Figure 3

Detection of stable virions isolated from N. benthamiana leaves inoculated with transcripts of various mutants. (A) Electron micrographs depicting virions isolated from leaves inoculated with mock, WT BBSV, and BM3 and BM4 mutants. Bar = 200 nm. (B) Detection of mutated CP in the purified virions. Purified virions were separated on an SDS-PAGE gel and stained with Coomassie brilliant blue R250 staining (top panel), or subjected to Western blot analysis using a BBSV CP antibody to confirm the CP identity (bottom panel). (C) Detection of protected viral RNAs in virions underwent an incubation period that permitted the degradation of unprotected RNAs, with RT-PCR (top panel). The bottom panel shows the RT-PCR results with total RNA extracted from the same inoculated leaves (without 30-minute inoculation), serving as controls for viral RNAs available for virion assembly.