Fig 1: Dopamine concentrations mediated by DcDop2 are involved in the coevolution between CLas and D. citri resulting in increased fecundity.(A) Temporal expression patterns of DcDop2 between the ovaries of CLas- and CLas+ females. (B) Effects of RNAi on DcDop2 expression at protein and mRNA levels in CLas- and CLas+ females treated with dsDcDop2 for 48 h. (C, D) Comparison of TAG and glycogen levels in fat bodies of CLas- and CLas+ females treated with dsDcDop2 for 48 h. (E) Lipid droplets stained with Nile red in fat bodies dissected from CLas+ females treated with dsDcDop2 for 48 h. Scale bar = 40 μm. (F) Ovary phenotypes in dsDcDop2-treated CLas+ females at 48 h. Scale bar = 200 μm. o: ovary, s: spermathecae. (G–I) Comparison of preoviposition period, oviposition period, and fecundity of CLas- and CLas+ adults treated with dsDcDop2, Dop2 inhibitor (Pimozide, HY-12987, MedChemExpress), and dopamine rescue. (J) CLas titer in ovaries of CLas+ females at 7 DAE treated with dsDcDop2, Dop2 inhibitor (Pimozide, HY-12987, MedChemExpress), and DA rescue. (K) Representative confocal images of the reproductive system of CLas+ females treated with dsDcDop2, Dop2 inhibitor (Pimozide, HY-12987, MedChemExpress), and dopamine rescue. Scale bar = 200 μm. DAPI: cell nuclei were stained with DAPI and visualized in blue. DcDop2-FITC: DcDop2 signal visualized in green by staining with FITC. CLas-Cy3: CLas signal visualized in red by staining with Cy3. Merge: merged imaging of co-localization of cell nuclei, DcDop2 and CLas. Results in (A–D) and (J) are displayed as means ± SEM with three independent biological replicates, each with three technical replicates. Data in (G–I) are shown as means ± SEM with 30 independent biological replicates. Significant differences between treatments and controls are indicated by asterisks in (A–D) (Student’s t-test, *p<0.05, **p<0.01, ***p<0.001). For (G–J), significant differences among different treatments are indicated by lowercase letters based on one-way ANOVA followed by Tukey’s Honest Significant Difference tests at p<0.05. Figure 3—source data 1.Original file for the western blot analysis in Figure 3B (anti-DcDop2). Figure 3—source data 2.Original file for the western blot analysis in Figure 3B (anti-β-ACT).