Fig 1.
Blue D. citri body color originates from sub-cellular blue pigmentation of the fat body cells as shown during live dissection imaging.
Dissections occurred under natural light in 1xPBS and are not fixed. (A) Ventral view of a non-blue male D. citri. (B) Fat body cells from a non-blue adult. (C) Single fat body cell from non-blue adult showing droplet-like composition of cell and transparency. (D) Ventral view of blue, adult, female D. citri. (E) Ventral view of the blue, adult female from (D), after separating the abdomen from the thorax and beginning to remove internal organs. (F) Loose cloud of fat body cells from the blue, adult female in (D), showing higher intensity of blue color in areas with higher concentration of cells, as well as delicate nature of the fat body organ and ephemeral nature of individual cell cohesion. (G) Single fat body cell from blue, adult female in (D), showing droplet-like composition of cells where some droplets are pigmented bright blue, and cells vary in size.
Fig 2.
D. citri body mass by sex and color morph.
Blue color morph females have 40% more body mass compared to non-blue color females. Color morph had no effect on male body mass.
Fig 3.
The titer of the bacterial endosymbionts Profftella, Carsonella, and Wolbachia in CLas-non-exposed, blue and non-blue adult D. citri, represented as natural logarithm (endosymbiont titer/insect equivalent).
Titer of Profftella and Wolbachia were higher in non-blue D. citri as compared to blue D. citri (P <0.05, n = 10 biological replicates).
Fig 4.
CLas titer in blue and non-blue color morphs of D. citri adults and in citrus leaves inoculated by blue and non-blue insects.
(A) CLas copy number per insect equivalent is ~8-fold greater in non-blue compared to blue D. citri (P <0.01). (B) CLas copy number in midrib DNA samples from citrus leaves inoculated with non-blue D. citri is ~6 fold greater than in leaves inoculated with blue D. citri (P <0.01).
Fig 5.
Western blot analysis of hemocyanin protein expression in D. citri.
Twenty μg of protein extracted from each of the four D. citri samples was used in western blot analysis. Blue: blue color morph adults reared on healthy sweet orange. Non-blue: non-blue color morph adults reared on healthy sweet orange. Healthy: non-exposed adults reared on healthy citron. Infected: CLas-exposed adults reared on infected citron.
Fig 6.
Comparison of copper mass between color morph D. citri adults using ICP mass spectrometry.
Fig 7.
Hemocyanin-1 expression 10-days post injection in blue color morph D. citri adults.
qPCR quantification of hemocyanin-1 (relative to α-tublin) in D. citri adults revealed significant reduction of hemocyanin-1 expression in dsRNA microinjected insects compared to control dsRNA injected insects.
Fig 8.
CLas titer 10-days post injection in blue color morph adult D. citri.
(A) The copy number of CLas in hemocyanin silenced blue color morph reared on sweet orange and citron compared to non-silenced D. citri. Reduction in CLas titer in hemocyanin silenced insects was not statistically significant when data from different host plants are considered separately but was significant when normalized values of both host plants were considered together (B) Box plot analysis of normalized Ct values in control and hemocyanin dsRNA treated insects from both host plants. The increase in CLas qPCR Ct values in hemocyanin dsRNA treated insects represents a reduction in CLas titer. Sweet orange D. citri: n = 20; citron D. citri: n = 20.