By John A. Lindbo (auth.), John M. Watson, Ming-Bo Wang (eds.)
Studies with regards to pathogen-mediated virus resistance in vegetation have been instrumental in supplying many of the historic observations which eventually ended in the important discovery of double-stranded RNA (dsRNA)-induced gene silencing or RNA interference (RNAi), which has considering the fact that revolutionized examine on plant-virus interactions. In Antiviral Resistance in vegetation: tools and Protocols, specialist researchers within the box aspect some of the tools that are now everyday to check the phenomenon of RNA silencing relating to viral infections of vegetation. those contain equipment and methods for the isolation and quantitative/qualitative analyses of plant small 21-24 nucleotide RNAs equivalent to small interfering RNAs (siRNAs) and microRNAs (miRNAs) in addition to the research and manipulation of virus-induced gene silencing (VIGS) in either monocotyledonous and dicotyledenous vegetation and using hairpin RNA (hpRNA) transgenes. Written within the hugely winning Methods in Molecular Biology™ sequence layout, chapters contain introductions to their respective themes, lists of the required fabrics and reagents, step by step, effectively reproducible laboratory protocols, and key pointers on troubleshooting and heading off identified pitfalls.
Authoritative and sensible, Antiviral Resistance in crops: tools and Protocols seeks to help scientists within the extra examine of this crucially vital botanical trait.
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Additional info for Antiviral Resistance in Plants: Methods and Protocols
The SWI2/ SNF2-like chromatin remodeling factor DRD1 (38),—a homologue of CLASSY1, is also required for the physical interaction of RNA pol V with the target loci (82). AGO4, RNA pol V, and the methyltransferase DRM2 have all been found to localize in Cajal bodies, which are found adjacent to the nucleolus (91, 92). These bodies appear to centralize the DNA methylation machinery. However, AGO4 displays dynamic localization as it is found throughout the nucleus, presumably at methylation sites (92).
This appears counterintuitive as siRNAs, processed from a dsRNA precursor molecule, should contain relatively equal amount of plus- and minus-strand species. To account for this strand bias, Molnár and colleagues (93), based on their analysis of siRNA distribution from Cymbidium ringspot tombusvirus, proposed a model where certain regions of the viral ssRNAs may form partially double-stranded secondary structures that are processed by a DCL into siRNAs. This local base-pairing model has also been suggested to account for siRNA production from the translational leader sequence of the cauliflower mosaic DNA virus that forms extensive secondary structures (94).
The 2b protein dramatically reduces the accumulation of all three size classes of viral siRNAs in Arabidopsis (22), possibly because its inhibition on AGO1 activity reduces the amount of viral RNA cleavage product that serves as template for RDRs. A more destructive mode of action by silencing suppressors has recently been reported for the Polerovirusencoded P0 protein. The P0 protein contains an F-box motif and targets the PAZ domain and its adjacent upstream sequence in AGO1 and mediates its degradation (109, 110).
Antiviral Resistance in Plants: Methods and Protocols by John A. Lindbo (auth.), John M. Watson, Ming-Bo Wang (eds.)