Fig. 5
Spacio-temporal accumulation pattern of RYMVMg1ΔP1ΔCP/CterPSA RNA. N. benthamiana leaves were infiltrated with RYMVMg1ΔP1ΔCP/CterPSA construct together with CP and P19 constructs under the optimal 1:2,5:5 amplicon:CP:P19 ratio. 35S::PSA construct in combination with P19/P1/P0 RNA silencing suppressor cocktail was used as control. Two independent experiments were performed. A- At 5 dpi, RNAs were prepared from infiltrated area (1), adjacent non-infiltrated area of the same leaves (2) and systemic leaves (3). Semi-quantitative RT-PCR was performed after reverse transcription using a mixture of oligodT and CterPSA specific primers. PCR amplification was performed with CterPSA specific primers. NbEF1-α expression was used as an internal control. For both RYMVMg1ΔP1ΔCP/CterPSA RNA B 35S::PSA C constructs, RNA accumulation was followed from 2 to 15 days post-inoculation (dpi) in the infiltrated area. q-RT-PCR was performed after RNA reverse transcription using a mixture of oligodT and CterPSA specific primers. PCR amplification was performed with CterPSA specific primers. Normalizations were performed using GAPDH used as an internal housekeeping control. Expressions were evaluated relative to the 2 dpi time point. The two independent experiments are represented by different colors