Silencing of <i>NbUBC2</i> and <i>NbHUB1</i> affected ChiLCV pathogenesis and deposition of H2B-ub and H3-K4me3 on the viral genome.

<p>(A-G) Phenotype of <i>N</i>. <i>benthamiana</i> plants infiltrated with pTRV-<i>NtPDS</i>, empty vector (pTRV: 00), with or without ChiLCV inoculated, pTRV-<i>NbUBC2</i>, pTRV-<i>NbHUB1</i> plants at 21 dpi. (H) Detection of <i>NbHUB1</i> specific siRNAs in three independent silenced plants (P1, P2, P3) of <i>N</i>. <i>benthamiana</i>. (I) Detection of <i>NbUBC2</i> specific siRNA in three different silenced plants of <i>N</i>. <i>benthamiana</i>. (J) Symptom severity graph showing disease development of ChiLCV on pTRV:00, <i>NbUBC2-</i>, and <i>NbHUB1</i>-silenced <i>N</i>. <i>benthamiana</i> plants. (K-L) Relative level of ChiLCV DNA accumulation in either mock-, or virus-inoculated silenced plants by Southern blot and qPCR assays. Total plant DNA loaded is indicated below the blot (K). The statistical significance between the mean values was calculated by <i>t</i>-test (***p<0.001, **p<0.01). (M) Expression analysis of C2 transcripts level in pTRV:00, <i>NBUBC2</i>-and<i>NbHUB1</i>-silenced plants inoculated with ChiLCV (21 dpi). The statistical significance of the differences between the mean values were calculated by performing <i>t</i>-test (***p<0.001, **p<0.01). (N) An analysis of the viral titer in the absence (WT) or in the presence of pTRV infiltrated <i>N</i>. <i>benthamiana</i> plants inoculated with ChiLCV at 21 dpi. (O) Detection of H3-K4me3 and H2B-ub in <i>NbUBC2</i>, and <i>NbHUB1</i> silenced <i>N</i>. <i>benthamiana</i> plants by immunoblotting assays.