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 crops have been instrumental in offering many of the old observations which finally resulted in the important discovery of double-stranded RNA (dsRNA)-induced gene silencing or RNA interference (RNAi), which has given that revolutionized study on plant-virus interactions. In Antiviral Resistance in vegetation: equipment and Protocols, specialist researchers within the box aspect the various equipment that are now standard to review the phenomenon of RNA silencing on the subject of viral infections of vegetation. those contain tools and strategies for the isolation and quantitative/qualitative analyses of plant small 21-24 nucleotide RNAs reminiscent of 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 profitable Methods in Molecular Biology™ sequence layout, chapters comprise introductions to their respective issues, lists of the required fabrics and reagents, step by step, effortlessly reproducible laboratory protocols, and key tips about troubleshooting and heading off identified pitfalls.
Authoritative and functional, Antiviral Resistance in vegetation: equipment and Protocols seeks to help scientists within the additional learn of this crucially very important botanical trait.
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Additional resources for Antiviral Resistance in Plants: Methods and Protocols
However, several pieces of recent evidence suggest that interference of host sRNA pathways by viral silencing suppressors is not the primary cause of disease symptoms. First, not all viral suppressors appear to affect the endogenous miRNA pathway in plants (122). Second, a recent study showed that systemic infection of dcl2 dcl4 double mutant plants with silencing suppressor-deficient TCV and CMV induces severe disease symptoms similar to those of wild-type viruses (22, 126). Also, a study based on ectopic expression of DCL1 indicated that developmental anomalies associated with transgenic expression of the silencing suppressor HC-Pro do not result from general impairments in sRNA pathways (127).
Cell 125: 1111–1124 Allen E, Xie Z, Gustafson AM, Carrington JC (2005) microRNA-directed phasing during trans-acting siRNA biogenesis in plants. Cell 121:207–221 Axtell MJ, Jan C, Rajagopalan R, Bartel DP (2006) A two-hit trigger for siRNA biogenesis in plants. Cell 127:565–577 Montgomery TA, Yoo SJ, Fahlgren N, Gilbert SD, Howell MD, Sullivan CM et al (2008) AGO1–miR173 complex initiates phased siRNA formation in plants. Proc Natl Acad Sci U S A 105:20055–20062 Peragine A, Yoshikawa M, Wu G, Albrecht HL, Poethig RS (2004) SGS3 and SGS2/ SDE1/RDR6 are required for juvenile development and the production of trans-acting siRNAs in Arabidopsis.
Binding to long dsRNAs or siRNA duplexes appears to be a predominant strategy adopted by the silencing suppressors to inhibit RNA silencing. The P14 protein of aureusvirus and the coat protein (CP) of Turnip crinkle carmovirus bind dsRNA without size selection (104), and these suppressors may inhibit silencing by sequestering long dsRNA precursors from processing by DCLs as well as siRNAs from incorporation into RISC. Many silencing suppressors bind specifically to 2 RNA Silencing and Antiviral Defense in Plants 29 21-nucleotide siRNA duplexes, such as P15 of Peanut clump pecluvirus, γB of Barley stripe mosaic hordeivirus, P21 of Beet yellows closterovirus, HC-Pro of Tobacco etch virus potyvirus, p19 of Tomato bushy stunt tombusvirus, 2b of Tomato aspermy Cucumovirus, and CP of Pelargonium flower break tombusvirus (104–107).