Changes in both trans- and cis-regulatory elements mediate insecticide resistance in a lepidopteron pest, Spodoptera exigua
Fig 6
A cis-acting mutation in the CYP321A8 promoter of the resistant strain facilitate binding of nuclear receptor, Knirps and contributes to upregulation of CYP321A8.
(A) Alignment of a region of the CYP321A8 promoter of the resistant and susceptible strains. Mutations at five positions are indicated above the alignment. (B) Activity of mutant constructs of the CYP321A8 promoter from the resistant strain, where mutations in the region from -385 to -142 were reverted to wild-type. Error bars display SD. Letters to the right of bars denote significant difference at p < 0.05 (ANOVA with post-hoc Tukey HSD). (C) Activity of the promoter constructs P(-385/-1)S (susceptible strain) and P(-385/-1)R (resistant strain) in reporter gene assays in the presence and absence of Knirps. PIB represents the empty vector pIB/V5-His. Error bars display SD. Letters above bars denote significant difference at p < 0.05 (ANOVA with post-hoc Tukey HSD). (D) Relative expression of Knirps in the susceptible and resistant strains of S. exigua as determined by RT-qPCR. Error bars display SD. (E) The activity of the promoter constructs (DNA5’-R and DNA5’-S) in the presence and absence of CncC/Maf constructs. Error bars display SD. Letters above bars denote a significant difference at p < 0.05 (ANOVA with post-hoc Tukey HSD).