FIGURE 4.

The changes in CP localization induced by TYLCV do not depend on a single viral protein. N. benthamiana leaves were co-infiltrated with A. tumefaciens carrying constructs to express CP-GFP or GFP-CP, and constructs to express each other virus protein independently (Rep, C2, C3, C4, and V2) or empty vector (EV) as control. (A) The subcellular localization of CP-GFP (upper panels) or GFP-CP (lower panels) was observed under the confocal microscope 2 days after infiltration. This experiment was repeated three times with similar results; more than 20 cells were observed per sample and replicate. Scale bar: 10 μm. (B) Expression of viral genes in the samples in (A), measured by quantitative reverse transcription PCR (qRT-PCR) 2 days after infiltration. The samples were collected after observation under the confocal microscope. The 25S ribosomal DNA interspacer (ITS) was used as normalizer. The expression of viral genes is represented relative to ITS. Gray columns show the virus gene expression level when each viral gene is independently co-expressed with CP-GFP (left) or GFP-CP (right) from a binary vector. Blue columns show the virus gene expression level when a TYLCV infectious clone is co-infiltrated with constructs to express CP-GFP (left) or GFP-CP (right). Green columns represent the EV control. (C) Expression of the CP gene in N. benthamiana leaves transiently expressing CP-GFP, GFP-CP, TYLCV, or empty vector, measured by qRT-PCR 2 days after infiltration. The expression of the CP gene is presented relative to normalizer ITS. The average values (±standard deviation) from three technical repeats of qRT-PCR are shown. This experiment was repeated three times with similar results.