Becker A. (Ed.) Virus-Induced Gene Silencing: Methods and Protocols

Humana Press, 2013. — 235 p. — (Methods in Molecular Biology № 975).Plants are amazing organisms to study, some are important sources for pharmaceuticals, and others can help to elucidate molecular mechanisms required for a plant’s development and its interactions with the biotic or abiotic environment. Functional genomics is vastly lagging behind the speed of genome sequencing as high-throughput gene function assays are difficult to design, specifically for non-model plants. Bioinformatics tools are useful for gene identification and annotation but are of limited value for predictions concerning gene functions as gene functions are uncovered best by experimental approaches. Virus-Induced-Gene-Silencing (VIGS) is an easy to use, fast, and reliable method to achieve down regulation of target gene expression. The book provides detailed protocols for VIGS experiments in several plant species including model and non-model plants. Also included in this book are recently developed protocols for VIGS-derived microRNA production in the plant or protein over expression, as well as chapters devoted to summarizing the molecular mechanisms of VIGS action and the vector systems developed so far. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.