Combined expression of p20 and p23 proteins from Citrus tristeza virus show enhanced local silencing suppressor activity

Abstract

Viruses developed a strategy to counter-defence the posttranscriptional gene silencing mechanism (PTGS) based on the activity of silencing suppressor proteins. Citrus tristeza virus (CTV), a member of the genus Closterovirus, has two suppressor proteins (p20 and p23) that target the local RNA silencing response of the host. In GFP transient co-expression assays performed on Nicotiana benthamiana 16C plants, local suppressor activity of p23 and p20 was similar. Co-expression of both proteins from a mild or a stem pitting CTV isolate showed stronger local suppression activity than either suppressor alone, with an increased GFP transcript level six- (for Gp M) to nine-fold (for Gp 3a) higher than non-inoculated 16C plants, in parallel with low accumulation of siRNAs. Further, GFP brightness of leaves infiltrated with Agrobacterium cultures at an OD₆₀₀ of 0.5 was comparable to those infiltrated with OD₆₀₀ 0.25. These findings indicate that combined action of p20 and p23 proteins results in enhanced suppressor activity.