Figure 5.

Analysis of the local and systemic silencing of GFP mRNA in transgenic N. benthamiana line 16c using ectopic expression of the MaYMV P0 in an agro-coinfiltration assay. (A) amino acid sequences of the P0s encoded by polereoviruses were aligned using the MUSCLE algorithm. The aligned sequences were used for inferring evolutionary relationships using the neighbor-joining method. The resulting tree, drawn to scale, is shown with the bootstrap of 1000 replicates; (B) multiple alignment of the polerovirus P0 amino acid sequences encompassing a conserved F-box motif LPxxL/I; (C) suppression of local silencing by MaYMV P0 protein. Images of infiltrated leaves were taken under long wave length UV light at 5 dpi; (D) Western blot analyses of GFP accumulation (in top panels) or Northern blot analyses of GFP mRNA levels (in bottom panels) in the extracts from the agro-infiltrated leaves are shown in (C); (E) suppression of systemic silencing by MaYMV P0 protein. The agro-infiltrated plants were imaged at 20 dpi. The infiltration combinations of sGFP with either empty vector or P19 were used as negative and positive cocontrols, respectively; and (F) immunoblot analyses of GFP accumulation (in top panels) or Northern blot analyses of GFP mRNA levels (in bottom panels) in the extracts from the systemic leaves are shown in (E).