Figure 5.

Replication and trafficking rates for PSTVd G/U double mutants. For each stem region containing two G/U pairs, G/U to U/U and G/U to G/C single mutants were separately combined to generate double mutants. The nucleotide coordinates for the coupled G/U pairs are given below the diagram of the canonical PSTVd secondary structure. Ten N. benthamiana plants were inoculated with each mutant and RNA from inoculated (local) and systemically infected leaves was evaluated for the presence of PSTVd by RNA blot analysis. For each mutation type, 10 boxes within a grid represent 10 inoculated plants. Lower grids show data for local inoculated leaves, where positive signals indicate replication. Upper grids show data for systemically infected leaves, where positive signals indicate potential systemic trafficking. Colored circles represent infected plants: double G/U to U/U (red); double G/U to G/C (blue). Symbols above the diagram represent results of progeny sequencing from systemically infected leaves. Red cross: retained one introduced mutation, one reverted to G/U. Red square: lost both introduced mutations (reverted to G/U) and acquired a new mutation. For single mutant data, see Figure 4.