Figure 3.

Per-knockout affects the DH content and the diapause metabolism of the female pupal hemolymph. The parental silkworm eggs were incubated at 25LD, and the larval and pupal stage maintained a 25LD environment. (A) DH mass spectrum. DH std, standard product of DH; WT and Per^−/− represents the 72 h pupal hemolymph of wild-type and mutant silkworm, respectively. (B) DH content in pupal hemolymph. In the multiple comparison of significant differences in the figure, there is no significant difference between the same letters, and there are significant differences between different letters (p ≤ 0.05, n = 3). (C) The diapause rescue effect of DH supplementation by female pupae on the eggs laid by mutant adults (n = 13–15 batches). The parental eggs were incubated at 25LD, and the pupa was injected 10 µL DH at age 72 h (±2 h). DH concentration (w:v) was 0.5 µg/µL and 1.0 µg/µL. The control was injected with the same volume of sterile water. The transcription levels of DHR (D) and Treh-2 (E) in the pupal ovaries were measured by qRT-PCR and with the reference gene was Rp49 (n = 3). (F) Glycogen content in the ovary (n = 4). (G) The content of trehalose in hemolymph (n = 6). In Figure (B–G), data were expressed as the mean ± SEM, the difference between Per^−/− and WT is: *, p < 0.05; **, p < 0.01; ***, p < 0.001.