Effects of virus induced gene silencing (VIGS) for pepper <i>CaMlo1</i> and <i>CaMlo2</i>.

<p><b>A.</b> Quantification of <i>CaMlo1</i> and <i>CaMlo2</i> gene expression in pepper plants (cv. A) subjected to VIGS. Two silencing constructs for each gene were developed (VIGS:CaMlo1-a and -b; VIGS:CaMlo2-a and -b). Columns refer to transcripts fold change with respect to plants inoculated with an empty TRV2 vector (EV plants). RNAs were isolated from pooled tissues of the 4^th, 5^th and 6^th leaf for each plant. Relative quantification was performed by using the ΔΔCt method and the reference gene <i>CaActin</i>. Bars refer to standard errors of the mean of three biological replicates. Asterisks refer to significant differences with respect to expression levels of EV plants, inferred by means comparison with a Student's t-test. <b>B</b>. Quantification of <i>L. taurica</i> colonization levels on EV, VIGS:CaMlo1 (a and b) and VIGS:CaMlo2 (a and b) pepper plants (cv. A) by counting the average colony number on the 4^th, 5^th and 6^th leaf for each plant. Bars refer to the standard error of the mean of seven plants for each treatment. Asterisks refer to significant differences with respect to EV plants, inferred by means comparison by Student's t-test. <b>C</b>. Quantification of <i>L. taurica</i> colonization levels on EV, VIGS:CaMlo1 (a and b) and VIGS:CaMlo2 (a and b) pepper plants (cv. A) by of fungal biomass by real time qPCR. DNAs were isolated from pooled 4^th, 5^th and 6^th infected leaves per plant. In total, seven plants were tested for each VIGS vector. The ΔΔCt method with <i>CaActin</i> as the reference gene was used for normalization. Columns refer to relative quantification with respect to EV plants. Bars refer to standard errors referred to seven biological replicates. Asterisks refer to significant differences with respect to EV plants, inferred by means comparison by Student's t-test.