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Abstract

A large body of evidence has lead to the suggestive proposal of a potential interplay between viroid-induced pathogenesis and RNA silencing regulatory mechanisms. A variety of techniques have been used to examine this interaction. This chapter outlines the use of a green fluorescent protein (GFP) sensor, containing viroid-specific sequence targets to study this phenomenon. In addition, a symptomatic transgenic line of Nicotiana benthamiana that expresses and processes Hop stunt viroid (HSVd) is used as stocks in grafting assays with the rdr6i-Nb lines, in which the RNA-dependent RNA polymerase 6 (NbRDR6) is constitutively silenced. The combination of the use of a viroid-specific GFP-sensor, the grafting assays with plants silenced for the RDR6 and the analysis of siRNAs allows to address how to monitor viroid-specific RNA silencing and how to associate this regulatory pathway with symptom expression.

Published in

Methods in Molecular Biology
2012, volume: 894, pages: 323-343
Title: Antiviral Resistance in Plants : methods and protocols
Publisher: Humana Press, Totowa, NJ

SLU Authors

UKÄ Subject classification

Agricultural Science

Publication identifier

  • DOI: https://doi.org/10.1007/978-1-61779-882-5_22
  • ISBN: 978-1-61779-881-8
  • eISBN: 978-1-61779-882-5

Permanent link to this page (URI)

https://res.slu.se/id/publ/84472