Figure 5.

Local and systemic green fluorescent protein (GFP) silencing suppression activity of 2b and 2b mutants in the transient expression systems. (A) Fluorescence in agroinfiltrated leaves of Nicotiana benthamiana for co‐expression of GFP from 2b and 2b mutants at 3 and 6 days post‐infiltration (dpi). The empty vector and p19 were used as negative and positive controls, respectively. (B) Northern and Western blot analysis with samples extracted from co‐infiltrated leaves at 3 and 6 dpi. A GFP DNA probe labelled with [α‐³² P]dCTP was used for the detection of GFP mRNA. Anti‐GFP and anti‐flag antibodies were used to detect the accumulation of GFP and 2b, respectively, in the infiltrated leaves. Methylene blue‐stained rRNA and Coomassie brilliant blue (CBB)‐stained protein were used as loading controls for RNA and protein, respectively. V, vector; wt, wild‐type. (C) Systemic silencing of GFP transgene in 16c plants co‐infiltrated with GFP and 2b or various 2b mutants. The empty vector and p19 were used as negative and positive controls, respectively. The number ratios (S/T) of plants exhibiting systemic silencing (S) among the total number of infiltrated plants (T) in three repeats are shown at the bottom of each panel.