PVS RNA transcription from pPVS-H-FL-β and inoculation of plants. a Formaldehyde-denaturing 1.2% agarose gel electrophoresis of RNA transcribed from pPVS-H-FL-β. M: ssRNA marker, lanes 1–4: capped RNA (0.5 μg) transcribed in vitro from pPVS-H-FL-β linearized using SpeI restriction endonuclease. b Upper leaves of Nicotiana occidentalis plants inoculated with mock at 15 days post inoculation (dpi) (left), PVS-H00 at 10 dpi (middle), and capped RNA transcripts from pPVS-H-FL-β at 15 dpi (right). c Northern blot hybridization of total RNA extracted from systemically infected leaves of N. occidentalis plants 14 or 15 dpi with mock (lane 1) and capped RNA transcripts from pPVS-H-FL-β (lanes 2–5) and PVS-H00 (lanes 6 and 7). Capped RNA transcripts (1 ng; lane 8, 10 ng; lane 9) were used as a positive control. The position of PVS genomic RNA (gRNA) is indicated by an arrowhead on right. The lower panel shows ethidium-bromide stained gel with ribosomal RNA (rRNA) as a loading control from corresponding total RNA samples. d Chenopodium quinoa plants inoculated with PVS-H00 (top) and the sap of N. occidentalis infected with capped RNA transcripts derived from pPVS-H-FL-β (bottom) at 17 dpi. Inoculated leaves are shown in left, and upper leaves are shown in right.