Plant Reverse Genetics

With contributions from leading researchers, this is an ideal guide to cutting-edge methods for isolating gene functions in plants. The book includes both high-throughput and genome-wide techniques, using Arabidopsis and rice, among other plants, as exemplars.

After the generation of genome sequence data from a wide variety of plants, databases are filled with sequence information of genes with no known biological function, and while bioinformatics tools can help analyze genome sequences and predict gene structures, experimental approaches to discover gene functions need to be widely implemented. In Plant Reverse Genetics: Methods and Protocols, leading researchers in the field describe cutting-edge methods, both high-throughput and genome-wide, involving the models Arabidopsis and rice as well as several other plants to provide comparative functional genomics information. With chapters on the analysis of high-throughput genome sequence data, the identification of non-coding RNA from sequence information, the comprehensive analysis of gene expression by microarrays, and metabolomic analysis, the thorough methods of the book are fully supported by scripts to aid their computational use. Written in the highly successful Methods in Molecular Biology™ series format, the chapters contain introductions to their respective topics, lists of the necessary materials, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and essential, Plant Reverse Genetics: Methods and Protocols is an ideal guide for researchers seeking an understanding of how the complex web of plant genes work together in a systems biology view.

Presents an experimental framework to ascribe functions to plant genes in model and crop plants Describes methods for generating mutants in model and specific target plants Includes detailed protocols for the analysis of transcriptome, metabolome, and ultra high-throughput sequencing data Examines high-throughput analysis supported by simple computer scripts to automate the process Includes supplementary material: sn.pub/extras

Inhalt

Analysis of High-Throughput Sequencing Data.- Identification of Plant MicroRNAs Using Expressed Sequence Tag Analysis.- Microarray Data Analysis.- Setting Up Reverse Transcription Quantitative-PCR Experiments.- Virus-Induced Gene Silencing in Nicotiana benthamiana and Other Plant Species.- Agroinoculation and Agroinfiltration: Simple Tools for Complex Gene Function Analyses.- Full-Length cDNA Overexpressor Gene Hunting System (FOX Hunting System).- Activation Tagging with En/Spm-I/dSpm Transposons in Arabidopsis.- Activation Tagging and Insertional Mutagenesis in Barley.- Methods for Rice Phenomics Studies.- Development of an Efficient Inverse PCR Method for Isolating Gene Tags from T-DNA Insertional Mutants in Rice.- Transposon Insertional Mutagenesis in Rice.- Reverse Genetics in Medicago truncatula Using Tnt1 Insertion Mutants.- Screening Arabidopsis Genotypes for Drought Stress Resistance.- Protein Tagging for Chromatin Immunoprecipitation (ChIP) from Arabidopsis.- Yeast One-Hybrid Screens for Detection of Transcription Factor DNA Interactions.- Plant Metabolomics by GC-MS and Differential Analysis.- Gramene Database: A Hub for Comparative Plant Genomics.