Figure 2

The mt-miR159b-mediated amiR-silencing is efficient in tobacco leaves. A) DsRED fluorescence is strongly reduced after infiltration of the pRed-35Spro::amiR-dsred (A) construct (a) as compared to empty vectors (ev). Bright filed image (bf), DsRED fluorescence (dsRED) and an overlay of both channels are shown. Infiltration site are indicated by dotted lines. Scale bar: 5 mm. B) Relative levels of mature amiR-dsred determined by stem loop qRT-PCR, relative transcript levels of dsred measured by qRT-PCR and transcript levels of nptII (kanamycin resistance) are shown. vc: vector controls, a: amiR samples sm: size marker, amplicon size of 243 bp is indicated. C) Western blot demonstrating reduced expression of dsRED after amiR-dsred mediated silencing of dsRED in tobacco leaves. Shown are three replicates of amiR-silencing (a) and vector controls (vc) represented by proteins of three different infiltration spots on the same leaf, for each construct. Reduced expression of dsRED after amiR-dsred infiltration was visualized using an anti-dsRED antibody. Equal sample loading is shown by anti-RUBISCO activase.