Geminiviruses Methods and Protocols

This book provides a comprehensive list of the methods used in basic and applied geminivirus research. Chapters guide readers through foundational methods, advanced techniques in cloning, sequencing, phylogenetic, recombination analysis, diagnostics,  and virus-host interaction studies. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

Authoritative and cutting-edge, Geminiviruses: Methods and Protocols aims to be a practical laboratory guide and serve as a valuable "one-stop-shop" for researchers new to geminiviruses, facilitating the establishment of a robust geminivirus research program.  

Extraction of geminivirus and associated satellite DNA from leaf samples.- Mechanical sap transmission of geminiviruses.- Cloning of complete geminivirus genomes by rolling circle amplification (RCA).- Biolistic inoculation of geminiviruses.- Agroinoculation of plants with geminiviruses.- Begomovirus transmission by the insect vector, the whitefly Bemisia tabaci.- Mastrevirus transmission by the insect vectors, leafhoppers.- Transmission of grabloviruses by insect vectors.- Sequence Demarcation Tool (SDT), a free user-friendly computer program using pairwise genetic identity calculations to classify nucleotide or amino acid sequences.- Phylogenetic analysis of geminiviruses.- Recombination analysis of geminiviruses using Recombination Detection Program (RDP).- Detection of tomato leaf curl New Delhi virus-ES by real-time quantitative PCR.- Detection of geminiviruses using loop-mediated isothermal amplification (LAMP).- Tissue printing hybridization using digoxigen-labeled DNA probes for the detection of geminiviruses and associated DNA satellites.- High throughput sequencing (HTS)-based diagnosis of geminiviruses.- CRISPR-Cas-based diagnosis of geminiviruses.- Full-length sequencing of circular DNA viruses using CIDER-Seq.- Subcellular localization of geminivirus proteins by laser scanning confocal microscopy.- Identification of interaction between geminivirus and plant proteins by bimolecular fluorescence complementation (BiFC).- Exploring the effect of tomato yellow leaf curl virus (TYLCV) infection on whitefly probing and feeding behaviour by Electrical Penetration Graph (EPG) technique.