Figure 2.

A female-specific lethal yeast interfering RNA larvicide targeting lncRNA genes. Significant female larval mortality resulted from oral consumption of yeast interfering RNA larvicide strain #478 [(a), *** = p < 0.001, Chi-square]. No significant death was observed in males following treatments with larvicide [(a); p > 0.05, Chi-square], and a control interfering RNA strain did not significantly impact survival of male or female larvae [(a) p > 0.05, Chi-square; data are represented as mean survival based on adult emergence following treatment of 180 total larvae, and error bars denote SEM]. (b–d) display quantification of the identical AAEL020379, AAEL020813, and AAEL022952 transcripts, with mRNA levels normalized to levels of the rpS17 housekeeping gene; error bars denote standard deviations. The #478 larvicide target transcripts are expressed throughout larval development in untreated first (L1), second (L2), third (L3), and fourth (L4) instar larvae (b), with no significant differences in expression levels noted between the various larval stages (p > 0.05, ANOVA; the expression levels are shown relative to L4). No significant differences in transcript levels were noted between third instar male and female larvae [(c), t-test, p > 0.05]. Silencing of these lncRNA targets of larvicide #478 (d) was confirmed through qRT-PCR (*** = p < 0.001 vs. target gene levels in control interfering RNA-fed larvae, Student’s t-test; error bars denote standard deviation).