Antiviral Resistance in Plants: Methods and Protocols by John A. Lindbo (auth.), John M. Watson, Ming-Bo Wang (eds.)

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 delivering many of the ancient observations which finally ended in the important discovery of double-stranded RNA (dsRNA)-induced gene silencing or RNA interference (RNAi), which has because revolutionized examine on plant-virus interactions. In Antiviral Resistance in vegetation: tools and Protocols, specialist researchers within the box aspect the various tools that are now accepted to review the phenomenon of RNA silencing with regards to viral infections of vegetation. those contain equipment and methods for the isolation and quantitative/qualitative analyses of plant small 21-24 nucleotide RNAs comparable 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 structure, chapters comprise introductions to their respective themes, lists of the required fabrics and reagents, step by step, easily reproducible laboratory protocols, and key tips about troubleshooting and heading off recognized pitfalls.

Authoritative and functional, Antiviral Resistance in vegetation: equipment and Protocols seeks to assist scientists within the extra research of this crucially very important botanical trait.

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EMBO J 17:170–180 25. Hutvagner G, Simard MJ (2008) Argonaute proteins: key players in RNA silencing. Nat Rev Mol Cell Biol 9:22–32 26. Parker JS, Barford D (2006) Argonaute: a Scaffold for the function of short regulatory RNAs. Trends Biochem Sci 31:622–630 27. Song J, Smith SK, Hannon GJ, Joshua-Tor L (2004) Crystal structure of Argonaute and its implications for RISC Slicer activity. Science 305:1434–1437 28. Vaucheret H (2008) Plant ARGONAUTES. Cell 13:350–358 29. Peters L, Meister G (2007) Argonaute proteins: mediators of RNA silencing.

Thaliana is said to have two primary roles: regulating gene expression and protecting the genome from “selfish DNA elements” (73). It has been suggested that some 30% of cytosine methylation in A. thaliana may be directed by siRNAs (39). The effective siRNA species in RdDM are ra-siRNAs, which direct the methylation of specific sequences that largely correspond to the region of complementarity between the siRNA and the target DNA (74). These 24-nucleotide siRNAs have their origin in methylated DNA and function to reinforce methylation on specific loci of repetitive DNA and retroelements (70).

Development 133: 2973–2981 73. Chan SW-L, Henderson IR, Jacobsen SE (2005) Gardening the genome: DNA methylation in Arabidopsis thaliana. Nat Rev Genet 6:351–360 74. Matzke MA, Birchler JA (2005) RNAimediated pathways in the nucleus. Nat Rev Genet 6:24–35 75. Kanno T, Huettel B, Mette MF, Aufsatz W, Jaligot E, Daxinger L et al (2005) Atypical RNA polymerase subunits required for RNAdirected DNA methylation. Nature 37: 761–765 76. Onodera Y, Haag JR, Ream T, Nunes PC, Pontes O, Pikaard CS (2005) Plant nuclear RNA polymerase IV mediates siRNA and DNA methylation-dependent heterochromatin formation.

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