Fig. 5.

Accumulation of viral RNA and siRNA in N. benthamiana plants before and after challenging inoculation. a Accumulation of siRNA derived from WT PVX, mutants E46A, N863A, N968A, E1001A at 10 dpai. b Accumulation of virus-derived RNA and siRNA after challenge inoculation with pCaPVX440GFP in the same plants. The protective interval was 10 days. The RNA and siRNA were detected by Northern blotting hybridization at 10 days after challenge with saps from pCaPVX440GFP infected plants. c Genomic organization and approximate sizes of genomic and subgenomic RNAs derived from pCaPVX440GFP. The red rectangles indicated the PVX CP subgenomic promoter (SGP), green rectangle indicated the position of TMV CP SGP. The experiments were repeated three times independently. Total siRNAs were used as loading control