Suppression of local RNA silencing is not sufficient to promote cell-to-cell movement of Turnip crinkle virus in Nicotiana benthamiana

The biological relationship between suppression of RNA silencing and virus movement poses an intriguing question in virus-plant interactions. Here, we have used a local RNA silencing assay, based on a movement-deficient Turnip crinkle virus TCV/GFPDeltaCP, to investigate the influence of silencing suppression by three different viral suppressors: the TCV 38K coat protein (CP), the 126K protein of Tobacco mosaic virus (TMV), and P19 of Tomato bushy stunt virus (TBSV) on cell-to-cell movement and long-distance spread of TCV/GFPDeltaCP. First, we found that TCV CP blocked the induction of local RNA silencing, but failed to support virus trafficking in silencing-suppressed transgenic plants, although it acted as a functional movement protein in non-transformed plants. Second, we demonstrated that the TMV 126K suppressor inhibited TCV/GFPDeltaCP-mediated RNA silencing, but did not facilitate intercellular spread of the chimaeric carmovirus. However, TMV and TMVDeltaCP prevented the initiation of RNA silencing by TCV/GFPDeltaCP and caused TCV/GFPDeltaCP to move between cells, although only TMV supported its long-distance spread. Third, TBSV P19 functioned as a movement protein for TCV/GFPDeltaCP and as a silencing suppressor in non-transformed and silencing-suppressed transgenic plants. We further identified three types of mutant P19 proteins that possessed no or varied functionality in silencing suppression and in the facilitation of carmovirus movement. These results suggest that, although suppression of local RNA silencing is essential for the maintenance of viral RNA, recovery of cell-to-cell movement and long-distance spread of movement-deficient carmoviruses is not a direct consequence of such silencing suppression.