RNAi and Plant Gene Function Analysis

With RNAi technology now essential to most plant science researchers, this book introduces the central topics, lists the necessary materials and reagents, offers reproducible laboratory protocols, and provides notes on troubleshooting and avoiding pitfalls.

The use of RNAi technology is essential for most plant science researchers. As DNA sequence information increases, so the need for functional annotation of target genes also increases. Authoritative and accessible, RNAi and Plant Gene Function Analysis: Methods and Protocols seeks to serve as an ideal guide to scientists of all backgrounds looking to further their knowledge of this fundamental science. Written in the highly successful Methods in Molecular Biology ™ series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

In addition to the functional annotation of unknown genes, RNAi technology has been applied to the genetic engineering of important plant metabolites including starches, oils, and storage proteins. RNAi has been used to engineer plants resistant to plant viruses and also to nematodes and insects. RNAi and Plant Gene Function Analysis: Methods and Protocols will provide both professionals and novices of plant biotechnology and breeding research with essential tips for designing experiments meant to explore plant gene function.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt

Overview of Plant RNAi .- Caveat of RNAi in Plants: the Off-Target Effect .- Plant Gateway Vectors for RNAi as a Tool for Functional Genomic Studies .- Heat-Inducible RNAi for Gene Functional Analysis in Plants .- Gene Function Analysis by Artificial MicroRNAs in Physcomitrella patens .- Virus-Induced Gene Silencing in Ornamental Plants .- Local RNA Silencing Mediated by Agroinfiltration .- Direct Transfer of Synthetic Double-Stranded RNA into Protoplasts of Arabidopsis thaliana .- Detection of Long and Short Double-Stranded RNAs .- Quantitative Stem-Loop RT-PCR for Detection of MicroRNAs .- Large Scale Sequencing of Plant Small RNAs .- Computational Prediction of Plant miRNA Targets.- Bisulfite S equencing for C ytosine- M ethylation A nalysis in Plants .- Using Nuclear Run-On Transcription Assays in RNAi Studies .- Proteomic Analysis of RNA-Interference Induced Knockdown Plant .- Comparative Analysis of Phosphoprotein Expression using 2D-DIGE