Table 1.
Primers used in this study.
Fig 1.
Multiple alignment of OmSpo with orthologs determined for other arthropods.
Location of a P/G rich domain, Helixes (Helix-C and Helix-K), a PERF motif and a Heme binding domain are indicated
Fig 2.
Multiple alignment of OmShd with orthologs determined for other arthropods.
Location of a P/G rich domain, Helixes (Helix-C, Helix-I and Helix-K), a PEFR motif and a Heme binding domain are indicated.
Fig 3.
Phylogenic tree of CYPs associated with ecdysteroidogenesis.
A phylogenetic tree of CYPs was constructed by the Neighbor-joining method. The bootstrap values (1000 replications) are represented at the branches. The scale bar indicates the numbers of amino acid substitutions per site. Gene numbers used for this analysis are described in the materials and methods.
Table 2.
Homologies of OmSpo and OmShd to their orthologs in other arthropods.
Table 3.
Effects of dsRNA injection on ecdysis of final instar nymphs.
Table 4.
Effects of 20E injection of nymphs after dsSpo injection.
Fig 4.
Chromatograms of 20-hydroxyecdysone (20E), produced by enzymatic assay of OmShd, analyzed with LC-MS/MS.
20E chromatograms from the medium of pIZT-OmShd vector (A) or pIZT vector (negative control) (B) were transfected to S2 cells. Intensity was indicated as counts per second (cps). MS/MS spectra of 20E standard (C) and 20E produced by OmShd (D).
Fig 5.
Expression sites of OmSpo and OmShd in nymphs and adult females of O. moubata.
OmSpo (A) and OmShd (B) expression in final instar nymphs determined 9 days after engorgement by qRT-PCR (n = 3–4, mean±se). OmSpo (C) and OmShd (D) expression in mated females determined 2 days after engorgement (n = 3–4, mean±se). n. d. indicates no detection.
Fig 6.
OmSpo expression in specific cells of ovaries from nymphs and adult females determined by WISH.
An overall ventral view of tick tissues in a final instar nymph (A). OmSpo localization in dissected reproductive tissues from the nymph in Fig 6A (B). OmSpo localization in small cells of the ovary with a band-like configuration (C, magnification of Fig 6B). Ovary of a final instar nymph stained with the sense probe (D). OmSpo localization in the ovary of a mated female (E). Ovary of mated female stained with the sense probe (F). Arrows indicate staining of OmSpo with the antisense probe. Ticks have a single ovary (Ov) connected to oviducts (Ovd) from both sides. Oviducts open into a uterus (Ut). Developing oocytes (Oo) are attached outside the ovary. Scale bars indicate 1 mm.
Fig 7.
Developmental changes in OmSpo and OmShd expression during nymphal development or egg maturation in adult females.
OmSpo (A) and OmShd (B) expression in nymphs was determined for ovaries and midgut, respectively. OmSpo (C) and OmShd (D) expression in adult females determined for ovaries and midgut, respectively. (n = 3–4, mean±se)