FIGURE 11.

The effect of iron/heme-related genes silencing in the insect physiology. Starved adult females were injected with 1 μg of dsRNA of Ferritin (Fer), IRP (IRP1), Heme Oxygenase (HO) and FLVCR. After 48 h, the animals were fed on blood. The adults survival curves (A) oviposition (D) and eggs viability (E) were monitored for 21 days (n = 24). Nymphs of the first instar (N1) were fed on blood supplemented with dsRNAs of the same genes. The nymphs survival curves (G) and the molting to the N2 instar (J) were monitored daily for 18 days (n = 30). The survival curves data were replotted as % of total mortality of adults (B) and nymphs (H) to show the time course of silenced-insects mortality. The mean day of death of adults and nymphs are shown in (C,I), respectively. Data shown in graphics are mean ± SE (F) Representative images of the phenotypic morphology of eggs laid by dsMal and dsFer-injected females (scale bar = 1,5 mm). dsMal-injected or fed insects were used as a control. Log-rank (Mantel-Cox) test was used for survival curves comparison, p < 0.0001 in (A) and p = 0,0001 in (G). One-way analysis of variance followed by Tukey’s multiple comparison test was used to evaluate differences in the adults and nymphs mean death days, ^∗p < 0.01, ^∗∗p < 0.05, ^∗∗∗p < 0.001. Two-way analysis of variance followed by Tukey’s multiple comparison test was used to compare molting curves of nymphs fed with iron and heme-related dsRNAs and control group (dsMAL) ^∗∗∗∗p < 0.0001. All the experiments described above were independently performed at least three times.