Role In Insect Behavior Research Articles

Three odorant-binding proteins of small hive beetles, Aethina tumida, participate in the response of bee colony volatiles

Aethina tumida (small hive beetle, SHB) is a rapidly spreading invasive parasite of bee colonies. The olfactory system plays a key role in insect behavior, and odorant-binding proteins (OBPs) are involved in the first step of the olfactory signal transduction pathway and the detection of host volatiles. However, the olfactory mechanism of OBPs in SHB-localized bee colonies is unclear. In this study, electroantennogram (EAG) and behavioral bioassay showed that only three compounds (2-heptanone, ocimene, and ethyl palmitate) from bee colonies triggered high electrophysiological and behavioral responses. Three antenna-specific OBP genes (OBP6, OBP11, and OBP19) were identified, and they were significantly expressed on adult days 6–7. Furthermore, by combining RNA interference (RNAi) with EAG, olfactometer bioassay, competitive fluorescence binding assays, and molecular docking, we found that these three OBP genes were involved in the recognition of 2-heptanone and ethyl palmitate, and AtumOBP6 is also involved in the recognition of ocimene. These data indicate that AtumOBP6, AtumOBP11, and AtumOBP19 play an important role in the olfactory response to bee colony volatiles. Our results provide new insights into the functions of the OBP families in A. tumida and help to explore more potential target genes for environmentally friendly pest control strategies.

Sand Fly larvae are capable of positive chemotaxis: a proof of concept study using Phlebotomus papatasi (Diptera: Psychodidae) as a model species.

Phlebotomine sand flies are important vectors of medical and veterinary importance, transmitting pathogens, such as the Leishmania parasites, responsible for 700,000 to 1 million new cases of leishmaniasis every year. The vast majority of the current sand fly surveillance and control tools are tailored against the adult stages, due to the limited knowledge on the ecology of the larval stages. Since vector control is primarily an ecological problem, an in-depth understanding of the behavior of the target insect pests across all the different life stages of their development is required prior to the development of effective control strategies. It is well known that chemical cues play an important role in insect behavior. While there are numerous studies investigating the behavior of adult sand flies in response to chemical sources, there is currently no information available on the response of their larval stages. In this study, novel bioassays were constructed to investigate the effect of chemical cues (gustatory and olfactory) on the behavior of Phlebotomus papatasi (Scopoli) sand fly larvae. The larvae exhibited a clear food preference within a few hours of exposure in a 2-choice bioassay, while, also, demonstrated positive chemotaxis in response to volatile stimuli emitted from their preferred food source. Identification of the specific chemical compounds (or the combination thereof) eliciting attractance response to sand fly immature stages could lead to the development of innovative, and targeted (larval-specific) tools for the surveillance, and management of these important public health pests.

Role of odorant binding protein C12 in the response of Tribolium castaneum to chemical agents

Tribolium castaneum is a worldwide pest of stored grain that mainly damages flour, and not only causes serious loss of flour quality but also leads to deterioration of flour quality. Chemical detection plays a key role in insect behavior, and the role of odorant-binding proteins (OBPs) in insect chemical detection has been widely studied. OBPs can interact with small molecule compounds and thereby modulate variation in insecticide susceptibility in insects. In this study, a total of 65 small molecule compounds are selected to investigate the bound effect with TcOBP C12. The molecular docking results showed that β-caryophyllene, (−)-catechin, butylated hydroxytoluene, diphenyl phthalate and quercetin were the top five compounds, with docking binding energies of −6.11, −5.25, −5.09, −5.05, and − 5.03 Kcal/mol, respectively. Molecular dynamics analysis indicated that odorant binding protein C12 (TcOBP C12) exhibited high binding affinity to all five tested chemical ligands, evidenced by fluorescence quenching assay in vitro. In addition, the contact toxicity assay results suggested that these chemical agents caused a dose-dependent increase in mortality rate for T. castaneum adults. The TcOBP C12 gene was upregulated >2 times after a 24-h exposure, indicating that OBP C12 may play an important role for T. castaneum in response to these chemical agents. In conclusion, our results provide a theoretical basis for future insecticide experiments and pest management.

Foreleg Transcriptomic Analysis of the Chemosensory Gene Families in Plagiodera versicolora (Coleoptera: Chrysomelidae).

Simple SummaryThe chemosensory system plays an important role in insect behavior, such as foraging, mating, host-seeking, oviposition, and so on. In contrast with the transcriptome of the antennae, the transcriptome of the forelegs studied in Coleoptera are much less understood. In this study, we performed a transcriptome analysis of adult forelegs in Plagiodera versicolora and 53 candidate chemosensory genes encoding 4 chemosensory proteins (CSPs), 19 odorant binding proteins (OBPs), 10 odorant receptors (ORs), 10 gustatory receptors (GRs), 6 ionotropic receptors (IRs), and 4 sensory neuron membrane proteins (SNMPs) were identified. Then, the expression of the chemosensory genes was examined using real-time quantitative PCR. These results will provide valuable information to help explore the chemoreception mechanism in P. versicolora.Plagiodera versicolora (Coleoptera: Chrysomelidae) is a worldwide leaf-eating forest pest in salicaceous trees. The forelegs play important roles in the chemoreception of insects. In this study, we conducted a transcriptome analysis of adult forelegs in P. versicolora and identified a total of 53 candidate chemosensory genes encoding 4 chemosensory proteins (CSPs), 19 odorant binding proteins (OBPs), 10 odorant receptors (ORs), 10 gustatory receptors (GRs), 6 ionotropic receptors (IRs), and 4 sensory neuron membrane proteins (SNMPs). Compared with the previous antennae transcriptome data, 1 CSP, 4 OBPs, 1 OR, 3 IRs, and 4 GRs were newly identified in the forelegs. Subsequently, the tissue expression profiles of 10 P. versicolora chemosensory genes were performed by real-time quantitative PCR. The results showed that PverOBP25, PverOBP27, and PverCSP6 were highly expressed in the antennae of both sexes. PverCSP11 and PverIR9 are predominately expressed in the forelegs than in the antennae. In addition, the expression levels of PverGR15 in female antennae and forelegs were significantly higher than those in the male antennae, implying that it may be involved in some female-specific behaviors such as oviposition site seeking. This work would greatly further the understanding of the chemoreception mechanism in P. versicolora.

Identification and functional analysis of a chemosensory protein from Bactrocera minax (Diptera: Tephritidae).

Olfaction has an indispensable role in insect behavior, enabling location of suitable host plants and oviposition sites, finding mates and evasion of natural enemies. Chemosensory proteins (CSPs) function to screen external odorants and transport them to olfactory receptor neurons, thereby increasing the sensitivity of the olfactory system. At present, CSP genes have been identified in many insect species, but there are relatively few studies on the function of CSP, especially in Tephritidae. In this study, we sequenced and analyzed 12 transcriptomes of Bactrocera minax and identified five CSP genes. The results of polymerase chain reactions with reverse transcription showed that BminCSP3 was highly expressed only in antennae. Results from competitive binding experiments showed that BminCSP3 has good binding ability to citral compared with 23 other volatile organic compounds. The docking model with citral showed hydrogen bond formation with residues (ARG97); however, no hydrogen bonds were formed in the docking of five other ligands (furfuryl alcohol, linalool, cis-3-hexenyl acetate, (R)-(+)-limonene and (+)-carvone). Electroantennogram (EAG) analyses revealed that citral was active in B. minax at the antennal level, and the EAG response value of female adults was significantly higher than that of male adults. Furthermore, the results of behavioral bioassays showed that females were significantly attracted to citral. Our results suggest that BminCSP3 plays an important role in the recognition of citral by B. minax adults. © 2022 Society of Chemical Industry.

Odorant binding protein C12 is involved in the defense against eugenol in Tribolium castaneum

Tribolium castaneum (T. castaneum) is a worldwide pest of stored grain that mainly harms flour, and not only causes serious loss of flour quality but also leads to deterioration of flour quality. Chemical detection plays a key role in insect behavior, and the role of odorant-binding proteins (OBPs) in insect chemical detection has been widely studied. However, the mechanism of OBPs in insect defense against exogenous toxic substances is still unclear. In this study, biochemical analysis showed that eugenol, the active component of A. vulgaris essential oil, significantly induced the expression of the OBP gene OBPC12 from T. castaneum (TcOBPC12). The mortality of late larvae treated with eugenol was higher than that of the control group after RNA interference (RNAi) against TcOBPC12, which indicates that the OBP gene is involved in the eugenol defense mechanism and leads to a decrease in sensitivity to eugenol. Tissue expression profiling showed that the expression of TcOBPC12 in the epidermis, hemolymph, and intestine was higher than in other larval tissues, and TcOBPC12 was expressed mainly in the epidermis, head, and fat body of adults. The developmental expression profile showed that the expression of TcOBPC12 in late eggs, early and late larval stages, and late adult stages was higher than in other developmental stages. These data suggest that TcOBPC12 may be involved in the absorption of exogenous toxic substances by the larvae from T. castaneum. Our results provide a theoretical basis for the metabolism and degradation mechanism of exogenous toxic substances and help explore more potential target genes of insect pests.

Identification and Expression Profile of Chemosensory Genes in the Small Hive Beetle Aethina tumida.

Simple SummaryThe small hive beetle is a destructive pest of honeybees, causing severe economic damage to the apiculture industry. Chemosensory genes play key roles in insect behavior, such as foraging and mating partners. However, the chemosensory genes are lacking in the small hive beetle. In order to better understand its chemosensory process at the molecular level, a total of 130 chemosensory genes, including 38 odorant receptors, 24 ionotropic receptors, 14 gustatory receptors, 3 sensory neuron membrane proteins, 29 odorant binding proteins, and 22 chemosensory proteins were identified from the transcriptomic data of antennae and forelegs. Reverse-transcription PCR showed that 3 OBPs (AtumOBP3, 26 and 28) and 3 CSPs (AtumCSP7, 8 and 21) were highly expressed in antennae. Overall, this study could provide a basis for elucidating functions of these chemosensory genes at the molecular level.Aethina tumida is a parasite and predator of honeybee causing severe loss to the bee industry. No effective and environmentally friendly methods are available to control this pest at present. Chemosensory genes play key roles in insect behavior which can potentially be used as targets for developing environmentally friendly pest control agents. In this study, the putative chemosensory genes in antennae and forelegs of A. tumida involved in olfaction or contact chemical communication of adults were investigated using RNA transcriptome sequencing and PCR methods. Based on transcriptomic data, unigenes encoding 38 odorant receptors (ORs), 24 ionotropic receptors (IRs), 14 gustatory receptors (GRs), 3 sensory neuron membrane proteins (SNMPs), 29 odorant binding proteins (OBPs), and 22 chemosensory proteins (CSPs) were identified. The analyses of tissue expression profiles revealed that genes encoding 38 ORs, 13 antennal IRs, 11 GRs, 1 SNMP, 24 OBPs, and 12 CSPs were predominately expressed in antennae. No significant differences in expression levels of these genes were found between males and females. Genes encoding 5 non-NMDA iGluRs, 3 GRs, 2 SNMPs, 5 OBPs, and 12 CSPs were predominately expressed in forelegs. RT-PCR assays for SNMPs, OBPs and CSPs further revealed that 3 OBPs (AtumOBP3, 26 and 28) and 3 CSPs (AtumCSP7, 8 and 21) were highly expressed in antennae. Our results enrich the gene inventory of A. tumida and facilitate the discovery of potential novel targets for developing new pest control measures.

Bacterial phylogeny predicts volatile organic compound composition and olfactory response of an aphid parasitoid

There is increasing evidence that microorganisms emit a wide range of volatile compounds (mVOCs, microbial volatile organic compounds) that act as insect semiochemicals, and therefore play an important role in insect behaviour. Although it is generally believed that phylogenetically closely related microbes tend to have similar phenotypic characteristics and therefore may elicit similar responses in insects, currently little is known about whether the evolutionary history and phylogenetic relationships among microorganisms have an impact on insect–microbe interactions. In this study, we tested the hypothesis that phylogenetic relationships among 40 Bacillus strains isolated from diverse environmental sources predicted mVOC composition and the olfactory response of the generalist aphid parasitoid Aphidius colemani. Results revealed that phylogenetically closely related Bacillus strains emitted similar blends of mVOCs and elicited a comparable olfactory response of A. colemani in Y‐tube olfactometer bioassays, varying between attraction and repellence. Analysis of the chemical composition of the mVOC blends showed that all Bacillus strains produced a highly similar set of volatiles, but often in different concentrations and ratios. Benzaldehyde was produced in relatively high concentrations by strains that repel A. colemani, while attractive mVOC blends contained relatively higher amounts of acetoin, 2,3‐butanediol, 2,3‐butanedione, eucalyptol and isoamylamine. Overall, these results indicate that bacterial phylogeny had a strong impact on mVOC compositions and as a result on the olfactory responses of insects.

Volatiles of bacteria associated with parasitoid habitats elicit distinct olfactory responses in an aphid parasitoid and its hyperparasitoid

Abstract To locate mating partners and essential resources such as food, oviposition sites and shelter, insects rely to a large extent on chemical cues. While most research has focused on cues derived from plants and insects, there is mounting evidence that indicates that micro‐organisms emit volatile compounds that may play an important role in insect behaviour. In this study, we assessed how volatile compounds emitted by phylogenetically diverse bacteria affected the olfactory response of the primary parasitoid Aphidius colemani and one of its secondary parasitoids, Dendrocerus aphidum. Olfactory responses were evaluated for volatile blends emitted by bacteria isolated from diverse sources from the parasitoid's habitat, including aphids, aphid mummies and honeydew, and from the parasitoids themselves. Results revealed that A. colemani showed a wide variation in response to bacterial volatiles, ranging from significant attraction over no response to significant repellence. Our results further showed that the olfactory response of A. colemani to bacterial volatile emissions was different from that of D. aphidum. Gas chromatography‐mass spectrometry analysis of the volatile blends revealed that bacterial strains repellent to A. colemani produced significantly higher amounts of esters, organic acids, aromatics and cycloalkanes than attractive strains. Strains repellent to D. aphidum produced significantly higher amounts of alcohols and ketones, whereas the strains attractive to D. aphidum produced higher amounts of the monoterpenes limonene, linalool and geraniol. Overall, our results indicate that bacterial volatiles can have an important impact on insect olfactory responses, and should therefore be considered as an additional, so far often overlooked factor in studying multitrophic interactions between plants and insects. A free Plain Language Summary can be found within the Supporting Information of this article.

Floral vibrations by buzz-pollinating bees achieve higher frequency, velocity and acceleration than flight and defence vibrations.

Vibrations play an important role in insect behaviour. In bees, vibrations are used in a variety of contexts including communication, as a warning signal to deter predators and during pollen foraging. However, little is known about how the biomechanical properties of bee vibrations vary across multiple behaviours within a species. In this study, we compared the properties of vibrations produced by Bombus terrestris audax (Hymenoptera: Apidae) workers in three contexts: during flight, during defensive buzzing, and in floral vibrations produced during pollen foraging on two buzz-pollinated plants (Solanum, Solanaceae). Using laser vibrometry, we were able to obtain contactless measures of both the frequency and amplitude of the thoracic vibrations of bees across the three behaviours. Despite all three types of vibrations being produced by the same power flight muscles, we found clear differences in the mechanical properties of the vibrations produced in different contexts. Both floral and defensive buzzes had higher frequency and amplitude velocity, acceleration and displacement than the vibrations produced during flight. Floral vibrations had the highest frequency, amplitude velocity and acceleration of all the behaviours studied. Vibration amplitude, and in particular acceleration, of floral vibrations has been suggested as the key property for removing pollen from buzz-pollinated anthers. By increasing frequency and amplitude velocity and acceleration of their vibrations during vibratory pollen collection, foraging bees may be able to maximise pollen removal from flowers, although their foraging decisions are likely to be influenced by the presumably high cost of producing floral vibrations.

Chemosensory Gene Families in the Oligophagous Pear Pest Cacopsylla chinensis (Hemiptera: Psyllidae).

Chemosensory systems play an important role in insect behavior, and some key associated genes have potential as novel targets for pest control. Cacopsylla chinensis is an oligophagous pest and has become one of the main pests of pear trees, but little is known about the molecular-level means by which it locates its hosts. In this study, we assembled the head transcriptome of C. chinensis using Illumina sequencing, and 63,052 Unigenes were identified. A total of 36 candidate chemosensory genes were identified, including five different families: 12 odorant binding proteins (OBPs), 11 chemosensory proteins (CSPs), 7 odorant receptors (ORs), 4 ionotropic receptors (IRs), and 2 gustatory receptors (GRs). The number of chemosensory gene families is consistent with that found in other Hemipteran species, indicating that our approach successfully obtained the chemosensory genes of C. chinensis. The tissue expression of all genes using quantitative real-time PCR (qRT-PCR) found that some genes displayed male head, female head, or nymph-biased specific/expression. Our results enrich the gene inventory of C. chinensis and provide valuable resources for the analysis of the functions of some key genes. This will help in developing molecular targets for disrupting feeding behavior in C. chinensis.

A Tyrosine-Hydroxylase Characterization of Dopaminergic Neurons in the Honey Bee Brain.

Dopamine (DA) plays a fundamental role in insect behavior as it acts both as a general modulator of behavior and as a value system in associative learning where it mediates the reinforcing properties of unconditioned stimuli (US). Here we aimed at characterizing the dopaminergic neurons in the central nervous system of the honey bee, an insect that serves as an established model for the study of learning and memory. We used tyrosine hydroxylase (TH) immunoreactivity (ir) to ensure that the neurons detected synthesize DA endogenously. We found three main dopaminergic clusters, C1–C3, which had been previously described; the C1 cluster is located in a small region adjacent to the esophagus (ES) and the antennal lobe (AL); the C2 cluster is situated above the C1 cluster, between the AL and the vertical lobe (VL) of the mushroom body (MB); the C3 cluster is located below the calyces (CA) of the MB. In addition, we found a novel dopaminergic cluster, C4, located above the dorsomedial border of the lobula, which innervates the visual neuropils of the bee brain. Additional smaller processes and clusters were found and are described. The profuse dopaminergic innervation of the entire bee brain and the specific connectivity of DA neurons, with visual, olfactory and gustatory circuits, provide a foundation for a deeper understanding of how these sensory modules are modulated by DA, and the DA-dependent value-based associations that occur during associative learning.

Comparative transcriptome analysis of chemosensory genes in two sister leaf beetles provides insights into chemosensory speciation

Divergence in chemosensory traits has been posited as an important component of chemosensory speciation in insects. In particular, chemosensory genes expressed in the peripheral sensory neurons are likely to influence insect behaviors such as preference for food, oviposition sites, and mates. Despite their key role in insect behavior and potentially speciation, the underlying genetic basis for divergence in chemosensory traits remains largely unexplored. One way to ascertain the role of chemosensory genes in speciation is to make comparisons of these genes across closely related species to detect the genetic signatures of divergence. Here, we used high throughput transcriptome analysis to compare chemosensory genes of the sister leaf beetles species Pyrrhalta maculicollis and P. aenescens, whose sexual isolation and host plant preference are mediated by divergent chemical signals. Although there was low overall divergence between transcriptome profiles, there were a number of genes that were differentially expressed between the species. Furthermore, we also detected two chemosensory genes under positive selection, one of which that was also differentially expressed between the species, suggesting a possible role for these genes in chemical-based premating reproductive isolation and host use. Combined with the available chemical and ecological work in this system, further studies of the divergent chemosensory genes presented here will provide insight into the process of chemosensory speciation among Pyrrhalta beetles.

Molecular identification and sex distribution of two chemosensory receptor families in Athetis lepigone by antennal transcriptome analysis

Chemosensory receptors play central roles in insect behavior and can potentially be used as new targets for pest control. Athetis lepigone Möschler (Lepidoptera: Noctuidae) is a polyphagous insect pest found in many countries in Europe and Asia. However, systemic identification of the chemosensory receptors in the antennae has not been reported. In the present study, we obtained the antennal transcriptome of A. lepigone using Illumina sequencing technology and identified 80 candidate chemosensory receptors, including 61 transcripts encoding for odorant receptors (ORs) and 19 for ionotropic receptors (IRs). This result is similar to that reported for other lepidopteran species, suggesting that our approach successfully identified ORs and IRs of A. lepigone. RT-PCR was used to determine the sex distribution patterns of all OR and IR genes. Some OR and IR genes showed male- or female-specific/predominant expression. Our results help understand the functions of chemosensory receptor genes in A. lepigone as well as other insects, and suggest that these genes could be potential targets for developing environment-friendly behavioral antagonists and pesticides against A. lepigone.

Molecular Characterization and Sex Distribution of Chemosensory Receptor Gene Family Based on Transcriptome Analysis of Scaeva pyrastri.

Chemosensory receptors play key roles in insect behavior. Thus, genes encoding these receptors have great potential for use in integrated pest management. The hover fly Scaeva pyrastri (L.) is an important pollinating insect and a natural enemy of aphids, mainly distributed in the Palearctic and Nearctic regions. However, a systematic identification of their chemosensory receptor genes in the antennae has not been reported. In the present study, we assembled the antennal transcriptome of S. pyrastri by using Illumina sequencing technology. Analysis of the transcriptome data identified 60 candidate chemosensory genes, including 38 for odorant receptors (ORs), 16 for ionotropic receptors (IRs), and 6 for gustatory receptors (GRs). The numbers are similar to those of other Diptera species, suggesting that we were able to successfully identify S. pyrastri chemosensory genes. We analyzed the expression patterns of all genes by using reverse transcriptase PCR (RT-PCR), and found that some genes exhibited sex-biased or sex-specific expression. These candidate chemosensory genes and their tissue expression profiles provide information for further studies aimed at fully understanding the molecular basis behind chemoreception-related behaviors in S. pyrastri.

Genetic polymorphism at an odorant receptor gene (Or39) among mosquitoes of the Anopheles gambiae complex in Senegal (West Africa).

BackgroundOlfaction plays a significant role in insect behavior during critical steps of their life-cycle, such as host-seeking during foraging or the search for a mate. Here, we explored genetic polymorphism within and divergence between sibling species of the African malaria mosquito, Anopheles gambiae sensu lato in the gene sequence and encoded peptides of an odorant receptor, Or39. This study included sympatric specimens of An. gambiae sensu stricto, An. coluzzii and An. arabiensis sampled together in the village of Dielmo, Senegal.ResultsA 1,601 bp genomic sequence composed of 6 exons and 5 introns was obtained for Or39 from 6–8 mosquitoes in each of the 3 species. DNA sequence analysis revealed a high level of molecular polymorphism (π = 0.0154; Haplotype diversity = 0.867) and high overall genetic differentiation between taxa (Fst > 0.92, P < 0.01). In total, 50 parsimony informative sites were recorded. Throughout the whole dataset, there were 13 non-synonymous mutations resulting in aminoacid changes in the encoded protein. Each of the 6 different identified peptides was species-specific and none was shared across species. Most aminoacid changes were located on the intracellular domains of the protein. However, intraspecific polymorphisms in An. gambiae and An. arabiensis as well as species-specific mutations also occurred in the first extracellular domain.ConclusionsAlthough obtained from a limited number of specimens, our results point towards genetic differences between cryptic species within the An. gambiae complex in a gene of biological relevance that might be of evolutionary significance when exposed to disruptive selective forces.

Identification of Olfactory Volatiles using Gas Chromatography-Multi-unit Recordings (GCMR) in the Insect Antennal Lobe

All organisms inhabit a world full of sensory stimuli that determine their behavioral and physiological response to their environment. Olfaction is especially important in insects, which use their olfactory systems to respond to, and discriminate amongst, complex odor stimuli. These odors elicit behaviors that mediate processes such as reproduction and habitat selection(1-3). Additionally, chemical sensing by insects mediates behaviors that are highly significant for agriculture and human health, including pollination(4-6), herbivory of food crops(7), and transmission of disease(8,9). Identification of olfactory signals and their role in insect behavior is thus important for understanding both ecological processes and human food resources and well-being. To date, the identification of volatiles that drive insect behavior has been difficult and often tedious. Current techniques include gas chromatography-coupled electroantennogram recording (GC-EAG), and gas chromatography-coupled single sensillum recordings (GC-SSR)(10-12). These techniques proved to be vital in the identification of bioactive compounds. We have developed a method that uses gas chromatography coupled to multi-channel electrophysiological recordings (termed 'GCMR') from neurons in the antennal lobe (AL; the insect's primary olfactory center)(13,14). This state-of-the-art technique allows us to probe how odor information is represented in the insect brain. Moreover, because neural responses to odors at this level of olfactory processing are highly sensitive owing to the degree of convergence of the antenna's receptor neurons into AL neurons, AL recordings will allow the detection of active constituents of natural odors efficiently and with high sensitivity. Here we describe GCMR and give an example of its use. Several general steps are involved in the detection of bioactive volatiles and insect response. Volatiles first need to be collected from sources of interest (in this example we use flowers from the genus Mimulus (Phyrmaceae)) and characterized as needed using standard GC-MS techniques(14-16). Insects are prepared for study using minimal dissection, after which a recording electrode is inserted into the antennal lobe and multi-channel neural recording begins. Post-processing of the neural data then reveals which particular odorants cause significant neural responses by the insect nervous system. Although the example we present here is specific to pollination studies, GCMR can be expanded to a wide range of study organisms and volatile sources. For instance, this method can be used in the identification of odorants attracting or repelling vector insects and crop pests. Moreover, GCMR can also be used to identify attractants for beneficial insects, such as pollinators. The technique may be expanded to non-insect subjects as well.

Parasitoids Modify Their Oviposition Behavior According to the Sexual Origin of Conspecific Cuticular Hydrocarbon Traces

Hydrocarbons play a crucial role in insect behavior in general and in sexual recognition in particular. Parasitoids often modify their oviposition behavior according to hydrocarbons left by conspecifics on the reproductive patch, such as oviposition markers left by females after oviposition, or cuticular hydrocarbon (CHC) traces left by individuals by walking or rubbing. This study determined whether Eupelmus vuilleti females are able to distinguish CHCs left by male or female conspecifics on seeds. The results show that the cuticular profile of E. vuilleti differs according to its gender, and that females are able to detect the sexual origin of these CHCs. Moreover, they adjust their oviposition behavior according to the nature of these traces. Although females lay fewer eggs on hosts when confronted with female CHCs, they lay more daughters when confronted with male CHCs, thus changing the sex ratio.

Neuroanatomy and neurophysiology of the locust hypocerebral ganglion

The insect stomatogastric ganglia control foregut movements. Most previous work on the system has concentrated on the frontal ganglion (FG), including research into the role of the FG in feeding as well as molting-related behavior, mostly in locusts, but also in other insect species. The stomatogastric system exerts its physiological actions by way of careful interaction and coordination between its different neural centers and pattern-generating circuits. One such hitherto unstudied neural center is the hypocerebral ganglion (HG), which is connected to the FG via the recurrent nerve. It sends two pairs of nerves along the esophagus and to the posterior region of the crop, terminating in the paired ingluvial ganglia. Very little is known about the neuroanatomy and neurophysiology of the insect HG. Here we investigate, for the first time, the neuronal composition of the locust HG, as well as its motor output. We identify rhythmic patterns endogenous to the isolated HG, demonstrating the presence of a central pattern-generating network. Our findings suggest interactions between the HG and FG rhythm-generating circuits leading to complex physiological actions of both ganglia. This work will serve as a basis for future investigation into the physiology of the HG and its role in insect behavior.

Roles and Effects of Environmental Carbon Dioxide in Insect Life

Carbon dioxide (CO(2)) is a ubiquitous sensory cue that plays multiple roles in insect behavior. In recent years understanding of the well-known role of CO(2) in foraging by hematophagous insects (e.g., mosquitoes) has grown, and research on the roles of CO(2) cues in the foraging and oviposition behavior of phytophagous insects and in behavior of social insects has stimulated interest in this area of insect sensory biology. This review considers those advances, as well as some of the mechanistic bases of the modulation of behavior by CO(2) and important progress in our understanding of the detection and CNS processing of CO(2) information in insects. Finally, this review briefly addresses how the ongoing increase in atmospheric CO(2) levels may affect insect life.