Turnip mosaic potyvirus resistance in Nicotiana benthamiana derived by post-transcriptional gene silencing

The coat protein (CP) gene of turnip mosaic potyvirus isolate ESC8 (TuMV-ESC8) was cloned and sequenced. Comparisons of the 867-nucleotide (nt) CP region with those of 11 TuMV isolates showed 86.7-89.3% nucleotide identity and 92.4-95.5% amino acid identity. The CP gene was cloned into a plant expression vector and transformed into Nicotiana benthamiana plants via Agrobacterium tumefaciens-mediated leaf disk transformation. Progeny from R0 lines was screened for resistance to TuMV-ESC8. Five of 29 tested lines showed TuMV protection in more than 50% of their progeny. Interestingly, some of the resistant plants transformed with the CP gene of TuMV displayed mild mosaicism in the new growing leaves at the later stages of evaluation; but these mosaic symptoms disappeared when the leaves were fully expanded. Collective data from steady-state RNA analysis and nuclear run-on assay of a line showed that the resistance was RNA-mediated through the post-transcriptional gene silencing mechanism.