Figure 5

Analysis of RNAi-mediated silencing in stable GUS expressing banana. (A) GUS enzyme activity of control (pSTARGATE transformed) and transgenic (ihpRNA vector transformed) lines at different developmental stages after transformation of a stable GUS expressing ECS line (pFAJ3000 transformed). Bars represent the mean ± SD. Mean values of undifferentiated cell clusters (6–35 days after transformation) were calculated from six independent biological replicates (n = 6); those at proliferating cell colonies (embryo induction) stage were measured from 10 independent lines (n = 10); and those at in vitro plantlet stage were the average of 12 independent lines per construct (n = 12) with two independent measurements per line. Significant differences with the control were analyzed using one-way ANOVA (*, P < 0.05). (B) Expression level of gusA^INT was determined by RT-qPCR. RNA was isolated from a portion of the leaves used for the GUS enzyme activity assay in (A). Gene expression analyses were performed using two technical replicates per line and the same number of biological replicates or independent lines as in A. Bars represented the mean ± SD. *: P < 0.05; Significant differences with the control was analyzed using one-way ANOVA.