Establishing VIGS of FWA. (A, Upper) Northern blot with TRV RNA species (Fig. S1B) and (Lower) sRNA Northern blot with FWAtr sRNAs in TRV:FWAtr-, TRV (T)-, or mock (M)-infected Col-0(FWAC) plants, 14 dpi. FDH and U6 were probed as loading controls, respectively. TRV:FWAtr is marked with a red asterisk. The black asterisks represent the 20-nt and 30-nt sRNA markers. For the sRNA Northern blot analysis, TRV:FWAtr-infected samples 7–12 were run on a separate gel as indicated by the white separating line between samples 6 and 7. (B, Upper) RT-PCR of TRV and TRV:FWAcds in TRV:FWAcds-, TRV (T)-, or mock (M)-infected Col-0(FWAC) plants, 14 dpi. FDH was assayed as an internal control. (Lower) sRNA Northern blot with FWAcds sRNAs in the same plants. U6 was probed as a loading control. The black asterisks represent the 20-nt and 30-nt sRNA markers. (C) FWA expression in TRV:FWAtr- (tr), TRV:FWAcds- (cds), TRV- (T), or mock (M)-infected Col-0(FWAC) plants, 14 dpi and 45 dpi. Each sample is represented by a black diamond and the average of all samples per treatment is represented by the red horizontal line. A two-tailed Student t test suggested FWA expression was repressed by TRV:FWAtr (*P < 0.05) and TRV:FWAcds (**P < 0.01), 14dpi. (D) Proportion of early- (black), intermediate- (dark gray), and late- (light gray) flowering V1 progeny from TRV:FWAtr-, TRV (T)-, or mock (M)-infected Col-0(FWAC) plants (A), compared with Col-0(FWACme) and Col-0(FWAC). Lines with a ratio of <1 (early):3 (late) are marked with a black asterisk.
