, as well as examples of how to use protoplasts for genome editing and gene function analysis in a number of major crop or model plant species. It continues by exploring protoplast automation, large scale functional genomics, and synthetic biology. 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.
Authoritative and practical, Protoplast Technology: Methods and Protocols aims to inspire the new generation of researchers to further improve their protocols and apply this technology to accelerate the field of plant genomic study.
Sommario
Physcomitrium patens Protoplasting and Protoplast Transfection.- Protoplast Isolation and Transfection in the Single-Cell C4 Species Bienertia sinuspersici.- Protoplast for Gene Functional Analysis in Arabidopsis.- Application of Protoplast Regeneration to CRISPR/Cas9 Mutagenesis in Nicotiana tabacum.- CRISPR DNA- and RNP-Mediated Genome Editing via Nicotiana benthamiana Protoplast Transformation and Regeneration.- Protoplast Isolation and Transfection in Rice.- Protoplast Isolation and Transfection in Maize.- Isolation and Transfection of Maize Endosperm Protoplasts.- Protoplast Isolation from Undifferentiated Maize Seedling Shoot Tissue.- Protoplast Isolation and Transfection in Wheat.- Protoplast Isolation, Transformation, and Regeneration for Forage and Turf Grasses.- Efficient Protoplast Isolation and DNA Transfection for Winter Oilseed Crops, Pennycress (Thlaspi arvense) and Camelina (Camelina sativa).- Protoplast Isolation,Transfection, and Gene Editing for Soybean (Glycine max).- Protoplast Isolation and Transformation in Oil Palm.- Protoplast Preparation and Fluorescence-Activated Cell Sorting for the Evaluation of Targeted Mutagenesis in Plant Cells.- Characterization of Programmable Transcription Activators in the Model Monocot Setaria viridis via Protoplast Transfection.- High-Throughput Transfection and Analysis of Soybean (Glycine max) Protoplasts.
Riassunto
The book begins with chapters that focus on basic protoplast techniques and their utilities, such as protocols on protoplast isolation, transfection, and regeneration, as well as examples of how to use protoplasts for genome editing and gene function analysis in a number of major crop or model plant species.
