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Front Physiol
2018 Jan 01;9:1188. doi: 10.3389/fphys.2018.01188.
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Proceeding From in vivo Functions of Pheromone Receptors: Peripheral-Coding Perception of Pheromones From Three Closely Related Species, Helicoverpa armigera, H. assulta, and Heliothis virescens.
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Three closely related species, Helicoverpa armigera, H. assulta, and Heliothis virescens from Lepidoptera Noctuidae, are used as a model system for exploring sexual communication and species isolation. Pheromone receptors (PRs) previously discovered in model moth species include seven in H. armigera, six in H. assulta, and six in H. virescens. PRs named OR6, OR13, and OR16 among these species were found to be functional, characterized by an in vitro Xenopus oocytes system. Using an in vivo transgenic fly system, functional assays of OR6, OR13, and OR16 clades from three closely related Noctuidae species showed that OR13 function was highly conserved, whereas OR6 and OR16 exhibited functional divergence. Similar results were produced from assays in the Xenopus oocytes system. Combined with earlier behavioral results and electrophysiological recordings, we found corresponding relationships among pheromones, PRs, and neurons at the periphery sensory system of each species. Our results provide vital information at the neuronal and molecular level, shedding insight into the sexual communication of closely related species in Lepidoptera.
FIGURE 1. The phylogeny of pheromone receptors (PRs) from seven moth species. (A) Phylogeny and number of PRs identified by species, including Helicoverpa armigera, H. assulta, Heliothis virescens, Bombyx mori, Spodoptera exigua, S. litura and S. littoralis.
(B) A phylogenetic tree of PR genes in different Lepidoptera species. Six clades (I to VI) are shown in this tree representing Orco, OR13, OR11, OR14/15, OR16, and OR6 clades, respectively.
FIGURE 2. Motif analysis of pheromone receptors (PRs) identified from three closely related Lepidoptera species, and the alignment of amino acid sequence of three set of PRs. (A) The eight motif-pattern discovered in nine PRs from Helicoverpa armigera, H. assulta, and Heliothis virescens. (B) The locations of each motif-pattern on the predicted protein sequence from N-terminal to C-terminal. Smaller numbers indicate higher conservation. (C) The alignment of amino acid sequence of clade OR6 from H. armigera, H. assulta, and H. virescens. TM1-TM7 indicates seven transmembrane domains. Harm: H. armigera; Hass: H. assulta; Hvir: H. virescens. (D) The alignment of amino acid sequence of clade OR13 from H. armigera, H. assulta, and H. virescens. (E) The alignment of amino acid sequence of clade OR16 from H. armigera, H. assulta, and H. virescens.
FIGURE 3. Odor coding of pheromone receptors (PRs) from three closely related species. (A) Responses of OR6-expressing neurons in at1 sensilla of transgenic flies. (B) Responses of OR13-expressing neurons in at1 sensilla of transgenic flies. (C) Responses of OR16-expressing neurons in at1 sensilla of transgenic flies. (D) Heatmap of response spectra of PR-expressing neurons in at1 sensilla of transgenic flies.
FIGURE 4. Odor coding of pheromone receptors from three closely related species across concentrations. (A) Dose-responses of OR6-expressing neurons in at1 sensilla of transgenic flies. (B) Dose-responses of OR13-expressing neurons in at1 sensilla of transgenic flies. (C) Dose-responses of OR16-expressing neurons in at1 sensilla of transgenic flies.
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