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      Identification and Functional Characterization of Two Putative Pheromone Receptors in the Potato Tuber Moth, Phthorimaea operculella

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          Abstract

          Pheromones are a kind of signal produced by an animal that evoke innate responses in conspecifics. In moth, pheromone components can be detected by specialized olfactory receptor neurons (OSNs) housed in long sensilla trichoids on the male antennae. The pheromone receptors (PRs) located in the dendrite membrane of OSNs are responsible for pheromone sensing in most Lepidopteran insects. The potato tuber moth Phthorimaea operculella is a destructive pest of Solanaceae crops. Although sex attractant is widely used in fields to monitor the population of P. operculella, no study has been reported on the mechanism the male moth of P. operculella uses to recognize sex pheromone components. In the present study, we cloned two pheromone receptor genes PopeOR1 and PopeOR3 in P. operculella. The transcripts of them were highly accumulated in the antennae of male adults. Functional analysis using the heterologous expression system of Xenopus oocyte demonstrated that these two PR proteins both responded to ( E, Z)-4,7–13: OAc and ( E, Z, Z)-4,7,10–13: OAc, the key sex pheromone components of P. operculella, whilst they responded differentially to these two ligands. Our findings for the first time characterized the function of pheromone receptors in gelechiid moth and could promote the olfactory based pest management of P. operculella in the field.

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          Most cited references38

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
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            MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods.

            Comparative analysis of molecular sequence data is essential for reconstructing the evolutionary histories of species and inferring the nature and extent of selective forces shaping the evolution of genes and species. Here, we announce the release of Molecular Evolutionary Genetics Analysis version 5 (MEGA5), which is a user-friendly software for mining online databases, building sequence alignments and phylogenetic trees, and using methods of evolutionary bioinformatics in basic biology, biomedicine, and evolution. The newest addition in MEGA5 is a collection of maximum likelihood (ML) analyses for inferring evolutionary trees, selecting best-fit substitution models (nucleotide or amino acid), inferring ancestral states and sequences (along with probabilities), and estimating evolutionary rates site-by-site. In computer simulation analyses, ML tree inference algorithms in MEGA5 compared favorably with other software packages in terms of computational efficiency and the accuracy of the estimates of phylogenetic trees, substitution parameters, and rate variation among sites. The MEGA user interface has now been enhanced to be activity driven to make it easier for the use of both beginners and experienced scientists. This version of MEGA is intended for the Windows platform, and it has been configured for effective use on Mac OS X and Linux desktops. It is available free of charge from http://www.megasoftware.net.
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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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                Author and article information

                Contributors
                Journal
                Front Physiol
                Front Physiol
                Front. Physiol.
                Frontiers in Physiology
                Frontiers Media S.A.
                1664-042X
                25 January 2021
                2020
                : 11
                : 618983
                Affiliations
                [1] 1Institute of Insect Sciences, Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province, Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Ministry of Agriculture, State Key Laboratory of Rice Biology , Hangzhou, China
                [2] 2State Key Laboratory for Biology of Plant Disease and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences , Beijing, China
                Author notes

                Edited by: Peng He, Guizhou University, China

                Reviewed by: Ya-Nan Zhang, Huaibei Normal University, China; Dan-Dan Zhang, Lund University, Sweden; Arthur de Fouchier, EA4443 Laboratoire d’Éthologie Expérimentale et Comparée (LEEC), France

                *Correspondence: Wenwu Zhou, wenwuzhzou@ 123456zju.edu.cn

                This article was submitted to Invertebrate Physiology, a section of the journal Frontiers in Physiology

                Article
                10.3389/fphys.2020.618983
                7868389
                33569012
                f53c5d4d-9a74-4b49-a151-f2f5e6a812bd
                Copyright © 2021 He, Cai, Zhu, Zhang, Zhang, Ge, Zhu, Zhou, Wang and Gao.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 19 October 2020
                : 18 December 2020
                Page count
                Figures: 4, Tables: 0, Equations: 0, References: 40, Pages: 9, Words: 5892
                Funding
                Funded by: National Nature Science Foundation of China
                Award ID: 31701798
                Funded by: Key Research and Development Program of Zhejiang Province
                Award ID: 2018C04G2011264
                Funded by: National Key Research and Development Program 10.13039/501100012166
                Award ID: 2018YFD0200802
                Categories
                Physiology
                Original Research

                Anatomy & Physiology
                phthorimaea operculella,pheromone receptor,xenopus oocytes,pheromone communication,sexual pheromone

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