Encapsidation of genomic but not subgenomic Turnip yellow mosaic virus RNA by coat protein provided in trans

We have studied the encapsidation requirements of Turnip yellow mosaic virus (TYMV) genomic and subgenomic RNA using an “agroinfiltration” procedure involving transient expression of RNAs and coat protein (CP) in Nicotiana benthamiana leaves. Although N. benthamiana is a nonhost, expression of TYMV RNA in its leaves by agroinfiltration resulted in efficient local infection and production of the expected virions containing genomic and subgenomic RNAs together with empty capsids. A nonreplicating genomic RNA with a mutation in the polymerase domain was efficiently encapsidated by CP provided in trans, even though RNA levels were a thousand-fold lower than in normal infections. In contrast, encapsidation of CP mRNA was not observed under these conditions, even when the CP mRNA had authentic 5′- and 3′-termini. Deletion of the 3′-tRNA-like structure from the genomic RNA did not alter the encapsidation behavior, suggesting that this feature does not play a role in the encapsidation of TYMV RNA. Our results indicate differences in the encapsidation process between genomic and subgenomic RNAs, and suggest an interaction between RNA replication and the packaging of subgenomic RNA.