Figure 4

(A) Symptoms observed in the tertiary leaves of the PLRV infected potato plants. (1) PLRV infected control without agroinfiltration; (2) agroinfiltrated with the empty vector pART27; (3) agroinfiltrated with the plasmid containing only antisense sequence (pART27-antisense MP); (4) agroinfiltrated with the plasmid containing only sense sequence (pART27-sense MP); (5) agroinfiltrated with the pART27-MP siRNA construct; (B) Confirmation of expression of siRNA using Northern blotting analysis. Higher expression of siRNA in the leaves at 10 dpi (lane 1) was observed as compared to 5 dpi (lane 2), whereas empty vector (mock) agro-infected leaves did not show any siRNA (lane 3). (C) Detection of PLRV RNA by RT-PCR. Amplified cDNA fragments (627 bp) were analyzed by electrophoresis on 0.8% agarose gel. M: 100 bp marker; lane 1: PLRV was observed in control plant; lane 2: PLRV from tertiary leaves of the plant containing the empty vector pART26; lane 3: PLRV from tertiary leaves of the plant containing the antisense construct (pART27-antisense MP); lane 4: PLRV from tertiary leaves of the plant containing the sense construct (pART27-sense MP); lane 5: no PLRV in tertiary leaves of the plant containing MP siRNA (pART27-MP). (D) Actin PCR was performed as an internal control.