Plant Cell Electroporation and Electrofusion Protocols

About this book

Leading experts and innovators describe in detail plant cell electroporation and electrofusion techniques for many types of plants. Their protocols cover a diverse set of plant cell types, including different cell types from a single organism (i.e., leaf cells and pollen) and the most important model plants, including maize and tobacco. Each protocol also contains detailed instructions for growth and growth optimization, protoplast generation, and protoplast regeneration for specific plant cell types, along with extensive troubleshooting advice and descriptions of expected results to simplify study design.

Contents

Part I. Theory And Instrumentation. Electroporation Theory: Concepts and Mechanisms. Effects of Pulse Length and Strength on Electroporation Efficiency. Instrumentation. Part II. Electroporation Protocols. Electroporation of Agrobacterium tumefaciens. Electroporation of DNA into the Unicellular Green Alga Chlamydomonas reinhardtii. Pollen Electrotransformation in Tobacco. Electroporation of Tobacco Leaf Protoplasts Using Plasmid DNA or Total Genomic DNA. Electroporation of Brassica. Transformation of Maize by Electroporation of Embryos. Transient Gene Expression Analysis in Electroporated Maize Protoplasts. Reporter Genes and Transient Assays for Plants. Part III. Electrofusion Protocols. Electrofusion of Plant Protoplasts: Selection and Screening for Somatic Hybrids of Nicotiana. Protoplast Electrofusion and Regeneration in Potato. Polymer- Supported Electrofusion of Protoplasts: A Novel Method and a Synergistic Effect. Index.

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