Replication, encapsidation, and translation of PLRV RNA in agrobacterium-infiltrated Nicotiana benthamiana tissue. (A) Northern blot analysis of total RNA extracted from 0.1 to 0.2 g of tissue infiltrated with Agrobacterium tumefaciens carrying plasmid pBPY, encoding mutations in the CP of PLRV. (B) Detection of viral RNA from the same tissue used in panel A except that total RNA was extracted after exposing the RNA to conditions that allow only encapsidated RNA to remain intact. The positions of the genomic (g) and subgenomic (sg) RNAs are indicated in panels A and B. (C) Detection of PLRV structural proteins from total protein extracts of agrobacterium-infiltrated tissue that were separated by sodium dodecyl sulfate-12.5% polyacrylamide gel electrophoresis, blotted to nitrocellulose, and probed with antibodies prepared against purified PLRV. The positions of the 22-kDa CP, the full-length RT, and a 40-kDa intermediate protein are indicated. Several minor bands are apparent RT degradation products. ΔRTO (abbreviated ΔRTD on figure) does not produce the RT, and the minor bands are not present in this sample. Minor bands below the CP are assumed to be CP degradation products.