The interaction between plant viroid-induced symptoms and RNA silencing.
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.
Instituto de Biologia Moleculaire y Celular de Plantas, Valencia, Spain. vpallas@ibmcp.upv.es
Vicente
V
Pallas
German
G
Martinez
Gustavo
G
Gomez
D018553:Cucumis sativus / Q000235:genetics / Q000821:virology; D049452:Green Fluorescent Proteins / Q000235:genetics; D010935:Plant Diseases / Q000821:virology*; D010942:Plant Viruses / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering; D012367:RNA, Viral / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase / Q000037:antagonists & inhibitors / Q000235:genetics; D014026:Nicotiana / Q000235:genetics* / Q000821:virology*; D014772:Viroids / Q000235:genetics*
D034741:RNA, Small Interfering; D012367:RNA, Viral; D049452:Green Fluorescent Proteins; D012324:RNA-Dependent RNA Polymerase