Analysis and Identification of Chemosensory Genes in the Transcriptome of Adult Rhoptroceros cyatheae

Analysis and Identification of Chemosensory Genes in the Transcriptome of Adult Rhoptroceros cyatheae | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Analysis and Identification of Chemosensory Genes in the Transcriptome of Adult Rhoptroceros cyatheae mengqing zhou, yu Jiang, xiaona Zhang, gaoyin wu, tao peng, sheng Liang, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7729120/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract Rhoptroceros cyatheae (Hymenoptera) belongs to the genus Rhopographus of Selandriidae. It mainly causes large-scale infestations during the sprouting period of Alsophila spinulosa , and is among the herbivorous insects that harm this plant.The body of R. cyatheae contains chemosensory genes that detect and transduce chemical signals related to host location, feeding, mating, and oviposition. However, to date, no reports on the chemosensory genes of R. cyatheae have been published. Thus,on the basis of a the tran-scriptome database of male and female adult individuals of R. cyatheae , a total of 30,296 unigenes were identified, with an N50 length of 3,286 bp. Through comparisons with six major public databases, namely NR, Swiss-Prot, Pfam, eggNOG, GO, and KEGG, a total of 11,109 unigenes were annotated, accounting for 36.67%. Among these, the number of unigenes annotated in the NR database was the largest, reaching 10,774, whereas the number of unigenes annotated in the KEGG database was the smallest, at 6,300. An analysis of the annotation information, 90 candidate chemosensory genes of R. cyatheae , including 11 OBPs, 10 CSPs, 6 NPC2s, 24 ORs (comprising 23 typical OR genes and 1 Orco gene), 20 IRs, 15 GRs and 4 SNMPs. A phylogenetic tree of chemosensory genes was subsequently constructed to investigate the homology between the chemosensory genes of the R. cyatheae and those of other insect species. Furthermore, 13 chemosensory genes differentially expressed between males and females, and their tissue expression profiles were verified via RT‒qPCR. These findings lay a molecular foundation for further research on the gene functions and olfactory perception mechanisms of R. cyatheae . Rhoptroceros cyatheae adult transcriptome transcriptome sequencing chemosensory genes Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 1. Introduction The olfactory chemosensory system plays a crucial role in determining the life activities of insects. It enables the detection of olfactory information carried by odor molecules, which is associated with specific behaviors such as host location, mating, foraging, and evading predators [ 1 ].When hydrophobic molecules enter the sensillar lymph through chemosensilla, odorant-binding proteins (OBPs), chemosensory proteins (CSPs), and Niemann-Pick type C2 proteins (NPC2s) in the lymph bind and transport these molecules, which are subsequently detected by olfactory receptor neurons (ORNs). Odorant receptors (ORs), ionotropic receptors (IRs), gustatory receptors (GRs), or sensory neuron membrane proteins (SNMPs) on ORN membranes convert chemical signals into electrical signals. These electrical signals are then transduced by ORNs to the central nervous system in the insect brain [ 2 ]. After processing by the central nervous system, these signals trigger corresponding physiological and behavioral responses [ 3 ]. In the process of odor detection, the initial step in the olfactory recognition of hydrophobic odorants involves the binding and transport of odor molecules by OBPs, CSPs, and NPC2s present in the sensillar lymph [ 4 ].OBPs, CSPs, and NPC2s are all small molecular-weight water-soluble proteins. Classical OBPs possess six conserved cysteine residues, whereas minus-C OBPs contain four conserved cysteine residues, and atypical OBPs feature only two conserved cysteine residues. Compared with OBPs, which are more evolutionarily conserved ,CSPs typically carry four conserved cysteine residues. Initially identified in mammals for their role in shaping cholesterol transport, NPC2s were later found to participate in olfactory responses in insects [ 5 – 7 ].OBPs primarily recognize and transport volatile compounds; CSPs mainly bind nonvolatile substances; whereas NPC2s are responsible for binding and transporting hydrophobic compounds[ 8 ]. Transmembrane receptor proteins (ORs, IRs, GRs, and SNMPs) on olfactory neurons transmit olfactory signals across the membrane to the central nervous system subsequently. Insect odorant receptors can be categorized into ORs and odorant receptor co-receptors (Orco). Orco facilitates the precise localization of OR proteins to the dendritic membranes of olfactory neurons [ 9 ].IRs evolved from a class of ionotropic glutamate receptors (iGluRs) and primarily function in sensing olfactory cues, gustatory signals, temperature and humidity [ 10 , 11 ]. GRs primarily mediate the perception of bitterness, sweetness, and carbon dioxide (CO₂). Additionally, accumulating evidence suggests their involvement in light and temperature sensing[ 12 , 13 ].SNMPs are transmembrane structural proteins that are categorized into SNMP1 and SNMP2, which play critical roles in the detection of plant volatiles and insect sex pheromones [ 14 ].These chemosensory genes play crucial roles in determining insect olfactory recognition and represent the foundational step in study of insect olfactory mechanisms. However, research on the olfactory mechanisms of Rhoptroceros cyatheae adults remains limited. Elucidating the olfactory mechanisms of both female and male R. cyatheae is significant for understanding how they recognize semiochemicals and rely on chemical communication and sensory mechanisms to locate hosts and mates. Concurrently, the precise identification of olfactory-related genes in R. cyatheae lays the groundwork for subsequent investigations into its olfactory mechanisms. R. cyatheae , which belongs the family Selandriidae and genus Rhopographus , is distributed in regions such as Sichuan and Guizhou. It is among the primary herbivorous insects that infest Alsophila spinulosa (a tree fern species) [ 15 ].This insect completes four to five generations annually, with its occurrence period spanning from April to June each year. During outbreaks, R. cyatheae can cause severe infestations, where the young leaves of extensive A. spinulosa stands are completely defoliated by its larvae [ 16 ].The tree fern A. spinulosa faces challenges due to its lengthy reproductive cycle via spores and difficult natural regeneration. Outbreaks of the R. cyatheae drastically reduce spore production, severely impacting the population sustainability of this species [ 17 ]. A. spinulosa is a Class II protected plant in China, hailed as the 'living fossil' of terrestrial plants, and possesses extremely high research and pharmacological value [ 18 ].However, in recent years, the population of R. cyatheae has continued to increase, posing a significant threat to the survival and reproduction of A. spinulosa [ 19 , 20 ].Therefore, it is crucial to develop novel biological control methods that are safer, more environmentally friendly, and less ecologically damaging. In this study, RNA-Seq technology was used to obtain adults transcriptome data from of female and male of R. cyatheae . We identified chemosensory-related genes including OBPs, CSPs, NPC2s, ORs, GRs, IRs, and SNMPs. The findings not only provide a solid theoretical foundation for future research on olfactory genes and mechanisms in R. cyatheae , but they also hold profound significance for advancing ecological regulation strategies and developing green control technologies targeting this species. 2. Materials and Methods 2.1Insect Rearing and Sample Collection The test insects, R. cyatheae (Hymenoptera: Selandriidae), were collected from December 2024 to May 2025 in the Chishui Alsophila National Nature Reserve, Guizhou Province, China. Larvae sampled from the field were reared in an intelligent biochemical incubator (model GZL-P8000-C3, Hefei DuskTec Biological Technology Co., Ltd.) under controlled conditions: a temperature of 25 ± 1°C, a relative humidity of75%±10%, and a 14h light:10h dark cycle. After eclosion, the adult pecimens R. cyatheae were provided with a 10% honey solution as a dietary supplement and maintained under the same environmental conditions described above. Adults of R. cyatheae (4 days post-eclosion) were rinsed with RNase-free and DNase-free water, transferred into 5 ml cryovials, immediately frozen in liquid nitrogen, and stored in a − 80°C ultralow temperature freezer (MDF-86V180E) for transcriptome sequencing. Three biological replicates were established for both sexes. To investigate the expression levels of chemosensory genes in different tissues of R. cyatheae , twelve tissue types were dissected from adult male and female individuals under a laminar flow hood (model LJ-SZM, Leica, Germany): female antennae (Fant), male antennae (Mant), female heads without antennae (Fhea), male heads without antennae (Mhea), female thoraces (Ftho), male thoraces (Mtho), female abdomens (Fabd), male abdomens (Mabd), female wings (Fwin), male wings (Mwin), female legs (Fleg), and male legs (Mleg) A total of 150 individuals per sex were obtained. All the samples were placed in labeled 5 ml prechilled cryovials, rapidly frozen in liquid nitrogen, and stored at − 80°C until RNA extraction. Each sample was processed with three biological replicates, and each biological replicate included three technical replicates. 2.2 Total RNA Extraction and Detection, cDNA Library Construction and Sequencing Total RNA was extracted from female and male adult samples of R. cyatheae via QIAzol Lysis Reagent (Qiagen, Germany). The integrity of the total RNA was assessed through 1% agarose gel electrophoresis, while the concentration and purity were measured using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, USA). The RNA samples were then precisely evaluated for purity and integrity using an Agilent 5300 Bioanalyzer (Agilent Technologies, USA). This study included three biological replicates. Approximately 1 µg of total RNA from each sample was used for cDNA library construction. The libraries were sequenced on an Illumina NovaSeq X platform, with both library preparation and sequencing performed in collaboration with Shanghai Majorbio Technology. 2.3 Transcriptome Assembly and Functional Annotation The raw reads obtained from sequencing were first processed using Fastp to remove adapter sequences and low-quality reads, yielding clean reads [ 21 , 22 ].Next, Trinity ( https://github.com/trinityrnaseq/trinityrnaseq/wiki ) was used to perform de novo assembly for all the clean reads [ 23 ].The assembly results were filtered and optimized using TransRate ( http://hibberdlab.com/transrate/ ), and then the assembly completeness of the unigenes was evaluated with BUSCO (Benchmarking Universal Single-Copy Orthologs, http://busco.ezlab.org).Additionall y, the Q20, Q30, GC content, error rate, and nucleotide base composition of the clean data were calculated(S2).Finally, to obtain comprehensive functional annotations, all unigenes derived from this transcriptome sequence were subjected to alignment-based annotation against six major databases: the Non-redundant Protein Database (NR), Pfam, Clusters of Orthologous Groups (COG), Swiss-Prot, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Ontology (GO). 2.4 Identification and Expression Analysis of Chemosensory Genes To identify putative chemosensory genes (OBPs, CSPs, NPC2s, ORs, GRs, IRs, and SNMPs), we searched the transcriptome annotation results obtained in Section 1.4 using olfactory gene keywords (OBP and odorant binding protein, CSP and chemosensory protein, NPC2 and Niemann–Pick type C2 protein, OR and odorant receptor, GR and gustatory receptor, IR and ionotropic receptor, SNMP and sensory neuron membrane protein). The initial identification was further validated through manual BLASTX and BLASTP alignments, with an E-value threshold of < 10⁻⁵ set for screening candidate olfactory-related genes. The ORF finder ( https://www.ncbi.nlm.nih.gov/orffinder/ ) was used to predict the open reading frames (ORFs) of all the putative chemosensory genes. For the candidate OBP, CSP, and NPC2 genes, the N-terminal signal peptides were predicted using the SignalP-6.0 online program ( https://services.healthtech.dtu.dk/services/SignalP-6.0/ ; Almagro Armenteros et al.). Molecular weights (MWs) and isoelectric points (pI) were obtained using the ExPASy proteomics server ( https://www.expasy.org/resources/compute-pi-mw ). For candidate OR, IR, and SNMP genes, the transmembrane domains (TMDs) of the annotated proteins were predicted with the online program DeepTMHMM-1.0 ( https://services.healthtech.dtu.dk/services/DeepTMHMM-1.0/ ). Multiple sequence alignment of chemosensory gene amino acid sequences was performed via the ClustalX algorithm in MEGA 12.0 software[ 24 , 25 ], with subsequent color coding implemented through ESPript 3.0 ( https://espript.ibcp.fr/ESPript/ESPript/index.php).Phylogeneti c analysis of candidate the chemosensory genes was conducted using amino acid sequences from R. cyatheae and other insects. The Maximum likelihood Method (ML) with the Poisson model was employed in MEGA12.0 to infer phylogenetic relationships, incorporating 1,000 bootstrap replicates and retaining branches supported by ≥ 95% bootstrap values[ 26 ].Following the preliminary reconstruction of the relevant phylogenetic trees, visualization and editing were performed using the Interactive Tree of Life tool (iTOL v7) ( https://itol.embl.de/ ). 2.5 Screening and Analysis of Differentially Expressed Genes To compare differences in olfactory gene expression differences between female and male adults of R. cyathea , RSEM was employed for expression quantification, with Fragments Per Kilobase of transcript per Million mapped reads (FPKM) values employed to assess gene expression levels[ 27 ].Differential expression analysis of read counts was performed using DESeq2 on basis of the negative binomial distribution model. Differentially expressed genes were identified using a threshold of fold change (FC) ≥ 1 and an adjusted P-value < 0.05[ 28 ]. 2.6 RT-qPCR Analysis of Expression Levels of Candidate Chemosensory Genes On the basis of the differential gene expression analysis results, chemosensory genes with sex-biased expression (female- or male-biased) were preliminarily screened under the threshold conditions of q-value 1 as candidate genes. These candidate genes were subsequently validated for their quantitative expression levels across the 12 distinct tissues using real-time quantitative polymerase chain reaction (RT-qPCR).To further remove genomic DNA (gDNA) and synthesize cDNA, TransScript All-in-One First-Strand cDNA Synthesis SuperMix (Transgen Biotech, Beijing, China) was used to reverse transcribe 1 µg of total RNA per sample into cDNA RcyaRPL32 and RcyaRPS18 were used as reference genes (internal controls) to normalize the expression data. Specific primers were designed using Primer Premier 5.0 ( https://bioinfo.ut.ee/primer3-0.4.0/ ) and are listed in Table 3 . qPCR measurements were subsequently performed using 2×Universal Blue SYBR Green qPCR Master Mix (Servibio, Wuhan, China) and SweScript All-in-One RT SuperMix for qPCR (Servibio, Wuhan, China). Each qPCR was performed according to the manufacturer's instructions under the following conditions: initial denaturation at 95°C for 30 s, followed by 40 cycles of denaturation at 95°C for 15 s, annealing/extension at 60°C for 30 s. Subsequently, a melting curve analysis was generated by continuous fluorescence measurement from 65°C to 95°C with increments of 0.5°C. All qPCR assays were conducted with three technical replicates and three biological replicates. Table 3 The information of primers used for RT-qPCR assays 引物名称 Forward primer (5’-3’) Reverse primer (5’-3’) RcyaRPL32 GACAAGCTCAAGCGTAACTGGC ACGATAGCTTTGCGTTTCTTGC RcyaRPS18 TGCCTCGGGTTGGACATAATC CAAGGGTGTCGGTCGTCGTT RcyaOBP7 CCGAAAGACAAGATGACGGAGAT TGATCCTTATGCTCGTGGAAACA RcyaOBP8 GCTTCCTTCATTGCGTTCTCAC AGCGGTCCTGGCCTCTTCTT RcyaCSP2 TTCATCAATCACCTCCGCCC GATTCAAGCGGCACCTACAGG RcyaCSP10 CCTCTGGGTCGATCTTGGTC ACTGAAAGATGTGCTGGGTGAG RcyaOR19 AACAATGGCGTCCGTGGTAC AGATCGTGCGTCTATGAACAGG RcyaOR14 CCAAGTAAGCGAATGTTGACGAT TTGGAGAGGTTGACTAACACAGCC RcyaOR20 AACATGACGACCAGTGCTGAAA AGGAGTCGGTTATCGGCTTTCA RcyaOR17 CCTAACGACCGACTTGACCAAC GCTCAGAACTTATCGCACCTACA RcyaGR5a CTGGCAACGAGAAATATGAACG CCTGAGCGGATTGGTGAAAC RcyaiGluR4 GCGAAACCGATTCCTATACCTT ATTCAAGACCTCCGAACACCTAC RcyaSNMP2a ACGCACGAAAGGAATTACAGG TGGCGATGTAGAAATGAGGTAATG RcyaNPC2d GGTCAAATGAGATAAACCTCCAGAT CGTGGGTCAAGGCGTAGATAG RcyaNPC2e CTGACCGACCACGAAGTTGTTT CGGTGCAGGTGAAGATAGAAGATG 2.7 Statistical Analysis The relative expression levels of chemosensory genes in each sample were calculated via the 2^(-ΔΔCt) method, with the tissue showing the lowest expression level as the reference. Subsequently, logarithmic transformation was performed on the data. If the data failed to meet the requirements for normality and homogeneity of variance, the nonparametric Kruskal-Wallis test was used to compare differences; otherwise, one-way analysis of variance (ANOVA) followed by Duncan's test was applied for difference testing. All the data were statistically analyzed using SPSS Statistics 25 and Origin 2021 Pro, and are presented as the mean ± standard error of the mean (SEM). 3. Results 3.1 Transcriptome Sequencing Statistics and Sequence Assembly The transcriptomes of female and male adults of R. cyatheae were sequenced on a second-generation high-throughput sequencing platform. A total of 40.94 Gb of clean reads were obtained, with each sample yielding at least 6.34 Gb of clean data. The Q20 and Q30 base percentages exceeded 98% and 96%, respectively, while the GC content across samples varied between 41.48% and 43.32%( Table 2 .in the Supplementary Materials). The average base error rate for all quality-controlled data was less than 0.02%, indicating high-quality sequencing data.Sequence assembly was performed using Trinity software, and the results revealed that 44,396 transcripts were obtained, with an average length of 1393.21 bp and an N50 length of 4356 bp (Table 1 ).Further assembly analysis of the transcripts yielded a total of 30,296 unigenes, with an average length of 1,393.21 bp and an N50 length of 3,286 bp. In terms of the length distribution of the unigenes and transcripts, those with lengths ranging from 200 to 500 bp were the most abundant, accounting for 14,157 unigenes (47%) and 15,698 transcripts (35%). In contrast, those with lengths ranging from 4,001 to 4,500 bp were the least abundant, with only 486 unigenes (2%) and 1,123 transcripts (3%) (Fig. 1 ). For the candidate OR, IR, and SNMP genes, the transmembrane domains (TMDs) of the annotated proteins were predicted using the online program DeepTMHMM-1.0 ( https://services.healthtech.dtu.dk/services/DeepTMHMM-1.0/ ). Multiple sequence alignment of chemosensory gene amino acid sequences was performed using the ClustalX algorithm in MEGA 12.0 software.The assembly completeness was assessed using BUSCO, which yielded a score exceeding 95%, indicating high-quality assembly. The raw reads of R. cyatheae were subsequently submitted to the NCBI Short Read Archive database under accession number PRJNA1309270. Table 2 Information for 6 samples used for transcriptome analysis Sample Raw reads Raw bases Clean reads Clean bases Error rate(%) Q20(%) Q30(%) GC content(%) RF1 42,742,146 6,454,064,046 42,440,672 6,344,273,982 0.0119 98.76 96.18 43.23 RF2 44,087,188 6,657,165,388 43,765,508 6,559,551,741 0.0119 98.74 96.14 42.19 RF3 51,216,586 7,733,704,486 50,908,962 7,641,273,693 0.0119 98.81 96.3 43.32 RM1 43,398,736 6,553,209,136 43,123,866 6,460,256,582 0.0119 98.77 96.25 42.17 RM2 44,020,738 6,647,131,438 43,726,486 6,549,862,099 0.012 98.72 96.1 42.19 RM3 49,702,156 7,505,025,556 49,365,188 7,385,834,680 0.0119 98.74 96.17 41.48 Table 1 Number and length of transcripts and unigenes in R. cyatheae. Type Unigene Transcript Total number 30296 44396 Total base 42208633 88696954 Largest length (bp) 33119 33119 Smallest length (bp) 201 201 Average length (bp) 1393.21 1997.86 N50 length (bp) 3286 4356 E90N50 length (bp) 4853 4215 Fragment mapped percent(%) 81.988 93.204 GC percent (%) 38.14 39.02 TransRate score 0.46504 0.51992 BUSCO score C:95.5% C:98.0 3.2 Functional Annotation and Classification of Transcriptome The functional annotation results indicated that 10,716 unigenes (35.94%) were matched in the NCBI NR protein database with an E-value threshold of 10⁻⁵ (Fig. 2 A).Species classification indicated that the highest sequence homology was observed in Athalia rosae (32.57%), followed by Diprion similis (21.35%), Neodiprion lecontei (10.20%), Neodiprion fabricii (7.99%), Neodiprion virginianus (5.10%), and Neodiprion pinetum (4.53%). The species that exhibited the highest homology with R. cyatheae were exclusively hymenopteran insects. In the KEGG annotation, a total of 7,163 unigenes were classified into six KEGG functional groups (Fig. 2 B): Human Diseases,with 1,745 unigenes (24.36%);Environmental Information Processing, with 861 unigenes (12.02%);Genetic Information Processing with 1,161 unigenes (16.21%); Cellular Processes, with 898 unigenes (14.54%);Organismal Systems ,with 1,225 unigenes (17.10%), and Metabolism, with 1,273 unigenes (17.78%) For GO annotation, the transcripts were functionally categorized according to their GO classifications (Fig. 2 C). A total of 6,619 genes (22.20%) were assigned to three GO categories, namely, molecular function, cellular component, and biological process, encompassing 51 specific terms. Within the biological process category, the terms “cellular process” and “metabolic process” exhibited the greatest representation. With respect to cellular components, “cell part” constituted the most dominant group. Regarding molecular function, the terms “binding” and “catalytic activity” were the most abundant groups. On the basis of the results of the GO analysis, genes associated with “binding” were significantly enriched in R.cyatheae, followed by “cell part” and “catalytic activity” which ranked second and third, respectively. 3.3 Identified Candidate Water-Soluble Proteins Candidate OBPs Eleven candidate RcyaOBPs exceeding 600 bp in length were identified in R. cyatheae and designated RcyaOBP1 through RcyaOBP11. These genes encoded proteins ranging from 136 to 273 amino acid residues, with molecular weights of 4.32 to 9.20 kDa and isoelectric points (pI) ranging from 4.20 to 9.16. Sequence alignment via BLASTP revealed that the candidate RcyaOBPs share sequence identities ranging from 26.71% to 63.16% with other hymenopteran OBPs. Among these candidates, only RcyaOBP8 was predicted to lack a signal peptide, whereas RcyaOBP3 and RcyaOBP5 lacked conserved cysteine residues (Table 4 .in the Supplementary Materials). Multiple sequence alignment revealed that nine candidate RcyaOBPs (RcyaOBP1 and RcyaOBP4–11) contained six conserved cysteine residues with the pattern X11–37-Cys1-X20–27-Cys2-X3-Cys3-X26–49-Cys4-X8–11-Cys5-X8-Cys6-X5–18, which were characterized as classical OBPs. RcyaOBP2, harboring four conserved cysteine residues, was a Minus-COBPs. RcyaOBP3 was classified as atypical OBPs with 2 cysteines (Fig. 3 ). A Maximum likelihood (ML) tree was constructed using the OBPs of R. cyatheae and nine other Hymenopteran species. The results revealed that many RcyaOBPs were highly divergent into different clades, while the OBPs of species such as BdioOBPs ( Baryscapus dioryctriae ), NvitOBPs ( Nasonia vitripennis ), and CcunOBPs ( Chouioia cunea ) were closely clustered in multiple clades (Fig. 7 ). Table 4 Bioinformatics analysis of identified R. cyatheae OBP genes Gene ID Gene Name ORF (aa) Signal Peptide Group PI MW(Kd) BLAST Annotation Description Query E value Identify(%) TRINITY_DN10134_c0_g2 RcyaOBP1 136 21 complete 4.32 15.07 XP_046625102.1, general odorant-binding protein 83a-like isoform X2 [Neodiprion virginianus] 99% 2E-56 63.16 TRINITY_DN16238_c0_g1 RcyaOBP2 214 16 complete 4.53 23.57 QHN69060.1, odorant binding protein 3 [Sirex nitobei] 53% 1E-16 41.88 TRINITY_DN16731_c0_g1 RcyaOBP3 145 30 5prime_partial 9.2 15.97 QHN69068.1, odorant binding protein 11 [Sirex nitobei] 90% 4E-33 43.61 TRINITY_DN17528_c0_g1 RcyaOBP4 138 19 complete 4.77 15.50 XP_046663889.1, general odorant-binding protein 19d-like [Homalodisca vitripennis] 91% 7E-06 27.69 TRINITY_DN19984_c0_g1 RcyaOBP5 279 50 5prime_partial 7.66 31,15 XP_058448369.1, general odorant-binding protein 56d-like [Malaya genurostris] 26% 9E-10 45.95 TRINITY_DN20475_c0_g1 RcyaOBP6 151 22 complete 6.35 17.00 QHN69059.1, odorant binding protein 2 [Sirex nitobei] 88% 6E-45 52.24 TRINITY_DN3522_c0_g2 RcyaOBP7 141 19 complete 5.22 15.66 XP_033217025.1, general odorant-binding protein 56a-like [Belonocnema kinseyi] 87% 2E-27 48 TRINITY_DN3640_c0_g1 RcyaOBP8 273 0 complete 5.46 30.70 XP_033342464.1, general odorant-binding protein 56a-like [Megalopta genalis] 43% 1E-10 27.87 TRINITY_DN4219_c0_g3 RcyaOBP9 143 20 complete 4.71 16.04 XP_046431779.1, general odorant-binding protein 83a-like [Neodiprion fabricii] 97% 2E-36 46.43 TRINITY_DN4317_c0_g1 RcyaOBP10 133 17 complete 5.46 14.20 XP_046625102.1, general odorant-binding protein 83a-like isoform X2 [Neodiprion virginianus] 99% 2E-56 63.16 TRINITY_DN923_c0_g5 RcyaOBP11 142 21 complete 5.17 15.80 XP_012256027.2, general odorant-binding protein 83a-like [Athalia rosae] 99% 2E-60 61.97 Note: †ORF: open reading frame. Amino acid sequences of R. cyatheae OBP genes are listed in the supplementary data. Candidate CSPs Through bioinformatic analysis, ten candidate RcyaCSPs exceeding 600 bp in length were identified in the adult transcriptome of R. cyatheae . These genes encode proteins ranging from 118 to 157 amino acid residues, with molecular weights of 4.77 to 9.41 kDa and isoelectric points (pI) ranging from 4.36 to 7.70. The candidate RcyaCSP genes exhibit 46.15% to 69.17% homology with other hymenopteran CSPs. Among these candidates, only RcyaCSP8 was predicted to lack a signal peptide, while RcyaCSP3 and RcyaCSP5 lacked five conserved cysteine residues (Table 5 .in the Supplementary Materials).The multiple sequence alignment results showed that ten candidate RcyaCSPs (RcyaCSP1 and RcyaCSP4–11) contained four conserved cysteine residues with the pattern X38–75-Cys1-X6–8-Cys2-X18–19-Cys3-X2-Cys4-X37–51 (Fig. 4 ).An ML tree was constructed using CSPs from R. cyatheae and 12 other hymenopteran species. The results revealed that many RcyaCSPs were highly divergent, forming distinct clades, while CSPs from the other 12 hymenopteran species clustered closely within multiple branches (Fig. 8 ). Table 5 Bioinformatics analysis of identified R. cyatheae CSP genes Gene ID Gene Name Complete ORF (aa) Signal Peptide Group Pl MW (Kd) BLAST Annotation Description Query Cover E value Identify(%) >TRINITY_DN2639_c0_g1 RcyaCSP1 123 19 complete 7.61 14.31 QGW50256.1, chemosensory protein 9 [Chouioia cunea] 94% 6E-33 46.15 >TRINITY_DN14066_c0_g1 RcyaCSP2 139 25 5prime_partial 5.74 15.52 XP_046492842.1, chemosensory protein 4 [Neodiprion pinetum] 90% 3E-55 67.46 >TRINITY_DN20688_c0_g1 RcyaCSP3 138 24 complete 9.41 15.87 QHN69075.1, chemosensory protein 3 [Sirex noctilio] 99% 6E-59 63.31 >TRINITY_DN14055_c0_g1_ RcyaCSP4 118 20 complete 5.11 13.74 BAS29775.1, chemosensory protein [Camponotus japonicus] 95% 2E-37 53.1 >TRINITY_DN3807_c0_g1_ RcyaCSP5 118 25 complete 9.11 13.36 XP_072754896.1, chemosensory protein 2 [Anoplolepis gracilipes] 99% 6E-54 69.17 >TRINITY_DN8223_c0_g1 RcyaCSP6 124 16 complete 9.32 13.93 ALG36159.1, chemosensory protein 6 [Sclerodermus sp. MQW-2015] 99% 7E-63 75 >TRINITY_DN1049_c0_g1 RcyaCSP7 154 0 complete 9.01 17.54 ALG36159.1, chemosensory protein 6 [Sclerodermus sp. MQW-2015] 54% 8.E-15 67.92 >TRINITY_DN9812_c0_g1 RcyaCSP8 120 18 complete 4.89 13.80 BFW56612.1, Chemosensory protein [Polyrhachis lamellidens] 79% 9E-49 60.16 >TRINITY_DN6262_c0_g1 RcyaCSP9 157 41 5prime_partial 4.77 17.70 ALG36155.1, chemosensory protein 2 [Sclerodermus sp. MQW-2015] 95% 2E-39 52.14 >TRINITY_DN17317_c0_g1_ RcyaCSP10 129 18 complete 5.46 14.32 UEN71179.1, chemosensory protein 3 [Gregopimpla kuwanae] 71% 8E-41 61.61 Note: †ORF: open reading frame; ND: not detected. Amino acid sequences of R. cyatheae OBP genes are listed in the supplementary data. Candidate NPC2s Six full-length putative RcyaNPC2 proteins encoding 144–161 amino acid residues with molecular weights ranging from 4.21 kDa to 8.10 kDa were identified in R. cyatheae. These RcyaNPC2s shared 26.71–66.44% amino acid sequence identity with other known insect NPC2s (Table 6 .in Supplementary Materials).The multiple alignment revealed revealed that six NPC2s contained six conserved cysteine residues with the pattern X20–24-Cys1-X13–14-Cys2-X4–5-Cys3-X43–45-Cys4-X6–13-Cys5-X40–42-Cys6-X8–33 (Fig. 5 ). A Maximum likelihood (ML) tree was constructed using the CSPs of R. cyatheae and ten other Hymenopteran species. The results revealed that many RcyaCSPs were highly divergent into distinct clades, while the NPC2s of B.dioryctriae (BdioNPC2s), N. vitripennis (NvitNPC2s), and the CSPs of C. cunea (CcunCSPs) were closely clustered in multiple clades. Specifically, RcyaNPC2a and RcyaNPC2e formed a monophyletic clade, whereas RcyaNPC2f and BdioNPC2a aggregated into an independent clade separated from the other branches (Fig. 9 ). Table 6 Bioinformatics analysis of identified R. cyatheae NPC2 genes Gene ID Gene Name Complete ORF (aa) Signal Peptide Group PI MW (Kd) BLAST Annotation Description Query E value Identify(%) TRINITY_DN8475_c0_g1 RcyaNPC2a 151 19 complete 5.22 16.34 XP_015509554.1, NPC intracellular cholesterol transporter 2 homolog a [Neodiprion lecontei] 97% 7E-66 63.51 TRINITY_DN10314_c0_g4 RcyaNPC2b 161 28 complete 4.21 17.56 XP_046433474.1, NPC intracellular cholesterol transporter 2 homolog a-like [Neodiprion fabricii] 91% 1E-67 66.44 TRINITY_DN437_c3_g1 RcyaNPC2c 156 19 complete 7.56 17.19 XP_015514132.1, NPC intracellular cholesterol transporter 2 [Neodiprion lecontei] 99% 4E-54 52.56 TRINITY_DN10489_c0_g1 RcyaNPC2d 154 22 complete 8.10 17.37 XP_046626566.1, MD-2-related lipid-recognition protein-like [Neodiprion virginianus] 98% 4E-48 49.06 TRINITY_DN3106_c0_g2 RcyaNPC2e 153 18 complete 4.99 16.55 XP_032685878.1, NPC intracellular cholesterol transporter 2 homolog a-like [Odontomachus brunneus] 94% 1E-5 26.71 TRINITY_DN3972_c0_g1 RcyaNPC2f 144 0 complete 7.69 15.86 XP_046489940.1NPC intracellular cholesterol transporter 2 [Neodiprion pinetum] 86% 7E-28 44.8 Note: †ORF: open reading frame; ND: not detected. Amino acid sequences of R. cyatheae OBP genes are listed in the supplementary data. 3.4 Identified Candidate Transmembrane Proteins Candidate ORs In R. cyatheae , a total of 24 putative RcyaORs encoding 57–411 amino acid residues were identified. The putative RcyaORs exhibited 25.00–81.82% similarity with other orthologous genes in Hymenoptera species. Among them, 79 RcyaORs were full-length and encoded 0–6 transmembrane domains (TMDs) (Table 7 .in the Supplementary Materials). The full-length RcyaOrco gene was successfully identified, encoding 114 amino acid residues with 3 TMDs,and showing 81.82% similarity to ArosOrco of Athalia rosae. As expected, RcyaOrco clustered with Orco sequences from other hymenopteran species, forming a distinct clade in the maximum likelihood (ML) tree constructed for ORs. Five ORs were segregated into the Orco clade, where they clustered with Orco orthologs from other species. Furthermore, most RcyaORs demonstrated close phylogenetic relationships with ORs from Microplitis mediator and B.dioryctriae (Fig. 10 ). Table 7 Bioinformatics analysis of identified R. cyatheae OR genes Gene ID Gene Name Complete ORF (aa) Group TMD BLAST Annotation Description Query E value Identify(%) TRINITY_DN10140_c0_g1 RcyaOrco 114 internal 3 XP_012253637.2, odorant receptor coreceptor [Athalia rosae] 87% 2E-49 81.82 TRINITY_DN9632_c0_g5 RcyaOR 1 109 5prime_partial 2 XP_048516116.1, odorant receptor 43a [Athalia rosae] 97% 1e-24 46.73 TRINITY_DN9632_c0_g1 RcyaOR 2 102 3prime_partial 1 XP_048516116.1, odorant receptor 43a [Athalia rosae] 91% 1e-19 56.99 TRINITY_DN9666_c0_g6 RcyaOR 3 119 5prime_partial 2 XP_015595561.1, odorant receptor Or1 isoform X1 [Cephus cinctus] 81% 1e-24 50.50 TRINITY_DN9081_c0_g1 RcyaOR 4 98 internal 2 XP_046626501.1, odorant receptor 49b-like [Neodiprion virginianus] 98% 3e-23 50.00 TRINITY_DN8388_c0_g1 RcyaOR 5 299 5prime_partial 4 XP_012266577.3, odorant receptor 4-like isoform X1 [Athalia rosae] 74% 6e-56 42.53 TRINITY_DN8388_c0_g2 RcyaOR 6 63 complete 0 UEN71227.1, olfactory receptor 44 [Gregopimpla kuwanae] 95% 5e-10 47.54 TRINITY_DN21124_c0_g1 RcyaOR 7 75 5prime_partial 0 XP_046735418.1, odorant receptor Or2-like [Diprion similis] 82% 3e-16 62.90 TRINITY_DN7376_c0_g2 RcyaOR 8 75 3prime_partial 2 XP_046738658.1, odorant receptor 13a-like [Diprion similis] 99% 8e-31 75.68 TRINITY_DN7376_c0_g3 RcyaOR 9 211 internal 0 XP_046738674.1, odorant receptor 22c-like [Diprion similis] 26% 3e-07 48.21 TRINITY_DN10220_c0_g1 RcyaOR 10 154 complete 2 XP_048513569.1, odorant receptor 4-like isoform X2 [Athalia rosae] 70% 4e-35 62.73 TRINITY_DN2585_c0_g1 RcyaOR 11 157 complete 0 XP_046751421.1, gustatory and odorant receptor 24 [Diprion similis] 79% 9e-24 44.70 TRINITY_DN7376_c0_g1 RcyaOR 12 57 5prime_partial 0 XP_046738658.1, odorant receptor 13a-like [Diprion similis] 91% 4e-14 60.38 TRINITY_DN9666_c0_g7 RcyaOR 13 144 internal 1 KYM98886.1, Odorant receptor 46a, isoform A [Cyphomyrmex costatus] 63% 1e-34 61.54 TRINITY_DN3654_c0_g2 RcyaOR 14 121 complete 2 XP_046751356.1, odorant receptor Or1-like [Diprion similis] 98% 5e-24 39.53 TRINITY_DN9081_c0_g2 RcyaOR 15 143 internal 2 XP_046749494.1, odorant receptor 43a-like [Diprion similis] 100% 7e-35 44.76 TRINITY_DN20577_c0_g1 RcyaOR 16 114 5prime_partial 1 XP_046749183.1, putative odorant receptor 92a isoform X1 [Diprion similis] 90% 2e-26 57.28 TRINITY_DN6018_c0_g1 RcyaOR 17 411 complete 6 XP_046416448.1, odorant receptor 83a-like isoform X1 [Neodiprion fabricii] 99% 1e-136 45.70 TRINITY_DN7376_c0_g5 RcyaOR 18 153 internal 3 XP_046612742.1, odorant receptor 49a-like [Neodiprion virginianus] 98% 2e-44 52.98 TRINITY_DN2418_c0_g1 RcyaOR 19 388 complete 6 XP_048504947.1, odorant receptor 46a-like [Athalia rosae] 96% 3e-140 53.00 TRINITY_DN5024_c0_g1 RcyaOR 20 292 complete 5 XP_048510426.1, odorant receptor 49b-like isoform X4 [Athalia rosae] 98% 5e-106 53.77 TRINITY_DN20952_c0_g1 RcyaOR 21 71 5prime_partial 0 XP_046736951.1, odorant receptor 83a-like [Diprion similis] 96% 3e-25 53.77 TRINITY_DN4315_c0_g1 RcyaOR 22 109 complete 1 XP_048505323.1, odorant receptor 13a-like [Athalia rosae] 99% 1e-43 70.64 TRINITY_DN9632_c0_g2 RcyaOR 23 234 internal 3 XP_028050636.1, odorant receptor 67a [Monomorium pharaonis] 86% 8e-04 25.00 Note: †ORF: open reading frame; ND: not detected. Amino acid sequences of R. cyatheae OBP genes are listed in the supplementary data. Candidate IRs In R cyatheae, a total of 21 putative RcyaIR genes were identified, encoding 79–962 amino acid residues. These putative RcyaIRs showed 45.16–100.00% similarity to orthologous genes in other Hymenoptera species. Among them, five RcyaIRs were full-length and predicted to contain 0–3 transmembrane domains (TMDs) (Table 8 .in the Supplementary Materials). Phylogenetic analysis revealed that RcyaIRs diverged into several distinct subfamilies. A subset of RcyaIRs clustered within the same clades as the conserved IR25a, IR64a, IR75x, IR93a, and IR76b gene families, while the remaining RcyaIRs were distributed across other distinct phylogenetic branches (Fig. 11 ). Table 8 Bioinformatics analysis of identified R. cyatheae GR genes Gene ID Gene Name Complete ORF (aa) Group TMD BLAST Annotation Description Query E value Identify(%) TRINITY_DN12238_c0_g1 GR23a 197 3prime_partial 4 XP_046587885.1, putative gustatory receptor 28a [Neodiprion lecontei] 93% 8E-11 30.77 TRINITY_DN18654_c0_g1 GR28a1 104 5prime_partial 2 XP_046467114.2, putative gustatory receptor 28a [Neodiprion pinetum] 84% 2E-16 51.14 TRINITY_DN19044_c0_g1 GR28a2 105 internal 2 SP:Q9VM09, Putative gustatory receptor 28a [Neodiprion lecontei] 60.4 3.9E + 00 21.7 TRINITY_DN14445_c0_g1 GR28a3 118 internal 2 XP_021922466.1, putative gustatory receptor 28a [Zootermopsis nevadensis] 71% 5E-19 47.62 TRINITY_DN5103_c0_g1 GR28b1 394 complete 7 XP_020706207.2, putative gustatory receptor 28b [Athalia rosae] 99% 9E-177 61.36 TRINITY_DN2953_c0_g1 GR28b2 197 complete 4 XP_020706207.2, putative gustatory receptor 28b [Athalia rosae] 99% 3E-78 58.37 TRINITY_DN19790_c0_g1 GR28b3 99 5prime_partial 0 XP_020706667.2, putative gustatory receptor 28b [Athalia rosae] 87% 3E-29 68.18 TRINITY_DN21105_c0_g1 GR28b4 62 5prime_partial 1 XP_048510231.1, putative gustatory receptor 28b [Athalia rosae] 79% 3E-09 58 TRINITY_DN20930_c0_g1 GR28b5 74 internal 0 XP_020706667.2, putative gustatory receptor 28b [Athalia rosae] 81% 2E-19 63.33 TRINITY_DN20385_c0_g1 GR43a 276 5prime_partial 3 XP_046738260.1, gustatory receptor for sugar taste 43a-like isoform X3 [Diprion similis] 98% 7E-180 90.04 TRINITY_DN1720_c0_g1 GR5a 492 complete 7 XP_046409931.1, gustatory receptor 5a for trehalose-like isoform X2 [Neodiprion fabricii] 100% 0 67.21 TRINITY_DN10840_c0_g1 GR68a1 179 5prime_partial 3 XP_046490075.2, gustatory receptor 68a-like [Neodiprion pinetum] 93% 6.00E-25 34.71 TRINITY_DN14093_c0_g1 GR68a2 191 5prime_partial 3 XP_046490075.2, gustatory receptor 68a-like [Zootermopsis nevadensis] 90% 1E-05 25.67 TRINITY_DN7972_c0_g1 GR2 68 internal 1 ALG36126.1, gustatory receptor 2 [Sclerodermus sp. MQW-2015] 94% 3E-04 35.06 TRINITY_DN12794_c0_g3 GR68a3 87 5prime_partial 1 XP_059490989.1, gustatory receptor 68a-like [Neocloeon triangulifer] 93% 2.00E-05 35.37 Note: †ORF: open reading frame;ND: not detected. Amino acid sequences of R. cyatheae OBP genes are listed in the supplementary data. Candidate GRs Fifteen putative RcyaGRs encoding proteins ranging from 68 to 492 amino acid residues were identified in R. cyatheae . These candidate genes exhibited 21.70% to 90.04% sequence similarity to known insect GRs. Among them, four RcyaGRs were full-length, while the others lacked either one or both terminal regions. The predicted number of transmembrane domains (TMDs) ranged from 0 to 7 (Table 9 .in Supplementary Materials). Phylogenetic analysis revealed that RcyaGR43a and RcyaGR5a clustered within the fructose receptor subfamily and trehalose receptor subfamily, thus these two genes were tentatively predicted to function as sweet GRs (sugar receptors)18. Additionally, twelve RcyaGRs were distributed across different clades with GRs from ten other species, specifically within the GR28, GR62a, GR68f, and GR43a gene family clades. These clades were grouped on the basis of the GR database sequences employed in this study (Fig. 12 ) Table 9 Bioinformatics analysis of identified R. cyatheae IR genes gene ID Gene Name Complete ORF (aa) Group TMD BLAST Annotation Description Query E value Identify(%) TRINITY_DN11180_c0_g1 RcyaiGluR1 574 internal 2 XP_048513708.1, glutamate receptor ionotropic, kainate 2 isoform X3 [Athalia rosae] 100% 0 98.95 TRINITY_DN1947_c0_g1 RcyaIR93a 634 complete 3 XP_020712212.2, ionotropic receptor 93a [Athalia rosae] 100% 0 76.37 TRINITY_DN2322_c0_g1 RcyaiGluR2 193 5prime_partial 1 XP_012251025.2, glutamate receptor ionotropic, kainate 2-like isoform X2 [Athalia rosae] 99% 4E-88 70.31 TRINITY_DN3458_c0_g1 RcyaiGluR3 974 complete 3 XP_020712340.2, glutamate receptor ionotropic, kainate 2-like isoform X2 [Athalia rosae] 100% 0 94.15 TRINITY_DN3557_c0_g1 RcyaIR25a1 930 complete 3 XP_046429951.1, ionotropic receptor 25a [Neodiprion fabricii] 96% 0 85.75 TRINITY_DN3557_c0_g2 RcyaIR25a2 372 5prime_partial 3 XP_046429951.1, ionotropic receptor 25a [Diprion similis] 99% 0 94.85 TRINITY_DN491_c6_g1 RcyaiGluR4 577 complete 3 XP_012265004.2, glutamate receptor ionotropic, delta-1 isoform X1 [Athalia rosae] 100% 0 68.28 TRINITY_DN6525_c0_g1 RcyaIR25a3 266 complete 0 XP_012263496.2, ionotropic receptor 25a [Athalia rosae] 96% 1E-141 78.12 TRINITY_DN6776_c0_g1 RcyaiGluR5 198 internal 1 XP_020711610.2, glutamate receptor ionotropic, kainate 2 isoform X9 [Athalia rosae] 100% 2E-137 100 TRINITY_DN745_c0_g1 RcyaiGluR6 300 complete 1 XP_048512589.1, glutamate receptor ionotropic, kainate 2-like [Athalia rosae] 100% 3E-145 69.97 TRINITY_DN7652_c0_g3 RcyaiGluR7 867 complete 3 XP_046435180.1, glutamate receptor ionotropic, kainate 2 isoform X1 [Neodiprion fabricii] 100% 0 98.85 TRINITY_DN8730_c0_g1 RcyaiGluR8 303 complete 2 XP_025602091.2, glutamate receptor ionotropic, kainate 2-like [Athalia rosae] 99% 9E-151 73.18 TRINITY_DN8827_c0_g2 RcyaIR2 955 complete 3 XP_048507714.1, glutamate receptor ionotropic, NMDA 2B isoform X2 [Athalia rosae] 100% 0 97.07 TRINITY_DN8926_c0_g1 RcyaiGluR9 369 3prime_partial 0 XP_012251025.2, glutamate receptor ionotropic, kainate 2-like isoform X2 [Athalia rosae] 99% 2E-178 68.02 TRINITY_DN8926_c0_g2 RcyaiGluR10 180 internal 0 XP_048512589.1, glutamate receptor ionotropic, kainate 2-like [Athalia rosae] 100% 5E-76 66.67 TRINITY_DN12985_c0_g2 RcyaIR1 84 internal 0 RLZ02219.1, Ionotropic receptor 122 [Cephus cinctus] 73% 5E-10 45.16 TRINITY_DN5799_c0_g3 RcyaIR75a1 179 internal 1 XP_048506765.1ionotropic receptor 75a-like, partial [Athalia rosae] 100% 6E-72 59.78 TRINITY_DN5186_c0_g1 RcyaIR75a2 86 3prime_partial 1 XP_048511101.1, ionotropic receptor 75a-like isoform X3 [Athalia rosae] 81% 1E-19 55.71 TRINITY_DN5799_c0_g4 RcyaIR75a3 148 internal 0 XP_046410795.1, ionotropic receptor 75a-like isoform X1 [Neodiprion fabricii] 100% 2E-53 61.49 TRINITY_DN12543_c0_g1 RcyaIR21a 79 internal 0 XP_048508018.1, ionotropic receptor 21a isoform X1 [Athalia rosae] 99% 3E-38 79.75 Note: †ORF: open reading frame; ND: not detected. Amino acid sequences of R. cyatheae OBP genes are listed in the supplementary data. Candidate SNMPs Four putative RcyaSNMPs encoding proteins ranging from 60 to 530 amino acids were identified in R. cyatheae . These genes exhibited 54.97% to 70.08% similarity to their orthologs in other hymenopteran species. Phylogenetic analysis revealed that two RcyaSNMPs belong to the SNMP1 family, whereas the other two cluster within the SNMP2 family (S10). Phylogenetic analysis revealed that two RcyaSNMP genes clustered within the SNMP1 family, whereas the other two grouped into the SNMP2 family (Table 10 .in the Supplementary Materials). Specifically, RcyaSNMP1a was predicted to contain two transmembrane domains (TMDs), whereas RcyaSNMP2b and RcyaSNMP1 each harbored one TMD, and RcyaSNMP2a lacked detectable TMDs (Table 10 )1. Additionally, a separate phylogenetic tree constructed using SNMPs from R. cyatheae and 14 other Hymenopteran species, together with 10 CD36 homologs, revealed that all RcyaSNMP orthologs fell exclusively within the SNMP family clade and were distinct from the CD36 protein family (Fig. 13 ) Table 10 Bioinformatics analysis of identified R. cyatheae SNMP genes Gene ID Gene Name Complete ORF (aa) Group TMD BLAST Annotation Description Query E value Identify(%) TRINITY_DN1927_c0_g3 RcyaSNMP2a 60 5prime_partial 0 XP_015517411.2, sensory neuron membrane protein 2 isoform X1 [Neodiprion lecontei] 97% 0 55.89 TRINITY_DN7126_c0_g1 RcyaSNMP2b 434 complete 1 XP_046587242.1, sensory neuron membrane protein 2 isoform X2 [Neodiprion lecontei] 98% 3E-177 57.48 TRINITY_DN5757_c0_g1 RcyaSNMP1a 530 complete 2 XP_012257194.1, sensory neuron membrane protein 1 isoform X1 [Athalia rosae] 99% 0 71.08 TRINITY_DN11233_c0_g1 RcyaSNMP1b 151 5prime_partial 1 XP_020708289.1, sensory neuron membrane protein 1 isoform X2 [Athalia rosae] 98% 5E-49 54.97 Note: †ORF: open reading frame; ND: not detected. Amino acid sequences of R. cyatheae OBP genes are listed in the supplementary data. 3.5 Analysis of Candidate Differentially Expressed Genes In this study, the expression levels of sex-biased differentially expressed genes in R. cyatheae were evaluated on the basis of their FPKM values, as shown in the heatmap (Fig. 6 . In the Supplementary Materials). Among these chemosensory genes, RcyaOBP7, RcyaOBP8, RcyaNPC2d, RcyaNPC2e, RcyaSNMP2a, and RcyaOR19 were expressed at significantly higher levels in male adults than in female adults, whereas the remaining genes were expressed at higher levels in female adults. 3.6 Profiling of Chemosensory Genes in R. cyatheae The tissue-specific expression profiles of 13 newly identified candidate chemosensory genes (2 RcyaCSPs, 4 RcyaORs, 2 RcyaIRs, 1 RcyaGR, 1 RcyaSNMP, and 2 RcyaNPC2) in R. cyatheae were investigated via RT‒qPCR (Fig. 14 ). (Fant and Mant: female and male antennae; Fhea 和 Mhea: female and male heads without an-tennae; Fab: female abdomens without ovipositors and digestive tracts, Fov: female ovipositors; Mge: male genitalia; Fabd and Mabd: male abdomens; Fwin and Mwin : male and female wings; Ftho and Mtho: male and female thorax;Fleg and Mleg: male and female legs).The error bars represents standard errors and the small letters above each bar indicate significant differences in transcript abundances (p < 0.05). 4. Discussion With the development of transcriptome and genome research technologies, it has become increasingly important to construct insect transcriptome database-based identification methods for chemosensory genes to facilitate the study of chemosensory mechanisms [ 29 ].To date, the olfactory mechanisms mediating host location and oviposition site selection in R. cyatheae remain unknown. Therefore, in this study identified 90 candidate genes encoding olfactory-related proteins from the adult transcriptome of R. cyatheae , including 11 OBPs, 10 CSPs, 24 ORs, 15 GRs, 20 IRs, 4 SNMPs, and 6 NPC2s. The expression profiles of 13 differentially expressed chemosensory genes were subsequently validated across different tissues using RT‒qPCR, providing insights for exploring the functions of these genes.It provides a molecular basis for the systematic study of the chemosensory mechanism in R cyatheae . OBPs are hydrophobic binding proteins that can bind to and transport odorant substances (including pheromones and plant volatiles) through the lymph of sensilla [ 30 ].Within hymenopteran insects, some species possess only a few odorant-binding proteins (OBPs). For instance, Scleroderma guani has as few as two OBPs [ 31 ];there are 4 OBPs in Macrocentrus cingulum [ 32 – 34 ].Eleven OBP genes were identified in the transcriptome of R. cyatheae, a relatively low number comparable to that observed in IkuwOBP [ 35 ].This number is significantly lower than the number identified in N. vitripennis (90 OBPs). The differences in the number of OBPs among insects may be influenced by variations in different species, sexes, habitats, types of odor molecules, and feeding ranges [ 36 ].It may also result from the limitations of transcriptome samples and differences in sequencing methods. For example, samples based solely on the adult antenna transcriptome may exclude some OBP genes expressed in other tissues or at different developmental stages [ 2 , 37 ].Phylogenetic analysis revealed that the 11 RcyaOBPs were distributed across different clades alongside OBPs from nine hymenopteran species, suggesting potential functional divergence among RcyaOBPs. Specifically, RcyaOBP7 clustered within the same clade as IkuwOBP and AmelOBP, whereas RcyaOBP8 formed a clade with CcunOBP and CvesOBP. The expression levels of these two differentially expressed OBP genes were further quantified via RT‒qPCR across various tissues. The highest expression of RcyaOBP7 was detected in female antennae. Numerous studies have demonstrated that OBPs with antenna-specific and highly abundant expression play critical roles in binding key volatile compounds during host location and foraging processes in hymenopteran insects[ 38 ].For example when the AmelOBP4 gene is knocked out, the preference for plant volatiles (such as linalool and 1-octen-3-ol) and sex pheromones (ethyl oleate, methyl palmitate, methyl linoleate, and β-ocimene) is significantly reduced [ 39 ]. The expression of RcyaOBP8 was found to be significantly greater in male heads than in other tissues, whereas its expression was markedly reduced in female heads compared with other tissues. These findings suggested that RcyaOBP8 might function in pheromone recognition or male-specific behaviors. For instance, SfruOBP31 is highly expressed in the abdomen, adult heads, and male reproductive organs of Spodoptera frugiperda . Knockout of the SfruOBP31 gene was demonstrated to disrupt larval phototaxis and male reproductive processes[ 40 , 41 ].The functions of these OBP genes merit further study. CSPs are soluble proteins that perform functions similar to those of odorant-binding proteins (OBPs). As the second category of binding proteins in the insect chemosensory system, they are more conserved than OBPs [ 42 ],(including heads, thoraxes, labial palps, tarsi, pheromone glands, and ejaculatory ducts)These proteins are widely distributed across both olfactory tissues (such as antennae, legs, wings, and proboscis) and non-olfactory tissues (including heads, thoraxes, labial palps, tarsi, pheromone glands, and ejaculatory ducts) [ 38 , 39 ]. For example, after silencing the NlugCSP8 gene in olfactory tissues, the response of Nilaparvata lugens to hexanal is significantly inhibited, and the attractiveness of nerol to N. lugen s is lost [ 43 ].In addition to their olfactory functions, CSPs may also be involved in insect development, nutrient absorption, and insecticide resistance [ 8 ].For instance, knockout of the ovary- specific gene OcomCSP12 leads to a significant reduction in ovarian specificity [ 44 ]; Additionally ,silencing of DlonCSP3 impairs the foraging behavior of Diachasmimorpha longicaudata [ 45 ].Ten CSP genes were identified in the adult transcriptome of R. cyatheae , with numbers is similar to those of BdioCSPs[ 46 ]、SspCSPs[ 47 ]、EforCSPs[ 48 ] among others.According to the RT‒qPCR results, we observed that RcyaCSP10 exhibitted female antennae-biased expression, whereas RcyaCSP2 demonstrates distinct tissue specificity with notably low expression in female heads. The detailed functions of these two RcyaCSPs in R. cyatheae require further investigation. NPC2 is considered a soluble small-molecule protein and a member of the MD-2-related lipid recognition (ML) protein superfamily, which includes myeloid differentiation factor-2 (MD-2) [ 45 ].It serves as a carrier for transporting semiochemicals and other hydrophobic compounds, such as cholesterol[ 49 , 50 ].In the NPC2 gene family, 6 NPC2 genes were identified, making up a larger number than that in B.dioryctriae (3 NPC2 genes) [ 46 ] and in D. longicaudata (4 NPC2 genes) [ 50 ].Previous studies on hymenopteran insects have demonstrated that NPC2 is highly expressed in both antennae and ovipositors [ 51 ] .In this study, RcyaNPC2d and RcyaNPC2e were expressed in antennae, head, legs, thorax, abdomen, and wings, but their expression levels were significantly higher in male heads than in female heads. This phenomenon might be attributed to the functional divergence of NPC2 proteins.RcyaNPC2d and RcyaNPC2e, which exhibit this distinct expression pattern, could serve as target genes to investigate their physiological functions in R. cyatheae . Insect ORs genes serve as crucial chemoreceptor organs within insect olfactory systems, evolutionarily derived from GRs [ 52 ]. These receptors are sensitive to many different types of compounds, including insect pheromone components and host plant compounds, such as esters, alcohols, and ketones[ 53 ]. Within the RcyaOR gene family, 23 OR genes and one Orco gene were identified, representing a relatively small number. In contrast, some Hymenopteran insects possess more than one hundred OR genes; for instance, Apis cerana has 113 OR genes. [ 54 ] and Apis florea has 180 OR genes [ 55 ].The variation in OR gene numbers may be influenced by factors such as interspecies variation, as well as the depth and breadth of sequencing methodologies [ 56 ].Within hymenopteran insect species, a single Orco gene is present, which functions as a highly conserved coreceptor [ 57 ].In many insect species, the absence of the Orco gene leads to reduced sex pheromone recognition, decreased oviposition preference, and impaired larval attraction to host plants [ 58 , 59 ].According to the RT‒qPCR results, OR14, OR17, OR19, and OR20 were expressed across all 12 examined tissues, with the lowest expression levels observed in female heads. This pattern may reflect functional differentiation among OR genes. Compared with those in Lepidoptera and Coleoptera, numerous OR genes have been identified in hymenopteran species in recent years; however, the functional characterization of these genes remains insufficient despite their quantitative abundance.Screening for specifically expressed ORs may be an effective method for narrowing the range of host localized target genes. Four RcyaOR genes with sex-biased expression in males and females were selected for subsequent studies. IRs are receptor genes that are evolutionarily derived from ionotropic glutamate receptors (iGluRs). They are expressed in gustatory receptor neurons, respond to diverse odorants, and represent a highly conserved family of ligand-gated ion channels [ 10 , 60 ].In IR gene family, 20 IR genes were identified, representing a number similar to those in Aphidius gifuensis (25 IRs)[ 29 ] and Bombus impatiens (23个IR)[ 61 ].In this study, ten iGluRs were identified. Given that iGluRs are ancestral precursors of IRs, the limited evolutionary expansion of iGluRs into IRs in R. cyatheae may be associated with its specialized feeding habits.iGluR4 was expressed in all 12 tissues but showed significant sex-specific expression differences in the head. iGluR4 may play a role in odor recognition and localization[ 62 ], but further experimental validation.Furthermore, as an ancient chemoreceptor family, insect IRs can be classified into several distinct subfamilies. Phylogenetic analysis revealed that the 20 RcyaIRs identified in R. cyatheae are distributed among typical subfamilies, including iGluR, IR25a, IR64a, IR75a, IR76b, and IR93a. Notably, three IR25a orthologs were identified in R. cyatheae . and three IR25a homologs were identified in R. cyatheae . However, compared with IR25a1 and IR25a2,IR25a3 clustered in a distinct phylogenetic clade, suggesting functional divergence of the IR25a gene. The coreceptor family of IR25a shares similar distribution patterns and functional profiles with the Orco family, although they are expressed in distinct types of sensilla[ 52 ].The IR25a gene mediates cold temperature and humidity changes in Drosophila melanogaster [ 63 , 64 ]. R. cyatheae survives prolonged winter months, suggesting that the RcyaIR25 gene may also be involved in thermosensation.The Ir75a gene arose through duplication in the ancestor of the family Drosophilidae (~ 30–35 million years ago) and exhibits differential sensitivity to carboxylic acids [ 65 , 66 ].Furthermore, the primary host plant of R. cyatheae is A. spinulosa , which releases volatile compounds such as acetic acid and ethyl acetate into the environment [ 67 ] .These odorants may be detected by Ir75a in R. cyatheae , thus, RcyIr75a likely participates in host localization in this species, but further experimental validation is needed. Insects rely on a unique family of GRs, which function as tetrameric ligand-gated cation channels [ 68 ].These receptors primarily mediate the perception of carbon dioxide, fructose, various sugars, bitter compounds, and other chemosensory stimuli [ 69 ].Fifteen GR genes were identified in the RcyaGR gene family of R. cyatheae , which is close to Atherigona orientalis (20 GRs) [ 70 ].In hymenopteran insects, the number of GR genes identified via transcriptomic techniques ranges from as few as 2 ( M. mediator ) [ 71 ], to 49 ( B.dioryctriae )[ 46 ].Three RcyaIR genes were identified in the transcriptome of R. cyatheae and predicted to function as sweet gustatory receptors (GRs).In D melanogaster , GR43a is crucial for detecting fructose and sucrose receptor crucial for detecting fructose levels in the brain. It is also expressed in neurons of the proventricular ganglion and uterus. Four BidoGR43a.1–4 genes and one BidoGR64f gene cluster are located within the fructose receptor and trehalose receptor subfamilies, respectively [ 46 ].In D.melanogaster , GR43a is crucial for detecting fructose levels in the brain. It is also expressed in neurons of the proventricular ganglion and uterus. Four BidoGR43a.1–4 genes and one BidoGR64f gene cluster within the fructose receptor and trehalose receptor subfamilies, respectively.Additionally, DmelGR5a encodes as a receptor for trehalose and clusters within the trehalose receptor subfamily alongside :DmelGR64f [ 72 , 73 ].Phylogenetic analysis revealed that RcyaGR5a clustered with RcyaGR64f and BidoGR64f in the same clade, indicating that RcyaGR5a and RcyaGR64f belonged to to the trehalose receptor subfamily. The GR68a protein is likely involved in pheromone-driven courtship behavior in Drosophila[ 74 ].In this study, RcyaGR68a clustered within a branch containing numerous GRs, suggesting potential functional divergence. No candidate GRs from the CO₂ receptor subfamily were identified in R. cyatheae , possibly because of the lack of a dedicated transcriptome database for the labial palps of the sawfly [ 75 , 76 ].These RcyaGR receptors in R. cyatheae may perceive sweet and bitter sensations, warranting further investigation. SNMPs are transmembrane structural proteins that were initially identified as membrane-bound proteins in the olfactory receptor neurons of lepidopteran insects. They are proposed to play a role in odor detection[ 77 , 78 ].Four SNMP genes were identified in R.cyatheae , which is more than in M.mediator (2) and Sclerodermus sp. (2) [ 47 , 79 ].Numerous studies have demonstrated that most insects possess two types of SNMPs, namely SNMP1 and SNMP2[ 80 ].The RcyaSNMPs identified in this study belong to both the SNMP1 and SNMP2 subfamilies.The paralogs of the same SNMP type in R. cyatheae are similar to those in B.dioryctriae and Iseropus kuwanae , with the former encoding six SNMP1 genes and the latter encoding two SNMP genes[ 35 ].In D.melanogaster , SNMP1 has been demonstrated to be critical for detecting the pheromone cis-vaccenyl acetate (CVA) [ 81 ].MmedSNMP1 is expressed primarily in the sensilla of the antennae and may be involved in the perception of plant volatiles and sex pheromones[ 75 ].Therefore, phylogenetic analysis revealed that MmedSNMP1 clustered with RcyaSNMP1a and RcyaSNMP1b in the same clade, suggesting that RcyaSNMP1a and RcyaSNMP1b might participate in the perception of plant volatiles and sex pheromones. However, functional validation is needed to confirm this hypothesis. The current t literature on SNMP2 remains limited, with no functional studies reported to date. In this study, RcyaSNMP2a was expressed across all the examined tissues[ 82 ], albeit at minimal levels. 5. Conclusions On the basis of the transcriptome database of female and male adults of R. cyatheae , this study identified 90 candidate chemosensory genes were identified in this study. RT‒qPCR revealed distinct expression patterns of 13 chemosensory genes. This work preliminarily narrows down the range of candidate chemosensory genes involved in recognizing key information during host localization and oviposition processes, laying a foundation for further exploration of the molecular mechanisms underlying chemosensory perception in R. cyatheae and the development of novel biological control strategies. Declarations Ethics approval and consent to participate:not applicable. Consent for publication: not applicable Funding The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was supported by “Pest resistance mechanisms of different Cyathea spinosa based on three-generation full-length transcriptomes, two-generation transcriptomes, and protein metabolomes” (Grant No. 11904–0522093),and the Science and Technology Innovation Talent Team Building Project of the Science and Technology Innovation Talent Team Building Project of Guizhou Province (Project No.: Qiankehepingtairencai-CXTD [2023]010). CRediT authorship contribution statement Xiao-Ming Li and Ya-Nan Zhang designed the experiments. Qiang Liu, Sai Ma, Mao-Zhu Yin, Nan Gu, and Li-Fu Qian performed the experiments. Xiao-Ming Li, Qiang Liu, and Ya-Nan Zhang analysed the data. Xiao-Ming Li, Qiang Liu, and Ya-Nan Zhang wrote and revised the article. Acknowledgement First and foremost, I sincerely thank my supervisor, Professor Yang Weicheng. Throughout this study, Professor Yang not only provided guidance on the development of the research design and experimental protocol but also assisted in revising the statistical methods during the data analysis phase. Additionally, he reviewed the initial draft of the paper on multiple occasions and offered important revision suggestions, which further refined the research framework.Secondly, I would like to express my gratitude to the Administration of Guizhou Chishui Alsophila National Nature Reserve for its strong support for this experiment.Furthermore, I appreciate the technical support provided by all teachers in the research group during the experiment, as well as the assistance from my fellow group members in sample processing.This study was funded by Pest resistance mechanisms of different Cyathea spinosa based on three-generation full-length transcriptomes, two-generation transcriptomes, and protein metabolomes (Project No.: 11904–0522093) and the Science and Technology Innovation Talent Team Building Project of the Science and Technology Innovation Talent Team Building Project of Guizhou Province (Project No.: Qiankehepingtairencai-CXTD [2023]010), and then I would like to extend my sincere thanks for this financial support.Finally, I am grateful to my family for their understanding and support throughout the research process. Disclosure The authors declare no conflict of interest. Availability of data and materials All data generated or analyzed during this study are included in this published article (and its supplementary information files). The raw sequence dataset is available at the National Center for Biotechnology Information (NCBI) under the SRA Bioproject number PRJNA1309270. References Tian Z, Sun L, Li Y, Quan L, Zhang H, Yan W, et al. Antennal transcriptome analysis of the chemosensory gene families in carposina sasakii (lepidoptera: Carposinidae). BMC Genomics. 2018;19:544. https://doi.org/10.1186/s12864-018-4900-x . Ma X, Lu X, Zhang P, Deng X, Bai J, Xu Z, et al. Transcriptome analysis of antennal chemosensory genes in curculio dieckmanni faust. (coleoptera: Curculionidae). Front Physiol. 2022;13. https://doi.org/10.3389/fphys.2022.896793 . Vosshall LB, Amrein H, Morozov PS, Rzhetsky A, Axel R. A spatial map of olfactory receptor expression in the drosophila antenna. Cell. 1999;96:725–36. https://doi.org/10.1016/s0092-8674(00)80582-6 . Dippel S, Oberhofer G, Kahnt J, Gerischer L, Opitz L, Schachtner J, et al. Tissue-specific transcriptomics, chromosomal localization, and phylogeny of chemosensory and odorant binding proteins from the red flour beetle tribolium castaneum reveal subgroup specificities for olfaction or more general functions. BMC Genomics. 2014;15:1141. https://doi.org/10.1186/1471-2164-15-1141 . Guo S, Liu P, Tang Y, Chen J, Zhang T, Liu H. Identification and expression profiles of olfactory-related genes in the antennal transcriptome of graphosoma rubrolineatum (hemiptera: Pentatomidae). PLoS ONE. 2024;19:e0306986. https://doi.org/10.1371/journal.pone.0306986 . Zhou H, Yan H, Wang E, Zhang B, Xu X. Expression and functional analysis of niemann-pick C2 gene in phytoseiulus persimilis. Exp Appl Acarol. 2023;89:201–13. https://doi.org/10.1007/s10493-023-00781-8 . Ko DC, Binkley J, Sidow A, Scott MP. The integrity of a cholesterol-binding pocket in niemann-pick C2 protein is necessary to control lysosome cholesterol levels. Proc Natl Acad Sci U S A. 2003;100:2518–25. https://doi.org/10.1073/pnas.0530027100 . Pelosi P, Iovinella I, Zhu J, Wang G, Dani FR. Beyond chemoreception: Diverse tasks of soluble olfactory proteins in insects. Biol Rev. 2018;93:184–200. https://doi.org/10.1111/brv.12339 . Wang JJ, Ma C, Yue Y, Yang J, Chen LX, Wang YT, et al. Identification of candidate chemosensory genes in bactrocera cucurbitae based on antennal transcriptome analysis. Front Physiol. 2024;15. https://doi.org/10.3389/fphys.2024.1354530 . Rimal S, Lee Y. The multidimensional ionotropic receptors of drosophila melanogaster. Insect Mol Biol. 2018;27:1–7. https://doi.org/10.1111/imb.12347 . Kozma MT, Schmidt M, Ngo-Vu H, Sparks SD, Senatore A, Derby CD. Chemoreceptor proteins in the caribbean spiny lobster, panulirus argus: Expression of ionotropic receptors, gustatory receptors, and TRP channels in two chemosensory organs and brain. PLoS ONE. 2018;13:e0203935. https://doi.org/10.1371/journal.pone.0203935 . Agnihotri AR, Roy AA, Joshi RS. Gustatory receptors in lepidoptera: Chemosensation and beyond. Insect Mol Biol. 2016;25:519–29. https://doi.org/10.1111/imb.12246 . Mang D, Shu M, Endo H, Yoshizawa Y, Nagata S, Kikuta S, et al. Expression of a sugar clade gustatory receptor, BmGr6, in the oral sensory organs, midgut, and central nervous system of larvae of the silkworm bombyx mori. Insect Biochem Mol Biol. 2016;70:85–98. https://doi.org/10.1016/j.ibmb.2015.12.008 . Kang Z-W, Tian H-G, Liu F-H, Liu X, Jing X-F, Liu T-X. Identification and expression analysis of chemosensory receptor genes in an aphid endoparasitoid aphidius gifuensis. Sci Rep. 2017;7:3939. https://doi.org/10.1038/s41598-017-03988-z . Zhang B-C, Yang W-C, Xiao J-X, Bai X-J, Chen H-D. Effects of different host plants on the growth,development and gut bacterial communi-ty of Rhoptroceros cyatheae larvae [J]. J Environ Entomol. 2024;46:886–96. Xu D-S, Yang W-C,Wong T, He Q-Q,Liang S, Zhang T-Y. Biological Characteristics and Larval Population Dynamics of Rhoptroceros cyatheae in Chishui Guizhou[J]. Chin J Biol Control. 2021;37(2):218–27. https://doi.org/10.16409/j.cnki.2095-039x.2020.06.001 . Yang J, Yang W-C,Wu G-Y, Che B-J, Liang H-F, Zhou B-B. Optimizationof Tissue Culture and Propagation System of Alsophila spinulosa Based on PGGB Path way[J]. Acta Bot Boreali-Occidentalia Sinica. 2023;43:1488–98. Liu Y-L, Zhang L-N, Liang L, Zheng Y-Q, et al. Diversity of endophytic fungi from Alsophila spinulosa in ChishuiAlsophila National Nature Reserve, Guizhou Province, Southwest China. Mycosystema. 2021;40:2673–84. https://doi.org/10.13346/j.mycosystema.210177 . Zhang B, Yang W-C, He Q-Q, Chen H-D, Che B-J, Bai X-J. Analysis of differential effects of host plants on the gut microbes of rhoptroceros cyatheae. Front Microbiol. 2024;15:1392586. https://doi.org/10.3389/fmicb.2024.1392586 . Wang X, Ren Y-Z, Huang Q,Deng X-B, Chen C-W. Deng H.-P.Habitat suitability assessment of endangered plant Alsophila spi-nulosa in Chishui River area based on GIS and Maxent model. Acta Ecol Sin. 2021;41:6123–33. Chen S, Zhou Y, Chen Y, Gu J. fastp: An ultra-fast all-in-one FASTQ preprocessor. Bioinforma Oxf Engl. 2018;34:i884–90. https://doi.org/10.1093/bioinformatics/bty560 . Chen S. Ultrafast one-pass FASTQ data preprocessing, quality control, and deduplication using fastp. iMeta. 2023;2:e107. https://doi.org/10.1002/imt2.107 . Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, et al. Full-length transcriptome assembly from RNA-seq data without a reference genome. Nat Biotechnol. 2011;29:644–52. https://doi.org/10.1038/nbt.1883 . Kumar S, Stecher G, Suleski M, Sanderford M, Sharma S, Tamura K. MEGA12: Molecular evolutionary genetic analysis version 12 for adaptive and green computing. Mol Biol Evol. 2024;41:msae263. https://doi.org/10.1093/molbev/msae263 . Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol. 2018;35:1547–9. https://doi.org/10.1093/molbev/msy096 . Liu F, Ye Z, Baker A, Sun H, Zwiebel LJ. Gene editing reveals obligate and modulatory components of the CO2 receptor complex in the malaria vector mosquito, anopheles coluzzii. Insect Biochem Mol Biol. 2020;127:103470. https://doi.org/10.1016/j.ibmb.2020.103470 . Chowdhury HA, Bhattacharyya DK, Kalita JK. Differential expression analysis of RNA-seq reads: Overview, taxonomy, and tools. IEEE/ACM Trans Comput Biol Bioinform. 2020;17:566–86. https://doi.org/10.1109/TCBB.2018.2873010 . Vaes E, Khan M, Mombaerts P. Statistical analysis of differential gene expression relative to a fold change threshold on NanoString data of mouse odorant receptor genes. BMC Bioinformatics. 2014;15:39. https://doi.org/10.1186/1471-2105-15-39 . Kang Z-W, Tian H-G, Liu F-H, Liu X, Jing X-F, Liu T-X. Identification and expression analysis of chemosensory receptor genes in an aphid endoparasitoid aphidius gifuensis. Sci Rep. 2017;7:3939. https://doi.org/10.1038/s41598-017-03988-z . Hu P, Tao J, Cui M, Gao C, Lu P, Luo Y. Antennal transcriptome analysis and expression profiles of odorant binding proteins in eogystia hippophaecolus (lepidoptera: Cossidae). BMC Genomics. 2016;17:651. https://doi.org/10.1186/s12864-016-3008-4 . Lu D, Li X, Liu X, Zhang Q. Identification and molecular cloning of putative odorant-binding proteins and chemosensory protein from the bethylid wasp, scleroderma guani xiao et wu. J Chem Ecol. 2007;33:1359–75. https://doi.org/10.1007/s10886-007-9310-5 . Ahmed T, Zhang T, Wang Z, He K, Bai S. C-terminus methionene specifically involved in binding corn odorants to odorant binding Protein4 in macrocentrus cingulum. Front Physiol. 2017;8:62. https://doi.org/10.3389/fphys.2017.00062 . Ahmed T, Zhang T, Wang Z, He K, Bai S. Three amino acid residues bind corn odorants to McinOBP1 in the polyembryonic endoparasitoid of macrocentrus cingulum brischke. PLoS ONE. 2014;9:e93501. https://doi.org/10.1371/journal.pone.0093501 . Ahmed T, Zhang T, Wang Z, He K, Bai S. Molecular cloning, expression profile, odorant affinity, and stability of two odorant-binding proteins in macrocentrus cingulum brischke (hymenoptera: Braconidae). Arch Insect Biochem Physiol. 2017;94:e21374. https://doi.org/10.1002/arch.21374 . Li Y, Chen H, Liang X, Wang S, Zhu H, Yan M, et al. Identification of candidate chemosensory genes by antennal transcriptome analysis in an ectoparasitoid wasp. J Appl Entomol. 2022;146:335–51. https://doi.org/10.1111/jen.12962 . Vieira FG, Forêt S, He X, Rozas J, Field LM, Zhou J-J. Unique features of odorant-binding proteins of the parasitoid wasp nasonia vitripennis revealed by genome annotation and comparative analyses. PLoS ONE. 2012;7:e43034. https://doi.org/10.1371/journal.pone.0043034 . Wu Z-R, Fan J-T, Tong N, Guo J-M, Li Y, Lu M, et al. Transcriptome analysis and identification of chemosensory genes in the larvae of plagiodera versicolora. BMC Genomics. 2022;23:845. https://doi.org/10.1186/s12864-022-09079-2 . Li F, Venthur H, Lin K, Zhang C, Chen Z, Zhou J-J. Insect chemosensory proteins as targets in insecticide resistance and development. New Plant Prot. 2025;2:e70008. https://doi.org/10.1002/npp2.70008 . Liu N-Y, Li Z-B, Zhao N, Song Q-S, Zhu J-Y, Yang B. Identification and characterization of chemosensory gene families in the bark beetle, tomicus yunnanensis. Comp Biochem Physiol Part D Genomics Proteom. 2018;25:73–85. https://doi.org/10.1016/j.cbd.2017.11.003 . Han W-K, Tang F-X, Yan Y-Y, Wang Y, Zhang Y-X, Yu N, et al. An OBP gene highly expressed in non-chemosensory tissues affects the phototaxis and reproduction of spodoptera frugiperda. Insect Mol Biol. 2024;33:81–90. https://doi.org/10.1111/imb.12880 . Jia C, Mohamed A, Cattaneo AM, Huang X, Keyhani NO, Gu M, et al. Odorant-binding proteins and chemosensory proteins in spodoptera frugiperda: From genome-wide identification and developmental stage-related expression analysis to the perception of host plant odors, sex pheromones, and insecticides. Int J Mol Sci. 2023;24:5595. https://doi.org/10.3390/ijms24065595 . Zhu J, Iovinella I, Dani FR, Pelosi P, Wang G. Chemosensory proteins: a versatile binding family. Olfactory Concepts of Insect Control - Alternative to Insecticides. Cham: Springer; 2019. pp. 147–69. https://doi.org/10.1007/978-3-030-05165-5_6 . Waris MI, Younas A, Ul Qamar MT, Hao L, Ameen A, Ali S, et al. Silencing of chemosensory protein gene NlugCSP8 by RNAi induces declining behavioral responses of nilaparvata lugens. Front Physiol. 2018;9:379. https://doi.org/10.3389/fphys.2018.00379 . Ma C, Cui S, Tian Z, Zhang Y, Chen G, Gao X, et al. OcomCSP12, a chemosensory protein expressed specifically by ovary, mediates reproduction in ophraella communa (coleoptera: Chrysomelidae). Front Physiol. 2019;10:1290. https://doi.org/10.3389/fphys.2019.01290 . Wulff JP, Segura DF, Devescovi F, Muntaabski I, Milla FH, Scannapieco AC, et al. Identification and characterization of soluble binding proteins associated with host foraging in the parasitoid wasp diachasmimorpha longicaudata. PLoS ONE. 2021;16:e0252765. https://doi.org/10.1371/journal.pone.0252765 . Zhu X, Yu Q, Gan X, Song L, Zhang K, Zuo T, et al. Transcriptome analysis and identification of chemosensory genes in baryscapus dioryctriae (hymenoptera: Eulophidae). Insects. 2022;13:1098. https://doi.org/10.3390/insects13121098 . Zhou C-X, Min S-F, Yan-Long T, Wang M-Q. Analysis of antennal transcriptome and odorant binding protein expression profiles of the recently identified parasitoid wasp, sclerodermus sp. Comp Biochem Physiol Part D Genomics Proteom. 2015;16:10–9. https://doi.org/10.1016/j.cbd.2015.06.003 . He Y, Wang K, Zeng Y, Guo Z, Zhang Y, Wu Q, et al. Analysis of the antennal transcriptome and odorant-binding protein expression profiles of the parasitoid wasp encarsia formosa. Genomics. 2020;112:2291–301. https://doi.org/10.1016/j.ygeno.2019.12.025 . Zhu J, Guo M, Ban L, Song L-M, Liu Y, Pelosi P, et al. Niemann-pick C2 proteins: A new function for an old family. Front Physiol. 2018;9:52. https://doi.org/10.3389/fphys.2018.00052 . Thambi PJ, Modahl CM, Kini RM. Niemann-pick type C2 proteins in aedes aegypti: Molecular modelling and prediction of their structure-function relationships. Int J Mol Sci. 2024;25:1684. https://doi.org/10.3390/ijms25031684 . Zheng Y, Wang S-N, Peng Y, Lu Z-Y, Shan S, Yang Y-Q, et al. Functional characterization of a niemann-pick type C2 protein in the parasitoid wasp microplitis mediator. Insect Sci. 2018;25:765–77. https://doi.org/10.1111/1744-7917.12473 . Wicher D, Miazzi F. Functional properties of insect olfactory receptors: Ionotropic receptors and odorant receptors. Cell Tissue Res. 2021;383:7–19. https://doi.org/10.1007/s00441-020-03363-x . Wicher D. Chapter two - olfactory signaling in insects. In: Glatz R, editor. Progress in Molecular Biology and Translational Science. Academic; 2015. pp. 37–54. https://doi.org/10.1016/bs.pmbts.2014.11.002 . Park D, Jung JW, Choi B-S, Jayakodi M, Lee J, Lim J, et al. Uncovering the novel characteristics of asian honey bee, apis cerana, by whole genome sequencing. BMC Genomics. 2015;16:1. https://doi.org/10.1186/1471-2164-16-1 . Karpe SD, Jain R, Brockmann A, Sowdhamini R. Identification of complete repertoire of apis florea odorant receptors reveals complex orthologous relationships with apis mellifera. Genome Biol Evol. 2016;8:2879–95. https://doi.org/10.1093/gbe/evw202 . Shan S, Song X, Khashaveh A, Wang S-N, Lu Z-Y, Hussain Dhiloo K, et al. A female-biased odorant receptor tuned to the lepidopteran sex pheromone in parasitoid microplitis mediator guiding habitat of host insects. J Adv Res. 2022;43:1–12. https://doi.org/10.1016/j.jare.2022.03.006 . Butterwick JA, del Mármol J, Kim KH, Kahlson MA, Rogow JA, Walz T, et al. Cryo-EM structure of the insect olfactory receptor orco. Nature. 2018;560:447–52. https://doi.org/10.1038/s41586-018-0420-8 . Zhang Q, Chen J, Wang Y, Lu Y, Dong Z, Shi W, et al. The odorant receptor co-receptor gene contributes to mating and host-searching behaviors in parasitoid wasps. Pest Manag Sci. 2023;79:454–63. https://doi.org/10.1002/ps.7214 . Fan X-B, Mo B-T, Li G-C, Huang L-Q, Guo H, Gong X-L, et al. Mutagenesis of the odorant receptor co-receptor (orco) reveals severe olfactory defects in the crop pest moth helicoverpa armigera. BMC Biol. 2022;20:214. https://doi.org/10.1186/s12915-022-01411-2 . Koh T-W, He Z, Gorur-Shandilya S, Menuz K, Larter NK, Stewart S, et al. The drosophila IR20a clade of ionotropic receptors are candidate taste and pheromone receptors. Neuron. 2014;83:850–65. https://doi.org/10.1016/j.neuron.2014.07.012 . Fisher K, Guillén BM, Dahanukar A, Yamanaka N, Woodard SH. Expression analyses of chemosensory genes provide insights into evolution of gustatory receptor genes in the bumble bee bombus impatiens. BMC Genomics. 2025;26:575. https://doi.org/10.1186/s12864-025-11710-x . Abuin L, Bargeton B, Ulbrich MH, Isacoff EY, Kellenberger S, Benton R. Functional architecture of olfactory ionotropic glutamate receptors. Neuron. 2011;69:44–60. https://doi.org/10.1016/j.neuron.2010.11.042 . Ni L, Klein M, Svec KV, Budelli G, Chang EC, Ferrer AJ, et al. The ionotropic receptors IR21a and IR25a mediate cool sensing in drosophila. Elife. 2016;5:e13254. https://doi.org/10.7554/eLife.13254 . Enjin A, Zaharieva EE, Frank DD, Mansourian S, Suh GSB, Gallio M, et al. Humidity sensing in drosophila. Curr Biol CB. 2016;26:1352–8. https://doi.org/10.1016/j.cub.2016.03.049 . Prieto-Godino LL, Schmidt HR, Benton R. Molecular reconstruction of recurrent evolutionary switching in olfactory receptor specificity. Elife 10:e69732. https://doi.org/10.7554/eLife.69732 Prieto-Godino LL, Rytz R, Cruchet S, Bargeton B, Abuin L, Silbering AF, et al. Evolution of acid-sensing olfactory circuits in drosophilids. Neuron. 2017;93:661–e6766. https://doi.org/10.1016/j.neuron.2016.12.024 . Xiao J-X. 2024. Study on the host selection mechanism of Phthonoloba virifasciata and Rhoptroceros cyatheae[D]. Guizhou Nor-mal University; 2024. https://doi.org/10.27048/d.cnki.ggzsu.2023.000365 Frank HM, Walujkar S, Walsh RM, Laursen WJ, Theobald DL, Garrity PA, et al. Structural basis of ligand specificity and channel activation in an insect gustatory receptor. Cell Rep. 2024;43:114035. https://doi.org/10.1016/j.celrep.2024.114035 . Gomes JV, Singh-Bhagania S, Cenci M, Chacon Cordon C, Singh M, Butterwick JA. The molecular basis of sugar detection by an insect taste receptor. Nature. 2024;629:228–34. https://doi.org/10.1038/s41586-024-07255-w . Zhou Z, Luo Y, Wang X, He J, Zhou Q. Identification and sex expression profiles of candidate chemosensory genes from atherigona orientalis via the antennae and leg transcriptome analysis. Comp Biochem Physiol Part D Genomics Proteom. 2024;50:101222. https://doi.org/10.1016/j.cbd.2024.101222 . Wang S-N, Peng Y, Lu Z-Y, Dhiloo KH, Gu S-H, Li R-J, et al. Identification and expression analysis of putative chemosensory receptor genes in microplitis mediator by antennal transcriptome screening. Int J Biol Sci. 2015;11:737–51. https://doi.org/10.7150/ijbs.11786 . Chyb S, Dahanukar A, Wickens A, Carlson JR. Drosophila Gr5a encodes a taste receptor tuned to trehalose. Proc Natl Acad Sci U S A. 2003;100(2 Suppl 2):14526–30. https://doi.org/10.1073/pnas.2135339100 . Aryal B, Dhakal S, Shrestha B, Lee Y. Molecular and neuronal mechanisms for amino acid taste perception in the drosophila labellum. Curr Biol CB. 2022;32:1376–e13864. https://doi.org/10.1016/j.cub.2022.01.060 . Bray S, Amrein H. A putative drosophila pheromone receptor expressed in male-specific taste neurons is required for efficient courtship. Neuron. 2003;39:1019–29. https://doi.org/10.1016/s0896-6273(03)00542-7 . Guerenstein PG, Christensen TA, Hildebrand JG. Sensory processing of ambient CO2 information in the brain of the moth manduca sexta. J Comp Physiol Neuroethol Sens Neural Behav Physiol. 2004;190:707–25. https://doi.org/10.1007/s00359-004-0529-0 . Kent KS, Harrow ID, Quartararo P, Hildebrand JG. An accessory olfactory pathway in lepidoptera: The labial pit organ and its central projections in manduca sexta and certain other sphinx moths and silk moths. Cell Tissue Res. 1986;245:237–45. https://doi.org/10.1007/BF00213927 . Nichols Z, Vogt RG. The SNMP/CD36 gene family in diptera, hymenoptera and coleoptera: Drosophila melanogaster, D. pseudoobscura, anopheles gambiae, aedes aegypti, apis mellifera, and tribolium castaneum. Insect Biochem Mol Biol. 2008;38:398–415. https://doi.org/10.1016/j.ibmb.2007.11.003 . Rogers ME, Krieger J, Vogt RG. Antennal SNMPs (sensory neuron membrane proteins) of lepidoptera define a unique family of invertebrate CD36-like proteins. J Neurobiol. 2001;49:47–61. https://doi.org/10.1002/neu.1065 . Shan S, Wang S-N, Song X, Khashaveh A, Lu Z-Y, Dhiloo KH, et al. Molecular characterization and expression of sensory neuron membrane proteins in the parasitoid microplitis mediator (hymenoptera: Braconidae). Insect Sci. 2020;27:425–39. https://doi.org/10.1111/1744-7917.12667 . Li Y, Chen H, Hong T, Yan M, Wang J, Shao Z, et al. Identification of chemosensory genes by antennal transcriptome analysis and expression profiles of odorant-binding proteins in parasitoid wasp aulacocentrum confusum . Comp Biochem Physiol Part D Genomics Proteom. 2021;40:100881. https://doi.org/10.1016/j.cbd.2021.100881 . Vogt RG, Miller NE, Litvack R, Fandino RA, Sparks J, Staples J, et al. The insect SNMP gene family. Insect Biochem Mol Biol. 2009;39:448–56. https://doi.org/10.1016/j.ibmb.2009.03.007 . Zhang J, Liu Y, Walker WB, Dong S-L, Wang G-R. Identification and localization of two sensory neuron membrane proteins from spodoptera litura (lepidoptera: Noctuidae). Insect Sci. 2015;22:399–408. https://doi.org/10.1111/1744-7917.12131 . Additional Declarations No competing interests reported. Supplementary Files SupplementaryMaterials1Theaminoacidsequences.docx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 24 Feb, 2026 Reviews received at journal 19 Feb, 2026 Reviewers agreed at journal 27 Jan, 2026 Reviews received at journal 17 Dec, 2025 Reviewers agreed at journal 26 Nov, 2025 Reviewers invited by journal 12 Nov, 2025 Editor assigned by journal 13 Oct, 2025 Submission checks completed at journal 11 Oct, 2025 First submitted to journal 11 Oct, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7729120","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":549265650,"identity":"44ac67fe-c1b3-4842-9a70-409564eb2906","order_by":0,"name":"mengqing zhou","email":"","orcid":"","institution":"Guizhou Normal University","correspondingAuthor":false,"prefix":"","firstName":"mengqing","middleName":"","lastName":"zhou","suffix":""},{"id":549265651,"identity":"f628cad7-e2ab-4526-a6c0-ec0e3b1fbf91","order_by":1,"name":"yu Jiang","email":"","orcid":"","institution":"Guizhou Normal University","correspondingAuthor":false,"prefix":"","firstName":"yu","middleName":"","lastName":"Jiang","suffix":""},{"id":549265652,"identity":"0044e1a3-16ae-4446-8c79-1568ca3874d8","order_by":2,"name":"xiaona Zhang","email":"","orcid":"","institution":"Guizhou Normal University","correspondingAuthor":false,"prefix":"","firstName":"xiaona","middleName":"","lastName":"Zhang","suffix":""},{"id":549265653,"identity":"8662d151-2c27-46c8-a607-bcbddda851b3","order_by":3,"name":"gaoyin wu","email":"","orcid":"","institution":"Guizhou Normal University","correspondingAuthor":false,"prefix":"","firstName":"gaoyin","middleName":"","lastName":"wu","suffix":""},{"id":549265654,"identity":"63b31641-e76f-466c-813b-58ace96afa4a","order_by":4,"name":"tao peng","email":"","orcid":"","institution":"Guizhou Normal University","correspondingAuthor":false,"prefix":"","firstName":"tao","middleName":"","lastName":"peng","suffix":""},{"id":549265655,"identity":"d0931ec1-50e3-4727-82a0-bc04dc973951","order_by":5,"name":"sheng Liang","email":"","orcid":"","institution":"Chishui Alsophila National Nature Reserve Management Bureau.","correspondingAuthor":false,"prefix":"","firstName":"sheng","middleName":"","lastName":"Liang","suffix":""},{"id":549265656,"identity":"ff247ddb-4372-4c28-bac0-64afaf5d64dd","order_by":6,"name":"bangyou Liu","email":"","orcid":"","institution":"Chishui Alsophila National Nature Reserve Management Bureau.","correspondingAuthor":false,"prefix":"","firstName":"bangyou","middleName":"","lastName":"Liu","suffix":""},{"id":549265657,"identity":"110ecfe3-b459-430d-bb32-79012829aec8","order_by":7,"name":"shulin Yang","email":"","orcid":"","institution":"Guizhou Normal University","correspondingAuthor":false,"prefix":"","firstName":"shulin","middleName":"","lastName":"Yang","suffix":""},{"id":549265658,"identity":"b491361a-8549-4116-a155-c68b2dc729d0","order_by":8,"name":"hangdan Chen","email":"","orcid":"","institution":"Guizhou Normal University","correspondingAuthor":false,"prefix":"","firstName":"hangdan","middleName":"","lastName":"Chen","suffix":""},{"id":549265659,"identity":"e8cd5210-1a79-45ad-a99f-daa78a9449cc","order_by":9,"name":"fen Liu","email":"","orcid":"","institution":"Guizhou Normal University","correspondingAuthor":false,"prefix":"","firstName":"fen","middleName":"","lastName":"Liu","suffix":""},{"id":549265660,"identity":"2c7041d8-9c1f-49fd-8183-d801ddeeb9fb","order_by":10,"name":"qi Sun","email":"","orcid":"","institution":"Guizhou Normal University","correspondingAuthor":false,"prefix":"","firstName":"qi","middleName":"","lastName":"Sun","suffix":""},{"id":549265661,"identity":"2e187929-17f5-4f71-8977-239873b5b3a0","order_by":11,"name":"yuehua Song,","email":"","orcid":"","institution":"Guizhou Normal University","correspondingAuthor":false,"prefix":"","firstName":"","middleName":"yuehua","lastName":"Song","suffix":""},{"id":549265662,"identity":"fd5fe79f-65b4-44eb-a895-d8a0ebb6088f","order_by":12,"name":"Weicheng Yang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABD0lEQVRIiWNgGAWjYPACCQYDEJVQISHHz8DYQIKWD2dsjCUbiNPCANbCOLMtLXHDAUIqj589/JqnwkLenL338GvetsPGxucPtz34wWAnp4vDMoMzeWnWPGckDHf2nAMyzh2WM7uR2G7Yw5BsbIbDOoMDOWbGuW0SCQY3gAyessPGZjcY2yR4GA4kbsOl5fwboJZ/QC33gQwetsOJm/sPtkn+waflRo7x49wGkC08xg9ngLzPkNgmjc8WyRtvzJj/HJMw3HAmxwwcyBI3gFpkDHD7he98jvHHGTV18gbHzxh/AEdl//Fnkm8q7ORwaVE4wMAmAWXDGQyQaMIB5BsYmD9A2XDGKBgFo2AUjAIUAACLAGStKttTCQAAAABJRU5ErkJggg==","orcid":"","institution":"Guizhou Normal University","correspondingAuthor":true,"prefix":"","firstName":"Weicheng","middleName":"","lastName":"Yang","suffix":""}],"badges":[],"createdAt":"2025-09-27 14:38:26","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7729120/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7729120/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":96595659,"identity":"6fb4fec7-ff34-478e-9f31-0fd2eb3d603e","added_by":"auto","created_at":"2025-11-24 07:24:51","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":85320508,"visible":true,"origin":"","legend":"","description":"","filename":"9.2412AnalysisandIdentificationofChemosensoryGenesintheTranscriptomeofAdultRhoptroceroscyatheae.docx","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/e408adaf3a8b7ffd7c6d5626.docx"},{"id":96595677,"identity":"71284360-2f65-4f2f-8a31-0dc941ccc302","added_by":"auto","created_at":"2025-11-24 07:24:55","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":51703,"visible":true,"origin":"","legend":"","description":"","filename":"Tables.docx","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/a5517834cece039f9ec3ba61.docx"},{"id":96595654,"identity":"40a5aa98-6ad7-4e51-8b76-9493723e5cda","added_by":"auto","created_at":"2025-11-24 07:24:51","extension":"json","order_by":3,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":12910,"visible":true,"origin":"","legend":"","description":"","filename":"9c612a482fa84ef592b2e5880d75c2ca.json","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/45fa85060b46582274290cd1.json"},{"id":96595615,"identity":"5bcbbec9-9b3d-4451-86f8-8da2f2bcbee6","added_by":"auto","created_at":"2025-11-24 07:24:43","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":330314,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryMaterials1Theaminoacidsequences.docx","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/8fb28d935151eb758cac8c38.docx"},{"id":96595666,"identity":"e8655081-7c86-4dd2-9df0-b513f2724b65","added_by":"auto","created_at":"2025-11-24 07:24:53","extension":"xml","order_by":5,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":317440,"visible":true,"origin":"","legend":"","description":"","filename":"9c612a482fa84ef592b2e5880d75c2ca1enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/5401346acf2e2431d257f57c.xml"},{"id":96595673,"identity":"b066781d-ef9c-403f-9b77-a01a84c8975f","added_by":"auto","created_at":"2025-11-24 07:24:55","extension":"jpeg","order_by":6,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":119140,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/64dc4d4065263baeae51aa36.jpeg"},{"id":96595610,"identity":"5e598b44-ab0f-4130-963c-92e7a95a17a5","added_by":"auto","created_at":"2025-11-24 07:24:43","extension":"jpeg","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":565300,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage10.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/3047d267314935b9e5d3fb1b.jpeg"},{"id":96605483,"identity":"18c3d80a-e413-4796-927f-f6cd35e70506","added_by":"auto","created_at":"2025-11-24 09:23:13","extension":"jpeg","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":582538,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage11.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/3397135d5f086e750c9b31bc.jpeg"},{"id":96595601,"identity":"b34ab398-010f-449e-843b-ad0e68af7674","added_by":"auto","created_at":"2025-11-24 07:24:41","extension":"jpeg","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":342470,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage12.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/dc320ccbd70bf6037bcf0a50.jpeg"},{"id":96595647,"identity":"103fb607-d9aa-4d09-ae3e-fc9153ba4a71","added_by":"auto","created_at":"2025-11-24 07:24:50","extension":"png","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":55110,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage13.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/c5b391858488e3605863f399.png"},{"id":96595663,"identity":"4a615a1e-9f2c-46d1-ac79-0c5b61f00eb6","added_by":"auto","created_at":"2025-11-24 07:24:52","extension":"jpeg","order_by":11,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":215622,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage14.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/d73715b90255a314b6838ef8.jpeg"},{"id":96595603,"identity":"3aff58bb-9943-401d-b9ed-d58f2e1607bc","added_by":"auto","created_at":"2025-11-24 07:24:41","extension":"jpeg","order_by":12,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":152374,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/bea889a68307157cb0e2a1fa.jpeg"},{"id":96595602,"identity":"82bbf3bb-645d-47cd-873b-975afb065b40","added_by":"auto","created_at":"2025-11-24 07:24:41","extension":"jpeg","order_by":13,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":80184044,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/b311199b13fe574ff098d1ca.jpeg"},{"id":96595670,"identity":"a2111747-bbb0-4ac8-82fa-98b7317e697b","added_by":"auto","created_at":"2025-11-24 07:24:55","extension":"jpeg","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1532616,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/88a0b1f2e615c09dc27b758c.jpeg"},{"id":96605723,"identity":"df769524-fa45-4794-b6b4-4f4564efb11a","added_by":"auto","created_at":"2025-11-24 09:23:56","extension":"jpeg","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":306514,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/2824d31949a205e29548de87.jpeg"},{"id":96595582,"identity":"cd7ca424-9356-4d88-b022-961907f356ad","added_by":"auto","created_at":"2025-11-24 07:24:40","extension":"jpeg","order_by":16,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":444850,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage6.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/5f16db08c3a0870e86e784c8.jpeg"},{"id":96595668,"identity":"fe46a40b-f73a-4279-9f4a-d81a6d8be7e5","added_by":"auto","created_at":"2025-11-24 07:24:54","extension":"jpeg","order_by":17,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":200410,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage7.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/4e4e39474848da27bfa869bc.jpeg"},{"id":96595657,"identity":"85b367b0-3063-4473-b1c0-648c0db31b70","added_by":"auto","created_at":"2025-11-24 07:24:51","extension":"jpeg","order_by":18,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":264324,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage8.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/6ca54240e027b52747bc1558.jpeg"},{"id":96595646,"identity":"22359864-da64-49f4-b95c-6610c83b67a0","added_by":"auto","created_at":"2025-11-24 07:24:50","extension":"jpeg","order_by":19,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":630568,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage9.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/c405d64b727dbc633749505a.jpeg"},{"id":96595656,"identity":"9b32978e-8837-498f-9ca9-a3d46237904c","added_by":"auto","created_at":"2025-11-24 07:24:51","extension":"png","order_by":20,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":12326,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/4b743d8cefcbe15d1c7f8c61.png"},{"id":96595664,"identity":"68812cd5-3c21-46a4-a074-839c4ecd8fd0","added_by":"auto","created_at":"2025-11-24 07:24:52","extension":"png","order_by":21,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":64601,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage10.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/40df354edf54c8c113f68c88.png"},{"id":96595640,"identity":"0b4f21d5-2a2b-4972-9073-d33d0e9b0f6a","added_by":"auto","created_at":"2025-11-24 07:24:49","extension":"png","order_by":22,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":59246,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage11.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/f5d2fcd5200fcc2dc328eaa5.png"},{"id":96605836,"identity":"ee498955-99f4-4b8e-a193-65e06c097a09","added_by":"auto","created_at":"2025-11-24 09:24:11","extension":"png","order_by":23,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":42402,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage12.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/53ca7dfaaf5833651b526030.png"},{"id":96595600,"identity":"cf4b9619-fa80-4ed3-934a-c5cdd3dfc400","added_by":"auto","created_at":"2025-11-24 07:24:40","extension":"png","order_by":24,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":16980,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage13.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/b5ea7bb92fa871e079e6bf13.png"},{"id":96595665,"identity":"9f2a9092-2653-41a9-8d54-b43b18d48827","added_by":"auto","created_at":"2025-11-24 07:24:53","extension":"png","order_by":25,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":30625,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage14.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/509490317fd6c2f57146f715.png"},{"id":96595669,"identity":"8aa2504b-db3f-421d-9508-5b19b24b298d","added_by":"auto","created_at":"2025-11-24 07:24:54","extension":"png","order_by":26,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":27666,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/a76447872a5543a8443d5d16.png"},{"id":96595629,"identity":"e793d500-5538-46f9-8452-ec54e44cbab4","added_by":"auto","created_at":"2025-11-24 07:24:44","extension":"png","order_by":27,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":787307,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/90ca18adc6c0b0b8dddbf2cb.png"},{"id":96605173,"identity":"b0c52078-3747-4b3d-9fd2-99f27b450a0e","added_by":"auto","created_at":"2025-11-24 09:20:36","extension":"png","order_by":28,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":302350,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/52a0afd175d6a658eef7d5d2.png"},{"id":96595658,"identity":"27adeee6-0f0d-4e08-9d55-f56377230a79","added_by":"auto","created_at":"2025-11-24 07:24:51","extension":"png","order_by":29,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":33014,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/5af68c75510cae11ed8f1d36.png"},{"id":96595650,"identity":"d714cb8e-1786-428e-9572-a9d5a5683fd1","added_by":"auto","created_at":"2025-11-24 07:24:50","extension":"png","order_by":30,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":38249,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/90ce047a414f66fc251470a5.png"},{"id":96605448,"identity":"17cd8a66-e1ff-4edb-aa87-a25b58e89084","added_by":"auto","created_at":"2025-11-24 09:23:04","extension":"png","order_by":31,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":22725,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage7.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/c6263d7b46832e017373f70a.png"},{"id":96595606,"identity":"9288db6f-9a66-4283-8e3a-6fe6844bb87f","added_by":"auto","created_at":"2025-11-24 07:24:41","extension":"png","order_by":32,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":25565,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage8.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/3c57c951a5feff8b10a9c77b.png"},{"id":96595648,"identity":"c116f0c1-d895-41ee-a502-df468fced482","added_by":"auto","created_at":"2025-11-24 07:24:50","extension":"png","order_by":33,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":67178,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage9.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/c8ecbb9fd35b12ebe5480c10.png"},{"id":96605648,"identity":"c2ca68ee-939c-4615-b755-c5e22a9ecf7d","added_by":"auto","created_at":"2025-11-24 09:23:44","extension":"xml","order_by":34,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":315353,"visible":true,"origin":"","legend":"","description":"","filename":"9c612a482fa84ef592b2e5880d75c2ca1structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/f28d6f982eaac0f89b92e494.xml"},{"id":96595653,"identity":"4a1780d7-a146-49f9-a0c0-869c6714b99f","added_by":"auto","created_at":"2025-11-24 07:24:51","extension":"html","order_by":35,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":330025,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/efa6d9d74863a8edcd2684db.html"},{"id":96595641,"identity":"55f94403-2df0-4ac0-9697-3561fa60cf9b","added_by":"auto","created_at":"2025-11-24 07:24:49","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":212283,"visible":true,"origin":"","legend":"\u003cp\u003eUnigene and transcript lengths distribution of adults transcriptome in R. cyatheae.\u003c/p\u003e","description":"","filename":"image1.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/fd2833fb84ef95b54c8d89a9.png"},{"id":96605786,"identity":"47566163-37a7-48e6-ba07-34fdb6f02252","added_by":"auto","created_at":"2025-11-24 09:24:02","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":602888,"visible":true,"origin":"","legend":"\u003cp\u003eNon Redundant Protein Database(NR)annotation of unigenes(A);B Kyoto Encyclope-dia of Genes and Genomes(KEGG)annotation of unigenes(B); C Gene Ontology (GO) annotation of unigenes(C).\u003c/p\u003e","description":"","filename":"image2.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/a51126fbc0ca36c2e8a4282e.png"},{"id":96595667,"identity":"56b356c7-acec-482b-865d-4045b13d47d1","added_by":"auto","created_at":"2025-11-24 07:24:54","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":3937520,"visible":true,"origin":"","legend":"\u003cp\u003eMultiple amino acid sequence alignment of RcyaOBPs.\u003c/p\u003e","description":"","filename":"image3.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/3b6ef9863ad142fae44779f6.png"},{"id":96595644,"identity":"4429ea3a-6511-49c5-88db-a533c5bc47a1","added_by":"auto","created_at":"2025-11-24 07:24:50","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":988968,"visible":true,"origin":"","legend":"\u003cp\u003eMultiple amino acid sequence alignment of RcyaCSPs.\u003c/p\u003e","description":"","filename":"image6.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/13eed404b95fd06a4baf48eb.png"},{"id":96595612,"identity":"6cacdb77-0103-4124-9363-76c32bf554ab","added_by":"auto","created_at":"2025-11-24 07:24:43","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":666452,"visible":true,"origin":"","legend":"\u003cp\u003eMultiple amino acid sequence alignment of RcyaNPC2s.\u003c/p\u003e","description":"","filename":"image8.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/fa7cc97cebd733d14370f461.png"},{"id":96605882,"identity":"9623a2a5-96f3-485c-bf76-0af84ae45487","added_by":"auto","created_at":"2025-11-24 09:24:18","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":55110,"visible":true,"origin":"","legend":"\u003cp\u003eHeatmap of 13 chemosensory genes\u003c/p\u003e","description":"","filename":"image14.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/c5dc2811bc9d1fc61b91c434.png"},{"id":96595652,"identity":"143e817f-eb47-49a9-b4aa-704234effb9b","added_by":"auto","created_at":"2025-11-24 07:24:50","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":1993890,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree of odorant-binding proteins (OBPs) from R cyatheae and 9 other Hymenoptera species. The protein sequences used in this phylogenetic analysis are listed in the Supplementary Materials Supplementary Materials File S1. The color of the dots reflect their bootstrap values. The color scale is shown on the left, with the greener the color, indicating larger bootstrap values.The bootstrap repeats 1,000 times .\u003c/p\u003e","description":"","filename":"image5.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/84c7b73cf98787afc985b5f4.png"},{"id":96595674,"identity":"55f8f1ea-a53f-4fce-ba80-ce3223041e2e","added_by":"auto","created_at":"2025-11-24 07:24:55","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":1308693,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree of chemosensory proteins (CSPs) from R cyatheae and 12 other Hy-menoptera species. The protein sequences used in this phylogenetic analysis are listed in the Supplementary Materials File S2. The color of the dots reflect their bootstrap values. The color scale is shown on the left, with the greener the color, indicating larger bootstrap values. The bootstrap repeats 1,000 times.\u003c/p\u003e","description":"","filename":"image7.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/f50a734b827173e2babdcf15.png"},{"id":96604984,"identity":"9f6ad13f-99a9-47f6-a96f-c2cdbb71059c","added_by":"auto","created_at":"2025-11-24 09:17:04","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":802765,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree of Niemann–Pick type C2 proteins (NPC2s) from R cyatheae and 10 other Hymenoptera species. The protein sequences used in this phylogenetic analysis are listed in the Supplementary Materials Supplementary Materials File S3. The color of the dots reflect their bootstrap values. The color scale is shown on the left, with the greener the color, indicating larger bootstrap values.The bootstrap repeats 1,000 times .\u003c/p\u003e","description":"","filename":"image9.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/de0200e2fad959f395192aa4.png"},{"id":96595676,"identity":"543cf2c9-ca58-4ecc-8066-3c627cce656c","added_by":"auto","created_at":"2025-11-24 07:24:55","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":2006992,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree of gustatory receptors (ORs) from R cyatheae and 3 other insect species. The protein sequences used in this phylogenetic analysis are listed in the Supplemen-tary Materials File S4. The color of the dots reflect their bootstrap values. The color scale is shown on the left, with the greener the color, indicating larger bootstrap values.The bootstrap repeats 1,000 times .\u003c/p\u003e","description":"","filename":"image10.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/3e640ba56c9698931c010a08.png"},{"id":96595662,"identity":"2566832e-6e23-4565-a305-9c143c2bcf00","added_by":"auto","created_at":"2025-11-24 07:24:51","extension":"png","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":1925000,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree of ionotropic receptors (IRs) from R cyatheae and 14 other Hyme-noptera species. The protein sequences used in this phylogenetic analysis are listed in the Sup-plementary Materials File S5. The color of the dots reflect their bootstrap values. The color scale is shown on the left, with the greener the color, indicating larger bootstrap values. The boot-strap repeats 1,000 times .\u003c/p\u003e","description":"","filename":"image11.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/683d24f892911879edf6f645.png"},{"id":96595599,"identity":"ac4bd332-7624-481a-8e96-801d71f6db00","added_by":"auto","created_at":"2025-11-24 07:24:40","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":1965687,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree of gustatory receptors (GRs) from R cyatheae and 10 other insect species. The protein sequences used in this phylogenetic analysis are listed in the Supplemen-tary Materials File S6. The size and color of the dots reflect their bootstrap values. The color of the dots reflect their bootstrap values. The color scale is shown on the left, with the greener the color, indicating larger bootstrap values. The bootstrap repeats 1,000 times .\u003c/p\u003e","description":"","filename":"image12.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/298d294675895b260fec6343.png"},{"id":96595675,"identity":"f807da55-e750-463d-a970-06be3a0e0aae","added_by":"auto","created_at":"2025-11-24 07:24:55","extension":"png","order_by":13,"title":"Figure 13","display":"","copyAsset":false,"role":"figure","size":1151622,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree of RcyaSNMPs from 15 Hymenoptera species and 10 CD36 homo-logs (including epithelial membrane protein, croquemort, peste, santa maria and debris buster) from Drosophila melanogaster, D. busckii, Tribolium castaneum, Culex quinquefasciatus and Anopheles aquasalis. The protein sequences used in this phylogenetic analysis are listed in the Supplementary Materials File S7. The color of the dots reflect their bootstrap values. The color scale is shown on the left, with the greener the color, indicating larger bootstrap values.The bootstrap repeats 1,000 times .\u003c/p\u003e","description":"","filename":"image13.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/247cf6248c0c5c06f8a044ab.png"},{"id":96595660,"identity":"f96b8dc7-cd83-4c2f-8065-ab33311f3ea8","added_by":"auto","created_at":"2025-11-24 07:24:51","extension":"png","order_by":14,"title":"Figure 14","display":"","copyAsset":false,"role":"figure","size":609979,"visible":true,"origin":"","legend":"\u003cp\u003eExpression profiles of the candidate genes in different R cyatheae tissues.\u003c/p\u003e\n\u003cp\u003e(Fant and Mant: female and male antennae; Fhea 和 Mhea: female and male heads without an-tennae; Fab: female abdomens without ovipositors and digestive tracts, Fov: female ovipositors; Mge: male genitalia; Fabd and Mabd: male abdomens; Fwin and Mwin : male and female wings; Ftho and Mtho: male and female thorax;Fleg and Mleg：male and female legs).The error bars represents standard errors and the small letters above each bar indicate significant differences in transcript abundances (p \u0026lt; 0.05).\u003c/p\u003e","description":"","filename":"image15.png","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/83c6f30e8701b630cddab0e8.png"},{"id":96608501,"identity":"d5ac154e-6224-41f8-a27e-a42bdd7b4ed6","added_by":"auto","created_at":"2025-11-24 09:28:40","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":20886630,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/67471d99-f998-4e41-b13a-8181cbbbfb61.pdf"},{"id":96595642,"identity":"950188d9-45d1-4a59-9adb-28df77f09569","added_by":"auto","created_at":"2025-11-24 07:24:50","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":330314,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryMaterials1Theaminoacidsequences.docx","url":"https://assets-eu.researchsquare.com/files/rs-7729120/v1/2aec0ec2f62b3a81eafc53b0.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":" Analysis and Identification of Chemosensory Genes in the Transcriptome of Adult Rhoptroceros cyatheae","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eThe olfactory chemosensory system plays a crucial role in determining the life activities of insects. It enables the detection of olfactory information carried by odor molecules, which is associated with specific behaviors such as host location, mating, foraging, and evading predators [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e].When hydrophobic molecules enter the sensillar lymph through chemosensilla, odorant-binding proteins (OBPs), chemosensory proteins (CSPs), and Niemann-Pick type C2 proteins (NPC2s) in the lymph bind and transport these molecules, which are subsequently detected by olfactory receptor neurons (ORNs). Odorant receptors (ORs), ionotropic receptors (IRs), gustatory receptors (GRs), or sensory neuron membrane proteins (SNMPs) on ORN membranes convert chemical signals into electrical signals. These electrical signals are then transduced by ORNs to the central nervous system in the insect brain [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. After processing by the central nervous system, these signals trigger corresponding physiological and behavioral responses [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn the process of odor detection, the initial step in the olfactory recognition of hydrophobic odorants involves the binding and transport of odor molecules by OBPs, CSPs, and NPC2s present in the sensillar lymph [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].OBPs, CSPs, and NPC2s are all small molecular-weight water-soluble proteins. Classical OBPs possess six conserved cysteine residues, whereas minus-C OBPs contain four conserved cysteine residues, and atypical OBPs feature only two conserved cysteine residues. Compared with OBPs, which are more evolutionarily conserved ,CSPs typically carry four conserved cysteine residues. Initially identified in mammals for their role in shaping cholesterol transport, NPC2s were later found to participate in olfactory responses in insects [\u003cspan additionalcitationids=\"CR6\" citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].OBPs primarily recognize and transport volatile compounds; CSPs mainly bind nonvolatile substances; whereas NPC2s are responsible for binding and transporting hydrophobic compounds[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Transmembrane receptor proteins (ORs, IRs, GRs, and SNMPs) on olfactory neurons transmit olfactory signals across the membrane to the central nervous system subsequently. Insect odorant receptors can be categorized into ORs and odorant receptor co-receptors (Orco). Orco facilitates the precise localization of OR proteins to the dendritic membranes of olfactory neurons [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].IRs evolved from a class of ionotropic glutamate receptors (iGluRs) and primarily function in sensing olfactory cues, gustatory signals, temperature and humidity [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. GRs primarily mediate the perception of bitterness, sweetness, and carbon dioxide (CO₂). Additionally, accumulating evidence suggests their involvement in light and temperature sensing[\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].SNMPs are transmembrane structural proteins that are categorized into SNMP1 and SNMP2, which play critical roles in the detection of plant volatiles and insect sex pheromones [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].These chemosensory genes play crucial roles in determining insect olfactory recognition and represent the foundational step in study of insect olfactory mechanisms. However, research on the olfactory mechanisms of \u003cem\u003eRhoptroceros cyatheae\u003c/em\u003e adults remains limited. Elucidating the olfactory mechanisms of both female and male \u003cem\u003eR. cyatheae\u003c/em\u003e is significant for understanding how they recognize semiochemicals and rely on chemical communication and sensory mechanisms to locate hosts and mates. Concurrently, the precise identification of olfactory-related genes in \u003cem\u003eR. cyatheae\u003c/em\u003e lays the groundwork for subsequent investigations into its olfactory mechanisms.\u003c/p\u003e\u003cp\u003e\u003cem\u003eR. cyatheae\u003c/em\u003e, which belongs the family Selandriidae and genus \u003cem\u003eRhopographus\u003c/em\u003e, is distributed in regions such as Sichuan and Guizhou. It is among the primary herbivorous insects that infest Alsophila spinulosa (a tree fern species) [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].This insect completes four to five generations annually, with its occurrence period spanning from April to June each year. During outbreaks, \u003cem\u003eR. cyatheae\u003c/em\u003e can cause severe infestations, where the young leaves of extensive \u003cem\u003eA. spinulosa\u003c/em\u003e stands are completely defoliated by its larvae [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].The tree fern \u003cem\u003eA. spinulosa\u003c/em\u003e faces challenges due to its lengthy reproductive cycle via spores and difficult natural regeneration. Outbreaks of the \u003cem\u003eR. cyatheae\u003c/em\u003e drastically reduce spore production, severely impacting the population sustainability of this species [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003cem\u003eA. spinulosa\u003c/em\u003e is a Class II protected plant in China, hailed as the 'living fossil' of terrestrial plants, and possesses extremely high research and pharmacological value [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].However, in recent years, the population of \u003cem\u003eR. cyatheae\u003c/em\u003e has continued to increase, posing a significant threat to the survival and reproduction of \u003cem\u003eA. spinulosa\u003c/em\u003e[\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].Therefore, it is crucial to develop novel biological control methods that are safer, more environmentally friendly, and less ecologically damaging.\u003c/p\u003e\u003cp\u003eIn this study, RNA-Seq technology was used to obtain adults transcriptome data from of female and male of \u003cem\u003eR. cyatheae\u003c/em\u003e. We identified chemosensory-related genes including OBPs, CSPs, NPC2s, ORs, GRs, IRs, and SNMPs. The findings not only provide a solid theoretical foundation for future research on olfactory genes and mechanisms in \u003cem\u003eR. cyatheae\u003c/em\u003e, but they also hold profound significance for advancing ecological regulation strategies and developing green control technologies targeting this species.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1Insect Rearing and Sample Collection\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eThe test insects, \u003cem\u003eR. cyatheae\u003c/em\u003e (Hymenoptera: Selandriidae), were collected from December 2024 to May 2025 in the Chishui Alsophila National Nature Reserve, Guizhou Province, China. Larvae sampled from the field were reared in an intelligent biochemical incubator (model GZL-P8000-C3, Hefei DuskTec Biological Technology Co., Ltd.) under controlled conditions: a temperature of 25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C, a relative humidity of75%\u0026plusmn;10%, and a 14h light:10h dark cycle. After eclosion, the adult pecimens \u003cem\u003eR. cyatheae\u003c/em\u003e were provided with a 10% honey solution as a dietary supplement and maintained under the same environmental conditions described above.\u003c/p\u003e\u003cp\u003eAdults of \u003cem\u003eR. cyatheae\u003c/em\u003e (4 days post-eclosion) were rinsed with RNase-free and DNase-free water, transferred into 5 ml cryovials, immediately frozen in liquid nitrogen, and stored in a \u0026minus;\u0026thinsp;80\u0026deg;C ultralow temperature freezer (MDF-86V180E) for transcriptome sequencing. Three biological replicates were established for both sexes. To investigate the expression levels of chemosensory genes in different tissues of \u003cem\u003eR. cyatheae\u003c/em\u003e, twelve tissue types were dissected from adult male and female individuals under a laminar flow hood (model LJ-SZM, Leica, Germany): female antennae (Fant), male antennae (Mant), female heads without antennae (Fhea), male heads without antennae (Mhea), female thoraces (Ftho), male thoraces (Mtho), female abdomens (Fabd), male abdomens (Mabd), female wings (Fwin), male wings (Mwin), female legs (Fleg), and male legs (Mleg) A total of 150 individuals per sex were obtained. All the samples were placed in labeled 5 ml prechilled cryovials, rapidly frozen in liquid nitrogen, and stored at \u0026minus;\u0026thinsp;80\u0026deg;C until RNA extraction. Each sample was processed with three biological replicates, and each biological replicate included three technical replicates.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2 Total RNA Extraction and Detection, cDNA Library Construction and Sequencing\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eTotal RNA was extracted from female and male adult samples of \u003cem\u003eR. cyatheae\u003c/em\u003e via QIAzol Lysis Reagent (Qiagen, Germany). The integrity of the total RNA was assessed through 1% agarose gel electrophoresis, while the concentration and purity were measured using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, USA). The RNA samples were then precisely evaluated for purity and integrity using an Agilent 5300 Bioanalyzer (Agilent Technologies, USA). This study included three biological replicates. Approximately 1 \u0026micro;g of total RNA from each sample was used for cDNA library construction. The libraries were sequenced on an Illumina NovaSeq X platform, with both library preparation and sequencing performed in collaboration with Shanghai Majorbio Technology.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.3 Transcriptome Assembly and Functional Annotation\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eThe raw reads obtained from sequencing were first processed using Fastp to remove adapter sequences and low-quality reads, yielding clean reads [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].Next, Trinity (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://github.com/trinityrnaseq/trinityrnaseq/wiki\u003c/span\u003e\u003cspan address=\"https://github.com/trinityrnaseq/trinityrnaseq/wiki\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) was used to perform de novo assembly for all the clean reads [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].The assembly results were filtered and optimized using TransRate (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://hibberdlab.com/transrate/\u003c/span\u003e\u003cspan address=\"http://hibberdlab.com/transrate/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), and then the assembly completeness of the unigenes was evaluated with BUSCO (Benchmarking Universal Single-Copy Orthologs, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://busco.ezlab.org).Additionall\u003c/span\u003e\u003cspan address=\"http://busco.ezlab.org).Additionall\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003ey, the Q20, Q30, GC content, error rate, and nucleotide base composition of the clean data were calculated(S2).Finally, to obtain comprehensive functional annotations, all unigenes derived from this transcriptome sequence were subjected to alignment-based annotation against six major databases: the Non-redundant Protein Database (NR), Pfam, Clusters of Orthologous Groups (COG), Swiss-Prot, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Ontology (GO).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003e2.4 Identification and Expression Analysis of Chemosensory Genes\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eTo identify putative chemosensory genes (OBPs, CSPs, NPC2s, ORs, GRs, IRs, and SNMPs), we searched the transcriptome annotation results obtained in Section 1.4 using olfactory gene keywords (OBP and odorant binding protein, CSP and chemosensory protein, NPC2 and Niemann\u0026ndash;Pick type C2 protein, OR and odorant receptor, GR and gustatory receptor, IR and ionotropic receptor, SNMP and sensory neuron membrane protein). The initial identification was further validated through manual BLASTX and BLASTP alignments, with an E-value threshold of \u0026lt;\u0026thinsp;10⁻⁵ set for screening candidate olfactory-related genes. The ORF finder (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ncbi.nlm.nih.gov/orffinder/\u003c/span\u003e\u003cspan address=\"https://www.ncbi.nlm.nih.gov/orffinder/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) was used to predict the open reading frames (ORFs) of all the putative chemosensory genes. For the candidate OBP, CSP, and NPC2 genes, the N-terminal signal peptides were predicted using the SignalP-6.0 online program (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://services.healthtech.dtu.dk/services/SignalP-6.0/\u003c/span\u003e\u003cspan address=\"https://services.healthtech.dtu.dk/services/SignalP-6.0/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e; Almagro Armenteros et al.). Molecular weights (MWs) and isoelectric points (pI) were obtained using the ExPASy proteomics server (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.expasy.org/resources/compute-pi-mw\u003c/span\u003e\u003cspan address=\"https://www.expasy.org/resources/compute-pi-mw\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). For candidate OR, IR, and SNMP genes, the transmembrane domains (TMDs) of the annotated proteins were predicted with the online program DeepTMHMM-1.0 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://services.healthtech.dtu.dk/services/DeepTMHMM-1.0/\u003c/span\u003e\u003cspan address=\"https://services.healthtech.dtu.dk/services/DeepTMHMM-1.0/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Multiple sequence alignment of chemosensory gene amino acid sequences was performed via the ClustalX algorithm in MEGA 12.0 software[\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e], with subsequent color coding implemented through ESPript 3.0 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://espript.ibcp.fr/ESPript/ESPript/index.php).Phylogeneti\u003c/span\u003e\u003cspan address=\"https://espript.ibcp.fr/ESPript/ESPript/index.php).Phylogeneti\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003ec analysis of candidate the chemosensory genes was conducted using amino acid sequences from \u003cem\u003eR. cyatheae\u003c/em\u003e and other insects. The Maximum likelihood Method (ML) with the Poisson model was employed in MEGA12.0 to infer phylogenetic relationships, incorporating 1,000 bootstrap replicates and retaining branches supported by \u0026ge;\u0026thinsp;95% bootstrap values[\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e].Following the preliminary reconstruction of the relevant phylogenetic trees, visualization and editing were performed using the Interactive Tree of Life tool (iTOL v7) (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://itol.embl.de/\u003c/span\u003e\u003cspan address=\"https://itol.embl.de/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003e2.5 Screening and Analysis of Differentially Expressed Genes\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eTo compare differences in olfactory gene expression differences between female and male adults of \u003cem\u003eR. cyathea\u003c/em\u003e, RSEM was employed for expression quantification, with Fragments Per Kilobase of transcript per Million mapped reads (FPKM) values employed to assess gene expression levels[\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e].Differential expression analysis of read counts was performed using DESeq2 on basis of the negative binomial distribution model. Differentially expressed genes were identified using a threshold of fold change (FC)\u0026thinsp;\u0026ge;\u0026thinsp;1 and an adjusted P-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05[\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003e2.6 RT-qPCR Analysis of Expression Levels of Candidate Chemosensory Genes\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eOn the basis of the differential gene expression analysis results, chemosensory genes with sex-biased expression (female- or male-biased) were preliminarily screened under the threshold conditions of q-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05 and |log2(fold change)| \u0026gt;1 as candidate genes. These candidate genes were subsequently validated for their quantitative expression levels across the 12 distinct tissues using real-time quantitative polymerase chain reaction (RT-qPCR).To further remove genomic DNA (gDNA) and synthesize cDNA, TransScript All-in-One First-Strand cDNA Synthesis SuperMix (Transgen Biotech, Beijing, China) was used to reverse transcribe 1 \u0026micro;g of total RNA per sample into cDNA RcyaRPL32 and RcyaRPS18 were used as reference genes (internal controls) to normalize the expression data. Specific primers were designed using Primer Premier 5.0 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://bioinfo.ut.ee/primer3-0.4.0/\u003c/span\u003e\u003cspan address=\"https://bioinfo.ut.ee/primer3-0.4.0/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) and are listed in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e3\u003c/span\u003e. qPCR measurements were subsequently performed using 2\u0026times;Universal Blue SYBR Green qPCR Master Mix (Servibio, Wuhan, China) and SweScript All-in-One RT SuperMix for qPCR (Servibio, Wuhan, China). Each qPCR was performed according to the manufacturer's instructions under the following conditions: initial denaturation at 95\u0026deg;C for 30 s, followed by 40 cycles of denaturation at 95\u0026deg;C for 15 s, annealing/extension at 60\u0026deg;C for 30 s. Subsequently, a melting curve analysis was generated by continuous fluorescence measurement from 65\u0026deg;C to 95\u0026deg;C with increments of 0.5\u0026deg;C. All qPCR assays were conducted with three technical replicates and three biological replicates.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eThe information of primers used for RT-qPCR assays\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003e引物名称\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eForward primer (5\u0026rsquo;-3\u0026rsquo;)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eReverse primer (5\u0026rsquo;-3\u0026rsquo;)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaRPL32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGACAAGCTCAAGCGTAACTGGC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eACGATAGCTTTGCGTTTCTTGC\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaRPS18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTGCCTCGGGTTGGACATAATC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCAAGGGTGTCGGTCGTCGTT\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaOBP7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCCGAAAGACAAGATGACGGAGAT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eTGATCCTTATGCTCGTGGAAACA\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaOBP8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGCTTCCTTCATTGCGTTCTCAC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAGCGGTCCTGGCCTCTTCTT\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaCSP2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTTCATCAATCACCTCCGCCC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eGATTCAAGCGGCACCTACAGG\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaCSP10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCCTCTGGGTCGATCTTGGTC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eACTGAAAGATGTGCTGGGTGAG\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaOR19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAACAATGGCGTCCGTGGTAC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAGATCGTGCGTCTATGAACAGG\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaOR14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCCAAGTAAGCGAATGTTGACGAT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eTTGGAGAGGTTGACTAACACAGCC\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaOR20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAACATGACGACCAGTGCTGAAA\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAGGAGTCGGTTATCGGCTTTCA\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaOR17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCCTAACGACCGACTTGACCAAC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eGCTCAGAACTTATCGCACCTACA\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaGR5a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCTGGCAACGAGAAATATGAACG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCCTGAGCGGATTGGTGAAAC\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaiGluR4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGCGAAACCGATTCCTATACCTT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eATTCAAGACCTCCGAACACCTAC\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaSNMP2a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eACGCACGAAAGGAATTACAGG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eTGGCGATGTAGAAATGAGGTAATG\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaNPC2d\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGGTCAAATGAGATAAACCTCCAGAT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCGTGGGTCAAGGCGTAGATAG\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRcyaNPC2e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCTGACCGACCACGAAGTTGTTT\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCGGTGCAGGTGAAGATAGAAGATG\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\u003ch2\u003e2.7 Statistical Analysis\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eThe relative expression levels of chemosensory genes in each sample were calculated via the 2^(-ΔΔCt) method, with the tissue showing the lowest expression level as the reference. Subsequently, logarithmic transformation was performed on the data. If the data failed to meet the requirements for normality and homogeneity of variance, the nonparametric Kruskal-Wallis test was used to compare differences; otherwise, one-way analysis of variance (ANOVA) followed by Duncan's test was applied for difference testing. All the data were statistically analyzed using SPSS Statistics 25 and Origin 2021 Pro, and are presented as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of the mean (SEM).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003e3.1 Transcriptome Sequencing Statistics and Sequence Assembly\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eThe transcriptomes of female and male adults of \u003cem\u003eR. cyatheae\u003c/em\u003e were sequenced on a second-generation high-throughput sequencing platform. A total of 40.94 Gb of clean reads were obtained, with each sample yielding at least 6.34 Gb of clean data. The Q20 and Q30 base percentages exceeded 98% and 96%, respectively, while the GC content across samples varied between 41.48% and 43.32%( Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e .in the Supplementary Materials). The average base error rate for all quality-controlled data was less than 0.02%, indicating high-quality sequencing data.Sequence assembly was performed using Trinity software, and the results revealed that 44,396 transcripts were obtained, with an average length of 1393.21 bp and an N50 length of 4356 bp (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e1\u003c/span\u003e).Further assembly analysis of the transcripts yielded a total of 30,296 unigenes, with an average length of 1,393.21 bp and an N50 length of 3,286 bp. In terms of the length distribution of the unigenes and transcripts, those with lengths ranging from 200 to 500 bp were the most abundant, accounting for 14,157 unigenes (47%) and 15,698 transcripts (35%). In contrast, those with lengths ranging from 4,001 to 4,500 bp were the least abundant, with only 486 unigenes (2%) and 1,123 transcripts (3%) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). For the candidate OR, IR, and SNMP genes, the transmembrane domains (TMDs) of the annotated proteins were predicted using the online program DeepTMHMM-1.0 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://services.healthtech.dtu.dk/services/DeepTMHMM-1.0/\u003c/span\u003e\u003cspan address=\"https://services.healthtech.dtu.dk/services/DeepTMHMM-1.0/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Multiple sequence alignment of chemosensory gene amino acid sequences was performed using the ClustalX algorithm in MEGA 12.0 software.The assembly completeness was assessed using BUSCO, which yielded a score exceeding 95%, indicating high-quality assembly. The raw reads of \u003cem\u003eR. cyatheae\u003c/em\u003e were subsequently submitted to the NCBI Short Read Archive database under accession number PRJNA1309270.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eInformation for 6 samples used for transcriptome analysis\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSample\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRaw reads\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eRaw bases\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eClean reads\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eClean bases\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eError rate(%)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eQ20(%)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eQ30(%)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eGC content(%)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRF1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e42,742,146\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6,454,064,046\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e42,440,672\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e6,344,273,982\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.0119\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e98.76\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e96.18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e43.23\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRF2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e44,087,188\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6,657,165,388\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e43,765,508\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e6,559,551,741\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.0119\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e98.74\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e96.14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e42.19\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRF3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e51,216,586\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e7,733,704,486\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e50,908,962\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e7,641,273,693\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.0119\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e98.81\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e96.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e43.32\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRM1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e43,398,736\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6,553,209,136\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e43,123,866\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e6,460,256,582\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.0119\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e98.77\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e96.25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e42.17\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRM2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e44,020,738\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6,647,131,438\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e43,726,486\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e6,549,862,099\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.012\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e98.72\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e96.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e42.19\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRM3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e49,702,156\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e7,505,025,556\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e49,365,188\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e7,385,834,680\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.0119\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e98.74\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e96.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e41.48\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eNumber and length of transcripts and unigenes in R. cyatheae.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eType\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eUnigene\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eTranscript\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTotal number\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e30296\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e44396\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTotal base\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e42208633\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e88696954\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLargest length (bp)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e33119\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e33119\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSmallest length (bp)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e201\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e201\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAverage length (bp)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1393.21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1997.86\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eN50 length (bp)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3286\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4356\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eE90N50 length (bp)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4853\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4215\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFragment mapped percent(%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e81.988\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e93.204\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGC percent (%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e38.14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e39.02\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTransRate score\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.46504\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.51992\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBUSCO score\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eC:95.5%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eC:98.0\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003e3.2 Functional Annotation and Classification of Transcriptome\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eThe functional annotation results indicated that 10,716 unigenes (35.94%) were matched in the NCBI NR protein database with an E-value threshold of 10⁻⁵ (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA).Species classification indicated that the highest sequence homology was observed in Athalia rosae (32.57%), followed by Diprion similis (21.35%), Neodiprion lecontei (10.20%), Neodiprion fabricii (7.99%), Neodiprion virginianus (5.10%), and Neodiprion pinetum (4.53%). The species that exhibited the highest homology with \u003cem\u003eR. cyatheae\u003c/em\u003e were exclusively hymenopteran insects.\u003c/p\u003e\u003cp\u003eIn the KEGG annotation, a total of 7,163 unigenes were classified into six KEGG functional groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB): Human Diseases,with 1,745 unigenes (24.36%);Environmental Information Processing, with 861 unigenes (12.02%);Genetic Information Processing with 1,161 unigenes (16.21%); Cellular Processes, with 898 unigenes (14.54%);Organismal Systems ,with 1,225 unigenes (17.10%), and Metabolism, with 1,273 unigenes (17.78%)\u003c/p\u003e\u003cp\u003eFor GO annotation, the transcripts were functionally categorized according to their GO classifications (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC). A total of 6,619 genes (22.20%) were assigned to three GO categories, namely, molecular function, cellular component, and biological process, encompassing 51 specific terms. Within the biological process category, the terms \u0026ldquo;cellular process\u0026rdquo; and \u0026ldquo;metabolic process\u0026rdquo; exhibited the greatest representation. With respect to cellular components, \u0026ldquo;cell part\u0026rdquo; constituted the most dominant group. Regarding molecular function, the terms \u0026ldquo;binding\u0026rdquo; and \u0026ldquo;catalytic activity\u0026rdquo; were the most abundant groups. On the basis of the results of the GO analysis, genes associated with \u0026ldquo;binding\u0026rdquo; were significantly enriched in R.cyatheae, followed by \u0026ldquo;cell part\u0026rdquo; and \u0026ldquo;catalytic activity\u0026rdquo; which ranked second and third, respectively.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003e3.3 Identified Candidate Water-Soluble Proteins\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eCandidate OBPs\u003c/p\u003e\u003cp\u003eEleven candidate RcyaOBPs exceeding 600 bp in length were identified in \u003cem\u003eR. cyatheae\u003c/em\u003e and designated RcyaOBP1 through RcyaOBP11. These genes encoded proteins ranging from 136 to 273 amino acid residues, with molecular weights of 4.32 to 9.20 kDa and isoelectric points (pI) ranging from 4.20 to 9.16. Sequence alignment via BLASTP revealed that the candidate RcyaOBPs share sequence identities ranging from 26.71% to 63.16% with other hymenopteran OBPs. Among these candidates, only RcyaOBP8 was predicted to lack a signal peptide, whereas RcyaOBP3 and RcyaOBP5 lacked conserved cysteine residues (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e.in the Supplementary Materials). Multiple sequence alignment revealed that nine candidate RcyaOBPs (RcyaOBP1 and RcyaOBP4\u0026ndash;11) contained six conserved cysteine residues with the pattern X11\u0026ndash;37-Cys1-X20\u0026ndash;27-Cys2-X3-Cys3-X26\u0026ndash;49-Cys4-X8\u0026ndash;11-Cys5-X8-Cys6-X5\u0026ndash;18, which were characterized as classical OBPs. RcyaOBP2, harboring four conserved cysteine residues, was a Minus-COBPs. RcyaOBP3 was classified as atypical OBPs with 2 cysteines (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). A Maximum likelihood (ML) tree was constructed using the OBPs of \u003cem\u003eR. cyatheae\u003c/em\u003e and nine other Hymenopteran species. The results revealed that many RcyaOBPs were highly divergent into different clades, while the OBPs of species such as BdioOBPs (\u003cem\u003eBaryscapus dioryctriae\u003c/em\u003e), NvitOBPs (\u003cem\u003eNasonia vitripennis\u003c/em\u003e), and CcunOBPs (\u003cem\u003eChouioia cunea\u003c/em\u003e) were closely clustered in multiple clades (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e7\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eBioinformatics analysis of identified R. cyatheae OBP genes\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"11\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGene ID\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGene Name\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eORF (aa)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eSignal Peptide\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eGroup\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003ePI\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eMW(Kd)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"4\" nameend=\"c11\" namest=\"c8\"\u003e\u003cp\u003eBLAST Annotation\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eDescription\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eQuery\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003eE value\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003eIdentify(%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN10134_c0_g2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOBP1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e136\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e4.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e15.07\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_046625102.1, general odorant-binding protein 83a-like isoform X2 [Neodiprion virginianus]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2E-56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e63.16\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN16238_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOBP2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e214\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e4.53\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e23.57\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eQHN69060.1, odorant binding protein 3\u003c/p\u003e\u003cp\u003e[Sirex nitobei]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e53%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e1E-16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e41.88\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN16731_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOBP3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e145\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e30\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e9.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e15.97\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eQHN69068.1, odorant binding protein 11\u003c/p\u003e\u003cp\u003e[Sirex nitobei]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e90%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e4E-33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e43.61\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN17528_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOBP4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e138\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e4.77\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e15.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_046663889.1, general odorant-binding protein 19d-like\u003c/p\u003e\u003cp\u003e[Homalodisca vitripennis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e91%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e7E-06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e27.69\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN19984_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOBP5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e279\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e7.66\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e31,15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_058448369.1, general odorant-binding protein 56d-like\u003c/p\u003e\u003cp\u003e[Malaya genurostris]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e26%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e9E-10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e45.95\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN20475_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOBP6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e151\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e6.35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e17.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eQHN69059.1, odorant binding protein 2\u003c/p\u003e\u003cp\u003e[Sirex nitobei]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e88%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e6E-45\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e52.24\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN3522_c0_g2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOBP7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e141\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e5.22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e15.66\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_033217025.1, general odorant-binding protein 56a-like\u003c/p\u003e\u003cp\u003e[Belonocnema kinseyi]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e87%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2E-27\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e48\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN3640_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOBP8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e273\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e5.46\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e30.70\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_033342464.1, general odorant-binding protein 56a-like\u003c/p\u003e\u003cp\u003e[Megalopta genalis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e43%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e1E-10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e27.87\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN4219_c0_g3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOBP9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e143\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e4.71\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e16.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_046431779.1, general odorant-binding protein 83a-like\u003c/p\u003e\u003cp\u003e[Neodiprion fabricii]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e97%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2E-36\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e46.43\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN4317_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOBP10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e133\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e5.46\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e14.20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_046625102.1, general odorant-binding protein 83a-like isoform X2 [Neodiprion virginianus]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2E-56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e63.16\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN923_c0_g5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOBP11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e142\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e5.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e15.80\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_012256027.2, general odorant-binding protein 83a-like\u003c/p\u003e\u003cp\u003e[Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2E-60\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e61.97\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"11\"\u003eNote: \u0026dagger;ORF: open reading frame.\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"11\"\u003eAmino acid sequences of \u003cem\u003eR. cyatheae\u003c/em\u003e OBP genes are listed in the supplementary data.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eCandidate CSPs\u003c/p\u003e\u003cp\u003eThrough bioinformatic analysis, ten candidate RcyaCSPs exceeding 600 bp in length were identified in the adult transcriptome of \u003cem\u003eR. cyatheae\u003c/em\u003e. These genes encode proteins ranging from 118 to 157 amino acid residues, with molecular weights of 4.77 to 9.41 kDa and isoelectric points (pI) ranging from 4.36 to 7.70. The candidate RcyaCSP genes exhibit 46.15% to 69.17% homology with other hymenopteran CSPs. Among these candidates, only RcyaCSP8 was predicted to lack a signal peptide, while RcyaCSP3 and RcyaCSP5 lacked five conserved cysteine residues (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e.in the Supplementary Materials).The multiple sequence alignment results showed that ten candidate RcyaCSPs (RcyaCSP1 and RcyaCSP4\u0026ndash;11) contained four conserved cysteine residues with the pattern X38\u0026ndash;75-Cys1-X6\u0026ndash;8-Cys2-X18\u0026ndash;19-Cys3-X2-Cys4-X37\u0026ndash;51 (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e4\u003c/span\u003e).An ML tree was constructed using CSPs from \u003cem\u003eR. cyatheae\u003c/em\u003e and 12 other hymenopteran species. The results revealed that many RcyaCSPs were highly divergent, forming distinct clades, while CSPs from the other 12 hymenopteran species clustered closely within multiple branches (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e8\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eBioinformatics analysis of identified R. cyatheae CSP genes\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"11\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGene ID\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGene Name\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eComplete ORF (aa)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eSignal Peptide\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eGroup\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003ePl\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eMW\u003c/p\u003e\u003cp\u003e(Kd)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"4\" nameend=\"c11\" namest=\"c8\"\u003e\u003cp\u003eBLAST Annotation\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eDescription\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eQuery Cover\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003eE value\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003eIdentify(%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u0026gt;TRINITY_DN2639_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaCSP1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e123\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e7.61\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e14.31\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eQGW50256.1, chemosensory protein 9 [Chouioia cunea]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e94%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e6E-33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e46.15\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u0026gt;TRINITY_DN14066_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaCSP2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e139\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e5.74\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e15.52\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_046492842.1, chemosensory protein 4 [Neodiprion pinetum]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e90%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3E-55\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e67.46\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u0026gt;TRINITY_DN20688_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaCSP3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e138\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e9.41\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e15.87\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eQHN69075.1, chemosensory protein 3 [Sirex noctilio]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e6E-59\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e63.31\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u0026gt;TRINITY_DN14055_c0_g1_\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaCSP4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e118\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e5.11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e13.74\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eBAS29775.1, chemosensory protein [Camponotus japonicus]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e95%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2E-37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e53.1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u0026gt;TRINITY_DN3807_c0_g1_\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaCSP5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e118\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e9.11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e13.36\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_072754896.1, chemosensory protein 2 [Anoplolepis gracilipes]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e6E-54\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e69.17\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u0026gt;TRINITY_DN8223_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaCSP6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e124\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e9.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e13.93\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eALG36159.1, chemosensory protein 6 [Sclerodermus sp. MQW-2015]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e7E-63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e75\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u0026gt;TRINITY_DN1049_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaCSP7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e154\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e9.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e17.54\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eALG36159.1, chemosensory protein 6 [Sclerodermus sp. MQW-2015]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e54%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e8.E-15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e67.92\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u0026gt;TRINITY_DN9812_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaCSP8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e120\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e4.89\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e13.80\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eBFW56612.1, Chemosensory protein [Polyrhachis lamellidens]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e79%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e9E-49\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e60.16\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u0026gt;TRINITY_DN6262_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaCSP9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e157\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e41\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e4.77\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e17.70\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eALG36155.1, chemosensory protein 2 [Sclerodermus sp. MQW-2015]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e95%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2E-39\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e52.14\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u0026gt;TRINITY_DN17317_c0_g1_\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaCSP10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e129\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e5.46\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e14.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eUEN71179.1, chemosensory protein 3 [Gregopimpla kuwanae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e71%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e8E-41\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e61.61\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"11\"\u003eNote: \u0026dagger;ORF: open reading frame; ND: not detected.\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"11\"\u003eAmino acid sequences of \u003cem\u003eR. cyatheae\u003c/em\u003e OBP genes are listed in the supplementary data.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eCandidate NPC2s\u003c/p\u003e\u003cp\u003eSix full-length putative RcyaNPC2 proteins encoding 144\u0026ndash;161 amino acid residues with molecular weights ranging from 4.21 kDa to 8.10 kDa were identified in R. cyatheae. These RcyaNPC2s shared 26.71\u0026ndash;66.44% amino acid sequence identity with other known insect NPC2s (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e.in Supplementary Materials).The multiple alignment revealed revealed that six NPC2s contained six conserved cysteine residues with the pattern X20\u0026ndash;24-Cys1-X13\u0026ndash;14-Cys2-X4\u0026ndash;5-Cys3-X43\u0026ndash;45-Cys4-X6\u0026ndash;13-Cys5-X40\u0026ndash;42-Cys6-X8\u0026ndash;33 (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e5\u003c/span\u003e). A Maximum likelihood (ML) tree was constructed using the CSPs of \u003cem\u003eR. cyatheae\u003c/em\u003e and ten other Hymenopteran species. The results revealed that many RcyaCSPs were highly divergent into distinct clades, while the NPC2s of \u003cem\u003eB.dioryctriae\u003c/em\u003e (BdioNPC2s), \u003cem\u003eN. vitripennis\u003c/em\u003e (NvitNPC2s), and the CSPs of C. cunea (CcunCSPs) were closely clustered in multiple clades. Specifically, RcyaNPC2a and RcyaNPC2e formed a monophyletic clade, whereas RcyaNPC2f and BdioNPC2a aggregated into an independent clade separated from the other branches (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e9\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eBioinformatics analysis of identified R. cyatheae NPC2 genes\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"11\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGene ID\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGene Name\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eComplete ORF (aa)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eSignal Peptide\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eGroup\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003ePI\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eMW\u003c/p\u003e\u003cp\u003e(Kd)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eBLAST Annotation\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eDescription\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eQuery\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003eE value\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003eIdentify(%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN8475_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaNPC2a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e151\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e5.22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e16.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_015509554.1, NPC intracellular cholesterol transporter 2 homolog a [Neodiprion lecontei]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e97%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e7E-66\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e63.51\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN10314_c0_g4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaNPC2b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e161\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e4.21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e17.56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_046433474.1, NPC intracellular cholesterol transporter 2 homolog a-like [Neodiprion fabricii]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e91%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e1E-67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e66.44\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN437_c3_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaNPC2c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e156\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e7.56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e17.19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_015514132.1, NPC intracellular cholesterol transporter 2 [Neodiprion lecontei]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e4E-54\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e52.56\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN10489_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaNPC2d\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e154\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e8.10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e17.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_046626566.1, MD-2-related lipid-recognition protein-like [Neodiprion virginianus]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e98%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e4E-48\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e49.06\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN3106_c0_g2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaNPC2e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e153\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e4.99\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e16.55\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_032685878.1, NPC intracellular cholesterol transporter 2 homolog a-like [Odontomachus brunneus]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e94%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e1E-5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e26.71\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN3972_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaNPC2f\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e144\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e7.69\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e15.86\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eXP_046489940.1NPC intracellular cholesterol transporter 2 [Neodiprion pinetum]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e86%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e7E-28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e44.8\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"11\"\u003eNote: \u0026dagger;ORF: open reading frame; ND: not detected.\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"11\"\u003eAmino acid sequences of \u003cem\u003eR. cyatheae\u003c/em\u003e OBP genes are listed in the supplementary data.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003e3.4 Identified Candidate Transmembrane Proteins\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eCandidate ORs\u003c/p\u003e\u003cp\u003eIn \u003cem\u003eR. cyatheae\u003c/em\u003e, a total of 24 putative RcyaORs encoding 57\u0026ndash;411 amino acid residues were identified. The putative RcyaORs exhibited 25.00\u0026ndash;81.82% similarity with other orthologous genes in Hymenoptera species. Among them, 79 RcyaORs were full-length and encoded 0\u0026ndash;6 transmembrane domains (TMDs) (Table\u0026nbsp;\u003cspan refid=\"Tab7\" class=\"InternalRef\"\u003e7\u003c/span\u003e.in the Supplementary Materials). The full-length RcyaOrco gene was successfully identified, encoding 114 amino acid residues with 3 TMDs,and showing 81.82% similarity to ArosOrco of Athalia rosae. As expected, RcyaOrco clustered with Orco sequences from other hymenopteran species, forming a distinct clade in the maximum likelihood (ML) tree constructed for ORs. Five ORs were segregated into the Orco clade, where they clustered with Orco orthologs from other species. Furthermore, most RcyaORs demonstrated close phylogenetic relationships with ORs from \u003cem\u003eMicroplitis mediator\u003c/em\u003e and \u003cem\u003eB.dioryctriae\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e10\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab7\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eBioinformatics analysis of identified R. cyatheae OR genes\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGene ID\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGene Name\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eComplete ORF (aa)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eGroup\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eTMD\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eBLAST Annotation\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eDescription\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eQuery\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eE value\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eIdentify(%)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN10140_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOrco\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e114\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_012253637.2, odorant receptor coreceptor [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e87%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2E-49\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e81.82\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN9632_c0_g5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e109\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048516116.1, odorant receptor 43a [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e97%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1e-24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e46.73\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN9632_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e102\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048516116.1, odorant receptor 43a [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e91%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1e-19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e56.99\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN9666_c0_g6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e119\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_015595561.1, odorant receptor Or1 isoform X1 [Cephus cinctus]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e81%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1e-24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e50.50\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN9081_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e98\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046626501.1, odorant receptor 49b-like [Neodiprion virginianus]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e98%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3e-23\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e50.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN8388_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e299\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_012266577.3, odorant receptor 4-like isoform X1 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e74%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e6e-56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e42.53\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN8388_c0_g2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eUEN71227.1, olfactory receptor 44 [Gregopimpla kuwanae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e95%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e5e-10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e47.54\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN21124_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e75\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046735418.1, odorant receptor Or2-like [Diprion similis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e82%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3e-16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e62.90\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN7376_c0_g2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e75\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046738658.1, odorant receptor 13a-like [Diprion similis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e8e-31\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e75.68\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN7376_c0_g3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e211\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046738674.1, odorant receptor 22c-like [Diprion similis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e26%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3e-07\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e48.21\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN10220_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e154\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048513569.1, odorant receptor 4-like isoform X2 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e70%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e4e-35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e62.73\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN2585_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e157\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046751421.1, gustatory and odorant receptor 24 [Diprion similis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e79%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e9e-24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e44.70\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN7376_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e57\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046738658.1, odorant receptor 13a-like [Diprion similis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e91%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e4e-14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e60.38\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN9666_c0_g7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e144\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eKYM98886.1, Odorant receptor 46a, isoform A [Cyphomyrmex costatus]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e63%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1e-34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e61.54\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN3654_c0_g2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e121\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046751356.1, odorant receptor Or1-like [Diprion similis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e98%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e5e-24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e39.53\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN9081_c0_g2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e143\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046749494.1, odorant receptor 43a-like [Diprion similis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e7e-35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e44.76\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN20577_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e114\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046749183.1, putative odorant receptor 92a isoform X1 [Diprion similis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e90%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2e-26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e57.28\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN6018_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e411\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046416448.1, odorant receptor 83a-like isoform X1 [Neodiprion fabricii]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1e-136\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e45.70\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN7376_c0_g5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e153\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046612742.1, odorant receptor 49a-like [Neodiprion virginianus]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e98%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2e-44\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e52.98\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN2418_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e388\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048504947.1, odorant receptor 46a-like [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e96%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3e-140\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e53.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN5024_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e292\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048510426.1, odorant receptor 49b-like isoform X4 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e98%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e5e-106\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e53.77\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN20952_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e71\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046736951.1, odorant receptor 83a-like [Diprion similis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e96%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3e-25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e53.77\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN4315_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e109\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048505323.1, odorant receptor 13a-like [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1e-43\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e70.64\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN9632_c0_g2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaOR 23\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e234\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_028050636.1, odorant receptor 67a [Monomorium pharaonis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e86%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e8e-04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e25.00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"9\"\u003eNote: \u0026dagger;ORF: open reading frame; ND: not detected.\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"9\"\u003eAmino acid sequences of \u003cem\u003eR. cyatheae\u003c/em\u003e OBP genes are listed in the supplementary data.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eCandidate IRs\u003c/p\u003e\u003cp\u003eIn R cyatheae, a total of 21 putative RcyaIR genes were identified, encoding 79\u0026ndash;962 amino acid residues. These putative RcyaIRs showed 45.16\u0026ndash;100.00% similarity to orthologous genes in other Hymenoptera species. Among them, five RcyaIRs were full-length and predicted to contain 0\u0026ndash;3 transmembrane domains (TMDs) (Table\u0026nbsp;\u003cspan refid=\"Tab8\" class=\"InternalRef\"\u003e8\u003c/span\u003e.in the Supplementary Materials). Phylogenetic analysis revealed that RcyaIRs diverged into several distinct subfamilies. A subset of RcyaIRs clustered within the same clades as the conserved IR25a, IR64a, IR75x, IR93a, and IR76b gene families, while the remaining RcyaIRs were distributed across other distinct phylogenetic branches (Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e11\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab8\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 8\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eBioinformatics analysis of identified R. cyatheae GR genes\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGene ID\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGene Name\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eComplete ORF (aa)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eGroup\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eTMD\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eBLAST Annotation\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eDescription\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eQuery\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eE value\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eIdentify(%)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN12238_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR23a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e197\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046587885.1, putative gustatory receptor 28a [Neodiprion lecontei]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e93%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e8E-11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e30.77\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN18654_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR28a1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e104\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046467114.2, putative gustatory receptor 28a [Neodiprion pinetum]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e84%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2E-16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e51.14\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN19044_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR28a2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e105\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eSP:Q9VM09, Putative gustatory receptor 28a\u003c/p\u003e\u003cp\u003e[Neodiprion lecontei]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e60.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3.9E\u0026thinsp;+\u0026thinsp;00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e21.7\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN14445_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR28a3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e118\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_021922466.1, putative gustatory receptor 28a [Zootermopsis nevadensis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e71%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e5E-19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e47.62\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN5103_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR28b1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e394\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_020706207.2, putative gustatory receptor 28b\u003c/p\u003e\u003cp\u003e[Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e9E-177\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e61.36\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN2953_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR28b2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e197\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_020706207.2, putative gustatory receptor 28b\u003c/p\u003e\u003cp\u003e[Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3E-78\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e58.37\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN19790_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR28b3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e99\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_020706667.2, putative gustatory receptor 28b\u003c/p\u003e\u003cp\u003e[Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e87%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3E-29\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e68.18\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN21105_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR28b4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e62\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048510231.1, putative gustatory receptor 28b\u003c/p\u003e\u003cp\u003e[Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e79%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3E-09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e58\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN20930_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR28b5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e74\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_020706667.2, putative gustatory receptor 28b\u003c/p\u003e\u003cp\u003e[Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e81%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2E-19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e63.33\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN20385_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR43a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e276\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046738260.1, gustatory receptor for sugar taste 43a-like isoform X3 [Diprion similis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e98%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e7E-180\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e90.04\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN1720_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR5a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e492\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046409931.1, gustatory receptor 5a for trehalose-like isoform X2 [Neodiprion fabricii]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e67.21\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN10840_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR68a1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e179\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046490075.2, gustatory receptor 68a-like\u003c/p\u003e\u003cp\u003e[Neodiprion pinetum]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e93%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e6.00E-25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e34.71\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN14093_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR68a2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e191\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046490075.2, gustatory receptor 68a-like\u003c/p\u003e\u003cp\u003e[Zootermopsis nevadensis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e90%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1E-05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e25.67\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN7972_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e68\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eALG36126.1, gustatory receptor 2\u003c/p\u003e\u003cp\u003e[Sclerodermus sp. MQW-2015]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e94%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3E-04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e35.06\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN12794_c0_g3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGR68a3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e87\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_059490989.1, gustatory receptor 68a-like [Neocloeon triangulifer]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e93%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.00E-05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e35.37\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"9\"\u003eNote: \u0026dagger;ORF: open reading frame;ND: not detected.\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"9\"\u003eAmino acid sequences of \u003cem\u003eR. cyatheae\u003c/em\u003e OBP genes are listed in the supplementary data.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eCandidate GRs\u003c/p\u003e\u003cp\u003eFifteen putative RcyaGRs encoding proteins ranging from 68 to 492 amino acid residues were identified in R. \u003cem\u003ecyatheae\u003c/em\u003e. These candidate genes exhibited 21.70% to 90.04% sequence similarity to known insect GRs. Among them, four RcyaGRs were full-length, while the others lacked either one or both terminal regions. The predicted number of transmembrane domains (TMDs) ranged from 0 to 7 (Table\u0026nbsp;\u003cspan refid=\"Tab9\" class=\"InternalRef\"\u003e9\u003c/span\u003e.in Supplementary Materials). Phylogenetic analysis revealed that RcyaGR43a and RcyaGR5a clustered within the fructose receptor subfamily and trehalose receptor subfamily, thus these two genes were tentatively predicted to function as sweet GRs (sugar receptors)18. Additionally, twelve RcyaGRs were distributed across different clades with GRs from ten other species, specifically within the GR28, GR62a, GR68f, and GR43a gene family clades. These clades were grouped on the basis of the GR database sequences employed in this study (Fig.\u0026nbsp;\u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e12\u003c/span\u003e)\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab9\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 9\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eBioinformatics analysis of identified R. cyatheae IR genes\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003egene ID\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGene Name\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eComplete ORF (aa)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eGroup\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eTMD\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eBLAST Annotation\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eDescription\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eQuery\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eE value\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eIdentify(%)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN11180_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaiGluR1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e574\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048513708.1, glutamate receptor ionotropic, kainate 2 isoform X3 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e98.95\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN1947_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaIR93a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e634\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_020712212.2, ionotropic receptor 93a\u003c/p\u003e\u003cp\u003e[Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e76.37\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN2322_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaiGluR2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e193\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_012251025.2, glutamate receptor ionotropic, kainate 2-like isoform X2 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e4E-88\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e70.31\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN3458_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaiGluR3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e974\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_020712340.2, glutamate receptor ionotropic, kainate 2-like isoform X2 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e94.15\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN3557_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaIR25a1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e930\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046429951.1, ionotropic receptor 25a\u003c/p\u003e\u003cp\u003e[Neodiprion fabricii]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e96%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e85.75\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN3557_c0_g2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaIR25a2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e372\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046429951.1, ionotropic receptor 25a\u003c/p\u003e\u003cp\u003e[Diprion similis]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e94.85\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN491_c6_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaiGluR4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e577\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_012265004.2, glutamate receptor ionotropic, delta-1 isoform X1 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e68.28\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN6525_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaIR25a3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e266\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_012263496.2, ionotropic receptor 25a\u003c/p\u003e\u003cp\u003e[Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e96%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1E-141\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e78.12\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN6776_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaiGluR5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e198\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_020711610.2, glutamate receptor ionotropic, kainate 2 isoform X9 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2E-137\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e100\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN745_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaiGluR6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e300\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048512589.1, glutamate receptor ionotropic, kainate 2-like [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3E-145\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e69.97\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN7652_c0_g3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaiGluR7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e867\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046435180.1, glutamate receptor ionotropic, kainate 2 isoform X1 [Neodiprion fabricii]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e98.85\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN8730_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaiGluR8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e303\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_025602091.2, glutamate receptor ionotropic, kainate 2-like [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e9E-151\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e73.18\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN8827_c0_g2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaIR2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e955\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048507714.1, glutamate receptor ionotropic, NMDA 2B isoform X2 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e97.07\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN8926_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaiGluR9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e369\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_012251025.2, glutamate receptor ionotropic, kainate 2-like isoform X2 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2E-178\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e68.02\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN8926_c0_g2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaiGluR10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e180\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048512589.1, glutamate receptor ionotropic, kainate 2-like [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e5E-76\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e66.67\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN12985_c0_g2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaIR1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e84\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eRLZ02219.1, Ionotropic receptor 122\u003c/p\u003e\u003cp\u003e[Cephus cinctus]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e73%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e5E-10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e45.16\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN5799_c0_g3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaIR75a1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e179\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048506765.1ionotropic receptor 75a-like, partial\u003c/p\u003e\u003cp\u003e[Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e6E-72\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e59.78\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN5186_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaIR75a2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e86\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048511101.1, ionotropic receptor 75a-like isoform X3 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e81%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1E-19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e55.71\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN5799_c0_g4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaIR75a3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e148\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046410795.1, ionotropic receptor 75a-like isoform X1\u003c/p\u003e\u003cp\u003e[Neodiprion fabricii]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e100%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2E-53\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e61.49\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN12543_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaIR21a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e79\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003einternal\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_048508018.1, ionotropic receptor 21a isoform X1\u003c/p\u003e\u003cp\u003e[Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3E-38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e79.75\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"9\"\u003eNote: \u0026dagger;ORF: open reading frame; ND: not detected.\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"9\"\u003eAmino acid sequences of \u003cem\u003eR. cyatheae\u003c/em\u003e OBP genes are listed in the supplementary data.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eCandidate SNMPs\u003c/p\u003e\u003cp\u003eFour putative RcyaSNMPs encoding proteins ranging from 60 to 530 amino acids were identified in \u003cem\u003eR. cyatheae\u003c/em\u003e. These genes exhibited 54.97% to 70.08% similarity to their orthologs in other hymenopteran species. Phylogenetic analysis revealed that two RcyaSNMPs belong to the SNMP1 family, whereas the other two cluster within the SNMP2 family (S10). Phylogenetic analysis revealed that two RcyaSNMP genes clustered within the SNMP1 family, whereas the other two grouped into the SNMP2 family (Table\u0026nbsp;\u003cspan refid=\"Tab10\" class=\"InternalRef\"\u003e10\u003c/span\u003e.in the Supplementary Materials). Specifically, RcyaSNMP1a was predicted to contain two transmembrane domains (TMDs), whereas RcyaSNMP2b and RcyaSNMP1 each harbored one TMD, and RcyaSNMP2a lacked detectable TMDs (Table\u0026nbsp;\u003cspan refid=\"Tab10\" class=\"InternalRef\"\u003e10\u003c/span\u003e)1. Additionally, a separate phylogenetic tree constructed using SNMPs from \u003cem\u003eR. cyatheae\u003c/em\u003e and 14 other Hymenopteran species, together with 10 CD36 homologs, revealed that all RcyaSNMP orthologs fell exclusively within the SNMP family clade and were distinct from the CD36 protein family (Fig.\u0026nbsp;\u003cspan refid=\"Fig12\" class=\"InternalRef\"\u003e13\u003c/span\u003e)\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab10\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 10\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eBioinformatics analysis of identified R. cyatheae SNMP genes\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGene ID\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGene Name\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eComplete ORF (aa)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eGroup\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eTMD\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eBLAST Annotation\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eDescription\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eQuery\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eE value\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eIdentify(%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN1927_c0_g3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaSNMP2a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e60\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_015517411.2, sensory neuron membrane protein 2 isoform X1 [Neodiprion lecontei]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e97%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e55.89\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN7126_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaSNMP2b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e434\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_046587242.1, sensory neuron membrane protein 2 isoform X2 [Neodiprion lecontei]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e98%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3E-177\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e57.48\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN5757_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaSNMP1a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e530\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ecomplete\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_012257194.1, sensory neuron membrane protein 1 isoform X1 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e99%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e71.08\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTRINITY_DN11233_c0_g1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRcyaSNMP1b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e151\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5prime_partial\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eXP_020708289.1, sensory neuron membrane protein 1 isoform X2 [Athalia rosae]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e98%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e5E-49\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e54.97\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"9\"\u003eNote: \u0026dagger;ORF: open reading frame; ND: not detected.\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"9\"\u003eAmino acid sequences of \u003cem\u003eR. cyatheae\u003c/em\u003e OBP genes are listed in the supplementary data.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\u003ch2\u003e3.5 Analysis of Candidate Differentially Expressed Genes\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eIn this study, the expression levels of sex-biased differentially expressed genes in \u003cem\u003eR. cyatheae\u003c/em\u003e were evaluated on the basis of their FPKM values, as shown in the heatmap (Fig.\u0026nbsp;\u003cspan refid=\"Fig13\" class=\"InternalRef\"\u003e6\u003c/span\u003e. In the Supplementary Materials). Among these chemosensory genes, RcyaOBP7, RcyaOBP8, RcyaNPC2d, RcyaNPC2e, RcyaSNMP2a, and RcyaOR19 were expressed at significantly higher levels in male adults than in female adults, whereas the remaining genes were expressed at higher levels in female adults.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\u003ch2\u003e3.6 Profiling of Chemosensory Genes in R. cyatheae\u003c/h2\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eThe tissue-specific expression profiles of 13 newly identified candidate chemosensory genes (2 RcyaCSPs, 4 RcyaORs, 2 RcyaIRs, 1 RcyaGR, 1 RcyaSNMP, and 2 RcyaNPC2) in \u003cem\u003eR. cyatheae\u003c/em\u003e were investigated via RT‒qPCR (Fig.\u0026nbsp;\u003cspan refid=\"Fig14\" class=\"InternalRef\"\u003e14\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003e(Fant and Mant: female and male antennae; Fhea 和 Mhea: female and male heads without an-tennae; Fab: female abdomens without ovipositors and digestive tracts, Fov: female ovipositors; Mge: male genitalia; Fabd and Mabd: male abdomens; Fwin and Mwin : male and female wings; Ftho and Mtho: male and female thorax;Fleg and Mleg: male and female legs).The error bars represents standard errors and the small letters above each bar indicate significant differences in transcript abundances (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eWith the development of transcriptome and genome research technologies, it has become increasingly important to construct insect transcriptome database-based identification methods for chemosensory genes to facilitate the study of chemosensory mechanisms [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e].To date, the olfactory mechanisms mediating host location and oviposition site selection in \u003cem\u003eR. cyatheae\u003c/em\u003e remain unknown. Therefore, in this study identified 90 candidate genes encoding olfactory-related proteins from the adult transcriptome of \u003cem\u003eR. cyatheae\u003c/em\u003e, including 11 OBPs, 10 CSPs, 24 ORs, 15 GRs, 20 IRs, 4 SNMPs, and 6 NPC2s. The expression profiles of 13 differentially expressed chemosensory genes were subsequently validated across different tissues using RT‒qPCR, providing insights for exploring the functions of these genes.It provides a molecular basis for the systematic study of the chemosensory mechanism in \u003cem\u003eR cyatheae\u003c/em\u003e .\u003c/p\u003e\u003cp\u003eOBPs are hydrophobic binding proteins that can bind to and transport odorant substances (including pheromones and plant volatiles) through the lymph of sensilla [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].Within hymenopteran insects, some species possess only a few odorant-binding proteins (OBPs). For instance, Scleroderma guani has as few as two OBPs [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e];there are 4 OBPs in Macrocentrus cingulum [\u003cspan additionalcitationids=\"CR33\" citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e].Eleven OBP genes were identified in the transcriptome of R. cyatheae, a relatively low number comparable to that observed in IkuwOBP [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e].This number is significantly lower than the number identified in N. vitripennis (90 OBPs). The differences in the number of OBPs among insects may be influenced by variations in different species, sexes, habitats, types of odor molecules, and feeding ranges [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].It may also result from the limitations of transcriptome samples and differences in sequencing methods. For example, samples based solely on the adult antenna transcriptome may exclude some OBP genes expressed in other tissues or at different developmental stages [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e].Phylogenetic analysis revealed that the 11 RcyaOBPs were distributed across different clades alongside OBPs from nine hymenopteran species, suggesting potential functional divergence among RcyaOBPs. Specifically, RcyaOBP7 clustered within the same clade as IkuwOBP and AmelOBP, whereas RcyaOBP8 formed a clade with CcunOBP and CvesOBP. The expression levels of these two differentially expressed OBP genes were further quantified via RT‒qPCR across various tissues. The highest expression of RcyaOBP7 was detected in female antennae. Numerous studies have demonstrated that OBPs with antenna-specific and highly abundant expression play critical roles in binding key volatile compounds during host location and foraging processes in hymenopteran insects[\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e].For example when the AmelOBP4 gene is knocked out, the preference for plant volatiles (such as linalool and 1-octen-3-ol) and sex pheromones (ethyl oleate, methyl palmitate, methyl linoleate, and β-ocimene) is significantly reduced [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. The expression of RcyaOBP8 was found to be significantly greater in male heads than in other tissues, whereas its expression was markedly reduced in female heads compared with other tissues. These findings suggested that RcyaOBP8 might function in pheromone recognition or male-specific behaviors. For instance, SfruOBP31 is highly expressed in the abdomen, adult heads, and male reproductive organs of \u003cem\u003eSpodoptera frugiperda\u003c/em\u003e. Knockout of the SfruOBP31 gene was demonstrated to disrupt larval phototaxis and male reproductive processes[\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e].The functions of these OBP genes merit further study.\u003c/p\u003e\u003cp\u003eCSPs are soluble proteins that perform functions similar to those of odorant-binding proteins (OBPs). As the second category of binding proteins in the insect chemosensory system, they are more conserved than OBPs [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e],(including heads, thoraxes, labial palps, tarsi, pheromone glands, and ejaculatory ducts)These proteins are widely distributed across both olfactory tissues (such as antennae, legs, wings, and proboscis) and non-olfactory tissues (including heads, thoraxes, labial palps, tarsi, pheromone glands, and ejaculatory ducts) [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. For example, after silencing the NlugCSP8 gene in olfactory tissues, the response of \u003cem\u003eNilaparvata lugens\u003c/em\u003e to hexanal is significantly inhibited, and the attractiveness of nerol to \u003cem\u003eN. lugen\u003c/em\u003es is lost [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e].In addition to their olfactory functions, CSPs may also be involved in insect development, nutrient absorption, and insecticide resistance [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].For instance, knockout of the ovary- specific gene OcomCSP12 leads to a significant reduction in ovarian specificity [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]; Additionally ,silencing of DlonCSP3 impairs the foraging behavior of \u003cem\u003eDiachasmimorpha longicaudata\u003c/em\u003e[\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e].Ten CSP genes were identified in the adult transcriptome of \u003cem\u003eR. cyatheae\u003c/em\u003e, with numbers is similar to those of BdioCSPs[\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]、SspCSPs[\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]、EforCSPs[\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e] among others.According to the RT‒qPCR results, we observed that RcyaCSP10 exhibitted female antennae-biased expression, whereas RcyaCSP2 demonstrates distinct tissue specificity with notably low expression in female heads. The detailed functions of these two RcyaCSPs in \u003cem\u003eR. cyatheae\u003c/em\u003e require further investigation.\u003c/p\u003e\u003cp\u003eNPC2 is considered a soluble small-molecule protein and a member of the MD-2-related lipid recognition (ML) protein superfamily, which includes myeloid differentiation factor-2 (MD-2) [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e].It serves as a carrier for transporting semiochemicals and other hydrophobic compounds, such as cholesterol[\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e].In the NPC2 gene family, 6 NPC2 genes were identified, making up a larger number than that in \u003cem\u003eB.dioryctriae\u003c/em\u003e (3 NPC2 genes) [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e] and in \u003cem\u003eD. longicaudata\u003c/em\u003e(4 NPC2 genes) [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e].Previous studies on hymenopteran insects have demonstrated that NPC2 is highly expressed in both antennae and ovipositors [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e] .In this study, RcyaNPC2d and RcyaNPC2e were expressed in antennae, head, legs, thorax, abdomen, and wings, but their expression levels were significantly higher in male heads than in female heads. This phenomenon might be attributed to the functional divergence of NPC2 proteins.RcyaNPC2d and RcyaNPC2e, which exhibit this distinct expression pattern, could serve as target genes to investigate their physiological functions in \u003cem\u003eR. cyatheae\u003c/em\u003e.\u003c/p\u003e\u003cp\u003eInsect ORs genes serve as crucial chemoreceptor organs within insect olfactory systems, evolutionarily derived from GRs [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e]. These receptors are sensitive to many different types of compounds, including insect pheromone components and host plant compounds, such as esters, alcohols, and ketones[\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e]. Within the RcyaOR gene family, 23 OR genes and one Orco gene were identified, representing a relatively small number. In contrast, some Hymenopteran insects possess more than one hundred OR genes; for instance, \u003cem\u003eApis cerana\u003c/em\u003e has 113 OR genes. [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e] and \u003cem\u003eApis florea\u003c/em\u003e has 180 OR genes [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e].The variation in OR gene numbers may be influenced by factors such as interspecies variation, as well as the depth and breadth of sequencing methodologies [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e].Within hymenopteran insect species, a single Orco gene is present, which functions as a highly conserved coreceptor [\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e].In many insect species, the absence of the Orco gene leads to reduced sex pheromone recognition, decreased oviposition preference, and impaired larval attraction to host plants [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e, \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e].According to the RT‒qPCR results, OR14, OR17, OR19, and OR20 were expressed across all 12 examined tissues, with the lowest expression levels observed in female heads. This pattern may reflect functional differentiation among OR genes. Compared with those in Lepidoptera and Coleoptera, numerous OR genes have been identified in hymenopteran species in recent years; however, the functional characterization of these genes remains insufficient despite their quantitative abundance.Screening for specifically expressed ORs may be an effective method for narrowing the range of host localized target genes. Four RcyaOR genes with sex-biased expression in males and females were selected for subsequent studies.\u003c/p\u003e\u003cp\u003eIRs are receptor genes that are evolutionarily derived from ionotropic glutamate receptors (iGluRs). They are expressed in gustatory receptor neurons, respond to diverse odorants, and represent a highly conserved family of ligand-gated ion channels [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e].In IR gene family, 20 IR genes were identified, representing a number similar to those in \u003cem\u003eAphidius gifuensis\u003c/em\u003e (25 IRs)[\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e] and \u003cem\u003eBombus impatiens\u003c/em\u003e(23个IR)[\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e].In this study, ten iGluRs were identified. Given that iGluRs are ancestral precursors of IRs, the limited evolutionary expansion of iGluRs into IRs in \u003cem\u003eR. cyatheae\u003c/em\u003e may be associated with its specialized feeding habits.iGluR4 was expressed in all 12 tissues but showed significant sex-specific expression differences in the head. iGluR4 may play a role in odor recognition and localization[\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e], but further experimental validation.Furthermore, as an ancient chemoreceptor family, insect IRs can be classified into several distinct subfamilies. Phylogenetic analysis revealed that the 20 RcyaIRs identified in \u003cem\u003eR. cyatheae\u003c/em\u003e are distributed among typical subfamilies, including iGluR, IR25a, IR64a, IR75a, IR76b, and IR93a. Notably, three IR25a orthologs were identified in \u003cem\u003eR. cyatheae\u003c/em\u003e. and three IR25a homologs were identified in \u003cem\u003eR. cyatheae\u003c/em\u003e. However, compared with IR25a1 and IR25a2,IR25a3 clustered in a distinct phylogenetic clade, suggesting functional divergence of the IR25a gene. The coreceptor family of IR25a shares similar distribution patterns and functional profiles with the Orco family, although they are expressed in distinct types of sensilla[\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e].The IR25a gene mediates cold temperature and humidity changes in \u003cem\u003eDrosophila melanogaster\u003c/em\u003e[\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e, \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e]. \u003cem\u003eR. cyatheae\u003c/em\u003e survives prolonged winter months, suggesting that the RcyaIR25 gene may also be involved in thermosensation.The Ir75a gene arose through duplication in the ancestor of the family \u003cem\u003eDrosophilidae\u003c/em\u003e (~\u0026thinsp;30\u0026ndash;35\u0026nbsp;million years ago) and exhibits differential sensitivity to carboxylic acids [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e, \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e].Furthermore, the primary host plant of \u003cem\u003eR. cyatheae\u003c/em\u003e is \u003cem\u003eA. spinulosa\u003c/em\u003e, which releases volatile compounds such as acetic acid and ethyl acetate into the environment [\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e] .These odorants may be detected by Ir75a in \u003cem\u003eR. cyatheae\u003c/em\u003e, thus, RcyIr75a likely participates in host localization in this species, but further experimental validation is needed.\u003c/p\u003e\u003cp\u003eInsects rely on a unique family of GRs, which function as tetrameric ligand-gated cation channels [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e].These receptors primarily mediate the perception of carbon dioxide, fructose, various sugars, bitter compounds, and other chemosensory stimuli [\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e].Fifteen GR genes were identified in the RcyaGR gene family of \u003cem\u003eR. cyatheae\u003c/em\u003e, which is close to \u003cem\u003eAtherigona orientalis\u003c/em\u003e (20 GRs) [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e].In hymenopteran insects, the number of GR genes identified via transcriptomic techniques ranges from as few as 2 (\u003cem\u003eM. mediator\u003c/em\u003e) [\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e], to 49 (\u003cem\u003eB.dioryctriae\u003c/em\u003e)[\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e].Three RcyaIR genes were identified in the transcriptome of R. cyatheae and predicted to function as sweet gustatory receptors (GRs).In \u003cem\u003eD melanogaster\u003c/em\u003e, GR43a is crucial for detecting fructose and sucrose receptor crucial for detecting fructose levels in the brain. It is also expressed in neurons of the proventricular ganglion and uterus. Four BidoGR43a.1\u0026ndash;4 genes and one BidoGR64f gene cluster are located within the fructose receptor and trehalose receptor subfamilies, respectively [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e].In \u003cem\u003eD.melanogaster\u003c/em\u003e, GR43a is crucial for detecting fructose levels in the brain. It is also expressed in neurons of the proventricular ganglion and uterus. Four BidoGR43a.1\u0026ndash;4 genes and one BidoGR64f gene cluster within the fructose receptor and trehalose receptor subfamilies, respectively.Additionally, DmelGR5a encodes as a receptor for trehalose and clusters within the trehalose receptor subfamily alongside :DmelGR64f [\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e, \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\u003c/span\u003e].Phylogenetic analysis revealed that RcyaGR5a clustered with RcyaGR64f and BidoGR64f in the same clade, indicating that RcyaGR5a and RcyaGR64f belonged to to the trehalose receptor subfamily. The GR68a protein is likely involved in pheromone-driven courtship behavior in Drosophila[\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e].In this study, RcyaGR68a clustered within a branch containing numerous GRs, suggesting potential functional divergence. No candidate GRs from the CO₂ receptor subfamily were identified in \u003cem\u003eR. cyatheae\u003c/em\u003e, possibly because of the lack of a dedicated transcriptome database for the labial palps of the sawfly [\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e, \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e76\u003c/span\u003e].These RcyaGR receptors in \u003cem\u003eR. cyatheae\u003c/em\u003e may perceive sweet and bitter sensations, warranting further investigation.\u003c/p\u003e\u003cp\u003eSNMPs are transmembrane structural proteins that were initially identified as membrane-bound proteins in the olfactory receptor neurons of lepidopteran insects. They are proposed to play a role in odor detection[\u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e77\u003c/span\u003e, \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e78\u003c/span\u003e].Four SNMP genes were identified in \u003cem\u003eR.cyatheae\u003c/em\u003e, which is more than in \u003cem\u003eM.mediator\u003c/em\u003e (2) and \u003cem\u003eSclerodermus\u003c/em\u003e sp. (2) [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e, \u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e79\u003c/span\u003e].Numerous studies have demonstrated that most insects possess two types of SNMPs, namely SNMP1 and SNMP2[\u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e80\u003c/span\u003e].The RcyaSNMPs identified in this study belong to both the SNMP1 and SNMP2 subfamilies.The paralogs of the same SNMP type in \u003cem\u003eR. cyatheae\u003c/em\u003e are similar to those in B.dioryctriae and \u003cem\u003eIseropus kuwanae\u003c/em\u003e, with the former encoding six SNMP1 genes and the latter encoding two SNMP genes[\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e].In \u003cem\u003eD.melanogaster\u003c/em\u003e, SNMP1 has been demonstrated to be critical for detecting the pheromone cis-vaccenyl acetate (CVA) [\u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e81\u003c/span\u003e].MmedSNMP1 is expressed primarily in the sensilla of the antennae and may be involved in the perception of plant volatiles and sex pheromones[\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e].Therefore, phylogenetic analysis revealed that MmedSNMP1 clustered with RcyaSNMP1a and RcyaSNMP1b in the same clade, suggesting that RcyaSNMP1a and RcyaSNMP1b might participate in the perception of plant volatiles and sex pheromones. However, functional validation is needed to confirm this hypothesis. The current t literature on SNMP2 remains limited, with no functional studies reported to date. In this study, RcyaSNMP2a was expressed across all the examined tissues[\u003cspan citationid=\"CR82\" class=\"CitationRef\"\u003e82\u003c/span\u003e], albeit at minimal levels.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e"},{"header":"5. Conclusions","content":"\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eOn the basis of the transcriptome database of female and male adults of \u003cem\u003eR. cyatheae\u003c/em\u003e, this study identified 90 candidate chemosensory genes were identified in this study. RT‒qPCR revealed distinct expression patterns of 13 chemosensory genes. This work preliminarily narrows down the range of candidate chemosensory genes involved in recognizing key information during host localization and oviposition processes, laying a foundation for further exploration of the molecular mechanisms underlying chemosensory perception in \u003cem\u003eR. cyatheae\u003c/em\u003e and the development of novel biological control strategies.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eEthics approval and consent to participate:not applicable.\u003c/p\u003e\n\u003cp\u003eConsent for publication: not applicable\u003c/p\u003e\n\u003cp\u003eFunding\u003c/p\u003e\n\u003cp\u003eThe author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was supported by \u0026ldquo;Pest resistance mechanisms of different Cyathea spinosa based on three-generation full-length transcriptomes, two-generation transcriptomes, and protein metabolomes\u0026rdquo; (Grant No. 11904\u0026ndash;0522093),and the Science and Technology Innovation Talent Team Building Project of the Science and Technology Innovation Talent Team Building Project of Guizhou Province (Project No.: Qiankehepingtairencai-CXTD [2023]010).\u003c/p\u003e\n\u003cp\u003eCRediT authorship contribution statement\u003c/p\u003e\n\u003cp\u003eXiao-Ming Li and Ya-Nan Zhang designed the experiments.\u003c/p\u003e\n\u003cp\u003eQiang Liu, Sai Ma, Mao-Zhu Yin, Nan Gu, and Li-Fu Qian performed the experiments.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eXiao-Ming Li, Qiang Liu, and Ya-Nan Zhang analysed the data.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eXiao-Ming Li, Qiang Liu, and Ya-Nan Zhang wrote and revised the article.\u003c/p\u003e\n\u003cp\u003eAcknowledgement\u003c/p\u003e\n\u003cp\u003eFirst and foremost, I sincerely thank my supervisor, Professor Yang Weicheng. Throughout this study, Professor Yang not only provided guidance on the development of the research design and experimental protocol but also assisted in revising the statistical methods during the data analysis phase. Additionally, he reviewed the initial draft of the paper on multiple occasions and offered important revision suggestions, which further refined the research framework.Secondly, I would like to express my gratitude to the Administration of Guizhou Chishui Alsophila National Nature Reserve for its strong support for this experiment.Furthermore, I appreciate the technical support provided by all teachers in the research group during the experiment, as well as the assistance from my fellow group members in sample processing.This study was funded by Pest resistance mechanisms of different Cyathea spinosa based on three-generation full-length transcriptomes, two-generation transcriptomes, and protein metabolomes (Project No.: 11904\u0026ndash;0522093) and the Science and Technology Innovation Talent Team Building Project of the Science and Technology Innovation Talent Team Building Project of Guizhou Province (Project No.: Qiankehepingtairencai-CXTD [2023]010), and then I would like to extend my sincere thanks for this financial support.Finally, I am grateful to my family for their understanding and support throughout the research process.\u003c/p\u003e\n\u003cp\u003eDisclosure\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e\n\u003cp\u003eAvailability of data and materials\u003c/p\u003e\n\u003cp\u003eAll data generated or analyzed during this study are included in this published article (and its supplementary information files). The raw sequence dataset is available at the National Center for Biotechnology Information (NCBI) under the SRA Bioproject number PRJNA1309270.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eTian Z, Sun L, Li Y, Quan L, Zhang H, Yan W, et al. Antennal transcriptome analysis of the chemosensory gene families in carposina sasakii (lepidoptera: Carposinidae). BMC Genomics. 2018;19:544. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12864-018-4900-x\u003c/span\u003e\u003cspan address=\"10.1186/s12864-018-4900-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMa X, Lu X, Zhang P, Deng X, Bai J, Xu Z, et al. Transcriptome analysis of antennal chemosensory genes in curculio dieckmanni faust. (coleoptera: Curculionidae). Front Physiol. 2022;13. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fphys.2022.896793\u003c/span\u003e\u003cspan address=\"10.3389/fphys.2022.896793\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVosshall LB, Amrein H, Morozov PS, Rzhetsky A, Axel R. A spatial map of olfactory receptor expression in the drosophila antenna. Cell. 1999;96:725\u0026ndash;36. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/s0092-8674(00)80582-6\u003c/span\u003e\u003cspan address=\"10.1016/s0092-8674(00)80582-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDippel S, Oberhofer G, Kahnt J, Gerischer L, Opitz L, Schachtner J, et al. Tissue-specific transcriptomics, chromosomal localization, and phylogeny of chemosensory and odorant binding proteins from the red flour beetle tribolium castaneum reveal subgroup specificities for olfaction or more general functions. BMC Genomics. 2014;15:1141. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/1471-2164-15-1141\u003c/span\u003e\u003cspan address=\"10.1186/1471-2164-15-1141\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGuo S, Liu P, Tang Y, Chen J, Zhang T, Liu H. Identification and expression profiles of olfactory-related genes in the antennal transcriptome of graphosoma rubrolineatum (hemiptera: Pentatomidae). PLoS ONE. 2024;19:e0306986. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0306986\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0306986\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhou H, Yan H, Wang E, Zhang B, Xu X. Expression and functional analysis of niemann-pick C2 gene in phytoseiulus persimilis. Exp Appl Acarol. 2023;89:201\u0026ndash;13. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10493-023-00781-8\u003c/span\u003e\u003cspan address=\"10.1007/s10493-023-00781-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKo DC, Binkley J, Sidow A, Scott MP. The integrity of a cholesterol-binding pocket in niemann-pick C2 protein is necessary to control lysosome cholesterol levels. Proc Natl Acad Sci U S A. 2003;100:2518\u0026ndash;25. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1073/pnas.0530027100\u003c/span\u003e\u003cspan address=\"10.1073/pnas.0530027100\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePelosi P, Iovinella I, Zhu J, Wang G, Dani FR. Beyond chemoreception: Diverse tasks of soluble olfactory proteins in insects. Biol Rev. 2018;93:184\u0026ndash;200. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/brv.12339\u003c/span\u003e\u003cspan address=\"10.1111/brv.12339\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWang JJ, Ma C, Yue Y, Yang J, Chen LX, Wang YT, et al. Identification of candidate chemosensory genes in bactrocera cucurbitae based on antennal transcriptome analysis. Front Physiol. 2024;15. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fphys.2024.1354530\u003c/span\u003e\u003cspan address=\"10.3389/fphys.2024.1354530\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRimal S, Lee Y. The multidimensional ionotropic receptors of drosophila melanogaster. Insect Mol Biol. 2018;27:1\u0026ndash;7. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/imb.12347\u003c/span\u003e\u003cspan address=\"10.1111/imb.12347\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKozma MT, Schmidt M, Ngo-Vu H, Sparks SD, Senatore A, Derby CD. Chemoreceptor proteins in the caribbean spiny lobster, panulirus argus: Expression of ionotropic receptors, gustatory receptors, and TRP channels in two chemosensory organs and brain. PLoS ONE. 2018;13:e0203935. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0203935\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0203935\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAgnihotri AR, Roy AA, Joshi RS. Gustatory receptors in lepidoptera: Chemosensation and beyond. Insect Mol Biol. 2016;25:519\u0026ndash;29. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/imb.12246\u003c/span\u003e\u003cspan address=\"10.1111/imb.12246\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMang D, Shu M, Endo H, Yoshizawa Y, Nagata S, Kikuta S, et al. Expression of a sugar clade gustatory receptor, BmGr6, in the oral sensory organs, midgut, and central nervous system of larvae of the silkworm bombyx mori. Insect Biochem Mol Biol. 2016;70:85\u0026ndash;98. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ibmb.2015.12.008\u003c/span\u003e\u003cspan address=\"10.1016/j.ibmb.2015.12.008\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKang Z-W, Tian H-G, Liu F-H, Liu X, Jing X-F, Liu T-X. Identification and expression analysis of chemosensory receptor genes in an aphid endoparasitoid aphidius gifuensis. Sci Rep. 2017;7:3939. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41598-017-03988-z\u003c/span\u003e\u003cspan address=\"10.1038/s41598-017-03988-z\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhang B-C, Yang W-C, Xiao J-X, Bai X-J, Chen H-D. Effects of different host plants on the growth,development and gut bacterial communi-ty of Rhoptroceros cyatheae larvae [J]. J Environ Entomol. 2024;46:886\u0026ndash;96.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eXu D-S, Yang W-C,Wong T, He Q-Q,Liang S, Zhang T-Y. Biological Characteristics and Larval Population Dynamics of Rhoptroceros cyatheae in Chishui Guizhou[J]. Chin J Biol Control. 2021;37(2):218\u0026ndash;27. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.16409/j.cnki.2095-039x.2020.06.001\u003c/span\u003e\u003cspan address=\"10.16409/j.cnki.2095-039x.2020.06.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eYang J, Yang W-C,Wu G-Y, Che B-J, Liang H-F, Zhou B-B. Optimizationof Tissue Culture and Propagation System of Alsophila spinulosa Based on PGGB Path way[J]. Acta Bot Boreali-Occidentalia Sinica. 2023;43:1488\u0026ndash;98.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLiu Y-L, Zhang L-N, Liang L, Zheng Y-Q, et al. Diversity of endophytic fungi from Alsophila spinulosa in ChishuiAlsophila National Nature Reserve, Guizhou Province, Southwest China. Mycosystema. 2021;40:2673\u0026ndash;84. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.13346/j.mycosystema.210177\u003c/span\u003e\u003cspan address=\"10.13346/j.mycosystema.210177\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhang B, Yang W-C, He Q-Q, Chen H-D, Che B-J, Bai X-J. Analysis of differential effects of host plants on the gut microbes of rhoptroceros cyatheae. Front Microbiol. 2024;15:1392586. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fmicb.2024.1392586\u003c/span\u003e\u003cspan address=\"10.3389/fmicb.2024.1392586\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWang X, Ren Y-Z, Huang Q,Deng X-B, Chen C-W. Deng H.-P.Habitat suitability assessment of endangered plant Alsophila spi-nulosa in Chishui River area based on GIS and Maxent model. Acta Ecol Sin. 2021;41:6123\u0026ndash;33.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChen S, Zhou Y, Chen Y, Gu J. fastp: An ultra-fast all-in-one FASTQ preprocessor. Bioinforma Oxf Engl. 2018;34:i884\u0026ndash;90. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/bioinformatics/bty560\u003c/span\u003e\u003cspan address=\"10.1093/bioinformatics/bty560\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChen S. Ultrafast one-pass FASTQ data preprocessing, quality control, and deduplication using fastp. iMeta. 2023;2:e107. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/imt2.107\u003c/span\u003e\u003cspan address=\"10.1002/imt2.107\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGrabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, et al. Full-length transcriptome assembly from RNA-seq data without a reference genome. Nat Biotechnol. 2011;29:644\u0026ndash;52. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/nbt.1883\u003c/span\u003e\u003cspan address=\"10.1038/nbt.1883\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKumar S, Stecher G, Suleski M, Sanderford M, Sharma S, Tamura K. MEGA12: Molecular evolutionary genetic analysis version 12 for adaptive and green computing. Mol Biol Evol. 2024;41:msae263. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/molbev/msae263\u003c/span\u003e\u003cspan address=\"10.1093/molbev/msae263\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol. 2018;35:1547\u0026ndash;9. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/molbev/msy096\u003c/span\u003e\u003cspan address=\"10.1093/molbev/msy096\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLiu F, Ye Z, Baker A, Sun H, Zwiebel LJ. Gene editing reveals obligate and modulatory components of the CO2 receptor complex in the malaria vector mosquito, anopheles coluzzii. Insect Biochem Mol Biol. 2020;127:103470. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ibmb.2020.103470\u003c/span\u003e\u003cspan address=\"10.1016/j.ibmb.2020.103470\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChowdhury HA, Bhattacharyya DK, Kalita JK. Differential expression analysis of RNA-seq reads: Overview, taxonomy, and tools. IEEE/ACM Trans Comput Biol Bioinform. 2020;17:566\u0026ndash;86. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1109/TCBB.2018.2873010\u003c/span\u003e\u003cspan address=\"10.1109/TCBB.2018.2873010\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVaes E, Khan M, Mombaerts P. Statistical analysis of differential gene expression relative to a fold change threshold on NanoString data of mouse odorant receptor genes. BMC Bioinformatics. 2014;15:39. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/1471-2105-15-39\u003c/span\u003e\u003cspan address=\"10.1186/1471-2105-15-39\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKang Z-W, Tian H-G, Liu F-H, Liu X, Jing X-F, Liu T-X. Identification and expression analysis of chemosensory receptor genes in an aphid endoparasitoid aphidius gifuensis. Sci Rep. 2017;7:3939. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41598-017-03988-z\u003c/span\u003e\u003cspan address=\"10.1038/s41598-017-03988-z\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHu P, Tao J, Cui M, Gao C, Lu P, Luo Y. Antennal transcriptome analysis and expression profiles of odorant binding proteins in eogystia hippophaecolus (lepidoptera: Cossidae). BMC Genomics. 2016;17:651. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12864-016-3008-4\u003c/span\u003e\u003cspan address=\"10.1186/s12864-016-3008-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLu D, Li X, Liu X, Zhang Q. Identification and molecular cloning of putative odorant-binding proteins and chemosensory protein from the bethylid wasp, scleroderma guani xiao et wu. J Chem Ecol. 2007;33:1359\u0026ndash;75. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10886-007-9310-5\u003c/span\u003e\u003cspan address=\"10.1007/s10886-007-9310-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAhmed T, Zhang T, Wang Z, He K, Bai S. C-terminus methionene specifically involved in binding corn odorants to odorant binding Protein4 in macrocentrus cingulum. Front Physiol. 2017;8:62. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fphys.2017.00062\u003c/span\u003e\u003cspan address=\"10.3389/fphys.2017.00062\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAhmed T, Zhang T, Wang Z, He K, Bai S. Three amino acid residues bind corn odorants to McinOBP1 in the polyembryonic endoparasitoid of macrocentrus cingulum brischke. PLoS ONE. 2014;9:e93501. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0093501\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0093501\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAhmed T, Zhang T, Wang Z, He K, Bai S. Molecular cloning, expression profile, odorant affinity, and stability of two odorant-binding proteins in macrocentrus cingulum brischke (hymenoptera: Braconidae). Arch Insect Biochem Physiol. 2017;94:e21374. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/arch.21374\u003c/span\u003e\u003cspan address=\"10.1002/arch.21374\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLi Y, Chen H, Liang X, Wang S, Zhu H, Yan M, et al. Identification of candidate chemosensory genes by antennal transcriptome analysis in an ectoparasitoid wasp. J Appl Entomol. 2022;146:335\u0026ndash;51. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/jen.12962\u003c/span\u003e\u003cspan address=\"10.1111/jen.12962\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVieira FG, For\u0026ecirc;t S, He X, Rozas J, Field LM, Zhou J-J. Unique features of odorant-binding proteins of the parasitoid wasp nasonia vitripennis revealed by genome annotation and comparative analyses. PLoS ONE. 2012;7:e43034. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0043034\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0043034\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWu Z-R, Fan J-T, Tong N, Guo J-M, Li Y, Lu M, et al. Transcriptome analysis and identification of chemosensory genes in the larvae of plagiodera versicolora. BMC Genomics. 2022;23:845. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12864-022-09079-2\u003c/span\u003e\u003cspan address=\"10.1186/s12864-022-09079-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLi F, Venthur H, Lin K, Zhang C, Chen Z, Zhou J-J. Insect chemosensory proteins as targets in insecticide resistance and development. New Plant Prot. 2025;2:e70008. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/npp2.70008\u003c/span\u003e\u003cspan address=\"10.1002/npp2.70008\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLiu N-Y, Li Z-B, Zhao N, Song Q-S, Zhu J-Y, Yang B. Identification and characterization of chemosensory gene families in the bark beetle, tomicus yunnanensis. Comp Biochem Physiol Part D Genomics Proteom. 2018;25:73\u0026ndash;85. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cbd.2017.11.003\u003c/span\u003e\u003cspan address=\"10.1016/j.cbd.2017.11.003\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHan W-K, Tang F-X, Yan Y-Y, Wang Y, Zhang Y-X, Yu N, et al. An OBP gene highly expressed in non-chemosensory tissues affects the phototaxis and reproduction of spodoptera frugiperda. Insect Mol Biol. 2024;33:81\u0026ndash;90. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/imb.12880\u003c/span\u003e\u003cspan address=\"10.1111/imb.12880\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eJia C, Mohamed A, Cattaneo AM, Huang X, Keyhani NO, Gu M, et al. Odorant-binding proteins and chemosensory proteins in spodoptera frugiperda: From genome-wide identification and developmental stage-related expression analysis to the perception of host plant odors, sex pheromones, and insecticides. Int J Mol Sci. 2023;24:5595. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/ijms24065595\u003c/span\u003e\u003cspan address=\"10.3390/ijms24065595\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhu J, Iovinella I, Dani FR, Pelosi P, Wang G. Chemosensory proteins: a versatile binding family. Olfactory Concepts of Insect Control - Alternative to Insecticides. Cham: Springer; 2019. pp. 147\u0026ndash;69. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/978-3-030-05165-5_6\u003c/span\u003e\u003cspan address=\"10.1007/978-3-030-05165-5_6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWaris MI, Younas A, Ul Qamar MT, Hao L, Ameen A, Ali S, et al. Silencing of chemosensory protein gene NlugCSP8 by RNAi induces declining behavioral responses of nilaparvata lugens. Front Physiol. 2018;9:379. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fphys.2018.00379\u003c/span\u003e\u003cspan address=\"10.3389/fphys.2018.00379\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMa C, Cui S, Tian Z, Zhang Y, Chen G, Gao X, et al. OcomCSP12, a chemosensory protein expressed specifically by ovary, mediates reproduction in ophraella communa (coleoptera: Chrysomelidae). Front Physiol. 2019;10:1290. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fphys.2019.01290\u003c/span\u003e\u003cspan address=\"10.3389/fphys.2019.01290\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWulff JP, Segura DF, Devescovi F, Muntaabski I, Milla FH, Scannapieco AC, et al. Identification and characterization of soluble binding proteins associated with host foraging in the parasitoid wasp diachasmimorpha longicaudata. PLoS ONE. 2021;16:e0252765. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0252765\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0252765\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhu X, Yu Q, Gan X, Song L, Zhang K, Zuo T, et al. Transcriptome analysis and identification of chemosensory genes in baryscapus dioryctriae (hymenoptera: Eulophidae). Insects. 2022;13:1098. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/insects13121098\u003c/span\u003e\u003cspan address=\"10.3390/insects13121098\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhou C-X, Min S-F, Yan-Long T, Wang M-Q. Analysis of antennal transcriptome and odorant binding protein expression profiles of the recently identified parasitoid wasp, sclerodermus sp. Comp Biochem Physiol Part D Genomics Proteom. 2015;16:10\u0026ndash;9. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cbd.2015.06.003\u003c/span\u003e\u003cspan address=\"10.1016/j.cbd.2015.06.003\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHe Y, Wang K, Zeng Y, Guo Z, Zhang Y, Wu Q, et al. Analysis of the antennal transcriptome and odorant-binding protein expression profiles of the parasitoid wasp encarsia formosa. Genomics. 2020;112:2291\u0026ndash;301. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ygeno.2019.12.025\u003c/span\u003e\u003cspan address=\"10.1016/j.ygeno.2019.12.025\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhu J, Guo M, Ban L, Song L-M, Liu Y, Pelosi P, et al. Niemann-pick C2 proteins: A new function for an old family. Front Physiol. 2018;9:52. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fphys.2018.00052\u003c/span\u003e\u003cspan address=\"10.3389/fphys.2018.00052\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eThambi PJ, Modahl CM, Kini RM. Niemann-pick type C2 proteins in aedes aegypti: Molecular modelling and prediction of their structure-function relationships. Int J Mol Sci. 2024;25:1684. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/ijms25031684\u003c/span\u003e\u003cspan address=\"10.3390/ijms25031684\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZheng Y, Wang S-N, Peng Y, Lu Z-Y, Shan S, Yang Y-Q, et al. Functional characterization of a niemann-pick type C2 protein in the parasitoid wasp microplitis mediator. Insect Sci. 2018;25:765\u0026ndash;77. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/1744-7917.12473\u003c/span\u003e\u003cspan address=\"10.1111/1744-7917.12473\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWicher D, Miazzi F. Functional properties of insect olfactory receptors: Ionotropic receptors and odorant receptors. Cell Tissue Res. 2021;383:7\u0026ndash;19. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00441-020-03363-x\u003c/span\u003e\u003cspan address=\"10.1007/s00441-020-03363-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWicher D. Chapter two - olfactory signaling in insects. In: Glatz R, editor. Progress in Molecular Biology and Translational Science. Academic; 2015. pp. 37\u0026ndash;54. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/bs.pmbts.2014.11.002\u003c/span\u003e\u003cspan address=\"10.1016/bs.pmbts.2014.11.002\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePark D, Jung JW, Choi B-S, Jayakodi M, Lee J, Lim J, et al. Uncovering the novel characteristics of asian honey bee, apis cerana, by whole genome sequencing. BMC Genomics. 2015;16:1. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/1471-2164-16-1\u003c/span\u003e\u003cspan address=\"10.1186/1471-2164-16-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKarpe SD, Jain R, Brockmann A, Sowdhamini R. Identification of complete repertoire of apis florea odorant receptors reveals complex orthologous relationships with apis mellifera. Genome Biol Evol. 2016;8:2879\u0026ndash;95. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/gbe/evw202\u003c/span\u003e\u003cspan address=\"10.1093/gbe/evw202\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eShan S, Song X, Khashaveh A, Wang S-N, Lu Z-Y, Hussain Dhiloo K, et al. A female-biased odorant receptor tuned to the lepidopteran sex pheromone in parasitoid microplitis mediator guiding habitat of host insects. J Adv Res. 2022;43:1\u0026ndash;12. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jare.2022.03.006\u003c/span\u003e\u003cspan address=\"10.1016/j.jare.2022.03.006\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eButterwick JA, del M\u0026aacute;rmol J, Kim KH, Kahlson MA, Rogow JA, Walz T, et al. Cryo-EM structure of the insect olfactory receptor orco. Nature. 2018;560:447\u0026ndash;52. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41586-018-0420-8\u003c/span\u003e\u003cspan address=\"10.1038/s41586-018-0420-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhang Q, Chen J, Wang Y, Lu Y, Dong Z, Shi W, et al. The odorant receptor co-receptor gene contributes to mating and host-searching behaviors in parasitoid wasps. Pest Manag Sci. 2023;79:454\u0026ndash;63. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/ps.7214\u003c/span\u003e\u003cspan address=\"10.1002/ps.7214\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFan X-B, Mo B-T, Li G-C, Huang L-Q, Guo H, Gong X-L, et al. Mutagenesis of the odorant receptor co-receptor (orco) reveals severe olfactory defects in the crop pest moth helicoverpa armigera. BMC Biol. 2022;20:214. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12915-022-01411-2\u003c/span\u003e\u003cspan address=\"10.1186/s12915-022-01411-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKoh T-W, He Z, Gorur-Shandilya S, Menuz K, Larter NK, Stewart S, et al. The drosophila IR20a clade of ionotropic receptors are candidate taste and pheromone receptors. Neuron. 2014;83:850\u0026ndash;65. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.neuron.2014.07.012\u003c/span\u003e\u003cspan address=\"10.1016/j.neuron.2014.07.012\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFisher K, Guill\u0026eacute;n BM, Dahanukar A, Yamanaka N, Woodard SH. Expression analyses of chemosensory genes provide insights into evolution of gustatory receptor genes in the bumble bee bombus impatiens. BMC Genomics. 2025;26:575. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12864-025-11710-x\u003c/span\u003e\u003cspan address=\"10.1186/s12864-025-11710-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAbuin L, Bargeton B, Ulbrich MH, Isacoff EY, Kellenberger S, Benton R. Functional architecture of olfactory ionotropic glutamate receptors. Neuron. 2011;69:44\u0026ndash;60. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.neuron.2010.11.042\u003c/span\u003e\u003cspan address=\"10.1016/j.neuron.2010.11.042\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNi L, Klein M, Svec KV, Budelli G, Chang EC, Ferrer AJ, et al. The ionotropic receptors IR21a and IR25a mediate cool sensing in drosophila. Elife. 2016;5:e13254. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.7554/eLife.13254\u003c/span\u003e\u003cspan address=\"10.7554/eLife.13254\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eEnjin A, Zaharieva EE, Frank DD, Mansourian S, Suh GSB, Gallio M, et al. Humidity sensing in drosophila. Curr Biol CB. 2016;26:1352\u0026ndash;8. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cub.2016.03.049\u003c/span\u003e\u003cspan address=\"10.1016/j.cub.2016.03.049\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePrieto-Godino LL, Schmidt HR, Benton R. Molecular reconstruction of recurrent evolutionary switching in olfactory receptor specificity. Elife 10:e69732. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.7554/eLife.69732\u003c/span\u003e\u003cspan address=\"10.7554/eLife.69732\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePrieto-Godino LL, Rytz R, Cruchet S, Bargeton B, Abuin L, Silbering AF, et al. Evolution of acid-sensing olfactory circuits in drosophilids. Neuron. 2017;93:661\u0026ndash;e6766. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.neuron.2016.12.024\u003c/span\u003e\u003cspan address=\"10.1016/j.neuron.2016.12.024\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eXiao J-X. 2024. Study on the host selection mechanism of Phthonoloba virifasciata and Rhoptroceros cyatheae[D]. Guizhou Nor-mal University; 2024. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.27048/d.cnki.ggzsu.2023.000365\u003c/span\u003e\u003cspan address=\"10.27048/d.cnki.ggzsu.2023.000365\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFrank HM, Walujkar S, Walsh RM, Laursen WJ, Theobald DL, Garrity PA, et al. Structural basis of ligand specificity and channel activation in an insect gustatory receptor. Cell Rep. 2024;43:114035. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.celrep.2024.114035\u003c/span\u003e\u003cspan address=\"10.1016/j.celrep.2024.114035\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGomes JV, Singh-Bhagania S, Cenci M, Chacon Cordon C, Singh M, Butterwick JA. The molecular basis of sugar detection by an insect taste receptor. Nature. 2024;629:228\u0026ndash;34. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41586-024-07255-w\u003c/span\u003e\u003cspan address=\"10.1038/s41586-024-07255-w\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhou Z, Luo Y, Wang X, He J, Zhou Q. Identification and sex expression profiles of candidate chemosensory genes from atherigona orientalis via the antennae and leg transcriptome analysis. Comp Biochem Physiol Part D Genomics Proteom. 2024;50:101222. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cbd.2024.101222\u003c/span\u003e\u003cspan address=\"10.1016/j.cbd.2024.101222\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWang S-N, Peng Y, Lu Z-Y, Dhiloo KH, Gu S-H, Li R-J, et al. Identification and expression analysis of putative chemosensory receptor genes in microplitis mediator by antennal transcriptome screening. Int J Biol Sci. 2015;11:737\u0026ndash;51. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.7150/ijbs.11786\u003c/span\u003e\u003cspan address=\"10.7150/ijbs.11786\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChyb S, Dahanukar A, Wickens A, Carlson JR. Drosophila Gr5a encodes a taste receptor tuned to trehalose. Proc Natl Acad Sci U S A. 2003;100(2 Suppl 2):14526\u0026ndash;30. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1073/pnas.2135339100\u003c/span\u003e\u003cspan address=\"10.1073/pnas.2135339100\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAryal B, Dhakal S, Shrestha B, Lee Y. Molecular and neuronal mechanisms for amino acid taste perception in the drosophila labellum. Curr Biol CB. 2022;32:1376\u0026ndash;e13864. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cub.2022.01.060\u003c/span\u003e\u003cspan address=\"10.1016/j.cub.2022.01.060\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBray S, Amrein H. A putative drosophila pheromone receptor expressed in male-specific taste neurons is required for efficient courtship. Neuron. 2003;39:1019\u0026ndash;29. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/s0896-6273(03)00542-7\u003c/span\u003e\u003cspan address=\"10.1016/s0896-6273(03)00542-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGuerenstein PG, Christensen TA, Hildebrand JG. Sensory processing of ambient CO2 information in the brain of the moth manduca sexta. J Comp Physiol Neuroethol Sens Neural Behav Physiol. 2004;190:707\u0026ndash;25. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00359-004-0529-0\u003c/span\u003e\u003cspan address=\"10.1007/s00359-004-0529-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKent KS, Harrow ID, Quartararo P, Hildebrand JG. An accessory olfactory pathway in lepidoptera: The labial pit organ and its central projections in manduca sexta and certain other sphinx moths and silk moths. Cell Tissue Res. 1986;245:237\u0026ndash;45. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF00213927\u003c/span\u003e\u003cspan address=\"10.1007/BF00213927\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNichols Z, Vogt RG. The SNMP/CD36 gene family in diptera, hymenoptera and coleoptera: Drosophila melanogaster, D. pseudoobscura, anopheles gambiae, aedes aegypti, apis mellifera, and tribolium castaneum. Insect Biochem Mol Biol. 2008;38:398\u0026ndash;415. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ibmb.2007.11.003\u003c/span\u003e\u003cspan address=\"10.1016/j.ibmb.2007.11.003\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRogers ME, Krieger J, Vogt RG. Antennal SNMPs (sensory neuron membrane proteins) of lepidoptera define a unique family of invertebrate CD36-like proteins. J Neurobiol. 2001;49:47\u0026ndash;61. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/neu.1065\u003c/span\u003e\u003cspan address=\"10.1002/neu.1065\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eShan S, Wang S-N, Song X, Khashaveh A, Lu Z-Y, Dhiloo KH, et al. Molecular characterization and expression of sensory neuron membrane proteins in the parasitoid microplitis mediator (hymenoptera: Braconidae). Insect Sci. 2020;27:425\u0026ndash;39. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/1744-7917.12667\u003c/span\u003e\u003cspan address=\"10.1111/1744-7917.12667\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLi Y, Chen H, Hong T, Yan M, Wang J, Shao Z, et al. Identification of chemosensory genes by antennal transcriptome analysis and expression profiles of odorant-binding proteins in parasitoid wasp \u003cem\u003eaulacocentrum confusum\u003c/em\u003e. Comp Biochem Physiol Part D Genomics Proteom. 2021;40:100881. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cbd.2021.100881\u003c/span\u003e\u003cspan address=\"10.1016/j.cbd.2021.100881\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVogt RG, Miller NE, Litvack R, Fandino RA, Sparks J, Staples J, et al. The insect SNMP gene family. Insect Biochem Mol Biol. 2009;39:448\u0026ndash;56. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ibmb.2009.03.007\u003c/span\u003e\u003cspan address=\"10.1016/j.ibmb.2009.03.007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhang J, Liu Y, Walker WB, Dong S-L, Wang G-R. Identification and localization of two sensory neuron membrane proteins from spodoptera litura (lepidoptera: Noctuidae). Insect Sci. 2015;22:399\u0026ndash;408. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/1744-7917.12131\u003c/span\u003e\u003cspan address=\"10.1111/1744-7917.12131\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-genomics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"gics","sideBox":"Learn more about [BMC Genomics](http://bmcgenomics.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/gics","title":"BMC Genomics","twitterHandle":"#BMCGenomics","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Rhoptroceros cyatheae, adult transcriptome, transcriptome sequencing, chemosensory genes","lastPublishedDoi":"10.21203/rs.3.rs-7729120/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7729120/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003eRhoptroceros cyatheae\u003c/em\u003e (Hymenoptera) belongs to the genus \u003cem\u003eRhopographus\u003c/em\u003e of Selandriidae. It mainly causes large-scale infestations during the sprouting period of \u003cem\u003eAlsophila spinulosa\u003c/em\u003e, and is among the herbivorous insects that harm this plant.The body of \u003cem\u003eR. cyatheae\u003c/em\u003e contains chemosensory genes that detect and transduce chemical signals related to host location, feeding, mating, and oviposition. However, to date, no reports on the chemosensory genes of \u003cem\u003eR. cyatheae\u003c/em\u003e have been published. Thus,on the basis of a the tran-scriptome database of male and female adult individuals of \u003cem\u003eR. cyatheae\u003c/em\u003e, a total of 30,296 unigenes were identified, with an N50 length of 3,286 bp. Through comparisons with six major public databases, namely NR, Swiss-Prot, Pfam, eggNOG, GO, and KEGG, a total of 11,109 unigenes were annotated, accounting for 36.67%. Among these, the number of unigenes annotated in the NR database was the largest, reaching 10,774, whereas the number of unigenes annotated in the KEGG database was the smallest, at 6,300. An analysis of the annotation information, 90 candidate chemosensory genes of \u003cem\u003eR. cyatheae\u003c/em\u003e, including 11 OBPs, 10 CSPs, 6 NPC2s, 24 ORs (comprising 23 typical OR genes and 1 Orco gene), 20 IRs, 15 GRs and 4 SNMPs. A phylogenetic tree of chemosensory genes was subsequently constructed to investigate the homology between the chemosensory genes of the \u003cem\u003eR. cyatheae\u003c/em\u003e and those of other insect species. Furthermore, 13 chemosensory genes differentially expressed between males and females, and their tissue expression profiles were verified via RT‒qPCR. These findings lay a molecular foundation for further research on the gene functions and olfactory perception mechanisms of \u003cem\u003eR. cyatheae\u003c/em\u003e.\u003c/p\u003e","manuscriptTitle":" Analysis and Identification of Chemosensory Genes in the Transcriptome of Adult Rhoptroceros cyatheae","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-24 07:24:10","doi":"10.21203/rs.3.rs-7729120/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-02-24T12:38:20+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-19T06:12:23+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"98007388891964246495376374471062752569","date":"2026-01-28T03:39:31+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-17T23:09:01+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"153296311169019857429810214822479029412","date":"2025-11-26T18:18:52+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-12T23:44:34+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-10-13T11:57:06+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-10-11T16:09:24+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Genomics","date":"2025-10-11T16:03:38+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-genomics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"gics","sideBox":"Learn more about [BMC Genomics](http://bmcgenomics.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/gics","title":"BMC Genomics","twitterHandle":"#BMCGenomics","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"bc4f1086-f5a9-43e6-9f32-bfb7cb7540e7","owner":[],"postedDate":"November 24th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-04-26T13:53:14+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-24 07:24:10","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7729120","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7729120","identity":"rs-7729120","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}