Insect Odorant-binding Proteins Research Articles

Odorant binding protein 18 increases the pathogen resistance of the imported willow leaf beetle, Plagiodera versicolora.

Insect odorant-binding proteins (OBPs) are a class of small molecular weight soluble proteins. In the past few years, OBPs had been found to work as carriers of ligands and play a crucial role in olfaction and various other physiological processes, like immunity. A subset of insect OBPs had been found to be expressed differently and play a function in immunity of fungal infection. However, there are few studies on the role of OBPs in immunity of bacterial infection. To identify the immune-related OBPs of Plagiodera versicolora after infected by Pseudomonas aeruginosa, we determined the mortality of P. versicolora to P. aeruginosa and selected the time point of 50% mortality of larvae to collect samples for RNA-seq. RNAi technology was used to investigate the function of immune-related OBPs after P. aeruginosa infection. RNA-seq data shows that PverOBP18 gene significantly up-regulated by 1.8-fold and further RT-qPCR affirmed its expression. Developmental expression profile showed that the expression of PverOBP18 was highest in the pupae, followed by the female adults, and lower in the 1st-3rd larvae and male adults with lowest in eggs. Tissue expression profiling showed that PverOBP18 was dominantly expressed in the epidermis. RNAi knockdown of PverOBP18 significantly reduced the expression of bacterial recognition receptor gene PGRP and antibacterial peptide gene Attacin and reduced the resistance of P. versicolora to P. aeruginosa infection. Our results indicated that PverOBP18 gene increased the pathogen resistance of P. versicolora by cooperating with the immune genes and provided valuable insights into using OBPs as targets to design novel strategies for management of P. versicolora.

Insight into insect odorant binding proteins: An alternative approach for pest management

Insects have enormous impact on public health as agricultural pests and disease vectors worldwide. Olfaction is the most crucial recognition process that mediates chemical communication in insects. Odorant binding proteins (OBP) are the small soluble proteins present in the sensory organs of the insects and are involved in many behavioural contexts, such as locating host, mating partners, escaping predators, flight, hygrosensation, immunity and identifying oviposition sites. OBPs are typically thought to function as a molecular carrier that bind, solubilize, and transport hydrophobic odorant molecules through the sensillar fluid and protect them from odorant degradation, thus maximizing olfactory system’s sensitivity. Due to their large structural and functional diversity, OBPs can be used as molecular targets for species, stage and sex specific insect pest management. In this review article, we will explore the structural and functional aspects of insect OBP and its potential applications in insect pest management. Antennal transcriptomics, ligand binding assay, olfactory repellents, pheromone-based disruption, RNA interference (RNAi) and genetic modifications can be effective management tactics for insect pests.

Isolation, characterization and docking analysis of insecticidal compound from Ocimum canum methanolic leaf extracts and its potential against three mosquito vectors

Vector management is an important challenge and burden to developing countries. Frequent and indiscriminate application of chemical insecticides for vector control has resulted in the development of resistance and undesirable effects on beneficial organisms. Hence an alternative approach is needed for the mosquito control programme. The present study investigated the insecticidal activity and spectral analysis of Ocimum canum methanol leaf extract against three mosquito species. One out of four fractions (F1-F4), the 4th fraction, revealed showed good insecticidal activity in larva and adult Cx. quinquefasciatus, Ae. aegypti and An. stephensi (0.114, 0.325 and 0.173 ppm) and (1.798, 1.061 and 0.871 ppm). The mortality was found to be dosage dependant. Spectral analysis of 1H NMR and 13C NMR indicates the presence of 6 protons at δH value of 7.45 (s) and 3 protons at δH value of 7.85 (s) ppm. FT-IR identified the possible band near the capping region at 3417.16 cm-1. LC-MS confirmed the eugenol compound with 99.79% purity at the molecular weight of 164. Further, eugenol docking analysis revealed high homology similarity with insect odorant binding protein 3OGN and AchE protein 2BG9. Overall, the result suggests that isolated eugenol compounds were found to be effective for the eco-friendly management of mosquito vectors. These findings could be useful for designing an efficient mosquitocidal compound in the future.

An OBP gene highly expressed in non-chemosensory tissues affects the phototaxis and reproduction of Spodoptera frugiperda.

Insect odorant binding proteins (OBPs) were initially regarded as carriers of the odorants involved in chemosensation. However, it had been observed that a growing number of OBP genes exhibited broad expression patterns beyond chemosensory tissues. Here, an OBP gene (OBP31) was found to be highly expressed in the larval ventral nerve cord, adult brain and male reproductive organ of Spodoptera frugiperda. An OBP31 knockout strain (OBP31-/- ) was generated by CRISPR/Cas9 mutagenesis. For OBP31-/- , the larvae needed longer time to pupate, but there was no difference in the pupal weight between OBP31-/- and wild type (WT). OBP31-/- larvae showed stronger phototaxis than the WT larvae, indicating the importance of OBP31 in light perception. For mating rhythm of adults, OBP31-/- moths displayed an earlier second mating peak. In the cross-pairing of OBP31-/- and WT moths, the mating duration was longer, and hatchability was lower in OBP31-/- group and OBP31+/- ♂ group than that in the WT group. These results suggested that OBP31 played a vital role in larval light perception and male reproductive process and could provide valuable insights into understanding the biological functions of OBPs that were not specific in chemosensory tissues.

Roles of insect odorant binding proteins in communication and xenobiotic adaptation.

Odorant binding proteins (OBPs) are small water-soluble proteins mainly associated with olfaction, facilitating the transport of odorant molecules to their relevant receptors in the sensillum lymph. While traditionally considered essential for olfaction, recent research has revealed that OBPs are engaged in a diverse range of physiological functions in modulating chemical communication and defense. Over the past 10 years, emerging evidence suggests that OBPs play vital roles in purifying the perireceptor space from unwanted xenobiotics including plant volatiles and pesticides, potentially facilitating xenobiotic adaptation, such as host location, adaptation, and pesticide resistance. This multifunctionality can be attributed, in part, to their structural variability and effectiveness in transporting, sequestering, and concealing numerous hydrophobic molecules. Here, we firstly overviewed the classification and structural properties of OBPs in diverse insect orders. Subsequently, we discussed the myriad of functional roles of insect OBPs in communication and their adaptation to xenobiotics. By synthesizing the current knowledge in this field, our review paper contributes to a comprehensive understanding of the significance of insect OBPs in chemical ecology, xenobiotic adaptation, paving the way for future research in this fascinating area of study.

Minus-C odorant binding protein TcasOBP7G contributes to reproduction and defense against phytochemical in the red flour beetle, Tribolium castaneum

Insect odorant binding proteins (OBPs) paly an important role in the recognition and transmission signals. One odorant binding protein gene, TcasOBP7G, was significantly up-regulated after exposure to eucalyptol. Nevertheless, the causal role of TcasOBP7G in Tribolium castaneum is still unclear. Here, TcasOBP7G was cloned and characterized from T. castaneum, aligning with the typical characteristic of Minus-C OBPs. The analysis of gene expression profile showed that TcasOBP7G is highly expressed at late larval and adult stage, with particularly high expression levels in ovary and testis of adults, suggesting that TcasOBP7G might be associated with reproduction of T. castaneum. Furthermore, RNAi of TcasOBP7G led to a decrease in spawning, while the development of gonads was normal, suggesting that TcasOBP7G may medicate the reproduction of T. castaneum. Additionally, the mRNA level of TcasOBP7G was significantly increased after exposure to eucalyptol, and knockdown of TcasOBP7G resulted in an increased susceptibility of T. castaneum to eucalyptol, indicating that TcasOBP7G is associated with the tolerance of T. castaneum to eucalyptol. Altogether, these results indicate that TcasOBP7G is required for both reproduction and defense against phytochemical in T. castaneum. This study will provide a theoretical basis for the development of novel control strategies based on the olfaction of insects.

Key Amino Acid Residues Involved in Binding Interactions between Bactrocera minax Odorant-Binding Protein 3 (BminOBP3) and Undecanol.

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.

Influence of pH on indole-dependent heterodimeric interactions between Anopheles gambiae odorant-binding proteins OBP1 and OBP4

Insect Odorant Binding Proteins (OBPs) constitute important components of their olfactory apparatus, as they are essential for odor recognition. OBPs undergo conformational changes upon pH change, altering their interactions with odorants. Moreover, they can form heterodimers with novel binding characteristics. Anopheles gambiae OBP1 and OBP4 were found capable of forming heterodimers possibly involved in the specific perception of the attractant indole.In order to understand how these OBPs interact in the presence of indole and to investigate the likelihood of a pH-dependent heterodimerization mechanism, the crystal structures of OBP4 at pH 4.6 and 8.5 were determined. Structural comparison to each other and with the OBP4-indole complex (3Q8I, pH 6.85) revealed a flexible N-terminus and conformational changes in the α4-loop-α5 region at acidic pH. Fluorescence competition assays showed a weak binding of indole to OBP4 that becomes further impaired at acidic pH. Additional Molecular Dynamic and Differential Scanning Calorimetry studies displayed that the influence of pH on OBP4 stability is significant compared to the modest effect of indole. Furthermore, OBP1-OBP4 heterodimeric models were generated at pH 4.5, 6.5, and 8.5, and compared concerning their interface energy and cross-correlated motions in the absence and presence of indole. The results indicate that the increase in pH may induce the stabilization of OBP4 by increasing its helicity, thereby enabling indole binding at neutral pH that further stabilizes the protein and possibly promotes the creation of a binding site for OBP1. A decrease in interface stability and loss of correlated motions upon transition to acidic pH may provoke the heterodimeric dissociation allowing indole release.Finally, we propose a potential OBP1-OBP4 heterodimer formation/disruption mechanism induced by pH change and indole binding.

IOBPdb A Database for Experimentally Determined Functional Characterization of Insect Odorant Binding Proteins

Odorant binding proteins (OBPs) are extra-cellular proteins that solubilize and transport volatile organic compounds (VOCs). Thousands of OBPs have been identified through genome sequencing, and hundreds have been characterized by fluorescence ligand binding assays in individual studies. There is a limited understanding of the comparative structure-function relations of OBPs, primarily due to a lack of a centralized database that relates OBP binding affinity and structure. Combining 181 functional studies containing 382 unique OBPs from 91 insect species, we present a database, iOBPdb, of OBP binding affinities for 622 individual VOC targets. This initial database provides powerful search and associative capabilities for retrieving and analyzing OBP-VOC binding interaction data. We have validated this dataset using phylogenetic mapping to determine the authenticity of the collected sequences and whether they cluster according to their assigned subfamilies. Potential applications include developing molecular probes for biosensors, novel bioassays and drugs, targeted pesticides that inhibit VOC/OBP interactions, and understanding odor sensing and perception in the brain.

A Highly Expressed Antennae Odorant-Binding Protein Involved in Recognition of Herbivore-Induced Plant Volatiles in Dastarcus helophoroides

Natural enemies such as parasitoids and parasites depend on sensitive olfactory to search for their specific hosts. Herbivore-induced plant volatiles (HIPVs) are vital components in providing host information for many natural enemies of herbivores. However, the olfactory-related proteins involved in the recognition of HIPVs are rarely reported. In this study, we established an exhaustive tissue and developmental expression profile of odorant-binding proteins (OBPs) from Dastarcus helophoroides, an essential natural enemy in the forestry ecosystem. Twenty DhelOBPs displayed various expression patterns in different organs and adult physiological states, suggesting a potential involvement in olfactory perception. In silico AlphaFold2-based modeling and molecular docking showed similar binding energies between six DhelOBPs (DhelOBP4, 5, 6, 14, 18, and 20) and HIPVs from Pinus massoniana. While in vitro fluorescence competitive binding assays showed only recombinant DhelOBP4, the most highly expressed in the antennae of emerging adults could bind to HIPVs with high binding affinities. RNAi-mediated behavioral assays indicated that DhelOBP4 was an essential functional protein for D. helophoroides adults recognizing two behaviorally attractive substances: p-cymene and γ-terpinene. Further binding conformation analyses revealed that Phe 54, Val 56, and Phe 71 might be the key binding sites for DhelOBP4 interacting with HIPVs. In conclusion, our results provide an essential molecular basis for the olfactory perception of D. helophoroides and reliable evidence for recognizing the HIPVs of natural enemies from insect OBPs' perspective.

An Overview of D7 Protein Structure and Physiological Roles in Blood-Feeding Nematocera.

Each time an insect bites a vertebrate host, skin and vascular injury caused by piercing triggers a series of responses including hemostasis, inflammation and immunity. In place, this set of redundant and interconnected responses would ultimately cause blood coagulation, itching and pain leading to host awareness, resulting in feeding interruption in the best-case scenario. Nevertheless, hematophagous arthropod saliva contains a complex cocktail of molecules that are crucial to the success of blood-feeding. Among important protein families described so far in the saliva of blood sucking arthropods, is the D7, abundantly expressed in blood feeding Nematocera. D7 proteins are distantly related to insect Odorant-Binding Proteins (OBP), and despite low sequence identity, observation of structural similarity led to the suggestion that like OBPs, they should bind/sequester small hydrophobic compounds. Members belonging to this family are divided in short forms and long forms, containing one or two OBP-like domains, respectively. Here, we provide a review of D7 proteins structure and function, discussing how gene duplication and some modifications in their OBP-like domains during the course of evolution lead to gain and loss of function among different hematophagous Diptera species.

Pleiotropic fitness effects of a Drosophila odorant-binding protein.

Insect odorant-binding proteins (OBPs) are members of a rapidly evolving multigene family traditionally thought to facilitate chemosensation. However, studies on Drosophila have shown that members of this family have evolved functions beyond chemosensation, as evident from their expression in reproductive tissues and the brain. Previous studies implicated diverse functions of Obp56h, a member of the largest gene cluster of the D. melanogaster Obp repertoire. Here, we examined the effect of CRISPR/Cas9-mediated deletion of Obp56h on 2 fitness phenotypes, on resistance to starvation stress and heat stress, and on locomotion and sleep phenotypes. Obp56h-/- mutants show a strong sexually dimorphic effect on starvation stress survival, with females being more resistant to starvation stress than the control. In contrast, Obp56h-/- females, but not males, are highly sensitive to heat stress. Both sexes show changes in locomotion and sleep patterns. Transcriptional profiling of RNA from heads of Obp56h-/- flies and the wildtype control reveals differentially expressed genes, including gene products associated with antimicrobial immune responses and members of the Turandot family of stress-induced secreted peptides. In addition, differentially expressed genes of unknown function were identified in both sexes. Genes encoding components of the mitochondrial electron transport chain, cuticular proteins, gene products associated with regulation of feeding behavior (Lst and CCHa2), ribosomal proteins, lncRNAs, snoRNAs, tRNAs, and snRNAs show changes in transcript abundances in Obp56h-/- females. These differentially expressed genes are likely to contribute to Obp56h-mediated effects on the diverse phenotypes that arise upon deletion of this OBP.

Identification and Expression Analysis of Odorant-Binding and Chemosensory Protein Genes in Virus Vector Nephotettix cincticeps.

The insect odorant binding proteins (OBPs) and chemosensory proteins (CSPs) are involved in the perception and discrimination of insects to host odor cues. Nephotettix cincticeps, one of the destructive pests of rice plants, not only directly damages hosts by sucking, but also indirectly transmits plant viruses in the field. Previous study found that two rice volatiles ((E)-β-caryophyllene and 2-heptanol) induced by rice dwarf virus (RDV) mediated the olfactory behavior of N. cincticeps, which may promote virus dispersal. However, the OBPs and CSPs in N. cincticeps are still unknown. In this study, to identify the OBP and CSP genes in N. cincticeps, transcriptomic analyses were performed. In total, 46,623 unigenes were obtained. Twenty putative OBP and 13 CSP genes were discovered and identified. Phylogenetic analyses revealed that five putative OBPs belonged to the plus-C OBP family, and the other classic OBPs and CSPs were distributed among other orthologous groups. A total of 12 OBP and 10 CSP genes were detected, and nine OBP and three CSP genes were highly expressed in N. cincticeps antennae compared with other tissues. This study, for the first time, provides a valuable resource to well understand the molecular mechanism of N. cincticeps in the perception and discrimination of the two volatiles induced by RDV infection.

Liposomal Formulations Loaded with a Eugenol Derivative for Application as Insecticides: Encapsulation Studies and In Silico Identification of Protein Targets.

A recently synthesized new eugenol derivative, ethyl 4-(2-methoxy-4-(oxiran-2-ylmethyl)phenoxy)butanoate, with a high insecticidal activity against Sf9 (Spodoptera frugiperda) insect cells, was encapsulated in the liposomal formulations of egg-phosphatidylcholine/cholesterol (Egg-PC:Ch) 70:30 and 100% dioleoylphosphatidylglycerol (DOPG), aiming at the future application as insecticides. Compound-loaded DOPG liposomes have sizes of 274 ± 12 nm, while Egg-PC:Ch liposomes exhibit smaller hydrodynamic diameters (69.5 ± 7 nm), high encapsulation efficiency (88.8 ± 2.7%), higher stability, and a more efficient compound release, thus, they were chosen for assays in Sf9 insect cells. The compound elicited a loss of cell viability up to 80% after 72 h of incubation. Relevantly, nanoencapsulation maintained the toxicity of the compound toward insect cells while lowering the toxicity toward human cells, thus showing the selectivity of the system. Structure-based inverted virtual screening was used to predict the most likely targets and molecular dynamics simulations and free energy calculations were used to demonstrate that this molecule can form a stable complex with insect odorant binding proteins and/or acetylcholinesterase. The results are promising for the future application of compound-loaded nanoliposome formulations as crop insecticides.

An odorant binding protein mediates Bactrocera dorsalis olfactory sensitivity to host plant volatiles and male attractant compounds

Odorant-binding proteins (OBPs) are believed to play critical roles in host-seeking behavior. However, little attention was paid to its different functions in male and female. The antenna-specific OBP gene from Bactrocera dorsalis, BdorOBP13, was cloned and its expression profile was examined. The results showed that BdorOBP13 was exclusively expressed in male and female adults, which exhibited a high transcript level in antennae. After injection of BdorOBP13 dsRNA, its transcript level in males and females decreased significantly. Electrophysiological responses of RNAi-injected flies to, methyl eugenol (male attractant) and γ-octalactone (female attractant) decreased significantly. However, no significant changes in the electrophysiological response were observed in RNAi-injected flies to benzothiazole, (+),dipentene, and ethyl tiglate. The behavioral bioassay showed that males treated with RNAi significantly reduced their preference to methyl eugenol, while RNAi-injected females showed a significantly lower preference to γ-octalactone, suggesting that BdorOBP13 may have different functions between males and females: it may be involved in the detection of methyl eugenol in males but is involved in the detection of γ-octalactone in females. These findings improve our understanding of insect OBPs and their roles in insect chemosensation, which may provide us with new molecular targets in the management of B. dorsalis.

Effects of carboxyl and acylamino linkers in synthetic derivatives of aphid alarm pheromone (E)-β-farnesene on repellent, binding and aphicidal activity

Insect odorant-binding proteins (OBPs) have significant roles in the transmission of semiochemical signals. In our previous study, a series of salicylate-substituted carboxyl (E)-β-farnesene (EBF) derivatives were synthesized with a carboxyl linker to bridge the EBF geranyl chain with salicylate aromatic group, and their binding mechanism with aphid OBPs was studied. In present study, to clarify the effects of linkers on the bioactivities of salicylate-substituted EBF derivatives against the aphid, a new series of salicylate-substituted EBF derivatives with an acylamino linker was designed and synthesized by the bioisosterism strategy. Their aphid-repellent activities, binding mechanisms with aphid OBPs, and aphicide activities were compared. The bioassay results indicated that salicylate-substituted EBF derivatives with a carboxyl linker showed higher aphid-repellent and OBP binding activities than the derivatives with an acylamino-linker. The molecular dynamic simulations and MM/GBSA calculation suggested that a more excellent hydrophobic linker such as carboxyl linker was essential for binding activity due to its stronger interactions with surrounding amino acids of the target proteins. Nevertheless, the salicylate substituted EBF derivatives with an acylamino linker exhibited significantly higher aphicidal activities than those with a carboxyl linker. In addition, the scaffold structure of EBF was essential to maintain the inherent aphid-repellent activity, target OBPs binding, and aphicidal activity. This work provides valuable guidelines for the rational design of different aphid control agents.

Spatial Expression Analysis of Odorant Binding Proteins in Both Sexes of the Aphid Parasitoid Aphidius gifuensis and Their Ligand Binding Properties.

In China, Aphidius gifuensis is one of the most common endoparasitoids of the green peach aphid Myzus persicae and grain aphid Sitobion miscanthi in the field. Insect odorant-binding proteins (OBPs) play vital roles in odor perception during feeding, host searching, mating and oviposition. In addition, some OBPs are involved in other physiological processes such as gustation and reproduction. In the present study, a comparative antennal transcriptomic analysis was applied between male and female A. gifuensis. The spatial expression patterns among antennae, heads, thoraxes, abdomens and legs of OBPs in both sexes were further profiled. Fifteen AgifOBPs were predicted, and 14 of them were identified by gene cloning, including 12 classic OBPs and 2 min-C OBPs. As expected, all OBPs were mainly expressed at high levels in antennae, heads or legs which are sensory organs and tissues. Finally, ligand binding properties of 2 OBPs (AgifOBP7 and AgifOBP9) were further evaluated. Female leg specifically expressed AgifOBP9 displays a broad and high binding property to aphid alarm pheromones, plant green volatiles and aphid sex pheromones (Ki < 10 μΜ). However, female leg specifically expressed AgifOBP7 displays poor affinity for all tested ligands except CAU-II-11 ((E)-3,7-dimethylocta-2,6-dien-1-yl-2-hydroxy-3-methoxybenzoate), a reported (E)-β-farnesene (EBF) analog with an exceptionally high binding affinity (Ki = 1.07 ± 0.08 μΜ). In summary, we reported the spatial expression pattern of the OBP repertoire in A. gifuensis, and further studied the binding properties of OBP7 and OBP9, which are mainly expressed in female legs, laying the foundation for the dissection of the contribution of OBPs to chemosensation in A. gifuensis.

A Comparative Evaluation of the Structural and Dynamic Properties of Insect Odorant Binding Proteins.

Insects devote a major part of their metabolic resources to the production of odorant binding proteins (OBPs). Although initially, these proteins were implicated in the solubilisation, binding and transport of semiochemicals to olfactory receptors, it is now recognised that they may play diverse, as yet uncharacterised, roles in insect physiology. The structures of these OBPs, the majority of which are known as “classical” OBPs, have shed some light on their potential functional roles. However, the dynamic properties of these proteins have received little attention despite their functional importance. Structural dynamics are encoded in the native protein fold and enable the adaptation of proteins to substrate binding. This paper provides a comparative review of the structural and dynamic properties of OBPs, making use of sequence/structure analysis, statistical and theoretical physics-based methods. It provides a new layer of information and additional methodological tools useful in unravelling the relationship between structure, dynamics and function of insect OBPs. The dynamic properties of OBPs, studied by means of elastic network models, reflect the similarities/dissimilarities observed in their respective structures and provides insights regarding protein motions that may have important implications for ligand recognition and binding. Furthermore, it was shown that the OBPs studied in this paper share conserved structural ‘core’ that may be of evolutionary and functional importance.

BarH1 regulates odorant-binding proteins expression and olfactory perception of Monochamus alternatus Hope

Insect odorant-binding proteins (OBPs) are a class of small soluble proteins that can be found in various tissues wherein binding and transport of small molecules are required. Thus, OBPs are not only involved in typical olfactory function by specific activities with odorants but also participate in other physiological processes in non-chemosensory tissues. To better understand the complex biological functions of OBPs, it is necessary to study the transcriptional regulation of their expression patterns. In this paper, an apparent gradient expression pattern of Obp19, that was highly and specifically expressed in antennae and played an essential role in the detection of camphene, was defined in the antennae of the Japanese pine sawyer. Further, the transcription factor BarH1, that also presented gradient expression pattern in antennae, was found to regulate expression of Obp19 directly through binding to its upstream DNA sequence. The condition of BarH1 gene silence, the gene expression levels of Obp19 significantly decreased. At the same time, additional olfactory genes also were regulated and thus influence camphene reception. These findings provide us an opportunity to incorporate Obps in the gene regulatory networks of insects, which contribute to a better understanding of the multiplicity and diversity of OBPs and the olfactory mediated behaviors.

Amino Alcohols from Eugenol as Potential Semisynthetic Insecticides: Chemical, Biological, and Computational Insights.

A series of β-amino alcohols were prepared by the reaction of eugenol epoxide with aliphatic and aromatic amine nucleophiles. The synthesized compounds were fully characterized and evaluated as potential insecticides through the assessment of their biological activity against Sf9 insect cells, compared with a commercial synthetic pesticide (chlorpyrifos, CHPY). Three derivatives bearing a terminal benzene ring, either substituted or unsubstituted, were identified as the most potent molecules, two of them displaying higher toxicity to insect cells than CHPY. In addition, the most promising molecules were able to increase the activity of serine proteases (caspases) pivotal to apoptosis and were more toxic to insect cells than human cells. Structure-based inverted virtual screening and molecular dynamics simulations demonstrate that these molecules likely target acetylcholinesterase and/or the insect odorant-binding proteins and are able to form stable complexes with these proteins. Encapsulation assays in liposomes of DMPG and DPPC/DMPG (1:1) were performed for the most active compound, and high encapsulation efficiencies were obtained. A thermosensitive formulation was achieved with the compound release being more efficient at higher temperatures.