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Odorant receptors (ORs) play an important role in insects to monitor and adapt to the external environment, such as host plant location, oviposition-site selection, mate recognition and natural enemy avoidance. In our study, we identified and characterized OR12 from three closely-related species, Helicoverpa armigera, Helicoverpa assulta, Heliothis virescens, sharing between 90 and 98% of their amino acids. The tissue expression pattern analysis in H. armigera showed that HarmOR12 was strongly expressed both in male and female antennae, but not in other tissues. Functional analysis performed in the heterologous Xenopus expression system showed that all three OR12 were tuned to six structurally related plant volatiles. Electroantennogram recordings from male and female antennae of H. armigera closely matched the data of in vitro functional studies. Our results revealed that OR12 has a conserved role in Heliothinae moths and might represent a suitable target for the control of these crop pests.
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27163122
???displayArticle.pmcLink???PMC4862629 ???displayArticle.link???PLoS One
Fig 1. Sequence alignment of OR12 of the three species studied in this work.Percent of identical amino acids ranges between 90 and 98%. The seven transmebrane domains are marked with black lines.
Fig 2. Phylogenetic tree of the ORs of the three species investigated in this work.Harm: H. armigera (red), Hass: H. assulta (blue), Hvir: H. virescens (green). The clade of PRs is highlighted in pink, that of Orco in gray and that of OR12 in light green.
Fig 3. Tissue expression pattern of OR12 of H. armigera.FA: female antenna; MA: male antenna; Fmp: female maxillary palps; Mmp: male maxillary palps; FP: female proboscises; MP: male proboscises; FH: female heads; MH: male heads; FT: female thoraxes; MT: male thoraxes; Fab: female abdomens; Mab: male abdomens; FL: female legs; ML: male legs; FG: female genitalia; MG: male genitalia.
Fig 4. Functional characterization of three OR12/Orco in Xenopus oocytes.(A), (C) and (E) Tuning curves of HarmOR12/Orco, HassOR12/Orco, and HvirOR12/Orco to an odor panel comprised of 61 plant volatile compounds, arranged along the x-axis according to the strength of the response they elicit, with the strongest stimuli near the center. (B), (D) and (F) Inward current responses of HarmOR12/Orco, HassOR12/Orco and Hvir12/Orco Xenopus oocytes in response to 10â4 M solution of tested compounds, respectively. (G) Orco-injected Xenopus oocytes fail to respond to all the tested compounds including the six volatiles. The holding potential was â80 mV.
Fig 5. EAG responses of male and female H. armigera to six volatile compounds.The relative EAG response values marked with significant differences (P<0.05) among tested chemicals were analyzed by the general linear model (PROC-GLM) and followed by the least-significant difference (LSD) method. Different letters mean significant differences between components. Values are the averages of 21 biological repetitions for both males and females. Error bars indicate SEM.
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