FIG 1.

In vivo visualization of dsRNA and RdRp in PVX infected N. benthamiana epidermal cells. (A) Visualization of the dsRNA during PVX infection at 36 (I), 48 (II), and 60 (III) hpi. The fluorescence from the dRBFC assay is shown in green, whereas the fluorescence from pGR.mCh is shown in red. The nucleus is indicated by a white asterisk and the peripheral dsRNA fluorescent foci are indicated by white arrowheads. Scale bar = 50 μm (I and II) or 10 μm (III). Note that the dRBFC signal in the nucleus represents endogenous d-bodies (28). (B and C) Subcellular localization of transiently expressed RdRp-YFP (green) in N. benthamiana epidermal cells in the absence of (B) or during (C) PVX infection at 48 hpi. The RdRp-YFP in panel C was expressed from pGR107/RdRp-YFP to ensure infection of PVX in the same cell. The cytoplasmic green fluorescent foci from RdRp-YFP are indicated by white arrowheads. The nuclei were labeled by a nuclear localization signal peptide (NLS)-tagged mRFP (Nul-mRFP). Scale bars = 50 μm. (D) Subcellular localization of perinuclear (I to III) or cytoplasmic (IV to VI) RdRp and dsRNA during PVX infection at 60 hpi; the RdRp-YFP (green) was expressed from pGR107/RdRp-YFP, and dsRNAs (red) were labeled by mRFP-based dRBFC plasmids. The nucleus is indicated by a white dashed line. All scale bars = 10 μm.