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      Antennal transcriptomic analysis of carboxylesterases and glutathione S-transferases associated with odorant degradation in the tea gray geometrid, Ectropis grisescens (Lepidoptera, Geometridae)

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          Abstract

          Introduction: Carboxylesterases (CXEs) and glutathione S-transferases (GSTs) can terminate olfactory signals during chemosensation by rapid degradation of odorants in the vicinity of receptors. The tea grey geometrid, Ectropis grisescens (Lepidoptera, Geometridae), one of the most devastating insect herbivores of tea plants in China, relies heavily on plant volatiles to locate the host plants as well as the oviposition sites. However, CXEs and GSTs involved in signal termination and odorant clearance in E. grisescens remains unknown.

          Methods: In this study, identification and spatial expression profiles of CXEs and GSTs in this major tea pest were investigated by transcriptomics and qRT-PCR, respectively.

          Results: As a result, we identified 28 CXEs and 16 GSTs from female and male antennal transcriptomes. Phylogenetic analyses clustered these candidates into several clades, among which antennal CXEs, mitochondrial and cytosolic CXEs, and delta group GSTs contained genes commonly associated with odorants degradation. Spatial expression profiles showed that most CXEs (26) were expressed in antennae. In comparison, putative GSTs exhibited a diverse expression pattern across different tissues, with one GST expressed specifically in the male antennae.

          Disscussion: These combined results suggest that 12 CXEs (EgriCXE1, 2, 4, 6, 8, 18, 20-22, 24, 26, and 29) and 5 GSTs (EgriGST1 and EgriGST delta group) provide a major source of candidate genes for odorants degradation in E. grisescens.

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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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            MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms.

            The Molecular Evolutionary Genetics Analysis (Mega) software implements many analytical methods and tools for phylogenomics and phylomedicine. Here, we report a transformation of Mega to enable cross-platform use on Microsoft Windows and Linux operating systems. Mega X does not require virtualization or emulation software and provides a uniform user experience across platforms. Mega X has additionally been upgraded to use multiple computing cores for many molecular evolutionary analyses. Mega X is available in two interfaces (graphical and command line) and can be downloaded from www.megasoftware.net free of charge.
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              Trinity: reconstructing a full-length transcriptome without a genome from RNA-Seq data

              Massively-parallel cDNA sequencing has opened the way to deep and efficient probing of transcriptomes. Current approaches for transcript reconstruction from such data often rely on aligning reads to a reference genome, and are thus unsuitable for samples with a partial or missing reference genome. Here, we present the Trinity methodology for de novo full-length transcriptome reconstruction, and evaluate it on samples from fission yeast, mouse, and whitefly – an insect whose genome has not yet been sequenced. Trinity fully reconstructs a large fraction of the transcripts present in the data, also reporting alternative splice isoforms and transcripts from recently duplicated genes. In all cases, Trinity performs better than other available de novo transcriptome assembly programs, and its sensitivity is comparable to methods relying on genome alignments. Our approach provides a unified and general solution for transcriptome reconstruction in any sample, especially in the complete absence of a reference genome.
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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
                04 April 2023
                2023
                : 14
                : 1183610
                Affiliations
                [1] 1 College of Agriculture , Xinyang Agriculture and Forestry University , Xinyang, China
                [2] 2 State Key Laboratory for Biology of Plant Diseases and Insect Pests , Institute of Plant Protection , Chinese Academy of Agricultural Sciences , Beijing, China
                [3] 3 College of Agriculture , Xinjiang Agricultural University , Urumqi, China
                [4] 4 Department of Entomology , University of Kentucky , Lexington, KY, United states
                Author notes

                Edited by: Fengqi Li, Guizhou University, China

                Reviewed by: Wen Xie, Insititute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, China

                Man Zhao, Henan Agricultural University, China

                *Correspondence: Xiangrui Li, xrli@ 123456ippcaas.cn

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

                Article
                1183610
                10.3389/fphys.2023.1183610
                10110894
                bac295ba-fe90-4f67-9335-1fe952d37d8d
                Copyright © 2023 Zhang, Chen, Zhao, Guo, Hong, Zhi, Zhang, Zhou, Zhang, Zhou and Li.

                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
                : 10 March 2023
                : 21 March 2023
                Funding
                This study was supported by the Natural Science Foundation of Henan Province (No. 212300410229), the Key Project for University Excellent Young Talents of Henan Province (No. 2020GGJS260), the Project of Science and Technology Innovation Team (Nos. XNKJTD-007 and KJCXTD-202001), special funds for Henan Province Scientific and Technological Development guided by the Central Government (Z20221341063), and Youth Foundation (Nos. 2019LG004 and 20200106), Xinyang Agriculture and Forestry University, P.R. China.
                Categories
                Physiology
                Original Research

                Anatomy & Physiology
                ectropis grisescens,antennal transcriptome,odorant-degrading enzyme,carboxylesterases,glutathione s-transferases

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