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      A moth odorant receptor highly expressed in the ovipositor is involved in detecting host-plant volatiles

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          Abstract

          Antennae are often considered to be the nostrils of insects. Here, we sequenced the transcriptome of the pheromone gland-ovipositor complex of Helicoverpa assulta and discovered that an odorant receptor (OR) gene, HassOR31, had much higher expression in the ovipositor than in antennae or other tissues. To determine whether the ovipositor was involved in odorant detection, we co-expressed HassOR31 and its co-receptor, HassORco, in a Xenopus oocyte model system, and demonstrated that the OR was responsive to 12 plant odorants, especially Z-3-hexenyl butyrate. These odorants elicited electrophysiological responses of some sensilla in the ovipositor, and HassOR31 and HassORco were co-expressed within ovipositor sensilla. Two oviposition preference experiments showed that female moths lacking antennae still preferentially selected oviposition sites containing plant volatiles. We suggest that the expression of HassOR31 in the ovipositor of H. assulta helps females to determine precise egg-laying sites in host plants.

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          When most insects reproduce they lay eggs that hatch into juveniles known as larvae. To provide good sources of food for the larvae, the adult insects have to carefully select where to lay the eggs. Host plants produce specific sets of chemicals known as odorants that the adult insects are able to smell using proteins called odorant receptors.

          It is generally thought that odorant receptors in the antennae on the head are responsible for guiding adult insects to good egg-laying sites. However, recent studies have reported that odorant receptors are also present in the egg-laying organs of several different species of moth. It remains unclear what role these odorant receptors may play in egg-laying.

          The oriental tobacco budworm ( Helicoverpa assulta) is considered a serious pest in agriculture. The adult moths lay their eggs on a narrow range of plants in the nightshade family including tobacco and hot pepper. Li et al. have now investigated the odorant receptors of H. assulta and found that one gene for an odorant receptor called HassOR31 was expressed much more in the egg-laying organs of the moths than in the antennae. Further experiments showed that this receptor was tuned to respond to 12 odorants that also stimulated responses in the egg-laying organ of H. assulta. Together these findings suggest that this odorant receptor in the egg-laying organ helps the moths find suitable host plants to lay their eggs on.

          The work of Li et al. may help us understand how H. assulta evolved to lay its eggs on specific members of the nightshade family and lead to new methods of controlling this pest. An insect’s sense of smell guides many other behaviors including finding food, mates and avoiding enemies. Therefore, these findings may inspire researchers to investigate whether odorant receptors in the antennae or other organs guide these behaviors.

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          Most cited references43

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          Insect olfactory receptors are heteromeric ligand-gated ion channels.

          In insects, each olfactory sensory neuron expresses between one and three ligand-binding members of the olfactory receptor (OR) gene family, along with the highly conserved and broadly expressed Or83b co-receptor. The functional insect OR consists of a heteromeric complex of unknown stoichiometry but comprising at least one variable odorant-binding subunit and one constant Or83b family subunit. Insect ORs lack homology to G-protein-coupled chemosensory receptors in vertebrates and possess a distinct seven-transmembrane topology with the amino terminus located intracellularly. Here we provide evidence that heteromeric insect ORs comprise a new class of ligand-activated non-selective cation channels. Heterologous cells expressing silkmoth, fruitfly or mosquito heteromeric OR complexes showed extracellular Ca2+ influx and cation-non-selective ion conductance on stimulation with odorant. Odour-evoked OR currents are independent of known G-protein-coupled second messenger pathways. The fast response kinetics and OR-subunit-dependent K+ ion selectivity of the insect OR complex support the hypothesis that the complex between OR and Or83b itself confers channel activity. Direct evidence for odorant-gated channels was obtained by outside-out patch-clamp recording of Xenopus oocyte and HEK293T cell membranes expressing insect OR complexes. The ligand-gated ion channel formed by an insect OR complex seems to be the basis for a unique strategy that insects have acquired to respond to the olfactory environment.
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            Drosophila odorant receptors are both ligand-gated and cyclic-nucleotide-activated cation channels.

            From worm to man, many odorant signals are perceived by the binding of volatile ligands to odorant receptors that belong to the G-protein-coupled receptor (GPCR) family. They couple to heterotrimeric G-proteins, most of which induce cAMP production. This second messenger then activates cyclic-nucleotide-gated ion channels to depolarize the olfactory receptor neuron, thus providing a signal for further neuronal processing. Recent findings, however, have challenged this concept of odorant signal transduction in insects, because their odorant receptors, which lack any sequence similarity to other GPCRs, are composed of conventional odorant receptors (for example, Or22a), dimerized with a ubiquitously expressed chaperone protein, such as Or83b in Drosophila. Or83b has a structure akin to GPCRs, but has an inverted orientation in the plasma membrane. However, G proteins are expressed in insect olfactory receptor neurons, and olfactory perception is modified by mutations affecting the cAMP transduction pathway. Here we show that application of odorants to mammalian cells co-expressing Or22a and Or83b results in non-selective cation currents activated by means of an ionotropic and a metabotropic pathway, and a subsequent increase in the intracellular Ca(2+) concentration. Expression of Or83b alone leads to functional ion channels not directly responding to odorants, but being directly activated by intracellular cAMP or cGMP. Insect odorant receptors thus form ligand-gated channels as well as complexes of odorant-sensing units and cyclic-nucleotide-activated non-selective cation channels. Thereby, they provide rapid and transient as well as sensitive and prolonged odorant signalling.
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              A spatial map of olfactory receptor expression in the Drosophila antenna.

              Insects provide an attractive system for the study of olfactory sensory perception. We have identified a novel family of seven transmembrane domain proteins, encoded by 100 to 200 genes, that is likely to represent the family of Drosophila odorant receptors. Members of this gene family are expressed in topographically defined subpopulations of olfactory sensory neurons in either the antenna or the maxillary palp. Sensory neurons express different complements of receptor genes, such that individual neurons are functionally distinct. The isolation of candidate odorant receptor genes along with a genetic analysis of olfactory-driven behavior in insects may ultimately afford a system to understand the mechanistic link between odor recognition and behavior.
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                Author and article information

                Contributors
                Role: Reviewing Editor
                Role: Senior Editor
                Journal
                eLife
                Elife
                eLife
                eLife
                eLife Sciences Publications, Ltd
                2050-084X
                21 May 2020
                2020
                : 9
                : e53706
                Affiliations
                [1 ]State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences BeijingChina
                [2 ]CAS Center for Excellence in Biotic Interactions, University of Chinese Academy of Sciences BeijingChina
                [3 ]Forestry College, Henan University of Science and Technology LuoyangChina
                Wageningen University Netherlands
                University of St Andrews United Kingdom
                Wageningen University Netherlands
                Max Planck Institute for Chemical Ecology Germany
                The Rockefeller University United States
                Author information
                https://orcid.org/0000-0002-4720-6255
                https://orcid.org/0000-0003-0418-8621
                Article
                53706
                10.7554/eLife.53706
                7308088
                32436842
                47b46ea9-47a0-4b39-b247-6626e32c4ea9
                © 2020, Li et al

                This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

                History
                : 18 November 2019
                : 20 May 2020
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 31830088
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 31772528
                Award Recipient :
                Funded by: National Key R and D Program of China;
                Award ID: 2017YFD0200400
                Award Recipient :
                The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
                Categories
                Research Article
                Ecology
                Custom metadata
                A moth can detect plant volatiles using an odorant receptor expressing in its ovipositor, and this odorant receptor has a much higher expression level in the ovipositor than antennae.

                Life sciences
                helicoverpa assulta,odorant receptor,pheromone gland-ovipositor,function,sensillum,oviposition,other

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