Figure 3.

Molecular characterization of VIGS of NbPAT and/or NbAsp5 genes in N. benthamiana leaves. A, Relative NbAsp5 gene expression in silenced plants compared with controls. At bottom, AAT activity is shown on native gels with protein extracts prepared from control and silenced plants. B, Relative NbPAT gene expression in control and silenced plants. At bottom, AAT activity and PAT activity are shown in protein extracts prepared from control and silenced plants resolved by native gel electrophoresis; PAT protein levels were analyzed by western blotting using anti-PAT specific antibodies (de la Torre et al., 2006). The expression levels for all genes were normalized to that of NbActin2. Error bars represent se. Asterisks indicate significant differences compared with control plants. AAT activity corresponding to NbASP5 and NbPAT on the gels was determined using extracts from N. benthamiana leaves (Fig. 2) according to a previously described protocol (de la Torre et al., 2007).