Symptoms elicited 14 days after the inoculation on <i>Nicotiana clevelandii</i> plants by the Rs-CMV and mutated Rs2-2a777 CMV virus (A).

<p>Northern blot hybridization analysis of total RNAs extracted from non-inoculated leaves 6 weeks after inoculation (B). The radiolabeled probe was specific for Rs-CMV RNA3. Ethidium bromide-stained rRNA from the same volume of each sample is shown below each lane.</p

Symptoms elicited 14 days after the inoculation on <i>Nicotiana clevelandii</i> plants by the Rs-2a777 CMV and the alanine-scanning mutated Rs2-CMV viruses.

<p>Name of the pictures taken from the plants indicate the consecutive amino acids of the exchange in the 2b protein sequence.</p

Symptoms elicited 5 days after the inoculation on <i>Chenopodium murale</i> plants inoculated by the Rs-2a777 CMV and the alanine-scanning mutant Rs2-CMV transcripts in the present of RNA1, 3 transcripts of Rs-CMV.

<p>The name of the constructs is indicated on the pictures.</p

Northern blot analysis of <i>Nicotiana clevelandii</i> plants 30 days after inoculation with Rs-CMV, and with the eight mutant caused altered phenotype on <i>N. clevelandii</i> plants.

<p>Total RNAs were extracted from non-inoculated leaves. The radiolabeled probe was specific for Rs-CMV RNA3. Ethidium bromide-stained rRNA from the same volume of each sample is shown below each lane.</p

Northern blot analysis of <i>Nicotiana clevelandii</i> plants 8 days after inoculation.

<p>Total RNAs were extracted from non-inoculated leaves. The radiolabeled probe was specific for Rs-CMV RNA3. Ethidium bromide-stained rRNA from the same volume of each sample is shown below each lane.</p

Detection of Rs-CMV and the eight mutants in inoculated leaves of <i>Nicotiana clevelandii</i> 3 days after inoculation.

<p>Samples were analyzed by RT-PCR using primers specific for RNA2 of CMV. M, DNA molecular size marker, 1000 bp and 1500 bp makers are indicated.</p

Suppression of RNA silencing in patch assays.

<p>A binary vector expressing the GFP reporter gene was agroinfiltred into <i>Nicotiana benthamiana</i> leaves, together with an empty binary vector or together with binary vectors expressing 2b protein and MEL/1-3/AAA, NVE/10-12/AAA, SPS/40-42/AAA, RHV/70-72/AAA, KKQ/22-24/AAA, QNR/31-33/AAA, RER/34-36/AAA and LPF/55-57/AAA 2b protein construct.</p

Cartoon representation of the siRNA-2b nucleoprotein complex with the localization of those eight mutants whose bearing altered phenotype on <i>Nicotiana clevelandii</i> plants.

<p>Aa triplets which are involved in the putative cell-to-cell movement related interactions are red while the other six mutations are colored green (A). Molecular surface representation of the siRNA-2b nucleoprotein complex. siRNA surfaces are colored light blue while 2b protein subunits are grey. Red protein surface regions indicate the three-dimensional localizations of those mutations which lead to movement-deficient behavior (B).</p

Immunoblot analyses of accumulation His-tagged 2b protein mutants in agroinfiltrated patches.

<p>Detection of the fluorescence of GFP proteins on SDS-PAGE by illuminating the gel with UV lamp. A penta-his antibody was detection of His-tagged 2b proteins. Coomassie staining was used to monitor the equivalence of protein loading and transfer.</p

X-ray structure of the PCV2 virion (PDB ID code: 3R0R).

<p>The last seven residues are not present in the X-ray structure but it is well visible that the C-terminal tail (red) of the PCV2 capsid protein is located at the edge of the CP pentamer.</p