CHC Profiles Research Articles

Male–female chemical interactions in a facultatively parthenogenetic stick insect

AbstractFacultative parthenogenesis is a form of reproduction in which females can either lay unfertilised eggs that typically develop into female offspring only or mate and lay fertilised eggs that develop into male and female offspring. Intriguingly, facultative parthenogens often occur in mixed‐sex populations where reproduction is mostly sexual and all‐female populations where reproduction is asexual. How all‐female populations avoid invasion by males remains unknown. Here, we explored the use of pheromones in male–female communication in a facultatively parthenogenetic stick insect, the peppermint stick insect (Megacrania batesii), and compared chemical signals between females descended from sexually versus parthenogenetically reproducing populations. If parthenogenetic females release less attractive pheromones, this could help explain the persistence of all‐female populations. We found that M. batesii exhibits slight sexual dimorphism in antenna morphology, and behavioural assays provided little evidence that males could locate females solely by volatile pheromones. However, CHC profiles differed substantially between different types of females. Analysis of CHC components indicated a clear genetic difference between females descended from all‐female versus mixed‐sex populations, as well as a maternal effect of female parthenogenetic versus sexual development. Together, our results suggest that males might rely more on close‐range chemical cues to differentiate females, and chemical communication could play a role in the persistence of all‐female populations.

Natural Diversity of Cuticular Pheromones in a Local Population of Drosophila after Laboratory Acclimation.

Experimental studies of insects are often based on strains raised for many generations in constant laboratory conditions. However, laboratory acclimation could reduce species diversity reflecting adaptation to varied natural niches. Hydrocarbons covering the insect cuticle (cuticular hydrocarbons; CHCs) are reliable adaptation markers. They are involved in dehydration reduction and protection against harmful factors. CHCs can also be involved in chemical communication principally related to reproduction. However, the diversity of CHC profiles in nature and their evolution in the laboratory have rarely been investigated. Here, we sampled CHC natural diversity in Drosophila melanogaster flies from a particular location in a temperate region. We also measured cis-Vaccenyl acetate, a male-specific volatile pheromone. After trapping flies using varied fruit baits, we set up 21 D. melanogaster lines and analysed their pheromones at capture and after 1 to 40 generations in the laboratory. Under laboratory conditions, the broad initial pheromonal diversity found in male and female flies rapidly changed and became more limited. In some females, we detected CHCs only reported in tropical populations: the presence of flies with a novel CHC profile may reflect the rapid adaptation of this cosmopolitan species to global warming in a temperate area.

Aphid-Ant Relationships: The Role of Cuticular Hydrocarbons and Different Chemical Stimuli in Triggering Mutualistic Behavior

In ant-aphid interactions, various aphid species offer honeydew to the ant partner and increase their density by ant attendance, whilst others never attend ant species, in which case ants tend to treat them as prey. In this regard, ants should have the ability to distinguish myrmecophile aphid species from non-mutualistic species, so that mutualistic aphids will be accepted as partners rather than prey. Although ant-aphid interactions are now the focus of chemical ecology studies, the role of the different chemical stimuli in determining mutualistic interactions has not been completely clarified. Therefore, we have investigated the cuticular hydrocarbon profile of four myrmecophiles aphid species using GC-MS. We also investigated the behavior of the worker-ants (Lasius niger L., Hymenoptera: Formicidae) to different chemical stimuli derived from aphids. We applied four treatments and found that the behavior of the ant workers varied depending on the source of the different treatments. In particular, the real aphid Aphis pomi and the sugar solution proved to be the most attractive to the ants, while the presence of pure extract of the individuals is enough to disturb the behavior of the ants. We provide evidence that the key stimuli of the tending behavior could be the CHC patterns of the aphids and the CHC profile of the aphids tends to be genus specific. This research will promote further investigations to test the behavior of ant workers towards other species of aphids and treatment combinations.

Cuticular Hydrocarbon Profiles of Himalayan Bumble Bees (Hymenoptera: Bombus Latreille) are Species-Specific and Show Elevational Variation.

Bumble bees are important pollinators in natural environments and agricultural farmlands, and they are in particular adapted to harsh environments like high mountain habitats. In these environments, animals are exposed to low temperature and face the risk of desiccation. The Eastern Himalayas are one of the recognized biodiversity hotspots worldwide. The area covers subtropical rainforest with warm temperature and high precipitation as well as high mountain ranges with peaks reaching up to 7,000m, shaping a diverse floral and faunal community at the different elevational zones. To identify possible adaptation strategies, we investigated the cuticular hydrocarbon profiles of four bumble bee species occurring at different elevational ranges in Arunachal Pradesh, the northeastern most state in India. At 17 locations along an elevational gradient, we collected workers of two species from lower elevations (B. albopleuralis and B. breviceps; ~ 100m - 3,000m asl) and two species from higher elevations (B. prshewalskyi and B. mirus; ~ 2,800m - 4,500m asl). The CHC profiles of all four species showed a significant degree of variation in the composition of hydrocarbons, indicating species specificity. We also found clear correlation with elevation. The weighted mean chain length of the hydrocarbons significantly differed between the low and high elevation species, and the proportion of saturated hydrocarbons in CHC profiles significantly increased with the elevational range of the bumble bee species. Our results indicate that bumble bees living at high elevations reduce the risk of water loss by adapting their CHC composition on their cuticle, a phenomenon that has also been found in other insects like ants and fruit flies.

Decoding the genetic and chemical basis of sexual attractiveness in parasitic wasps.

Attracting and securing potential mating partners is of fundamental importance for reproduction. Therefore, signaling sexual attractiveness is expected to be tightly coordinated in communication systems synchronizing senders and receivers. Chemical signaling has permeated through all taxa of life as the earliest and most widespread form of communication and is particularly prevalent in insects. However, it has been notoriously difficult to decipher how exactly information related to sexual signaling is encoded in complex chemical profiles. Similarly, our knowledge of the genetic basis of sexual signaling is very limited and usually restricted to a few case studies with comparably simple pheromonal communication mechanisms. The present study jointly addresses these two knowledge gaps by characterizing two fatty acid synthase genes, that most likely evolved by tandem gene duplication, simultaneously impacting sexual attractiveness and complex chemical surface profiles in parasitic wasps. Gene knock-down in female wasps dramatically reduces their sexual attractiveness coinciding with a drastic decrease in male courtship and copulation behavior. Concordantly, we found a striking shift of methyl-branching patterns in the female surface pheromonal compounds, which we subsequently demonstrate to be the main cause for the greatly reduced male response. Intriguingly, this suggests a potential coding mechanism for sexual attractiveness mediated by specific methyl-branching patterns in CHC profiles. So far, the genetic underpinnings of methyl-branched CHCs are not well understood despite their high potential for encoding information. Our study sheds light on how biologically relevant information can be encoded in complex chemical profiles and on the genetic basis of sexual attractiveness.

Humidity stress and its consequences for male pre‐ and post‐copulatory fitness traits in an insect

AbstractGlobal declines in insect abundance are of significant concern. While there is evidence that climate change is contributing to insect declines, we know little of the direct mechanisms responsible for these declines. Male fertility is compromised by increasing temperatures, and the thermal limit to fertility has been implicated as an important factor in the response of insects to climate change. However, climate change is affecting both temperature and hydric conditions, and the effects of water availability on male fertility have rarely been considered. Here we exposed male crickets Teleogryllus oceanicus to either low or high‐humidity environments while holding temperature constant. We measured water loss and the expression of both pre‐ and postmating reproductive traits. Males exposed to a low‐humidity environment lost more water than males exposed to a high‐humidity environment. A male's cuticular hydrocarbon profile (CHC) did not affect the amount of water lost, and males did not adjust the composition of their CHC profiles in response to hydric conditions. Males exposed to a low‐humidity environment were less likely to produce courtship song or produced songs of low quality. Their spermatophores failed to evacuate and their ejaculates contained sperm of reduced viability. The detrimental effects of low‐humidity on male reproductive traits will compromise male fertility and population persistence. We argue that limits to insect fertility based on temperature alone are likely to underestimate the true effects of climate change on insect persistence and that the explicit incorporation of water regulation into our modeling will yield more accurate predictions of the effects of climate change on insect declines.

Predicting the Weathering Time by the Empty Puparium of Sarcophaga peregrina (Diptera: Sarcophagidae) with the ANN Models.

Simple SummaryThe weathering time of cuticular hydrocarbons from the puparium could assist in estimating the postmortem interval (PMI) of decomposed corpses. However, the composition of cuticular hydrocarbons in the puparium is complicated and has not been well studied for sarcophagid species. Therefore, we examined the compounds of Sarcophaga peregrina (Robineau-Desvoidy, 1830) at varying temperatures and used various machine learning models to predict the weathering time. The artificial neural network (ANN) model may be optimal for weathering time estimation.Empty puparium are frequently collected at crime scenes and may provide valuable evidence in cases with a long postmortem interval (PMI). Here, we collected the puparium of Sarcophaga peregrina (Diptera: Sarcophagidae) (Robineau-Desvoidy, 1830) for 120 days at three temperatures (10 °C, 25 °C, and 40 °C) with the aim to estimate the weathering time of empty puparium. The CHC profiles were analyzed by gas chromatography-mass spectrometry (GC-MS). The partial least squares (PLS), support vector regression (SVR), and artificial neural network (ANN) models were used to estimate the weathering time. This identified 49 CHCs with a carbon chain length between 10 and 33 in empty puparium. The three models demonstrate that the variation tendency of hydrocarbon could be used to estimate the weathering time, while the ANN models show the best predictive ability among these three models. This work indicated that puparial hydrocarbon weathering has certain regularity with weathering time and can gain insight into estimating PMI in forensic investigations.

Similarities in Recognition Cues Lead to the Infiltration of Non-Nestmates in an Ant Species.

Chemical cues are among the most important information-sharing mechanisms in insect societies, in which cuticular hydrocarbons play a central role, e.g., from nestmate recognition to queen signaling. The nestmate recognition mechanism usually prevents intruders from taking advantage of the resources stored in the nest. However, nestmate recognition is not unconditionally effective, and foreign individuals can sometimes infiltrate unrelated nests and take advantage of the colony resources. In this study, we investigated the role of overall colony odor profiles on the ability of conspecific workers to drift into unrelated colonies. We hypothesized that drifters would have higher chances of success by infiltrating colonies with the odor profiles most similar to their own nest, avoiding being detected as non-nestmates. By performing a drifting bioassay, we found that workers of the ant Formica fusca infiltrated unrelated conspecific colonies at a rate of 2.4%, significantly infiltrating colonies displaying CHC profiles most similar to their natal nests. Notably, methyl branched hydrocarbons seem to play a role as recognition cues in this species. In addition, we show that environmental rather than genetic factors are responsible for most contributions on the CHC phenotype, presenting ca. of 50% and 27.5% of explained variation respectively, and playing a major role in how worker ants detect and prevent the infiltration of non-nestmates in the colony. Hence, relying on cuticular hydrocarbons similarities could be a profitably evolutionary strategy by which workers can identify conspecific colonies, evade detection by guards, and avoid competition with genetic relatives.

Social Behavior, Ovary Size, and Population of Origin Influence Cuticular Hydrocarbons in the Orchid Bee Euglossa dilemma.

AbstractCuticular hydrocarbons (CHCs) are waxy compounds on the surface of insects that prevent desiccation and frequently serve as chemical signals mediating social and mating behaviors. Although their function in eusocial species has been heavily investigated, little is known about the evolution of CHC-based communication in species with simpler forms of social organization lacking specialized castes. Here we investigate factors shaping CHC variation in the orchid bee Euglossa dilemma, which forms casteless social groups of two to three individuals. We first assess geographic variation, examining CHC profiles of males and females from three populations. We also consider CHC variation in the sister species, Euglossa viridissima, which occurs sympatrically with one population of E. dilemma. Next, we consider variation associated with female behavioral phases, to test the hypothesis that CHCs reflect ovary size and social dominance. We uncover a striking CHC polymorphism in E. dilemma spanning populations. In addition, we identify a separate set of CHCs that correlate with ovary size, social dominance, and expression of genes associated with social behavior, suggesting that CHCs convey reproductive and social information in E. dilemma. Together, our results reveal complex patterns of variation in which a subset of CHCs reflect the social and reproductive status of nestmates.

Cuticular hydrocarbons on old museum specimens of the spiny mason wasp, Odynerus spinipes (Hymenoptera: Vespidae: Eumeninae), shed light on the distribution and on regional frequencies of distinct chemotypes

The mason wasp Odynerus spinipes shows an exceptional case of intrasexual cuticular hydrocarbon (CHC) profile dimorphism. Females of this species display one of two CHC profiles (chemotypes) that differ qualitatively and quantitatively from each other. The ratio of the two chemotypes was previously shown to be close to 1:1 at three sites in Southern Germany, which might not be representative given the Palearctic distribution of the species. To infer the frequency of the two chemotypes across the entire distributional range of the species, we analyzed with GC–MS the CHC profile of 1042 dry-mounted specimens stored in private and museum collections. We complemented our sampling by including 324 samples collected and preserved specifically for studying their CHCs. We were capable of reliably identifying the chemotypes in 91% of dry-mounted samples, some of which collected almost 200 years ago. We found both chemotypes to occur in the Far East, the presumed glacial refuge of the species, and their frequency to differ considerably between sites and geographic regions. The geographic structure in the chemotype frequencies could be the result of differential selection regimes and/or different dispersal routes during the colonization of the Western Palearctic. The presented data pave the route for disentangling these factors by providing information where to geographically sample O. spinipes for population genetic analyses. They also form the much-needed basis for future studies aiming to understand the evolutionary and geographic origin as well as the genetics of the astounding CHC profile dimorphism that O. spinipes females exhibit.

Reproductive inhibition among nestmate queens in the invasive Argentine ant

In social species, the presence of several reproductive individuals can generate conflict. In social insects, as queen number increases, individual oviposition rate may decrease because of direct and indirect behavioural and/or chemical interactions. Understanding the factors that mediate differences in queen fecundity should provide insight into the regulation and maintenance of highly polygynous insect societies, such as those of the invasive Argentine ant (Linepithema humile). In this study, we investigated (1) whether differences in the oviposition rates of Argentine ant queens exposed to polygynous conditions could result from interactions among them; (2) whether such differences in fecundity stemmed from differences in worker attention; and (3) whether polygynous conditions affected the cuticular hydrocarbon profiles of queens (CHCs). We found that differences in queen fecundity and CHC profiles observed under polygynous conditions disappeared when queens were exposed to monogynous conditions, suggesting some form of reproductive inhibition may exist when queens cohabit. These differences did not seem to arise from variation in worker attention because more fecund queens were not more attractive to workers. Levels of some CHCs were higher in more fecund queens. These CHCs are associated with greater queen productivity and survival. Our findings indicate that such compounds could be multifunctional queen pheromones.

Perceived dominance status affects chemical signalling in the neriid fly Telostylinus angusticollis

Chemical communication mediates many social interactions in insects but is still less well understood than other forms of communication. In particular, chemical signalling of social dominance is believed to play an important role in competitive interactions in both sexes, but much of the evidence is correlational. Here we manipulated social dominance and examined its effect on CHC profiles in Telostylinus angusticollis, a fly with a resource defence polygyny mating system. Focal individuals' perception of their own dominance status was manipulated by placing them in an arena with larger or smaller competitors to render them ‘subordinate’ or ‘dominant.’ We found that social dominance treatment affected males' and females' social status (quantified as proximity to the larval medium/oviposition dish), as well as their CHC profiles. Dominant individuals tended to have CHC profiles less similar to those of the opposite sex. Moreover, dominant females exhibited an overall elevation of all CHC expression, relative to subordinate females, whereas males that perceived themselves as subordinate exhibited a near-significant downregulation of male-limited CHCs. Our findings suggest that T. angusticollis males and females alter their CHC profiles in response to their self-perceived social dominance status. These chemical signals could play a role in social interactions both within and between the sexes.

Effect of antibiotic treatment and gamma-irradiation on cuticular hydrocarbon profiles and mate choice in tsetse flies (Glossina m. morsitans)

BackgroundSymbiotic microbes represent a driving force of evolutionary innovation by conferring novel ecological traits to their hosts. Many insects are associated with microbial symbionts that contribute to their host’s nutrition, digestion, detoxification, reproduction, immune homeostasis, and defense. In addition, recent studies suggest a microbial involvement in chemical communication and mating behavior, which can ultimately impact reproductive isolation and, hence, speciation. Here we investigated whether a disruption of the microbiota through antibiotic treatment or irradiation affects cuticular hydrocarbon profiles, and possibly mate choice behavior in the tsetse fly, Glossina morsitans morsitans. Four independent experiments that differentially knock down the multiple bacterial symbionts of tsetse flies were conducted by subjecting tsetse flies to ampicillin, tetracycline, or gamma-irradiation and analyzing their cuticular hydrocarbon profiles in comparison to untreated controls by gas chromatography – mass spectrometry. In two of the antibiotic experiments, flies were mass-reared, while individual rearing was done for the third experiment to avoid possible chemical cross-contamination between individual flies.ResultsAll three antibiotic experiments yielded significant effects of antibiotic treatment (particularly tetracycline) on cuticular hydrocarbon profiles in both female and male G. m. morsitans, while irradiation itself had no effect on the CHC profiles. Importantly, tetracycline treatment reduced relative amounts of 15,19,23-trimethyl-heptatriacontane, a known compound of the female contact sex pheromone, in two of the three experiments, suggesting a possible implication of microbiota disturbance on mate choice decisions. Concordantly, both female and male flies preferred non-treated over tetracycline-treated flies in direct choice assays.ConclusionsWhile we cannot exclude the possibility that antibiotic treatment had a directly detrimental effect on fly vigor as we are unable to recolonize antibiotic treated flies with individual symbiont taxa, our results are consistent with an effect of the microbiota, particularly the obligate nutritional endosymbiont Wigglesworthia, on CHC profiles and mate choice behavior. These findings highlight the importance of considering host-microbiota interactions when studying chemical communication and mate choice in insects.

Learning Distinct Chemical Labels of Nestmates in Ants.

Colony coherence is essential for eusocial insects because it supports the inclusive fitness of colony members. Ants quickly and reliably recognize who belongs to the colony (nestmates) and who is an outsider (non-nestmates) based on chemical recognition cues (cuticular hydrocarbons: CHCs) which as a whole constitute a chemical label. The process of nestmate recognition often is described as matching a neural template with the label. In this study, we tested the prevailing view that ants use commonalities in the colony odor that are present in the CHC profile of all individuals of a colony or whether different CHC profiles are learned independently. We created and manipulated sub-colonies by adding one or two different hydrocarbons that were not present in the original colony odor of our Camponotus floridanus colony and later tested workers of the sub-colonies in one-on-one encounters for aggressive responses. We found that workers adjust their nestmate recognition by learning novel, manipulated CHC profiles, but still accept workers with the previous CHC profile. Workers from a sub-colony with two additional components showed aggression against workers with only one of the two components added to their CHC profile. Thus, additional components as well as the lack of a component can alter a label as “non-nestmate.” Our results suggest that ants have multiple-templates to recognize nestmates carrying distinct labels. This finding is in contrast to what previously has been proposed, i.e., a widening of the acceptance range of one template. We conclude that nestmate recognition in ants is a partitioned (multiple-template) process of the olfactory system that allows discrimination and categorization of nestmates by differences in their CHC profiles. Our findings have strong implications for our understanding of the underlying mechanisms of colony coherence and task allocation because they illustrate the importance of individual experience and task associated differences in the CHC profiles that can be instructive for the organization of insect societies.

Differences in the reliance on cuticular hydrocarbons as sexual signaling and species discrimination cues in parasitoid wasps

BackgroundCuticular hydrocarbons (CHC) have been documented to play crucial roles as species- and sex-specific cues in the chemical communication systems of a wide variety of insects. However, whether they are sufficient by themselves as the sole cue triggering sexual behavior as well as preference of con- over heterospecific mating partners is rarely assessed. We conducted behavioral assays in three representative species of parasitoid wasps (Hymenoptera: Pteromalidae) to determine their reliance on CHC as species-specific sexual signaling cues.ResultsWe found a surprising degree of either unspecific or insufficient sexual signaling when CHC are singled out as recognition cues. Most strikingly, the cosmopolitan species Nasonia vitripennis, expected to experience enhanced selection pressure to discriminate against other co-occurring parasitoids, did not discriminate against CHC of a partially sympatric species from another genus, Trichomalopsis sarcophagae. Focusing on the latter species, in turn, it became apparent that CHC are even insufficient as the sole cue triggering conspecific sexual behavior, hinting at the requirement of additional, synergistic sexual cues particularly important in this species. Finally, in the phylogenetically and chemically most divergent species Muscidifurax uniraptor, we intriguingly found both CHC-based sexual signaling as well as species discrimination behavior intact although this species is naturally parthenogenetic with sexual reproduction only occurring under laboratory conditions.ConclusionsOur findings implicate a discrepancy in the reliance on and specificity of CHC as sexual cues in our tested parasitioid wasps. CHC profiles were not sufficient for unambiguous discrimination and preference behavior, as demonstrated by clear cross-attraction between some of our tested wasp genera. Moreover, we could show that only in T. sarcophagae, additional behavioral cues need to be present for triggering natural mating behavior, hinting at an interesting shift in signaling hierarchy in this particular species. This demonstrates the importance of integrating multiple, potentially complementary signaling modalities in future studies for a better understanding of their individual contributions to natural sexual communication behavior.

On the Chemical Disguise of a Physogastric Termitophilous Rove Beetle

Inter-specific symbiotic links are often reinforced by morphological, physiological, or behavioural trait modification undergone by the associated species. In some cases, such as in physogastric termitophile staphylinids, such modifications do facilitate the social interaction. Here we inspect chemical traits of the physogastric staphylinid Corotoca melantho (Insecta: Coleoptera) and its termite host Constrictotermes cyphergaster (Insecta: Blattodea: Isoptera), aiming to verify whether staphylinids resemble their host. First, we compared CHC profiles of hosts and guests within and among termitaria, to gather evidence on the origin of such profiles in guests. Then, we examined nitrogen and carbon isotopic signatures of these cohabitants to inspect whether chemical disguise is achieved by predation of host workers by staphylinids. Beetles presented CHC more similar to the CHC of their cohabiting termites than to (i) their conspecifics and (ii) termites from another nest, thereby favouring the hypothesis on CHC acquisition by guests. Isotopic signatures revealed that such similarities could not be majorly determined by share nutrition between these cohabitants. In general, our results evidenced that chemical disguise in termitophiles may function as a strategy for social integration in morphological mimics.

Release from prey preservation behavior via prey switch allowed diversification of cuticular hydrocarbon profiles in digger wasps.

The cuticle of insects is covered by a layer of hydrocarbons (CHC), whose original function is the protection from desiccation and pathogens. However, in most insects CHC profiles are species specific. While this variability among species was largely linked to communication and recognition functions, additional selective forces may shape insect CHC profiles. Here, we show that in Philanthinae digger wasps (Crabronidae) the CHC profile coevolved with a peculiar brood-care strategy. In particular, we found that the behavior to embalm prey stored in the nest with hydrocarbons is adaptive to protect larval food from fungi in those species hunting for Hymenoptera. The prey embalming secretion is identical in composition to the alkene-dominated CHC profile in these species, suggesting that their profile is adaptively conserved for this purpose. In contrast, prey embalming is not required in those species that switched to Coleoptera as prey. Released from this chemical brood-care strategy, Coleoptera-hunting species considerably diversified their CHC profiles. Differential needs to successfully protect prey types used as larval food have thus driven the diversification of CHCs profiles of female Philanthinae wasps. To the best of our knowledge, this is the first evidence of a direct link between selection pressure for food preservation and CHC diversity.

Cuticle hydrocarbons in saline aquatic beetles.

Hydrocarbons are the principal component of insect cuticle and play an important role in maintaining water balance. Cuticular impermeability could be an adaptative response to salinity and desiccation in aquatic insects; however, cuticular hydrocarbons have been poorly explored in this group and there are no previous data on saline species. We characterized cuticular hydrocarbons of adults and larvae of two saline aquatic beetles, namely Nebrioporus baeticus (Dytiscidae) and Enochrus jesusarribasi (Hydrophilidae), using a gas chromatograph coupled to a mass spectrometer. The CHC profile of adults of both species, characterized by a high abundance of branched alkanes and low of unsaturated alkenes, seems to be more similar to that of some terrestrial beetles (e.g., desert Tenebrionidae) compared with other aquatic Coleoptera (freshwater Dytiscidae). Adults of E. jesusarribasi had longer chain compounds than N. baeticus, in agreement with their higher resistance to salinity and desiccation. The more permeable cuticle of larvae was characterized by a lower diversity in compounds, shorter carbon chain length and a higher proportion of unsaturated hydrocarbons compared with that of the adults. These results suggest that osmotic stress on aquatic insects could exert a selection pressure on CHC profile similar to aridity in terrestrial species.

Divergence of cuticular hydrocarbons in two sympatric grasshopper species and the evolution of fatty acid synthases and elongases across insects.

Cuticular hydrocarbons (CHCs) play a major role in the evolution of reproductive isolation between insect species. The CHC profiles of two closely related sympatric grasshopper species, Chorthippus biguttulus and C. mollis, differ mainly in the position of the first methyl group in major methyl-branched CHCs. The position of methyl branches is determined either by a fatty acid synthase (FAS) or by elongases. Both protein families showed an expansion in insects. Interestingly, the FAS family showed several lineage-specific expansions, especially in insect orders with highly diverse methyl-branched CHC profiles. We found five putative FASs and 12 putative elongases in the reference transcriptomes for both species. A dN/dS test showed no evidence for positive selection acting on FASs and elongases in these grasshoppers. However, one candidate FAS showed species-specific transcriptional differences and may contribute to the shift of the methyl-branch position between the species. In addition, transcript levels of four elongases were expressed differentially between the sexes. Our study indicates that complex methyl-branched CHC profiles are linked to an expansion of FASs genes, but that species differences can also mediated at the transcriptional level.

Sexual selection and population divergence I: The influence of socially flexible cuticular hydrocarbon expression in male field crickets (Teleogryllus oceanicus).

Debates about how coevolution of sexual traits and preferences might promote evolutionary diversification have permeated speciation research for over a century. Recent work demonstrates that the expression of such traits can be sensitive to variation in the social environment. Here, we examined social flexibility in a sexually selected male trait-cuticular hydrocarbon (CHC) profiles-in the field cricket Teleogryllus oceanicus and tested whether population genetic divergence predicts the extent or direction of social flexibility in allopatric populations. We manipulated male crickets' social environments during rearing and then characterized CHC profiles. CHC signatures varied considerably across populations and also in response to the social environment, but our prediction that increased social flexibility would be selected in more recently founded populations exposed to fluctuating demographic environments was unsupported. Furthermore, models examining the influence of drift and selection failed to support a role of sexual selection in driving population divergence in CHC profiles. Variation in social environments might alter the dynamics of sexual selection, but our results align with theoretical predictions that the role social flexibility plays in modulating evolutionary divergence depends critically on whether responses to variation in the social environment are homogeneous across populations, or whether gene by social environment interactions occur.