Inhibition of virus infection by transient expression of short hairpin RNA targeting the methyltransferase domain of Tobacco mosaic virus replicase

Abstract

One of the most efficient mechanisms by which plants protect themselves from viruses is the specific RNA-dependent silencing pathway termed post-transcriptional gene silencing (PTGS). Inhibition of Tobacco mosaicvirus in tobacco was demonstrated through transient expression of virus-derived short hairpin RNA encoding the methyltransferase domain of TMV replicase corresponding to the nucleotides 630 to 1,510 introduced through agro-infiltration into the leaves of tobacco plants. RT-PCR revealed that TMV infection was absent in hpRNA agro-infiltrated leaves while control leaves showed amplification of TMV. Protein analysis further confirmed the absence of TMV coat protein in hpRNA-containing leaves while it was prominent in the control leaves. Our data demonstrate that expression of hpRNA is an effective and predictable new approach to engineering resistance to TMV.