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Simple Summary: This study investigates the inhibitory effect of sex pheromone components of the plum fruit moth (PFM), Grapholita funebrana, loaded with different doses of (Z)-8-tetradecenyl acetate (Z8-14:Ac) or (Z)-10-tetradecenyl acetate (Z10-14:Ac) on the trapping of the closely related species, the male oriental fruit moth (OFM), G. molesta. Field tests showed that adding 5–30% of Z8-14:Ac to a mixture of Z8-12:Ac and E8-12:Ac did not significantly affect the trapping of PFM males but reduced OFM male trapping by more than 86%. In contrast, adding over 10% of Z10-14:Ac to a mixture of Z8-12:Ac and E8-12:Ac inhibited the trapping of both OFM and PFM males. GmolPBP2 from OFMs exhibited the strongest binding affinity for Z8-14:Ac compared to the other GmolPBP/GmolGOBP and was considered the most likely OBP for recognizing and transporting Z8-14:Ac. Mutating the Phe12 residue of GmolPBP2 to Ala12 resulted in a loss of affinity for Z8-14:Ac, suggesting that Phe12 was the key amino acid and that π–alkyl was the primary weak interaction maintaining the binding affinity for GmolPBP2. These findings enhance our understanding of the molecular mechanisms through which insects recognize the sex pheromones of closely related species and thereby contribute to the development of species-specific sex attractants for PFMs using secondary sex pheromones. The plum fruit moth (PFM), Grapholita funebrana, and the oriental fruit moth (OFM), G. molesta, are closely related fruit moth species that severely damage fruit trees in Rosaceae. Both species share common primary sex pheromone components Z8-12:Ac and E8-12:Ac. The secondary sex pheromone components of PFMs consist of Z8-12:OH, Z8-14:Ac, and Z10-14:Ac, while those of OFMs include Z8-12:OH and 12:OH. Previous researchers have proved that the inclusion of Z8-14:Ac and Z10-14:Ac did not augment PFM catches but inhibited OFM catches in orchards in Europe, thereby maintaining the species-specificity of the PFM sex attractant. However, which of these components, Z8-14:Ac or Z10-14:Ac, plays the major role in inhibiting OFM attraction remains unclear. In the current study, electroantennogram (EAG) assays indicated that both OFM and PFM males exhibited a moderate EAG response to Z8-14:Ac and Z10-14:Ac. Rubber septa loaded with varying ratios of Z8-14:Ac (1% to 30%) or Z10-14:Ac (5% to 110%) combined with a constant dose of Z8-12:Ac and E8-12:Ac produced diverse trapping effects. Sex attractants containing Z8-14:Ac did not significantly affect the trapping of PFM males but drastically reduced the capture of OFM males, with the reduction reaching up to 96.54%. Attractants containing more than 10% of Z10-14:Ac simultaneously reduced the number of OFM and PFM males captured. Z8-14:Ac was indispensable for maintaining the specificity of sex pheromones. Fluorescence competitive binding assays of recombinant GmolPBP2 showed the lowest Ki value (0.66 ± 0.02 μM) among the PBPs/GOBPs from OFMs, suggesting that it is the most likely target for Z8-14:Ac. Molecular dynamic simulation and site-directed mutagenesis assays confirmed that the Phe12 residue, which forms a π–alkyl interaction with Z8-14:Ac, was crucial for GmolPBP2 binding to Z8-14:Ac. In conclusion, Z8-14:Ac is vital to the specificity of PFM sex pheromones inhibiting OFM attractants when added to Z8-12:Ac and E8-12:Ac. This could be potentially used to develop species-specific sex attractants for the PFM. [ABSTRACT FROM AUTHOR]

Simple Summary: The leaf beetle Diorhabda rybakowi Weise poses a severe threat to desert grasslands in Northwest China with its third instar larvae and adults, and even to the local ecological environment, attributed to its outbreak. Green-control attractants or repellents have become popular in recent years, but the olfactory mechanism of D. rybakowi is still unclear. Therefore, we preliminarily screened the best reference genes under different conditions and determined the bioinformatics characteristics and tissue expression profiles of D. rybakowi olfactory target genes. The recommended reference genes, RPL13a and RPS18 for tissues and RPL19 and RPS18 for sexes, were determined. Notably, the transcriptional levels of DrybOBP3, DrybOBP6, DrybOBP7, DrybOBP10, DrybOBP11, DrybCSP2, and DrybCSP5 among eleven odorant-binding proteins (OBPs) and six chemosensory proteins (CSPs) were significantly higher in the antenna. In summary, our study provides a strong basis for deepening the research of olfaction molecular mechanisms in D. rybakowi. Diorhabda rybakowi Weise is one of the dominant pests feeding on Nitraria spp., a pioneer plant used for windbreaking and sand fixation purposes, and poses a threat to local livestock and ecosystems. To clarify the key olfactory genes of D. rybakowi and provide a theoretical basis for attractant and repellent development, the optimal reference genes under two different conditions (tissue and sex) were identified, and the bioinformatics and characterization of the tissue expression profiles of two categories of soluble olfactory proteins (OBPs and CSPs) were investigated. The results showed that the best reference genes were RPL13a and RPS18 for comparison among tissues, and RPL19 and RPS18 for comparison between sexes. Strong expressions of DrybOBP3, DrybOBP6, DrybOBP7, DrybOBP10, DrybOBP11, DrybCSP2, and DrybCSP5 were found in antennae, the most important olfactory organ for D. rybakowi. These findings not only provide a basis for further in-depth research on the olfactory molecular mechanisms of host-specialized pests but also provide a theoretical basis for the future development of new chemical attractants or repellents using volatiles to control D. rybakowi. [ABSTRACT FROM AUTHOR]

Simple Summary: Honeybees, Apis melllifera, are the most widely used bees in the world for pollination services. However, in recent years, continuous colony losses have been recorded worldwide. One of the factors behind these losses is associated with diseases specific to this species, such as those caused by viruses. One of the known viruses affecting honeybee populations is the deformed wing virus (DWV). DWV causes physical malformations, decreased olfactory sensitivity, learning difficulties, and behavioral alterations, which can compromise hive behaviors. Thus, we evaluated behavioral response, the expression of antenna-specific odorant-binding protein (OBP) genes, and brain genes related to bee behavior, especially nurse bees, in honeybees inoculated with the DWV variant A. We performed olfactory sensitivity analyses in beehives. We performed behavioral assays with the larvae-emitted alarm pheromone component to stimulate infected nurse bees. We found that high levels of viral replication in both the head and antennae altered the behavioral response, decreasing attraction to the pheromone component, and DWV-A infection decreased the gene expression of OBPs and brain genes. Thus, DWV-A infection in adult bees could compromise internal hive cohesion and A. mellifera nurse bee behaviors. Insect behavior is coordinated mainly by smell through the diverse odor-binding proteins (OBP) that allow them to identify and recognize their environment. Sensory information collected through smell is then analyzed and interpreted in the brain, allowing for correct insect functioning. The behavior of honeybees (Apis mellifera L.) can be affected by different pathogens, such as deformed wing virus (DWV). In particular, the DWV variant A (DWV-A) is capable of altering olfactory sensitivity and reducing the gene expression of different OBPs, including those associated with nursing behavior. The DWV is also capable of replicating itself in the sensory lobes of the brain, further compromising the processing of sensory information. This study evaluated the behavioral response of nurse honeybees exposed to a pheromone compound and the alterations in the gene expression of the pre- and post-synaptic neuronal genes neuroxins-1 and neurogilin-1 in the bee heads and OBP proteins in the antennae of nurse bees inoculated with DWV-A. The behavioral response of nurse bees exposed to the larval pheromone compound benzyl alcohol was analyzed using a Y-tube olfactometer. The viral load, the gene expression of OBP5 and OBP11 in antennae, and neuroxins-1 and neurogilin-1 in the bee heads were analyzed via qPCR. High viral loads significantly reduced the ability of 10- and 15-day-old nurse honeybees to choose the correct pheromone compound. Also, the gene expression of OBP5, OBP11, neuroxin-1, and neurogilin-1 in nurse honeybees decreased when they were highly infected with DWV-A. These results suggest that a DWV-A infection can disturb information processing and cause nursing honeybees to reduce their activity inside the hive, altering internal cohesion. [ABSTRACT FROM AUTHOR]

Simple Summary: Reference genes serve as a foundation for investigating the functionality of target genes in insects. Nevertheless, the selection of reference genes is highly dependent upon particular experimental circumstances. Recently, the bean bug, Riptortus pedestris (Hemiptera: Alydidae), has attracted increasing attention due to its damage on yield losses of soybean crops. However, there is a dearth of literature regarding the evaluation of reference genes for quantitative real-time PCR (qRT-PCR) in R. pedestris. The current study aimed to assess the stability of expression levels in 16 candidate reference genes in R. pedestris across six experimental conditions. This evaluation was conducted using ΔCt method, GeNorm, NormFinder, BestKeeper, and RefFinder. Our results indicate that the optimum reference genes were as follows: RPL7A and EF1 for the developmental stage, α−tubulin and EF1 for adult tissues, RPL32 and RPL7A for adult age, RPL32 and SOD for nymph age, RPS23 and RPL7A for temperature, and HSP70 and GAPDH for mating status. These findings will provide a basis for future investigations into the expression and function of target genes in bean bugs. Quantitative real-time PCR (qRT-PCR) is widely accepted as a precise and convenient method for quantitatively analyzing the expression of functional genes. The data normalization strongly depends upon stable reference genes. The bean bug, Riptortus pedestris (Hemiptera: Alydidae), is a significant pest of leguminous crops and broadly distributed across Southeast Asia. In this study, a total of 16 candidate reference genes (RPL32, RPS23, SDHA, UBQ, UCCR, GST, TATA−box, HSP70, GAPDH, RPL7A, SOD, RPS3, Actin, α−tubulin, AK, and EF1) were carefully chosen in R. pedestris, and their expression levels were assessed across various conditions, including different developmental stages, diverse tissues, temperature treatments, adult age, molting time, and mating status. Following this, the stability of these reference genes was evaluated using four algorithms (ΔCt, GeNorm, NormFinder, and BestKeeper). Ultimately, the comprehensive rankings were determined using the online tool RefFinder. Our results demonstrate that the reference gene for qRT-PCR analysis in R. pedestris is contingent upon the specific experimental conditions. RPL7A and EF1 are optimal reference genes for developmental stages. Furthermore, α−tubulin and EF1 exhibit the most stable expression across various adult tissues. RPL32 and RPL7A exhibit the most stable expression for adult age. For nymph age, RPL32 and SOD display the most stable expression. For temperature conditions, RPS23 and RPL7A were identified as the most suitable for monitoring gene expression. Lastly, we verified the practicability of evaluating expression levels of odorant-binding protein 37 (RpedOBP37) and cytochrome P450 6a2 (RpedCYP6) throughout developmental stages, tissues, and temperature conditions. These findings are a significant addition to the qRT-PCR analysis studies on R. pedestris, serving as a fundamental groundwork for future investigations on stable reference genes in R. pedestris as well as other organisms. [ABSTRACT FROM AUTHOR]

Simple Summary: Odorant-binding proteins (OBPs) play a crucial role in insect olfaction perception in which OBPs are believed to discriminate, capture, and ferry odors to olfactory receptors, and thus, OBP-based behavioral interference is thought to be a potential novel pest management measure. Our previous study demonstrated that Bactrocera minax OBP3 (BminOBP3) exhibited strong binding affinity only with undecanol, which is believed to be a main B. minax sex pheromone, among 13 ligands tested. Here, homology modeling and molecular docking were carried out to explore the interaction mechanism between BminOBP3 and undecanol. The results suggested that the C-terminus (I116-P122), especially the last three amino acids of the C-terminus (V120-P122), play an essential role in the ligand binding for BminOBP3, and this was further proven by mutagenesis studies and fluorescence binding assays. These findings have provided important insights into the molecular mechanism by which BminOBP3 interacts with undecanol. Moreover, this study could serve as a significant reference for developing potentially useful technology to control pests, although disturbing their olfaction judgments. Insect odorant-binding proteins (OBPs) are significant in binding and transporting odorants to specific receptors. Our previous study demonstrated that BminOBP3 exhibited a strong affinity with undecanol. However, the binding mechanism between them remains unknown. Here, using homology modeling and molecular docking, we found that the C-terminus (I116-P122), especially the hydrogenbonds formed by the last three amino acid residues (V120, F121, and P122) of the C-terminus, is essential for BminOBP3′s ligand binding. Mutant binding assays showed that the mutant T-OBP3 that lacks C-terminus (I116-P122) displayed a significant decrease in affinity to undecanol (Ki = 19.57 ± 0.45) compared with that of the wild-type protein BminOBP3 (Ki = 11.59 ± 0.51). In the mutant 3D2a that lacks F121 and P122 and the mutant V120A in which V120 was replaced by alanine, the bindings to undecanol were completely abolished. In conclusion, the C-terminus plays a crucial role in the binding interactions between BminOBP3 and undecanol. Based on the results, we discussed the ligand-binding process of BminOBP3. [ABSTRACT FROM AUTHOR]

Simple Summary: Globally, honey bees are exposed to many challenges, such as the Varroa destructor mite and various viruses, which lead to massive losses. It is generally believed that African honey bees are more resilient and better able to cope with these stressors. This study examined some molecular markers that may be associated with this resilience. Higher resistance to the varroa mite could be related to better olfactory sensing. Higher gene expression levels of the odorant binding protein OBP14 in the antennae of Ethiopian honey bees suggest that reproducing mites might be better detected and cleaned. Resistance or tolerance to viruses could be attributed to a better functioning antiviral RNAi system. Several genes involved in this pathway are upregulated and are positively correlated with the viral load in honey bees. Both mechanisms may contribute to the resilience of African bees to varroa infestation and viral infection. Varroa destructor mites and the viruses it vectors are two major factors leading to high losses of honey bees (Apis mellifera) colonies worldwide. However, honey bees in some African countries show resilience to varroa infestation and/or virus infections, although little is known about the mechanisms underlying this resilience. In this study, we investigated the expression profiles of some key molecular markers involved in olfactory sensing and RNA interference, as these processes may contribute to the bees' resilience to varroa infestation and virus infection, respectively. We found significantly higher gene expression of the odorant binding protein, OBP14, in the antennae of Ethiopian bees compared to Belgian bees. This result suggests the potential of OBP14 as a molecular marker of resilience to mite infestation. Scanning electron microscopy showed no significant differences in the antennal sensilla occurrence and distribution, suggesting that resilience arises from molecular processes rather than morphological adaptations. In addition, seven RNAi genes were upregulated in the Ethiopian honey bees and three of them—Dicer-Drosha, Argonaute 2, and TRBP2—were positively correlated with the viral load. We can conclude that the antiviral immune response was triggered when bees were experiencing severe viral infection and that this might contribute to the bees' resilience to viruses. [ABSTRACT FROM AUTHOR]

Simple Summary: Chemosensation is thought to be the primary sensory mechanism used by ticks to locate their host and partners, as they appear to lack functional eyes or hearing organs, in which their unique olfactory structure, namely Haller's organ, is considered to play a significant role, although little is known about its molecular biology. This review comprehended and discussed the existing molecular information on candidate chemoreceptors and potential binding proteins preferentially expressed in tick foreleg enriched Haller's organ, backed by pertinent bioinformatic data. Additionally, tick olfactory-related structural, behavioral, and neurological information was also given, and its limits were highlighted. Despite their expression in the forelegs of ticks, independent functional experiments should confirm the olfactory role of these chemosensory-related candidate molecules. Ticks are obligatory hematophagous ectoparasites and vectors of many animal and human pathogens. Chemosensation plays a significant role in tick communication with their environment, including seeking out blood meal hosts. Studies on the structure and function of Haller's organ and its components have improved our understanding regarding tick olfaction and its chemical ecology. Compared with the knowledge on insect olfaction, less is known about the molecular basis of olfaction in ticks. This review focused on the chemosensory-related candidate molecules likely involved in tick olfaction. Members of the ionotropic receptor family and a new class of odorant-binding proteins are now known to be involved in tick olfaction, which appear to differ from that of insects. These candidate molecules are more closely related to those of mites and spiders than to other arthropods. The amino acid sequences of candidate niemann–pick type C2 and microplusin-like proteins in ticks exhibit features indicating their potential role as binding proteins. In the future, more comprehensive pertinent research considering the existing shortcomings will be required to fully understand the molecular basis of tick olfactory chemoreception. This information may contribute to the development of new molecular-based control mechanisms to reduce tick populations and related disease transmission. [ABSTRACT FROM AUTHOR]

Simple Summary: The Asian longhorned beetle (ALB), Anoplophora glabripennis Motschulsky, is a destructive pest in its native habitat and one of the most serious invasive alien species in North America and Europe, causing substantial economic and ecological losses. In order to explore effective monitoring and management strategies, we summarize and create a comprehensive list of host plants, including 209 species (cultivars) that have been damaged by ALBs. Thus far, 143 olfactory protein genes have been found in ALBs. Host kairomones were preferentially bound to ALB recombinant odorant-binding proteins (OBPs), but the function of most OBPs is still unclear. Microbial communities may help ALBs degrade host plants. We analyzed the trapping effect of combined host kairomones and sex pheromones and found that trapping numbers are limited in the field. Therefore, we discussed host plant location behavioral processes from new perspectives and found that multiple cues are used to locate and recognize host plants. Overall, we suggest that further research should contribute to understanding the host resistance mechanism, microbial community influence mechanism, and visual cue recognition mechanism of host plants. This research may provide effective monitoring and management strategies for ALBs. The Asian longhorn beetle (ALB), Anoplophora glabripennis Motschulsky, is a polyphagous xylophage with dozens of reported host tree species. However, the mechanisms by which individuals locate and recognize host plants are still unknown. We summarize the current knowledge of the host plant list, host kairomones, odorant-binding proteins (OBPs) and microbial symbionts of this beetle and their practical applications, and finally discuss the host localization and recognition mechanisms. A total of 209 species (or cultivars) were reported as ALB host plants, including 101 species of higher sensitivity; host kairomones were preferentially bound to ALB recombinant OBPs, including cis-3-hexen-1-ol, δ-3-carene, nonanal, linalool, and β-caryophyllene. In addition, microbial symbionts may help ALB degrade their host. Complementarity of tree species with different levels of resistance may reduce damage, but trapping effectiveness for adults was limited using a combination of host kairomones and sex pheromones in the field. Therefore, we discuss host location behavior from a new perspective and show that multiple cues are used by ALB to locate and recognize host plants. Further research into host resistance mechanisms and visual signal recognition, and the interaction of sex pheromone synthesis, symbiont microbiota, and host plants may help reveal the host recognition mechanisms of ALBs. [ABSTRACT FROM AUTHOR]

Simple Summary: Orthaga achatina (Lepidoptera: Pyralidae) is one of the most serious pests of camphor trees. Insect olfaction play a crucial role in identification of host plants and oviposition sites. In this study, we identified the binding affinities of GOBPs to camphor volatiles and sex pheromone components using fluorescence competition binding assays. Moreover, key amino acid residues that bind to plant volatiles were identified in GOBPs using 3-D structure modeling and ligand molecular docking, predicting the interactions between the GOBPs and the host plant volatiles. Overall, OachGOBP2 showed a wider odorant binding spectrum and higher binding capacity than GOBP1. The olfactory system in insects are crucial for recognition of host plants and oviposition sites. General odorant binding proteins (GOBPs) are thought to be involved in detecting odorants released by host plants. Orthaga achatina (Lepidoptera: Pyralidae) is one of the most serious pests of camphor trees, Cinnamomum camphora (L.) Presl, an important urban tree species in southern China. In this study, we study the GOBPs of O. achatina. Firstly, two full-length GOBP genes (OachGOBP1 and OachGOBP2) were successfully cloned according to transcriptome sequencing results, and real-time quantitative PCR measurements showed that both GOBP genes were specifically expressed in the antennae of both sexes, proposing their important roles in olfaction. Then, both GOBP genes were heterologous expressed in Escherichia coli and fluorescence competitive binding assays were conducted. The results showed that OachGOBP1 could bind Farnesol (Ki = 9.49 μM) and Z11-16: OH (Ki = 1.57 μM). OachGOBP2 has a high binding affinity with two camphor plant volatiles (Farnesol, Ki = 7.33 μM; α-Phellandrene, Ki = 8.71 μM) and two sex pheromone components (Z11-16: OAc, Ki = 2.84 μM; Z11-16: OH, Ki = 3.30 μM). These results indicate that OachGOBP1 and OachGOBP2 differ in terms of odorants and other ligands. Furthermore, key amino acid residues that bind to plant volatiles were identified in GOBPs using 3-D structure modeling and ligand molecular docking, predicting the interactions between the GOBPs and the host plant volatiles. [ABSTRACT FROM AUTHOR]

Simple Summary: Ophraella communa Lesage is a specific biological control agent of the invasive weed Ambrosia artemisiifolia L. Understanding the molecular mechanism by which O. communa recognizes A. artemisiifolia will help improve its bio-control effect. Odorant-binding proteins (OBPs) play a vital role in insect olfactory perception. In this study, the sequence and expression characteristics of OcomOBP7 were analyzed. We obtained the pure protein of OcomOBP7 by prokaryotic expression and purification, and its binding characteristics were analyzed using a fluorescence competitive binding assay. Finally, we verified the function of OcomOBP7 in vivo using RNAi combined with an electroantennography (EAG) assay. The results showed that the binding ability of OcomOBP7 was broad-spectrum and was involved in the host plant localization of O. communa. The olfactory system plays a key role in various insect behaviors, and odorant-binding proteins participate in the first step of the olfactory process. Ophraella communa Lesage is an oligophagous phytophagous insect that is a specific biological control agent for Ambrosia artemisiifolia L. The leaf beetle must identify and locate A. artemisiifolia through olfaction; however, its odorant-binding protein (OBP) function has not yet been reported. In this study, OcomOBP7 was cloned, and its tissue expression profile and binding ability were analyzed using RT-qPCR and fluorescence binding assays, respectively. Sequence analysis demonstrated that OcomOBP7 belongs to the classical OBP family. The RT-qPCR results showed that OcomOBP7 was specifically expressed in the antennae, indicating that OcomOBP7 may be involved in chemical communication. The fluorescence binding assay showed that OcomOBP7 has an extensive binding ability to alkenes. The electroantennography experiments showed that O. communa antennal response to α-pinene and ocimene decreased significantly after interference because the two odors specifically bound to OcomOBP7. In summary, α-pinene and ocimene are odorant ligands corresponding to OcomOBP7, indicating that OcomOBP7 is involved in the chemical recognition of A. artemisiifolia. Our study lays a theoretical foundation for research into O. communa attractants, which is helpful for the better biological control of A. artemisiifolia by O. communa. [ABSTRACT FROM AUTHOR]

Simple Summary: Trichogramma wasps are minute egg parasitoids that are one of the most used biocontrol agents to manage lepidopteran pests. Given the great success in industrializing a mass-rearing method for the cost-effective production of Trichogramma dendrolimi, this parasitoid wasp has been used extensively to control agricultural and forestry pests in China. So far, while the biology and ecology of T. dendrolimi have been intensively studied to provide successful biological control, the limited genome information constrains the interpretation of the molecular mechanism underlying its host recognition and parasitism. Here, we assembled a high-quality reference genome for T. dendrolimi and identified the repetitive sequences and protein-coding genes. A phylogenomic tree constructed for T. dendrolimi and 24 hymenopteran species places the family Trichogrammatidae sister in the remaining Chalcidoidea species. We found that significantly expanded and contracted gene families are involved in the development, regulation and transport processes in T. dendrolimi, respectively. Additionally, 24 odorant binding proteins, 100 olfactory receptors, 27 gustatory receptors, 27 ionotropic receptors and 87 venom genes were identified in the genome of T. dendrolimi. Our study provides a foundation for future studies on the molecular mechanism underlying host recognition and parasitism in the Trichogramma species. Trichogramma dendrolimi is one of the most successfully industrialized Trichogramma species used to control agricultural and forestry pests in China. However, the molecular mechanisms underlying its host recognition and parasitism remain largely unknown, partially due to the limited genome information of this parasitoid wasp. Here, we present a high-quality de novo assembly of T. dendrolimi through a combination of Illumina and PacBio sequencing technologies. The final assembly had a length of 215.2 Mb and contains 316 scaffolds with a scaffold N50 size of 1.41 Mb. Repetitive sequences with a length of 63.4 Mb and 12,785 protein-coding genes were identified. Significantly expanded gene families were identified to be involved in the development and regulatory processes, while remarkably contracted gene families were involved in the transport processes in T. dendrolimi. The olfactory and venom-associated genes were identified in T. dendrolimi and 24 other hymenopteran species, using uniform methods combining BLAST and HMM profiling. The identified venom genes of T. dendrolimi were enriched in antioxidant activity, tricarboxylic acid cycle, response to oxidative stress and cell redox homeostasis. Our study provides an important resource for comparative genomics and functional studies to interpret the molecular mechanisms underlying host recognition and parasitism of Trichogramma species. [ABSTRACT FROM AUTHOR]

Simple Summary: During ontogeny, many insects change habitats, which are characterized by specific communities of opportunistic and pathogenic microorganisms. Therefore, different developmental stages may be subject to different directions of natural selection that cause alterations in immunity. We analyzed changes in humoral immunity during the intermolt period of last-instar larvae of the Colorado potato beetle under laboratory conditions. We observed an increase in basal expression of immunity- and stress-related genes at the stages of finishing feeding and migration into soil. Changes in gene expression in response to fungal infection were also examined. The results suggest that the larvae develop strong defenses against soil pathogens (especially against fungi) before migration into soil. The study is also promising in terms of a gene knockdown for enhancing the susceptibility of the Colorado potato beetle to native or introduced fungal pathogens in potato agrosystems. Different developmental stages of insects may be dissimilar in immunity functioning. Additionally, the stages often inhabit diverse environments with specific microbial communities. In the Colorado potato beetle, a strong increase in resistance to entomopathogenic fungi is observed during the intermolt period of last-instar larvae, but mechanisms of this change are insufficiently understood. We studied changes in the expression of immunity- and stress-related genes in the fat body and integument during this intermolt period by quantitative PCR. By the end of the instar, there was upregulation of transcription factors of Toll, IMD, and Jak–Stat pathways as well as genes encoding metalloprotease inhibitors, odorant-binding proteins, and heat shock proteins. Nonetheless, the expression of gene LdRBLk encoding β-lectin did not change during this period. Most of the aforementioned genes were upregulated in response to Metarhizium robertsii topical infection. The expression alterations were more pronounced in recently molted larvae than in finishing feeding larvae and in the integument compared to the fat body. We believe that upregulation of immune-system- and stress-related genes at the end of the intermolt period is an adaptation caused by migration of larvae into soil, where the probability of encountering entomopathogenic fungi is high. [ABSTRACT FROM AUTHOR]

Simple Summary: Dioryctria abietella (Lepidoptera: Pyralidae) is a destructive forest pest that feeds on the shoots and cones of Pinaceae plants. The reception of host volatiles and toxic compounds can be mediated via odorant binding proteins (OBPs) expressed in olfactory organs. Here, a number of DabiOBPs may be involved in reproduction, as indicated by the expression profiles. Further, DabiOBP5 and DabiOBP14 enriched in adult antennae could strongly interact with two plant volatiles and four insecticides. Of these, β-ionone derived from plant flowers possibly served as an feeding attractant of female moths, while a pear ester ethyl-(2E,4Z)-decadienoate identified as a non-host volatile may repel D. abietella. In addition, two DabiOBPs were possibly associated with insecticide resistance. Molecular simulations identified several key residues involved in ligand-binding, revealing different binding mechanisms of DabiOBP5 and/or DabiOBP14 with the best ligands. These findings provide potentially active compounds for the control of D. abietella and characterize the interaction mechanisms of two antennal DabiOBPs with plant odorants and insecticides. The management of forest pests has become a significant challenge, particularly for wood borers, because they spend most of the time in the trunks or cones. The coneworm, Dioryctria abietella, is a representative of cone borers as its larvae feed on the cones of Pinaceae plants. The molecular mechanisms underlying the interactions between this species and host plants or habitats can assist in developing strategies for pest control. In this study, we extended the expression profiles of 32 odorant binding proteins (OBPs) in the reproductive tissues of D. abietella, revealing the detectable transcription of 29 genes. Using two DabiOBPs highly expressed in antennae (DabiOBP5 and DabiOBP14) as targets, six compounds with high affinities (dissociation constants < 13 μM) were identified through a reverse chemical ecology strategy, including insecticides widely used for the control of lepidopteran pests. Of these compounds, a floral volatile β-ionone and a pear-produced ester ethyl-(2E,4Z)-decadienoate may serve as behaviorally active compounds in D. abietella. The strong binding of DabiOBPs to insecticides suggested their involvement in insecticide resistance, reflecting sophisticated detoxification mechanisms of this moth. In the molecular simulations, DabiOBP14 possessed stronger interactions with the six ligands compared to DabiOBP5, in which a few key residues within the binding pockets were involved in the formation of hydrogen bonds. This study provides some valuable reference active compounds for the development of lures or repellents in D. abietella and unravels the putative roles of two antenna-dominant DabiOBPs in the perception of plant-derived odorants and insecticides. [ABSTRACT FROM AUTHOR]

Simple Summary: The parasitic wasp Baryscapus dioryctriae (Hymenoptera: Eulophidae) was originally discovered in the pupae of Dioryctria insects. It can also parasitize many Pyralidae pests, such as Ostrinia furnacalis, Chilo suppressalis and Galleria mellonella, suggesting that this parasitic wasp has a great potential to serve as a natural enemy against agricultural and forest insect pests. The olfactory system plays an important role in this wasps' reception of chemical signals emitted from their surrounding environment, and various chemosensory genes are involved in this system. In this study, seven chemosensory gene families, namely, the odorant-binding proteins (OBPs), chemosensory proteins (CSPs), Niemann–Pick type C2 proteins (NPC2s), odorant receptors (ORs), ionotropic receptors (IRs), gustatory receptors (GRs) and sensory neuron membrane proteins (SNMPs) of B. dioryctriae were identified and analyzed using transcriptome sequencing and bioinformatic analyses. Moreover, the quantitative expression of the candidate chemosensory genes, which are female antennae- and ovipositor-biased expression was validated by RT-qPCR. These results lay a molecular foundation for further investigation of the mechanism of chemoreception by the olfactory system during host recognition, location, and oviposition site selection in B. dioryctriae. Baryscapus dioryctriae is a pupal endoparasitoid of many Pyralidae pests and has been used as a biocontrol agent against insect pests that heavily damage the cone and seed of the Korean pine. The olfactory system of wasps plays an essential role in sensing the chemical signals during their foraging, mating, host location, etc., and the chemosensory genes are involved in detecting and transducing these signals. Many chemosensory genes have been identified from the antennae of Hymenoptera; however, there are few reports on the chemosensory genes of Eulophidae wasps. In this study, the transcriptome databases based on ten different tissues of B. dioryctriae were first constructed, and 274 putative chemosensory genes, consisting of 27 OBPs, 9 CSPs, 3 NPC2s, 155 ORs, 49 GRs, 23 IRs and 8 SNMPs genes, were identified based on the transcriptomes and manual annotation. Phylogenetic trees of the chemosensory genes were constructed to investigate the orthologs between B. dioryctriae and other insect species. Additionally, twenty-eight chemosensory genes showed female antennae- and ovipositor-biased expression, which was validated by RT-qPCR. These findings not only built a molecular basis for further research on the processes of chemosensory perception in B. dioryctriae, but also enriched the identification of chemosensory genes from various tissues of Eulophidae wasps. [ABSTRACT FROM AUTHOR]

Cage assays show that all three olfactory organs, antennae, proboscis and maxillary palps, are involved in detecting and responding to ME over short distances, and the proboscis has a greater role than the maxillary palps. Olfactory Organs Detecting ME in B. dorsalis Our results showed that removal of any or all olfactory organs (antennae, proboscises and maxillary palps) had a significant effect on male response to ME over 6 h of exposure. Keywords: methyl eugenol; non-responsiveness; odorant binding protein; P450; trapping EN methyl eugenol non-responsiveness odorant binding protein P450 trapping 1004 12 11/17/22 20221101 NES 221101 1. [Extracted from the article]

However, the honey bees exhibited a reduction in consuming the sucrose containing not less than 1 ug/mL of CdCl SB 2 sb ; although non-significant, there exists the possibility that sucrose containing more than 1 ug/mL of Cd may affect honey bee sucrose responsiveness that is essential for olfactory learning in honey bees [[38]]. We, therefore, investigated whether honey bees exposed to chronic Cd exhibited altered olfactory learning, and we further compared the patterns of brain gene expression between cadmium-exposed bees and control bees using RNA-seq analysis. Our studies showed that the head weight was significantly lower in cadmium-exposed bees when compared to controls, suggesting chronic Cd exposure may have adverse effects on the total protein content of the mandibular and hypopharyngeal glands in the heads of adult honey bees, thus resulting in a decrease in head weight in Cd-exposed bees. [Extracted from the article]

Simple Summary: Starting with efforts to understand pheromone detection in moths, the ability of insects to localize food and mates has captivated researchers for over a century. Here we review recent advances in understanding the detection of volatiles at the level of olfactory receptors and odorant-binding proteins focusing on advances in biological understanding. Drosophila remains an important model system, but CRIPSR-mediated genome editing is opening the door to genetic analysis in a wide range of insects, including disease vectors and important agricultural pests. This review will spotlight new findings for the initial recognition and signaling events by odorant-binding proteins and olfactory receptors in responses to odorants. Finally, we discussed how some critical insect behaviors appear to have evolved multimodal mechanisms to maximize response robustness. Human and insect olfaction share many general features, but insects differ from mammalian systems in important ways. Mammalian olfactory neurons share the same overlying fluid layer in the nose, and neuronal tuning entirely depends upon receptor specificity. In insects, the olfactory neurons are anatomically segregated into sensilla, and small clusters of olfactory neurons dendrites share extracellular fluid that can be independently regulated in different sensilla. Small extracellular proteins called odorant-binding proteins are differentially secreted into this sensillum lymph fluid where they have been shown to confer sensitivity to specific odorants, and they can also affect the kinetics of the olfactory neuron responses. Insect olfactory receptors are not G-protein-coupled receptors, such as vertebrate olfactory receptors, but are ligand-gated ion channels opened by direct interactions with odorant molecules. Recently, several examples of insect olfactory neurons expressing multiple receptors have been identified, indicating that the mechanisms for neuronal tuning may be broader in insects than mammals. Finally, recent advances in genome editing are finding applications in many species, including agricultural pests and human disease vectors. [ABSTRACT FROM AUTHOR]

Simple Summary: In this study, we conducted antennal transcriptome analysis in Glenea cantor (Cerambycidae: Lamiinae) and identified 76 olfactory-related genes, including 29 odorant binding proteins (OBPs), 14 chemosensory proteins (CSPs), 13 odorant receptors (ORs), 18 ionotropic receptors (IRs) and 2 sensory neuron membrane proteins (SNMPs). We also verified the reliability of transcriptome differential genes by qRT-PCR, which indicated the reliability of the transcriptome. Based on the relative expression of 30 d adults, GcanOBP22 and GcanOBP25 were highly expressed not only in the antennae, but also in the wings and legs. In addition, GcanCSP4 was the highest expression on the female antennae at 12 d. These findings laid the foundation for further research on the mechanism of G. cantor olfactory mechanism at the molecular level. Glenea cantor Fabricius (Cerambycidae: Lamiinae) is a pest that devastates urban landscapes and causes ecological loss in southern China and Southeast Asian countries where its main host kapok trees are planted. The olfactory system plays a vital role in mating, foraging, and spawning in G. cantor as an ideal target for pest control. However, the olfactory mechanism of G. cantor is poorly understood at the molecular level. In this study, we first established the antennal transcriptome of G. cantor and identified 76 olfactory-related genes, including 29 odorant binding proteins (OBPs), 14 chemosensory proteins (CSPs), 13 odorant receptors (ORs), 18 ionotropic receptors (IRs) and 2 sensory neuron membrane proteins (SNMPs). Furthermore, the phylogenetic trees of olfactory genes were constructed to study the homology with other species of insects. We also verified the reliability of transcriptome differential genes by qRT-PCR, which indicated the reliability of the transcriptome. Based on the relative expression of 30 d adults, GcanOBP22 and GcanOBP25 were highly expressed not only in the antennae, but also in the wings and legs. In addition, GcanCSP4 was the highest expression on the female antennae at 12 d. These findings laid the foundation for further research on the mechanism of G. cantor olfactory mechanism at the molecular level. [ABSTRACT FROM AUTHOR]

Simple Summary: Insects use olfaction, i.e., their sense of smell, to detect odors that elicit behavioral responses, with structurally similar compounds eliciting different responses. The roles of specific recognition proteins, i.e., odorant-binding proteins (OBPs) and odorant receptors (ORs), located in insect antennae, in discriminating between structurally similar compounds are not fully understood. Here, we explore current research in understanding the role of OBPs and ORs in discriminating between enantiomers—mirror image structures—in insect chemical ecology and chemoperception. Olfaction is a complex recognition process that is critical for chemical communication in insects. Though some insect species are capable of discrimination between compounds that are structurally similar, little is understood about how this high level of discrimination arises. Some insects rely on discriminating between enantiomers of a compound, demonstrating an ability for highly selective recognition. The role of two major peripheral olfactory proteins in insect olfaction, i.e., odorant-binding proteins (OBPs) and odorant receptors (ORs) has been extensively studied. OBPs and ORs have variable discrimination capabilities, with some found to display highly specialized binding capability, whilst others exhibit promiscuous binding activity. A deeper understanding of how odorant-protein interactions induce a response in an insect relies on further analysis such as structural studies. In this review, we explore the potential role of OBPs and ORs in highly specific recognition, specifically enantiomeric discrimination. We summarize the state of research into OBP and OR function and focus on reported examples in the literature of clear enantiomeric discrimination by these proteins. [ABSTRACT FROM AUTHOR]

Simple Summary: In this study, we conducted a transcriptome analysis of adult antennae in Plagiodera versicolora (Coleoptera: Chrysomelidae) and identified a total of 98 candidate chemosensory genes, encoding 40 odorant receptors (ORs), 7 ionotropic receptors (IRs), 13 gustatory receptors (GRs), 10 chemosensory proteins (CSPs), 24 odorant binding proteins (OBPs), and 4 sensory neuron membrane proteins (SNMPs). The tissue expression profiles showed that almost all PverOBPs and PverORs were highly expressed in the antennae. In addition, the results revealed that PverOBP10, PverOBP12, PverOBP18, PverOR24, and PverOR35 showed female-biased expression profiles, indicating that these receptors may be involved in some female-specific behaviors such as oviposition site seeking. This work greatly promotes the understanding of the olfactory system and will help provide insight for functional studies of the chemoreception mechanism in P. versicolora. Insects can sense surrounding chemical signals by their accurate chemosensory systems. This system plays a vital role in the life history of insects. Several gene families participate in chemosensory processes, including odorant receptors (ORs), ionotropic receptors (IRs), gustatory receptors (GRs), chemosensory proteins (CSPs), odorant binding proteins (OBPs), and sensory neuron membrane proteins (SNMPs). Plagiodera versicolora (Coleoptera: Chrysomelidae), is a leaf-eating forest pest found in salicaceous trees worldwide. In this study, a transcriptome analysis of male and female adult antennae in P. versicolora individuals was conducted, which identified a total of 98 candidate chemosensory genes including 40 ORs, 7 IRs, 13 GRs, 10 CSPs, 24 OBPs, and 4 SNMPs. Subsequently, the tissue expression profiles of 15 P. versicolora OBPs (PverOBPs) and 39 ORs (PverORs) were conducted by quantitative real-time PCR. The data showed that almost all PverOBPs and PverORs were highly expressed in the male and female antennae. In addition, several OBPs and ORs (PverOBP10, PverOBP12, PverOBP18, PverOR24, and PverOR35) had higher expression levels in female antennae than those in the male antennae, indicating that these genes may be taking part in some female-specific behaviors, such as find mates, oviposition site, etc. This study deeply promotes further understanding of the chemosensory system and functional studies of the chemoreception genes in P. versicolora. [ABSTRACT FROM AUTHOR]

Simple Summary: Malaria fever kills millions of people annually in the tropical and subtropical countries of Africa and Asia. Because there is no effective vaccine, malaria prevention is exclusively dependent on avoiding human-vector interaction. The interaction of Vitex negundo essential oil constituents with Anopheles gambiae Odorant Binding Proteins (OBP), as well as its compositional variation, repellent efficacy, and toxicity profile, are investigated in this work. The oils were subjected to GC-MS analysis, a mosquito behavioral test, OBP-ligand interactions, Anopheles species authentication, and toxicity profile. Docking protocol validation was achieved by redocking the co-crystallized ligands and root mean square deviation (RMSD) calculation. The oil yields and compositions are climate–soil dependent with ≈71.39% monoterpenes and ≈16.32% sesquiterpene. Optimal repellency is achieved at 15 min at ED50 0.08–0.48% v/v while the RMSD was estimated to be within 0.24–1.35 Å. Strong affinities, −6.4 to −5.4 kcal/mol, were demonstrated by α-pinene, citronellal, linalool, and myrcene for OBP1, OBP7, OBP4, and OBP. respectively. The hydrophobic interactions involve Leu17, Cys35, ALA52, Leu73, Leu76, Ala88, Met91, Lys93, Trp114, Phe123, and Leu124 receptors on α-helixes 1–7 within the binding cavities, and may block the olfactory receptors resulting in disorientation. α-pinene, linalool, and myrcene are safe and suitable for use in the development of green and innovative repellents because their ligand efficiency metrics, ADME/tox, and repellency screening are all within the threshold values. (1) Background: Malaria fever affects millions of people yearly in Africa and Asia's tropical and subtropical areas. Because there is no effective vaccine, malaria prevention is solely dependent on avoiding human-vector interaction. (2) Aim: This study examines the interaction between the constituents of Vitex negundo essential oil and Anopheles gambiae Odorant Binding Proteins (OBP) as well as the compositional variation, repellent efficacy, and toxicity profile. (3) Methods: The oils were subjected to GC-MS and mosquito behavioral analysis. OBP–ligand interactions, Anopheles species authentication, and the toxicity profile were determined by molecular docking, PCR assay and in silico ADME/tox tool. Docking protocol validation was achieved by redocking the co-crystallized ligands into the protein binding pocket and root mean square deviation (RMSD) calculation. (4) Results: The oil yields and compositions are climate–soil dependent with ≈71.39% monoterpenes and ≈16.32% sesquiterpene. Optimal repellency is achieved at 15 min at ED50 0.08–0.48% v/v while the RMSD was estimated to be within 0.24–1.35 Å. Strong affinities were demonstrated by α-pinene (−6.4 kcal/mol), citronellal (−5.5 kcal/mol), linalool (−5.4 kcal/mol), and myrcene (−5.8 kcal/mol) for OBP1, OBP7, OBP4, and OBP; respectively. The hydrophobic interactions involve Leu17 (α-helix 1), Cys35 (α-helix 2), ALA52 (α-helix 3), Leu73, Leu76 (α-helix 4), Ala88, Met91, Lys93, Trp114 (α-helix 5), Phe123 (α-helix 6), and Leu124 (α-helix 7) receptors within the binding cavities, and may cause blocking of the olfactory receptors resulting in disorientation. (5) Conclusion: The ligand efficiency metrics, ADME/tox and repellency screening are within the threshold values; hence, α-pinene, linalool, and myrcene are safe and fit-to-use in the development of a green and novel repellent. [ABSTRACT FROM AUTHOR]

Simple Summary: Parasitic wasps are the major natural enemies of many organisms, and therefore they are broadly used in the biological control of numerous agricultural and horticultural pests. For example, Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) is a tiny natural egg parasitoid of various agricultural pest insects, including Plutella xylostella, Helicoverpa armigera, Spodoptera frugiperda and Ectomyelois ceratoniae. However, how T. pretiosum seek and localise host insect eggs is still not clear. The olfactory system is critical in guiding insect behaviours, including mating, feeding and oviposition, which play pivotal roles in the interactions between parasitoid wasps and their hosts. This project aimed to investigate T. pretiosum major olfactory tissue (antennae) and the olfactory genes, including odorant binding proteins (OBPs) and odorant receptors (ORs). T. pretiosum adult antennae were examined under scanning electron microscopy, and four types of olfactory sensilla were observed. Using T. pretiosum genome, 22 OBPs and 105 ORs were identified, which were further compared with olfactory genes of other Hymenoptera insect species. The expression patterns of OBPs between T. pretiosum male and female adults were examined to identify female- or male-specific OBPs. This study enriches our knowledge of T. pretiosum olfactory system and will help better use it in the integrated pest management (IPM) for many insect pest species. Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) is a tiny natural egg parasitoid of several agricultural pest insects, which has been widely used in the biological control for Plutella xylostella, Helicoverpa armigera, Spodoptera frugiperda and Ectomyelois ceratoniae. However, limited studies have been conducted on T. pretiosum olfactory system, which is critical in regulating insect behaviours. In this study, T. pretiosum adult antennae were investigated under ascanning electron microscopy (SEM). Four types of olfactory sensilla were observed, including chaetica sensilla (CS), trichoid sensilla (TS), faleate sensilla (FS) and placoid sensilla (PS). Using T. pretiosum genome, 22 putative odorant binding proteins (OBPs) and 105 odorant receptors (ORs) were identified, which were further compared with olfactory genes of Apis mellifera, Nasonia vitripennis and Diachasma alloeum. The expression patterns of OBPs between T. pretiosum male and female adults were examined by quantitative real time PCR (qRT-PCR) approaches. Three female-specific OBPs (TpreOBP19, TpreOBP15 and TpreOBP3) were identified, which may play crucial roles in T. pretiosum host-seeking and oviposition behaviours. This study enriches our knowledge of T. pretiosum olfactory genes and improves our understanding of its olfactory system. [ABSTRACT FROM AUTHOR]

Simple Summary: The olfactory sensillum, which is located in the antenna of insects, is the basic unit of the olfactory organ. Olfactory-related genes are expressed in the sensillum. It is believed that the process of olfaction recognition is mainly mediated by two gene families, odorant binding proteins (OBPs) and olfactory receptors (ORs). The honeybee possesses a large numbers of ORs, but few OBPs. Up to now, the function of OBPs in the honeybee has not yet been fully elucidated. In order to reveal the specific role of OBPs from Apis cerana cerana, we selected an OBP gene, AcerOBP6, which is highly expressed in the antennae of worker bees, acquired a purified protein via a prokaryotic expression system, and analyzed its function using bioinformatics, molecular biology, and electrophysiology. According to the result, AcerOBP6 was a protein with extensive binding affinity, and we speculated that its function was chiefly related to foraging. Overall, this research not only explains the essential role of OBPs in ligand binding, but also provides valuable resources to help researchers further understand the nature and mechanism of the olfactory system. Olfaction is essential in some behaviors of honeybee, such as nursing, foraging, attracting a mate, social communication, and kin recognition. OBPs (odorant binding proteins) play a key role in the first step of olfactory perception. Here, we focused on a classic OBP with a PBP-GOBP domain from the Asian honeybee, Apis cerana cerana. Beyond that, the mRNA expression profiles and the binding affinity of AcerOBP6 were researched. According to qRT-PCR analysis, AcerOBP6 transcripts were mainly expressed in the antennae of forager bees. In addition, we found that the expression level of AcerOBP6 was higher than that of AmelOBP6. The fluorescence competitive binding assay indicated that the AcerOBP6 protein had binding affinity with most of the tested odors, including queen pheromone, worker pheromone, and floral volatiles, among which the strongest one was linolenic acid (with a Ki value of 1.67). However, AcerOBP6 was not sensitive to the brood pheromones. A further study based on EAG assay revealed that the antennae had the strongest response to 2-heptanone. The EAG recording values of the selected ligands were all reduced after AcerOBP6 was silenced, with 8 of 14 declining significantly (p < 0.01) given that these odors could specifically bind to AcerOBP6. As revealed in our current study, AcerOBP6 might be a crucial protein involved in olfactory recognition for foraging. Overall, the research provides a foundation for exploring the olfactory mechanism of A. cerana cerana. [ABSTRACT FROM AUTHOR]

Simple Summary: The black soldier fly (BSF) is a saprophagous insect that is an efficient bioconverter of organic waste because of its extreme voracity and fast larval development. Specific chemical molecules from decaying organic substances can influence BSF behaviour; in particular, several are likely attractants for BSF and are able to induce female oviposition and larval feeding. In insects, the perception of volatile organic compounds (VOCs) is based on a wide arsenal of chemoreception proteins. As a first step in understanding this process, an assessment of chemoreception genes belonging to the larval and adult stages of Hermetia illucens was conducted together with candidate VOCs that potentially regulate adult females searching for oviposition sites and phagostimulants for their larval progeny. The genes encoding several odorant binding proteins (OBPs) of interest were identified and three-dimensional models of these proteins were created, allowing a preliminary investigation of how different VOCs may interact with their binding sites. The present study provides a road map for further analysis and correlation among insect olfactory proteins and VOCs indicative of organic decomposition, starting from a computational approach to establish a reliable correlation between them. The black soldier fly (BSF), Hermetia illucens (Diptera: Stratiomyidae), has considerable global interest due to its outstanding capacity in bioconverting organic waste to insect biomass, which can be used for livestock, poultry, and aquaculture feed. Mass production of this insect in colonies requires the development of methods concentrating oviposition in specific collection devices, while the mass production of larvae and disposing of waste may require substrates that are more palatable and more attractive to the insects. In insects, chemoreception plays an essential role throughout their life cycle, responding to an array of chemical, biological and environmental signals to locate and select food, mates, oviposition sites and avoid predators. To interpret these signals, insects use an arsenal of molecular components, including small proteins called odorant binding proteins (OBPs). Next generation sequencing was used to identify genes involved in chemoreception during the larval and adult stage of BSF, with particular attention to OBPs. The analysis of the de novo adult and larval transcriptome led to the identification of 27 and 31 OBPs for adults and larvae, respectively. Among these OBPs, 15 were common in larval and adult transcriptomes and the tertiary structures of 8 selected OBPs were modelled. In silico docking of ligands confirms the potential interaction with VOCs of interest. Starting from the information about the growth performance of H. illucens on different organic substrates from the agri-food sector, the present work demonstrates a possible correlation between a pool of selected VOCs, emitted by those substrates that are attractive for H. illucens females when searching for oviposition sites, as well as phagostimulants for larvae. The binding affinities between OBPs and selected ligands calculated by in silico modelling may indicate a correlation among OBPs, VOCs and behavioural preferences that will be the basis for further analysis. [ABSTRACT FROM AUTHOR]

Simple Summary: The odorant binding proteins (OBPs) interact with host chemical compounds to elicit olfactory responses. Transcriptome analysis of six different tissues of male and female Cacia cretifera thibetana was performed to unravel the interaction of OBPs with host compounds. In both sexes, differentially expressed genes were associated with the KEGG pathways such as cutin, suberine and wax biosynthesis, glycerophospholipid metabolism, choline metabolism in cancer, and the chemokine signaling pathway. The expression of 11 out of 31 OBPs were confirmed by quantitative RT-PCR and seven were found to be specifically expressed in antennae. CcreOBP6 and CcreOBP10 showed strong affinity for terpineol and trans-2-hexenal exhibiting their potential role as an attractant or repellent to control C. cretifera thibetana. This study characterized the transcriptome of Cacia cretifera thibetana and explored odorant binding proteins (OBPs) and their interaction with host-specific compounds. A total of 36 samples from six different organs including antennae, head, thorax, abdomen, wings, and legs (12 groups with 3 replicates per group) from both male and female insects were collected for RNA extraction. Transcriptomic analysis revealed a total of 89,897 transcripts as unigenes, with an average length of 1036 bp. Between male and female groups, 31,095 transcripts were identified as differentially expressed genes (DEGs). The KEGG pathway analysis revealed 26 DEGs associated with cutin, suberine, and wax biosynthesis and 70, 48, and 62 were linked to glycerophospholipid metabolism, choline metabolism in cancer, and chemokine signaling pathways, respectively. A total of 31 OBP genes were identified. Among them, the relative expression of 11 OBP genes (OBP6, 10, 12, 14, 17, 20, 22, 26, 28, 30, and 31) was confirmed by quantitative RT-PCR in different tissues. Seven OBP genes including CcreOBP6 and CcreOBP10 revealed antennae-specific expression. Further, we selected two OBPs (CcreOBP6 and CcreOBP10) for functional analysis to evaluate their binding affinity with 20 host odorant compounds. The CcreOBP6 and CcreOBP10 exhibited strong binding affinities with terpineol and trans-2-hexenal revealing their potential as an attractant or repellent for controlling C. cretifera thibetana. [ABSTRACT FROM AUTHOR]

Simple Summary: Odorant-binding proteins (OBPs) mediate chemical communication in insects and can provide a shortcut to deciphering their olfactory code. Although being less specific than olfactory receptors, they are easy to express, purify and characterize. Aphids, in particular, are endowed with a limited repertoire of only 10 OBPs, which are exceptionally well conserved across species. Therefore, using OBPs to study olfaction in aphids appears to be a very attractive strategy. Within our project to functionally characterize all OBPs of the pea aphid, Acyrthosiphon pisum, we have expressed OBP9 and found it to be narrowly tuned to the linear alcohols and aldehydes of 16 carbon atoms. 1-Hexadecanol has been reported as a strong feeding repellant produced by toxic fungi. On the other hand, 16-carbon linear aldehydes are components of several lepidopteran pheromones and could represent warning signals to avoid potential feeding competition. Aphid odorant-binding protein 9 is almost exclusively expressed in antennae and is well conserved between different aphid species. In order to investigate its function, we have expressed this protein and measured ligand-binding affinities to a number of common natural compounds. The best ligands are long-chain aldehydes and alcohols, in particular Z9-hexadecenal and Z11-hexadecenal, as well as 1-hexadecanol and Z11-1-hexadecenol. A model of this protein indicated Lys37 as the residue that is likely to establish strong interactions with the ligands, probably a Schiff base with aldehydes and a hydrogen bond with alcohols. Indeed, when we replaced this lysine with a leucine, the mutated protein lost its affinity to both long aldehydes and alcohols, while the binding of other volatiles was unaffected. Long-chain linear alcohols are common products of molds and have been reported as aphid antifeedants. Corresponding aldehydes, instead, are major components of sex pheromones for several species of Lepidoptera. We speculate that aphids might use OBP9 to avoid mold-contaminated plants as well as competition with lepidopteran larvae. [ABSTRACT FROM AUTHOR]

Simple Summary: The scarab beetle Holotrichia parallela is a serious underground pest and causes serious damages in China to a variety of crops. To reduce the use of pesticides, insect olfactory proteins attract more and more attention in the development of pollution-free control agents in plant protection. In this study, we evaluate the molecular mechanism in the scarab beetle to detect oviposition cues. We clone a leg biased gene HparOBP14 which encodes for an odorant-binding protein of the scarab beetle and demonstrate its involvement in binding, electrophysiological, and behavioral responses to the oviposition chemicals by the knockdown of HparOBP14 expression using RNA interference technique. Our study provides a strong theoretical basis for the development of environmentally acceptable strategies for H. parallela control. Holotrichia parallela is one of the agriculturally important scarab beetle pests in China. In this study, HparOBP14 was cloned, which is the most abundantly expressed among the OBP genes in the legs of female H. parallela adults. Sequence comparison and phylogenetic analysis showed that HparOBP14 has a Plus-C structure motif. The expression profile analysis revealed that HparOBP14 expression was the highest in the female antennae and then in the legs. The fluorescence competitive binding experiment of the recombinant HparOBP14 protein showed that HparOBP14 had an affinity with 6-methyl-5-heptene-2-one (plant volatile), 3-methylindole, p-cymene, methanol, formaldehyde, α-pinene, and geraniol (organic fertilizer volatile). Knockdown HparOBP14 expression decreased significantly the EAG response of the injected female adults to p-cymene, methanol, formaldehyde, α-pinene, and geraniol. Similarly, the injected female adults were significantly less attracted to geraniol and methanol. Therefore, HparOBP14 might bind organic matter volatiles during oviposition. These results are not only helpful to analyze the olfactory recognition mechanism of female adult H. parallela when choosing suitable oviposition sites, but also to provide target genes for green prevention and control of H. parallela in the future. [ABSTRACT FROM AUTHOR]

Simple Summary: The alfalfa plant bug, Adelphocoris lineolatus (Goeze) (Hemiptera: Miridae), is a serious agricultural pest in China. Previous studies have shown that olfaction plays a crucial role in the survival and reproduction of herbivorous insects, such as A. lineolatus, and OBPs (odorant-binding proteins) are involved in the initial steps of olfaction. Due to the complex intracellular environment in insects, detection of the expression of OBP genes by conventional methods is limited in vitro. In this study, we developed a PNA-GO- (peptide nucleic acid and graphene oxide) based mRNA biosensor that allows detection of the expression of the AlinOBP14 gene in A. lineolatus. We demonstrate that AlinOBP14 is expressed in the adult head, which is consistent with previous research on the expression profiles of AlinOBP14 in A. lineolatus using quantitative real-time PCR. The results demonstrate that the method we developed is both feasible and reliable. In addition, this method exhibited good stability and specificity in detection of chemically synthesized targets in vitro as well as the expression of the target OBP gene in vivo. OBPs play a crucial role in the recognition of ligands and are involved in the initial steps of semiochemical perception. The diverse expression of OBP genes allows them to participate in different physiological functions in insects. In contrast to classic OBPs with typical olfactory roles in A. lineolatus, the physiological functions of Plus-C OBPs remain largely unknown. In addition, detection of the expression of insect OBP genes by conventional methods is difficult in vitro. Here, we focused on AlinOBP14, a Plus-C OBP from A. lineolatus, and we developed a PNA-GO-based mRNA biosensor to detect the expression of AlinOBP14. The results demonstrated that AlinOBP14 plays dual roles in A. lineolatus. The AlinOBP14 is expressed beneath the epidermis of the vertex and gena in heads of A. lineolatus, and it functions as a carrier for three terpenoids, while AlinOBP14 is also expressed in the peripheral antennal lobe and functions as a carrier for endogenous compounds such as precursors for juvenile hormone (JH) and JHⅢ. Our investigation provides a new method to detect the expression of OBP genes in insects, and the technique will facilitate the use of these genes as potential targets for novel insect behavioral regulation strategies against the pest. [ABSTRACT FROM AUTHOR]