Insect Chemical Communication Research Articles

Method for disrupting the reproductive pheromone bond of the cotton bollworm on sunflower crops

Background: Synthetic analogs of sexual pheromones provide an effective method of biorational control of the number of phytophagous insects based on the directed disruption of reproductive functions in populations of the target pest on the protected crop without affecting the integrity of the agrocenosis. The work aims to determine the biological effectiveness of methods of mass trapping and disorientation of the cotton bollworm (Helicoverpa armigera) to protect sunflower (Helianthus annuus) crops.Methods: The experiments were carried out in 2022-2023 in the Central Zone of the Krasnodar Territory, Russia in the stationary scientific crop rotation at the Federal Research Center for Biological Plant Protection, Krasnodar (2.5 ha) and the Kononenko farm, Novovelichkovskaya village, Dinsky district (10 ha). The biological effectiveness of elimination and disorientation methods was judged by the degree of damage to plants and buds. The disorientation effect was calculated by the number of males captured at the disorientation sites compared to the control sites.Result: The decrease in males’ search activity (disorientation effect) reached 85.3%. The damage to buds by the pest in this variant was 2.0% compared to 10.8% in the control group. In the mass trapping variant, it equaled 1.5-3.0 and 7.5%, respectively, indicating the prospects of including these techniques in the H. armigera biorational control system.Conclusion: Our findings could aid future research on insect chemical communication to develop effective biological control methods.Keywords: Biological plant protection; Sunflower; Cotton bollworm; Synthetic sex pheromones; Elimination; Disorientation; Biological effectiveness

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.

Chemosensory input from mouthparts in response to sexually dimorphic cuticular wax mediates male sexual discrimination in Galerucella grisescens (Coleoptera: Chrysomelidae)

The surface of the insect body is covered with a hydrophobic layer called cuticular wax (CW). In addition to functioning as an anti-desiccation agent, CW is critical for chemical communication. It has been reported that in Chrysomelidae, males discriminate between sexes based on the sex-specific CW. However, little is known regarding the underlying sensory basis. Herein, we demonstrate that chemosensory input from mouthparts mediates sexual discrimination in male Galerucella grisescens (Chrysomelidae). Observations of mating behaviour, bioassays for CW, and chemical analyses revealed that G. grisescens possess qualitatively sexually dimorphic CW, and such compositional differences allow males to distinguish between sexes. Using electron microscopy, blocking male chemosensory organs, and electrophysiological experiments, we showed that male mouthparts bear chemosensory sensilla tuned to female CW components, and sensory input from them induces male aedeagal insertion, a common male behavioural response to females. Thus, detecting CW via mouthparts is essential for males to distinguish between sexes, consistent with the fact that males inspect conspecific individuals by licking their body surfaces. To our best knowledge, this is the first report describing the detailed functional roles of mouthparts in sexual discrimination in Coleoptera. We believe that this study will promote further studies on insect chemical communication.

Pathogen-Mediated Alterations of Insect Chemical Communication: From Pheromones to Behavior.

Pathogens can influence the physiology and behavior of both animal and plant hosts in a manner that promotes their own transmission and dispersal. Recent research focusing on insects has revealed that these manipulations can extend to the production of pheromones, which are pivotal in chemical communication. This review provides an overview of the current state of research and available data concerning the impacts of bacterial, viral, fungal, and eukaryotic pathogens on chemical communication across different insect orders. While our understanding of the influence of pathogenic bacteria on host chemical profiles is still limited, viral infections have been shown to induce behavioral changes in the host, such as altered pheromone production, olfaction, and locomotion. Entomopathogenic fungi affect host chemical communication by manipulating cuticular hydrocarbons and pheromone production, while various eukaryotic parasites have been observed to influence insect behavior by affecting the production of pheromones and other chemical cues. The effects induced by these infections are explored in the context of the evolutionary advantages they confer to the pathogen. The molecular mechanisms governing the observed pathogen-mediated behavioral changes, as well as the dynamic and mutually influential relationships between the pathogen and its host, are still poorly understood. A deeper comprehension of these mechanisms will prove invaluable in identifying novel targets in the perspective of practical applications aimed at controlling detrimental insect species.

Emergence of terpene chemical communication in insects: Evolutionary recruitment of isoprenoid metabolism.

Insects have evolved a chemical communication system using terpenoids, a structurally diverse class of specialized metabolites, previously thought to be exclusively produced by plants and microbes. Gene discovery, bioinformatics, and biochemical characterization of multiple insect terpene synthases (TPSs) revealed that isoprenyl diphosphate synthases (IDS), enzymes from primary isoprenoid metabolism, are their likely evolutionary progenitors. However, the mutations underlying the emergence of the TPS function remain a mystery. To address this gap, we present the first structural and mechanistic model for the evolutionary emergence of TPS function in insects. Through identifying key mechanistic differences between IDS and TPS enzymes, we hypothesize that the loss of isopentenyl diphosphate (IPP) binding motifs strongly correlates with the gain of the TPS function. Based on this premise, we have elaborated the first explicit structural definition of isopentenyl diphosphate-binding motifs (IBMs) and used the IBM definitions to examine previously characterized insect IDSs and TPSs and to predict the functions of as yet uncharacterized insect IDSs. Consistent with our hypothesis, we observed a clear pattern of disruptive substitutions to IBMs in characterized insect TPSs. In contrast, insect IDSs maintain essential consensus residues for binding IPP. Extending our analysis, we constructed the most comprehensive phylogeny of insect IDS sequences (430 full length sequences from eight insect orders) and used IBMs to predict the function of TPSs. Based on our analysis, we infer multiple, independent TPS emergence events across the class of insects, paving the way for future gene discovery efforts. This article is protected by copyright. All rights reserved.

Antennal sensory structures of Phenacoccus solenopsis (Hemiptera: Pseudococcidae)

BackgroundThe cotton mealybug Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) is one of the most devastating sap-sucking pests of cultivated plants. The success of P. solenopsis is attributable to its ecological resilience and insecticide resistance, making its control extremely difficult and expensive. Thus, alternative safe approaches are needed to prevent the pest population from reaching the economic threshold. One of these novel approaches is based on the fact that chemical communication via the olfactory system drives critical behaviors required for the survival and development of the species. This knowledge can be useful for controlling insect pests using traps based on semiochemicals. The antennae of insects are an invaluable model for studying the fundamentals of odor perception. Several efforts have been made to investigate the histological and ultrastructural organization of the olfactory organs, such as the antennae and maxillary palps, in many insect species. However, studies on the antennal sensory structures of Phenacoccus species are lacking. Furthermore, although enormous progress has been made in understanding the antennal structures of many mealybug species, the olfactory sensilla in the antennae of P. solenopsis have not yet been described. In this study, we describe, for the first time, the morphology and distribution of the antennal sensilla in male and female P. solenopsis using scanning electron microscopy.ResultsOur results revealed that the entire antennae length and the number of flagellar segments were different between the sexes. Eight morphological types of sensilla were identified on male antennae: trichoid sensilla, chaetic sensilla (three subtypes), basiconic sensilla (two subtypes), and campaniform sensilla (two subtypes). Six morphological types of sensilla were found on female antennae. Sensilla chaetica of subtype 2 and campaniform sensilla of subtype 1 were distributed only on male antennae, suggesting that these sensilla are involved in the recognition of female sex pheromones. The subtype 1 of sensilla chaetica was significantly more abundant on female antennae than on male ones, while subtype 3 was only located on the terminal flagellar segment of the antenna in both sexes.ConclusionsThis study provides insightful information for future electrophysiological and behavioral studies on chemical communication in insects, particularly the cotton mealybug, P. solenopsis that could help in developing new strategies for controlling this economically important insect species.

Herbicides and their potential to disrupt plant–insect chemical communication

AbstractEcological interactions between plants and insects are of paramount importance for the maintenance of biodiversity and ecosystem functioning. Herbicides have long been considered a threat to plant and insect populations, but global increases in intensive agriculture and availability of herbicide‐resistant crops have intensified concerns about their full impact on biodiversity. Here, we argue that exposure to sublethal herbicide doses has the potential to alter plant–insect interactions as a result of disruptions in their chemical communication. This is because herbicides interfere with biosynthetic pathways and phytohormones involved in the production of several classes of plant volatiles that mediate plant–insect chemical communication. Sublethal herbicide doses can modify the morphological and life‐history plant traits and affect interactions with insects. However, the potential changes in plant volatiles and their consequences for plant–insect chemical communication have not yet received as much attention. We discuss how target‐site (disruptors of primary metabolism) and non‐target‐site (synthetic auxins) herbicides could alter the production of plant volatiles and disrupt plant–insect chemical communication. We suggest research avenues to fill in the current gap in our knowledge that might derive recommendations and applied solutions to minimize herbicides' impacts on plant–insect interactions and biodiversity.

Plant–insect chemical communication in ecological communities: An information theory perspective

AbstractCross‐species communication, where signals are sent by one species and perceived by others, is one of the most intriguing types of communication that functionally links different species to form complex ecological networks. Global change and human activity can affect communication by increasing fluctuations in species composition and phenology, altering signal profiles and intensity, and introducing noise. So far, most studies on cross‐species communication have focused on a few specific species isolated from ecological communities. Scaling up investigations of cross‐species communication to the community level is currently hampered by a lack of conceptual and practical methodologies. Here, we propose an interdisciplinary framework based on information theory to investigate mechanisms shaping cross‐species communication at the community level. We use plants and insects, the cornerstones of most ecosystems, as a showcase and focus on chemical communication as the key communication channel. We first introduce some basic concepts of information theory, then we illustrate information patterns in plant–insect chemical communication, followed by a further exploration of how to integrate information theory into ecological and evolutionary processes to form testable mechanistic hypotheses. We conclude by highlighting the importance of community‐level information as a means to better understand the maintenance and workings of ecological systems, especially during rapid global change.

Enantiomeric Discrimination in Insects: The Role of OBPs and ORs.

Simple SummaryInsects 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.

Human Impacts on Insect Chemical Communication in the Anthropocene

The planet is presently undergoing dramatic changes caused by human activities. We are living in the era of the Anthropocene, where our activities directly affect all living organisms on Earth. Insects constitute a major part of the world’s biodiversity and currently, we see dwindling insect biomass but also outbreaks of certain populations. Most insects rely on chemical communication to locate food, mates, and suitable oviposition sites, but also to avoid enemies and detrimental microbes. Emissions of, e.g., CO2, NOx, and ozone can all affect the chemical communication channel, as can a rising temperature. Here, we present a review of the present state of the art in the context of anthropogenic impact on insect chemical communication. We concentrate on present knowledge regarding fruit flies, mosquitoes, moths, and bark beetles, as well as presenting our views on future developments and needs in this emerging field of research. We include insights from chemical, physiological, ethological, and ecological directions and we briefly present a new international research project, the Max Planck Centre for Next Generation Insect Chemical Ecology (nGICE), launched to further increase our understanding of the impact of human activities on insect olfaction and chemical communication.

Novel insights into insect chemical communication – an introduction

The topics of insect chemical communication in this issue can be subdivided into intra- and interspecific signaling mechanisms. For intraspecific chemical communication, several articles characterize sexual cues involved in mate recognition and mating behavior, others discuss mechanisms associated with the neurobiology of chemoreception. For interspecific chemical communication, one article links an olfactory recognition mechanism to a phoretic relationship, i.e., using a heterospecific organism for dispersal. Another identifies a parasitoid-derived mortality-inducing protein, and discusses its potential for biological pest control.

Expression Profiles and Functional Characterization of Chemosensory Protein 15 (HhalCSP15) in the Brown Marmorated Stink Bug Halyomorpha halys.

Chemosensory proteins (CSPs) have been identified in the sensory tissues of various insect species and are believed to be involved in chemical communication in insects. However, the physiological roles of CSPs in Halyomorpha halys, a highly invasive insect species, are rarely reported. Here, we focused on one of the antennal CSPs (HhalCSP15) and determined whether it was involved in olfactory perception. Reverse transcription PCR (RT-PCR) and quantitative real-time PCR (qRT-PCR) analysis showed that HhalCSP15 was enriched in nymph and male and female adult antennae, indicating its possible involvement in the chemosensory process. Fluorescence competitive binding assays revealed that three of 43 natural compounds showed binding abilities with HhalCSP15, including β-ionone (Ki=11.9±0.6μM), cis-3-hexen-1-yl benzoate (Ki=10.5±0.4μM), and methyl (2E,4E,6Z)-decatrienoate (EEZ-MDT; Ki=9.6±0.8μM). Docking analysis supported the experimental affinity for the three ligands. Additionally, the electrophysiological activities of the three ligands were further confirmed using electroantennography (EAG). EEZ-MDT is particularly interesting, as it serves as a kairomone when H. halys forages for host plants. We therefore conclude that HhalCSP15 might be involved in the detection of host-related volatiles. Our data provide a basis for further investigation of the physiological roles of CSPs in H. halys, and extend the olfactory function of CSPs in stink bugs.

Molecular ecology of plant volatiles in interactions with insect herbivores

Plant-derived volatile organic compounds (VOCs) play pivotal roles in interactions with insect herbivores. Individual VOCs can be directly toxic or deterrent, serve as signal molecules to attract natural enemies, and/or be perceived by distal plant tissues as a priming signal to prepare for expected herbivory. Environmental conditions, as well as the specific plant-insect interaction being investigated, strongly influence the observed functions of VOC blends. The complexity of plant-insect chemical communication via VOCs is further enriched by the sophisticated molecular perception mechanisms of insects, which can respond to one or more VOCs and thereby influence insect behavior in a manner that has yet to be fully elucidated. Despite numerous gaps in the current understanding of VOC-mediated plant-insect interactions, successful pest management strategies such as push-pull systems, synthetic odorant traps, and crop cultivars with modified VOC profiles have been developed to supplement chemical pesticide applications and enable more sustainable agricultural practices. Future studies in this field would benefit from examining the responses of both plants and insects in the same experiment to gain a more complete view of these interactive systems. Furthermore, a molecular evolutionary study of key genetic elements of the ecological interaction phenotypes could provide new insights into VOC-mediated plant communication with insect herbivores.

Aphid Odorant-Binding Protein 9 Is Narrowly Tuned to Linear Alcohols and Aldehydes of Sixteen Carbon Atoms.

Simple SummaryOdorant-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.

Behavioural and electrophysiological responses of Liothrips jatrophae (Thysanoptera: Phlaeothripidae) to conspecific extracts and some of its identified compounds

AbstractThis study investigated the behavioural and electrophysiological responses of the thrips Liothrips jatrophae to conspecific extracts and some of its identified compounds. We integrated four constituents of the insect's chemical communication: identification of the compounds from L. jatrophae extract, behavioural response to extracts and synthetic blends, morphology of the main olfactory receptors (antennae), and electrophysiological response of the main olfactory receptors of the antennae to extracts and to individual synthetic compounds and synthetic blends. Analysis by GC–MS revealed that the L. jatrophae extract contains a mixture of 11 compounds (perillene, tridecane, β‐acaridial, tetradecane, pentadecane, heptadecane, heptadecene, dodecyl isobutyrate, tridecyl isobutyrate, tetradecyl isobutyrate and hexadecyl isobutyrate). The major compounds were tridecane, pentadecane, tetradecyl isobutyrate and hexadecyl isobutyrate. When the thrips were exposed to the extracts, they exhibited an escape response, accompanied by other behavioural responses, such as rapid sideways movement and lifting of the abdomen and secretion of a drop at the tip of the abdomen. Morphology and distribution of the L. jatrophae antennal sensilla were examined with a scanning electron microscope (SEM). The antenna consists of a scape, pedicel and six flagella. The microphotographs showed two types of sensilla: basiconica and trichoidea. Single sensillum recording (SSR) showed that the sensilla basiconica located in segment IV of the antenna were the most sensitive to the L. jatrophae extracts and to a synthetic blend, whereas the sensilla trichoidea did not exhibit electrophysiological response.

Impacts of ionization radiation on the cuticular hydrocarbon profile and mating success of male house crickets (Acheta domesticus)

Purpose Ionizing radiation is well known to have drastic impacts on major life history features including survivorship, growth, fertility, and longevity. What is much less appreciated is how radiation stress can cause changes to more subtle traits, such as those associated with sexual signaling, an underappreciated but vital aspect of insect reproduction. In the House Cricket (Acheta domesticus) cuticular hydrocarbons are vital for sex and species recognition, as well as a possible indicator of stress, making them crucial for successful mating and reproduction. Materials and methods Here, we analyze the impacts of ionizing radiation on the cuticular hydrocarbons of male crickets and its subsequent impacts on mating success. We exposed juvenile (14-day, 4th instar) male crickets to a broad range of radiation doses (2 Gy − 2 Gy). Results We detected significant changes in individual cuticular hydrocarbons across a broad range of doses in mature male crickets using gas-liquid chromatography. Specifically, dose was identified as a significant contributing factor to hydrocarbon increases p < .0001. Mating success was significantly reduced in 12 Gy (p < .0001), 10 Gy (0.0001), and 7 Gy (0.0060) groups compared to non-irradiated controls. Conclusion Insect chemical communication can be species specific, and functionally specialized. Here, we show that radiation can alter the chemical signals used to attract mates in a large bodied insect and this may be a contributing factor to the described reduction in male mating success. Further research should be conducted to further analyze the various modes of communication employed by male crickets to attract mates i.e. acoustic signaling, and how this may also contribute to the reduction in mating success seen in irradiated males.

Functional and evolutionary characterization of chemosensory protein CSP2 in the whitefly, Bemisia tabaci.

Chemosensory proteins (CSPs) are thought to play essential roles in insect chemical communication, but their exact physiological functions remain unclear. In this study, we investigated the functions of the CSP2 gene in the whitefly Bemisia tabaci using protein expression and the binding affinity spectrum of CSP2 to different types of odor molecules. Moreover, the evolutionary characteristics of the CSP2 gene were studied. The data obtained using binding assay showed that the CSP2 protein can bind to a broad range of plant volatiles including the homoterpene (E)-3,8-dimethyl-1,4,7-nonatriene (DMNT) and its analogs. In addition, using a behavioral experimental approach we identified that DMNT can repel the selection and oviposition of B. tabaci. Furthermore, protein structure modeling, molecular docking analyses and a functional mutation experiment were carried out resulting in the final identification of key amino acid residue Y11, which displayed important roles in the binding of CSP2 to DMNT. The results also showed that Y11 is located in the pocket region where CSP2 has a pi-alkyl interaction with DMNT. Meanwhile, comparative and evolutionary analyses indicated that CSP2 shared a high sequence similarity with CSPs of other insect family members such as Sternorrhyncha and Auchenorrhyncha including aphids, whiteflies and planthoppers. These results suggested that CSP2 likely contributes to mediating responses of B. tabaci to plant volatiles, which may play a pivotal role in its feeding and oviposition preferences. Moreover, these findings could provide key information for exploring efficiency monitoring and integrated pest management strategies of B. tabaci.

Odorant receptor phylogeny confirms conserved channels for sex pheromone and host plant signals in tortricid moths.

The search for mates and food is mediated by volatile chemicals. Insects sense food odorants and sex pheromones through odorant receptors (ORs) and pheromone receptors (PRs), which are expressed in olfactory sensory neurons. Molecular phylogenetics of ORs, informed by behavioral and functional data, generates sound hypotheses for the identification of semiochemicals driving olfactory behavior. Studying orthologous receptors and their ligands across taxa affords insights into the role of chemical communication in reproductive isolation and phylogenetic divergence. The female sex pheromone of green budworm moth Hedya nubiferana (Lepidoptera, Totricidae) is a blend of two unsaturated acetates, only a blend of both elicits male attraction. Females produce in addition codlemone, which is the sex pheromone of another tortricid, codling moth Cydia pomonella. Codlemone also attracts green budworm moth males. Concomitantly, green budworm and codling moth males are attracted to the host plant volatile pear ester. A congruent behavioral response to the same pheromone and plant volatile in two tortricid species suggests co‐occurrence of dedicated olfactory channels. In codling moth, one PR is tuned to both compounds, the sex pheromone codlemone and the plant volatile pear ester. Our phylogenetic analysis finds that green budworm moth expresses an orthologous PR gene. Shared ancestry, and high levels of amino acid identity and sequence similarity, in codling and green budworm moth PRs offer an explanation for parallel attraction of both species to the same compounds. A conserved olfactory channel for a sex pheromone and a host plant volatile substantiates the alliance of social and habitat signals in insect chemical communication. Field attraction assays confirm that in silico investigations of ORs afford powerful predictions for an efficient identification of behavior‐modifying semiochemicals, for an improved understanding of the mechanisms of host plant attraction in insect herbivores and for the further development of sustainable insect control.

Characterization of the Expression and Functions of Two Odorant-Binding Proteins of Sitophilus zeamais Motschulsky (Coleoptera: Curculionoidea).

Odorant-binding proteins (OBPs) are important in insect chemical communication. The objective of this research was to identify the functions of two OBPs in Sitophilus zeamais. qRT-PCR and western blot (WB) were performed to investigate the expression profiles at the transcript and protein levels, respectively. Fluorescence competitive binding assays were used to measure the ability of the OBPs to bind to host volatiles, and a Y-tube olfactometer was used to verify the results (attraction/no response) via behavioral experiments. The RNAi was used to verify the function by knocking down the ability of proteins to bind odorants. qRT-PCR showed the highest expression SzeaOBP1 and SzeaOBP28 at the low-instar larva (LL) and eclosion adult (EA) stages, respectively. WB showed that both SzeaOBP1 and SzeaOBP28 were highly expressed in the EA stage. Fluorescence competitive binding assays indicated that SzeaOBP1 exhibited extremely high binding affinity with cetanol. SzeaOBP28 exhibited a pronounced binding affinity for 4-hydroxy-3-methoxybenzaldehyde. The behavioral experiment showed that the adult S. zeamais responded strongly to 4-hydroxy-3-methoxybenzaldehyde and valeraldehyde from Sorghum bicolor. The RNAi knockdown individuals displayed behavioral differences between normal insects and dsRNA (SzeaOBP1)-treated insects. We infer that they both have functions in perception and recognition of host volatiles, whereas SzeaOBP28 may also have other functions.

2-Alkyl-3-methoxypyrazines are potent attractants of florivorous scarabs (Melolonthidae, Cyclocephalini) associated with economically exploitable Neotropical palms (Arecaceae).

2-Alkyl-3-methoxypyrazines (MPs) are naturally occurring aromatic compounds involved in insect chemical communication as both pheromones and allelochemicals. Although rarely characterized in floral scents, they have been identified as major constituents in headspace samples from palm inflorescences and evidence pointed towards their function as attractive cues for scent-oriented pollinators, as well as florivores. In this study, we investigated the occurrence of MPs in economically exploitable palms belonging to Acrocomia and Attalea through headspace floral scent analysis and assessed their role in the selective attraction of flower-feeding cyclocephaline scarabs (Melolonthidae, Cyclocephalini) in field bioassays conducted in Brazil and Colombia. Three different MPs were identified among floral headspace samples of Acrocomia aculeata, Acr. intumescens, Attalea butyracea and Att. insignis: 2-isopropyl-3-methoxypyrazine (IPMP), 2-isobutyl-3-methoxypyrazine (IBMP) and 2-(sec-butyl)-3-methoxypyrazine (SBMP). Their combined estimated total scent emissions per inflorescence ranged from 246 μg h-1 to 6.2 mg h-1 . Scented traps, individually baited with either IPMP or SBMP, resulted in species-selective and high yield captures of Cyclocephala amazona and C. distincta, known florivores associated with over 20 different species of Neotropical palms. The identification of MPs as potent kairomones could prove instrumental in integrated pest management plans for these insects in emerging oil-yielding crops in South America, including those of the wine palm (Att. butyracea) and macaw palms (Acrocomia spp.). © 2018 Society of Chemical Industry.