Figure 4
From: Tuning LeSPL-CNR expression by SlymiR157 affects tomato fruit ripening
miRNA target – GFP reporter expression assay.
(a) PVX-based miRNA target – GFP reporter expression vectors. (b-e) Nicotinana benthamiana leaves were inoculated with PVX/GFP (GFP, b), PVX/SlymiR157Tar::GFP (miR157Tar::GFP, c), PVX/SlymiR157::GFP (miR157::GFP, d) or PVX/mSlymiR157::GFP (mmiR157::GFP, e). Photographs were taken under long-wavelength UV light at 7 days post inoculation. GFP fluorescence is green and leaf auto-fluorescence shows red. (f) Western blot detection of GFP and PVX coat protein (CP) in mock- or PVX-inoculated leaf tissues using antibody raised specifically against GFP and the viral CP. Positions of GFP and PVX CP are indicated. (g) Detection of recombinant PVX::miRNA target::GFP RNA in same leaf samples as used in Western blot by semi-qRT-PCR. The positions and sizes of amplified fragments and the number of PCR cycles are indicated. 18S rRNA was used as controls. (h-k) AC fruits were injected with PVX/GFP (h), PVX/SlymiR157::GFP (i), PVX/SlymiR157Tar::GFP (j) or PVX/mSlymiR157::GFP (k). Non-ripening sectors show green. Fruits were photographed at 7 days after breaker.
