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Insect olfactory sensing is crucial for finding food, mating, and oviposition preference. Odorant receptors (ORs) play a central role in the transmission of odorant signals into the environment by the peripheral olfactory system. Therefore, the identification and functional study of ORs are essential to better understand olfactory mechanisms in insects. OR studies on Diptera insects are primarily performed on Drosophila and mosquitoes, but few studies have been reported in Tephritidae. In this study, we examined three candidate ORs (BminOR3, BminOR12, and BminOR16) from Bactrocera minax. Our analysis of tissue expression revealed that the three BminORs were expressed in the antennae, with no difference between the male and female. In in vitro heterologous expression system of Xenopus oocytes. BminOR3/BminOrco responded strongly to 1-octen-3-ol, BminOR12/BminOrco responded to eight compounds [methyl salicylate, benzaldehyde, (Z)-3-hexenyl acetate, butyl acrylate, butyl propionate, 1-octanol, (S)-(+)-carvone and benzyl alcohol], and BminOR16/BminOrco slightly responded to undecanol. Our results concluded that BminOR3, BimOR12, and BminOR16 could play an important role in host-finding and oviposition positioning in B. minax.
FIGURE 1. Phylogenetic analysis of the three candidate ORs with candidate ORs from Dipteran. Bmin, B. minax; Csty, C. stygia; Mdom, M. domestica; Dmel, D. melanogaster; Bdor, B. dorsalis. The clade in green indicates the Orco co-receptor gene clade and the one in yellow is the pheromone receptor gene clade. The three BminORs are in red.
FIGURE 2. Alignment of the amino acid sequence of BminOR3, BminOR12, and BminOR16.
FIGURE 3. Expression profiles of BminOR3, BminOR12, and BminOR16 in different tissues of male and female B. minax. FA, female antennae; MA, male antennae; FH, female heads (without antennae); MH, male heads (without antennae); FT, female thoraxes; MT, male thoraxes; FAb, female abdomens; MAb, male abdomens; FL, female legs; ML, male legs.
FIGURE 4. Responses of Xenopus oocytes with co-expressed BminOR3/BminOrco to stimulation with volatile compounds. (A) Inward current responses of BminOR3/BminOrco Xenopus oocytes in response to 10–4 M of the volatile compounds. (B) BminOR3/BminOrco Xenopus oocytes stimulated with a range of 1-octen-3-ol concentrations. (C) Response profile of BminOR3/BminOrco Xenopus oocytes. Error bars indicate SEM (n = 6) (T-test, P < 0.05). (D) Dose-response curve of BminOR3/BminOrco Xenopus oocytes to 1-octen-3-ol. 1-Octen-3-ol EC50 = 3.467 × 10–7 M. Error bars indicates SEM (n = 6).
FIGURE 5. Responses of Xenopus oocytes with co-expressed BminOR12/BminOrco and BminOR16/BminOrco to stimulation with volatile compounds. Inward current responses of BminOR12/BminOrco (A) and BminOR16/BminOrco (C)
Xenopus oocytes in response to 10–4 M of the volatile compounds. Response profile of BminOR12/BminOrco (B) and BminOR16/BminOrco (D)
Xenopus oocytes. Error bars indicate SEM (n = 6) (T-test, P < 0.05).
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