Detection of CfMV infection in oat plants inoculated with CfMV. (A) RT-PCR–Reverse transcription PCR analysis of total RNA extracted from plants biolistically inoculated with CfMV (WT) and CfMV mutants (R3L, R5X, noCP). RNA from a non-inoculated plant (n.i.) was used as negative control and RNA from a plant previously known to be infected was used as positive control (p.c.). Samples were collected from the inoculated leaves (Inoculated) at 14 dpi and from upper leaves (Systemic) at 21 dpi. Primers amplifying the region of RdRp and CP genes were used. W.blot–Western blot analysis of plant total protein extracts with polyclonal anti CfMV CP antibody (CP-ab). An additional positive control was not used. Ribulose-1,5-bisphosphate carboxylase oxygenase large subunit (RuBP-L) was visualised with Ponceau S stain as loading control. (B) Detection of CfMV infection in sap-inoculated plants. Oat plants were sap-inoculated with CfMV and its mutants. Analysis and annotation as described in (A).