O gênero Polistes é considerado um grupo modelo para a compreensão da sociobiologia das interações de dominância/subordinação em animais invertebrados, devido à pequena diferenciação de castas que resulta em um alto potencial de conflitos reprodutivos entre os membros da colônia. A construção de ninhos satélites e os seus efeitos na estrutura colonial de algumas espécies de Polistes, constituem um campo de estudo de interesse para a área da evolução do comportamento social. Neste estudo foram quantificados os padrões sazonais do desenvolvimento e a composição colonial, bem como a relação desses padrões com a variação climática sazonal. Foram também comparadas as relações que determinam a hierarquia de dominância em colônias de P. satan em relação a um modelo teórico proposto para uma hierarquia linear de dominância. Além disso, mostramos os padrões de interações agonísticas entre indivíduos que compõem a hierarquia e suas inter-relações em colônias não manipuladas e manipuladas (remoção da dominante). Foram também analisados padrões de comunicação visual (marcas de dominância) e química (hidrocarbonetos cuticulares). Os resultados mostraram que fêmeas dominantes em colônias em pré-emergência apresentaram padrões de pigmentação cefálica e perfis de hidrocarbonetos cuticulares significativamente distintos das fêmeas subordinadas ou substitutas. Os aspectos relacionados à comunicação química e visual e suas implicações biológicas são discutidas. / The genus Polistes is an organism model to understand the sociobiology in invertebrates. This group presents low caste differentiation that result in potencial reproductive conflicts between nestmates. The construction os satellite nests and their effects on the colonial structure in some Polistes are an interesting topic to the evolution of socail behavior. In this study we quantify the seasonal patterns of nest development and thei composition. We analyzed the relationship between these patterns and climatic variation. The aspects related to the hierarchy of dominance in P. satan were compared to a theoretical model that predicts a dominance hierarchy im equilibrium. In addition, we presented the agressive interaction between individuals composing a hierarchy in experimental (dominant removal) and control colonies. Patterns of both visual (dominance labels) and chemical communication (cuticular hydrocarbons) were also verified in P. Satan. Dominant females from pre-emergence colonies present both distinct patterns of cephalic pigmentation and cuticular hydrocarbons than other females. These aspects involved with the sociobiology of Polistes and their biological implications are discussed in this study.

O gênero Polistes é considerado um grupo modelo para a compreensão da sociobiologia das interações de dominância/subordinação em animais invertebrados, devido à pequena diferenciação de castas que resulta em um alto potencial de conflitos reprodutivos entre os membros da colônia. A construção de ninhos satélites e os seus efeitos na estrutura colonial de algumas espécies de Polistes, constituem um campo de estudo de interesse para a área da evolução do comportamento social. Neste estudo foram quantificados os padrões sazonais do desenvolvimento e a composição colonial, bem como a relação desses padrões com a variação climática sazonal. Foram também comparadas as relações que determinam a hierarquia de dominância em colônias de P. satan em relação a um modelo teórico proposto para uma hierarquia linear de dominância. Além disso, mostramos os padrões de interações agonísticas entre indivíduos que compõem a hierarquia e suas inter-relações em colônias não manipuladas e manipuladas (remoção da dominante). Foram também analisados padrões de comunicação visual (marcas de dominância) e química (hidrocarbonetos cuticulares). Os resultados mostraram que fêmeas dominantes em colônias em pré-emergência apresentaram padrões de pigmentação cefálica e perfis de hidrocarbonetos cuticulares significativamente distintos das fêmeas subordinadas ou substitutas. Os aspectos relacionados à comunicação química e visual e suas implicações biológicas são discutidas. / The genus Polistes is an organism model to understand the sociobiology in invertebrates. This group presents low caste differentiation that result in potencial reproductive conflicts between nestmates. The construction os satellite nests and their effects on the colonial structure in some Polistes are an interesting topic to the evolution of socail behavior. In this study we quantify the seasonal patterns of nest development and thei composition. We analyzed the relationship between these patterns and climatic variation. The aspects related to the hierarchy of dominance in P. satan were compared to a theoretical model that predicts a dominance hierarchy im equilibrium. In addition, we presented the agressive interaction between individuals composing a hierarchy in experimental (dominant removal) and control colonies. Patterns of both visual (dominance labels) and chemical communication (cuticular hydrocarbons) were also verified in P. Satan. Dominant females from pre-emergence colonies present both distinct patterns of cephalic pigmentation and cuticular hydrocarbons than other females. These aspects involved with the sociobiology of Polistes and their biological implications are discussed in this study.

Chez les insectes sociaux, la capacité à reconnaître les compagnons du nid des autres a évolué du fait de l’intérêt à distinguer les individus apparentés. En effet, il est avantageux que les actes de coopération soient uniquement dirigés vers les individus apparentés. La faculté de discrimination repose sur la capacité à percevoir les signaux de reconnaissance (hydrocarbures cuticulaires) d'un individu rencontré et de le comparer avec une représentation de l'odeur de sa propre colonie. L'ontogénèse de la reconnaissance des compagnons du nid et les processus d'apprentissage qui pourraient les sous-tendre ont été peu explorés jusqu’à ce jour. Il a été suggéré que les insectes sociaux apprennent l'odeur de leur colonie pendant les premières étapes de la vie adulte. L'objectif de ce travail était de rechercher les principales caractéristiques de l'ontogénèse du processus de reconnaissance avec une approche comparative, en utilisant deux modèles : les fourmis et les guêpes sociales. Nous avons montré que le mécanisme général de reconnaissance proposé pour le genre Polistes ne s'appliquait pas à la guêpe P. dominula. Nous avons étudié les mécanismes alternatifs, suggérant l'apprentissage pré-imaginal et le mécanisme d'auto-référence. Nous avons mis en évidence pour la première fois que la fourmi Aphaenogaster senilis apprenait, durant la vie larvaire, des signaux l'importance possible de processus cognitifs précédemment négligés tels que chimiques importants pour la reconnaissance. Ainsi, l’existence et la fonctionnalité de la mémoire larvaire ont été révélées chez les fourmis, élargissant nos connaissances sur les mécanismes de reconnaissance des compagnons du nid, l'apprentissage et la mémoire. / In social insects the ability to recognize nestmates from non‐nestmates evolved as a result of the advantages of distinguishing related individuals. Indeed, are gained only if cooperative acts are directed towards realtives. This discrimination ability relies on the capacity of perceiving recognition cues (cuticular hydrocarbons) of an encountered individual and of comparing them with a neural representation of the own colony odour. Relatively little is known about the ontogeny of nestmate recognition, and the learning processes that might be involved. It has been suggested that social insects learn their colony odour during the early stages of adult life. The overall aim of the present work was to investigate the main features of the ontogeny of nestmate recognition process with a comparative approach, using two different models: ants and social wasps. In particular, I investigated wheter general recognition mechanism proposed for the Polistes genus applies to Polistes dominula and I found that this is not the case. I studied alternative mechanisms and provided novel insights into the ontogeny of nestmate recognition in social wasps, suggesting the possible importance of previously neglected cognitive processes, such as pre-imaginal learning and self-reference phenotype matching. In ants, I showed for the first time that Aphaenogaster senilis learn chemical cues important for adult nestmate recognition during the larval life, thus larval memory is something present and functional in ants. This result gives significant insight for the study of nestmate recognition, the mechanisms at the basis, learning and memory in ants, opening new questions concerning interesting phenomena as pre-imaginal learning.

Sexual conflict occurs when the fitness interests of males and females do not align with one another. The resolution of sexual conflict often depends on the level of control each sex has on the behavior in conflict. In Pogonomyrmex harvester ants with a genetically determined caste system, two separate lineages interbreed with one another during summer mating swarms. Diploid offspring sired by a single lineage develop into reproductive queens, while offspring sired by opposite-lineage parents develop into sterile workers. This results in sexual conflict, as males which mate with opposite lineage queens will produce only workers, resulting in no fitness benefit, while queens must mate with opposite-lineage males in order to obtain workers and survive. Despite these fitness differences, males do not discriminate between lineages prior to mating. One possible reason for the lack of male discrimination is that queens "mask" their identity cues, making discrimination difficult for males. In eusocial insects, identity cues are encoded by cuticular hydrocarbons (CHC’s) found on the exoskeleton of the insects. These cues contain information on the insect's reproductive status, sex, species, state, and nest membership. In addition to their communication functions, CHC’s also serve as desiccation-resistance molecules, preventing water from freely passing out of the cuticle. However, molecules that are best-suited for communication functions are poor desiccation resistance molecules, and molecules that are best-suited for waterproofing lack the diversity needed for communication; therefore, a tradeoff between these two functions is expected. In this dissertation, I explore sexual conflict in these ants and the chemical recognition cues that likely play a role in this conflict. To test for cryptic strategies in harvester ant mating swarms, I experimentally paired males and females from two interbreeding lineages of harvester ant with different fitness outcomes based on pairing, and measured the propensity to initiate copulation, pre-copulatory time, time in copula, and rate and amount of sperm transferred in each mating. Although females controlled copulation duration, males altered sperm transfer rates, resulting in no quantitative difference in total sperm transfer between lineages. To test for thermal constraints on the diversity and composition of cuticular hydrocarbon profiles, and changes in CHC profiles that occur in workers isolated from the queen, I surveyed the cuticular hydrocarbon profiles of a species complex of harvester ants. The CHC profiles of ants from more xeric environments showed evidence of constraints, while isolated workers differentiated from their queen-raised sisters, although not in queen-specific molecules. To test for queen identity masking and lack of discrimination ability in mating swarms, I tested for convergence in the CHC profiles of reproductives in two hybridizing lineages in response to the sexual conflict playing out in this species. Differences in CHC profiles were lost during the mating swarm, likely limiting male ability to discriminate between mates, limiting discrimination ability in mating swarms. To study the genetic regions that control CHC production, I created a physical linkage map of two of the interbreeding populations, and used that map to perform quantitative trait loci analysis on the cuticular hydrocarbon profile of recombinant males. One significant region associated with 13-methylnonacosane contained numerous odorant receptor genes, suggesting a link between that CHC production and the receptors that detect it, while a second region associated with n-pentacosane contained numerous genes that control expression levels. Overall, the genetic caste determination system in these ants leads to antagonistic coevolution between species. This coevolution is likely reinforced by the thermal constraints and exchange of recognition cues between species, lowering the ability of useful discrimination between lineages during mating swarms.

A integridade das sociedades de insetos depende da capacidade dos indivíduos em discriminar aqueles que não pertencem às suas colônias; essas interações de reconhecimento são mediadas por hidrocarbonetos presentes na camada de cera da cutícula e atuam como mensageiros que codificam informação sobre o meio interno (colônia) e externo (ambiente). Os hidrocarbonetos são produzidos por células dérmicas e o perfil desses compostos recebe influência tanto de fontes endógenas quanto exógenas e isso o torna suscetível às mudanças temporais. A caracterização desses compostos em insetos-praga é importante, pois provêm subsídios tanto para pesquisa básica como para o manejo e controle de pragas. Nesse contexto, o presente estudo abordou as variações dos hidrocarbonetos cuticulares em relação às subcastas morfológicas das operárias (condição endógena) e utilização do substrato foliar (variável exógena) na formiga-cortadeira Atta sexdens, conhecida como uma das principais pragas herbívoras da região Neotropical. Os compostos foram extraídos em solvente apolar hexano e as amostras foram analisadas em sistema de cromatografia a gás acoplado à espectrometria de massas (CG-MS). Foram identificados 30 hidrocarbonetos com cadeia carbônica entre 19 e 40 átomos distribuídos em três classes de compostos, sendo a dos hidrocarbonetos ramificados a mais representativa, especificamente os trimetialcanos. Foram encontradas variações significativas para todas as variáveis analisadas, sugerindo que as subcastas possuem assinaturas químicas semelhantes quanto à diversidade de compostos, mas com grandes diferenças na proporção relativa dos mesmos. Em relação ao substrato foliar, colônias que tiveram acesso ao mesmo recurso apresentam uma composição química cuticular mais semelhante em relação àquelas que tiveram acesso a um recurso diferente. Complementarmente, os testes comportamentais mostraram que, na presença de uma coespecífica intrusa, as operárias residentes são muito mais agressivas em relação àquelas que utilizaram outro tipo de substrato foliar. / The integrity of insect societies relies on the ability of individuals to discriminate between nesmates from non-nestmates. These interactions are mediated by hydrocarbons spread all over the cuticle that act as messengers coding information about the colony and external environment. Cuticular hydrocarbons are produced by dermic cells and its composition is influenced by both endogenous and exogenous sources which makes them subject to temporal changes. Thus, this study approached the variations on this chemical profile in relation to the worker subcastes and type of foliar substrate used by leafcutter ant Atta sexdens, this species is known as one of the major neotropical herbivore pests and this kind of study is important because it can provide subsides in researches related to both basic biology as to pest control methods. The compounds were extracted using apolar solvent (hexane) and the samples were analyzed using gas chromatography - mass spectrometry technique (GC-MS). It was found 30 hydrocarbons which carbon chains varies between 19 and 40 atoms separated in three classes of compounds and the branched hydrocarbons as the most abundant one, especially the trimethyalcanes. The statistical analysis revealed significant differences on all the variables analyzed suggesting that subcastes possess similar chemical signatures as to the variety of compounds but with great differences in relative proportions thereof. Furthermore, the results show that foliar substrate has influence on the composition of cuticular hydrocarbons and concomitantly affect the interspecific recognition, thus colonies that had access to the same substrate has similar cuticular composition comparing to those who do not. Additionally, behavior tests showed that in the presence of a conspecific intruder the resident workers tend to be much more aggressive in relation to those who had fed on a different foliar substrate.

Os hidrocarbonetos cuticulares têm importante papel no processo de reconhecimento dos membros da colônia de insetos sociais. Muitos estudos têm mostrado variações qualitativas e quantitativas nestes compostos entre os insetos adultos. Contudo, abordagens referentes à modulação do perfil destes compostos durante a formação da cutícula são escassas, e se restringem aos estágios larval de holometábolos e de ninfas de hemimetábolos. O principal objetivo dessa pesquisa foi caracterizar o perfil de hidrocarbonetos cuticulares e a expressão de genes potencialmente relacionados à sua biossíntese durante o processo de formação e maturação da cutícula adulta. Os perfis de hidrocarbonetos foram caracterizados por meio de GC/MS e mostraram diferenças quantitativas marcantes que significativamente discriminaram as cutículas pupal, adulta-farata e adulta. Em paralelo, sequências de enzimas que catalisam a desaturação (desaturases) ou elongação (elongases) de lipídeos, disponíveis no banco de dados do NCBI, foram utilizadas para o desenho de primers e estudo da expressão gênica por meio de RT-qPCR. Cinco genes de desaturases, e oito genes de elongases mostraram variação de expressão estatisticamente significante no tegumento de abelhas adultas em comparação com pupas e adultas-faratas. Testes de correlação entre os perfis de expressão gênica e de hidrocarbonetos cuticulares evidenciaram os genes potencialmente envolvidos com a biossíntese destes compostos para a formação e maturação da cutícula. Estes resultados corroboram a hipótese de que nos insetos sociais, a cutícula só amadurece completamente por ocasião do início da atividade de forrageamento. Associando estes dados a análises de evolução molecular das desaturases e elongases, pudemos sugerir as etapas da via de síntese de hidrocarbonetos catalisadas por estas enzimas, e assim eleger genes candidatos a futuro silenciamento mediado por RNA de interferência para pesquisa de função. / Cuticular hydrocarbons are important for recognition of nestmates in social insect colonies. Many studies have shown qualitative and quantitative variations in the cuticular hydrocarbons between adult insects. However, approaches on developmental profiles of these compounds during cuticle formation and differentiation are scarce, and restricted to larval stages of holometabolous and nymphs of hemimetabolous. The main objective of this work was to characterize the cuticular hydrocarbons profiles and the expression of genes potentially involved in the biosynthesis of these compounds during the synthesis and differentiation of the adult cuticle in the honeybee. The hydrocarbons profiles were characterized using GC/MS and showed remarkable quantitative differences, thus discriminating the pupal, pharate-adult and adult cuticles from each other. In parallel, we used annotated sequences of enzymes catalyzing lipid desaturation (desaturases) or elongation (elongases), available in NCBI data bank, for primers design and gene expression analysis using RT-qPCR. Five desaturase genes and eight elongase genes showed statistically significant expression changes in the integument of adult bees in comparison to pupae and pharate-adults. Correlation tests supported roles of some of the desaturase and elongase genes in hydrocarbons biosynthesis for incorporation into adult cuticle. In addition, these results go along with the hypothesis that in social insects the cuticle is just completed when the insect starts forager activity. Taken together, these data and an analysis on the molecular evolution of desaturases and elongases allowed suggesting the steps in the pathway of cuticular hydrocarbons biosynthesis that are catalyzed by these enzymes, and also allowed to elect candidate genes for further functional studies using gene silencing mediated by RNAi.

Regulation of reproduction is one central feature of social life. In particular, only a few individuals are in charge of producing offspring in eusocial species. This division of the reproductive labour is mainly mediated by pheromones emitted by the queens in social insects. These queen pheromones may signal the presence of a fertile queen so that workers react accordingly by taking care of her and not reproducing. Here I investigated two aspects of the reproduction of two polygynous ant species. The first one, Linepithema humile, is a unicolonial, highly polygynous and invasive species. It has been the focus of numerous studies about queen pheromones; in particular, it has been reported that queen cuticular hydrocarbons (CHC) profile is related to queen fertility. The other one, Tapinoma erraticum, is a multi-colonial, weakly polygynous and native species. Workers can lay haploid eggs in the absence of the queens which is impossible for Linepithema workers. The major part of my thesis dealt with the queen fertility signalling issue. In the first two chapters I demonstrate the link between queen fertility and queen pheromone output. I first study a queen releaser pheromone, the queen retrieval behaviour. This behaviour is performed by the workers who lay a chemical trail toward a queen located outside the nest. I successfully show this behaviour to be related to queen fertility, and not mating status, in L. humile and T. erraticum since only fertile queens (mated or not) induce such recruitment. I then highlight the role of queen fertility in the prevention of worker reproduction in T. erraticum. Again, mated fertile queens and unmated fertile queens are both able to induce such primer effect. In a third chapter I report that CHC profiles may discriminate female castes (workers, queens, virgin queens, and virgin egg-laying queens) in T. erraticum. Finally, chapter 4 summarizes my attempts to prove that CHC may be involved in queen retrieval or queen attraction. None of the various bioassays tested allows me to demonstrate the putative role of CHC as queen pheromone. In a second part, I was interested in the consequences of sex determination in T. erraticum. Chapter 5 presents the flow cytometry methodology (FC) which allowed me to score the number of sperm cells from spermathecae of several ant species, and to demonstrate that polygynous species (such as L. humile and T. erraticum) store less sperm than monogynous ones. FCM also allows determining the ploidy of sperm and adult somatic cells and chapter 6 which presents a large survey on the ploidy level within the species T. erraticum. This species displays diploid males that may produce diploid sperm which in turn can father a viable triploid female progeny. I report differences in the frequency of triploidy among female castes, the proportion of triploid workers being more important than triploid virgin queens whereas I never observed triploid mated fertile queens. Such results greatly suggest a putative regulatory mechanism involved in the rearing of triploid females. In the last chapter I investigated two populations that differ in the occurrence of triploid workers. I report these populations to vary in the number of queens and workers per nest. /La régulation de la reproduction est un aspect essentiel de la vie sociale. En particulier, chez les espèces eusociales, seuls quelques individus sont impliqués dans la production de la descendance. Dans les sociétés d’insectes, une telle division du travail reproducteur est principalement assurée par l’émission de phéromones par les reines. Ces phéromones royales renseignent les membres de la colonie sur la présence d’une reine fertile, de telle sorte que les ouvrières réagissent en s’occupant d’elle et en s’abstenant de se reproduire. Au cours de ce travail, je me suis intéressé à deux aspects de la reproduction au sein de deux espèces de fourmis polygynes. La première espèce, Linepithema humile, est invasive, unicoloniale et hautement polygyne dans les régions à climat méditerranéen. Elle a fait l’objet de nombreuses études portant notamment sur les phéromones royales. En particulier, il a été montré que le profil d’hydrocarbures cuticulaires (HCC) des reines est corrélé à leur fertilité. La seconde espèce, Tapinoma erraticum, est une espèce indigène, multi-coloniale et faiblement polygyne. Ses ouvrières sont capables de pondre des œufs mâles en absence de reines, ce dont sont incapables les ouvrières de Linepithema. Je me suis principalement intéressé à la question de la signalisation de la fertilité des reines. Dans les deux premiers chapitres, je démontre le lien existant entre la fertilité des reines et la production de phéromone royale. J’ai d’abord étudié une phéromone incitatrice (releaser) qui provoque un recrutement royal. Ce comportement collectif très caractéristique correspond à la mise en place d’une piste chimique en direction d’une reine découverte par les ouvrières en dehors du nid. Je montre que ce comportement est lié à la fertilité de la reine chez les espèces L. humile et T. erraticum car seules les reines fertiles (fécondées ou non) sont capables de d’induire le recrutement royal. Je mets ensuite en évidence le rôle de la fertilité des reines dans la régulation de la reproduction des ouvrières de T. erraticum. A nouveau, les reines fécondées fertiles et les reines vierges fertiles sont toutes deux capables d’induire un même effet déclencheur (primer), en l’occurrence, l’inhibition de la reproduction des ouvrières. Dans un troisième chapitre, je montre que les profils d’hydrocarbures (HCC) permettent de distinguer sans ambiguïté les différentes castes femelles (ouvrières, reines fertiles, reines vierges et reines vierges pondeuses) chez T. erraticum. Enfin, le chapitre 4 résume mes tentatives pour démontrer le rôle des HCC dans les phénomènes de recrutement royal ou d’attractivité des reines. Aucun des bio-essais réalisés ne me permet de démontrer l’implication des HCC dans la phéromone royale. Dans une seconde partie, je me suis intéressé aux conséquences du déterminisme du sexe chez T. erraticum. Le chapitre 5 présente cytométrie de flux (CF), une méthode qui me permet de compter les spermatozoïdes stockés dans les spermathèques de quelques espèces de fourmis et de montrer que les reines des espèces polygynes (telles que L. humile et T. erraticum) stockent moins de sperme que les espèces monogynes. La CF permet aussi de déterminer le niveau de ploïdie des cellules spermatiques ou somatiques chez l’adulte. Je me sers de cette application dans le chapitre 6 afin d’étudier le niveau de ploïdie au sein de l’espèce T. erraticum. Je montre que, dans les populations étudiées, il existe des mâles diploïdes et que ces mâles peuvent produire du sperme diploïde fertile, capable d’engendrer une descendance femelle triploïde. Je note des différences dans la fréquence des femelles triploïdes : la proportion d’ouvrières triploïdes est significativement plus importante que celle des reines vierges triploïdes. De plus, je n’ai jamais observé la présence de reines fécondées fertiles triploïdes. De tels résultats suggèrent fortement la présence d’un phénomène de régulation au cours de l’élevage du couvain triploïde. Dans le dernier chapitre, j’ai étudié deux populations de T. erraticum qui diffèrent au niveau de la proportion d’ouvrières triploïdes. Ces populations présentent des différences significatives dans le nombre de reines et d’ouvrières par nid.

Directional selection is prevalent in nature yet phenotypes tend to remain relatively constant, suggesting a limit to trait evolution. The genetic basis of evolutionary limits in unmanipulated populations, however, is generally not known. Given widespread pleiotropy, opposing selection on a focal trait may arise from the effects of the underlying alleles on other fitness components, generating net stabilizing selection on trait genetic variance and thus limiting evolution. Here, I look for the signature of stabilizing selection for a suite of cuticular hydrocarbons (CHCs) in Drosophila serrata. Despite strong directional sexual selection on CHCs, genetic variance differed between high and low fitness individuals and was greater among the low fitness males for seven of eight CHCs. Univariate tests of a difference in genetic variance were non-significant but have low power. My results implicate stabilizing selection, arising through pleiotropy, in generating a genetic limit to the evolution of CHCs in this species.

Directional selection is prevalent in nature yet phenotypes tend to remain relatively constant, suggesting a limit to trait evolution. The genetic basis of evolutionary limits in unmanipulated populations, however, is generally not known. Given widespread pleiotropy, opposing selection on a focal trait may arise from the effects of the underlying alleles on other fitness components, generating net stabilizing selection on trait genetic variance and thus limiting evolution. Here, I look for the signature of stabilizing selection for a suite of cuticular hydrocarbons (CHCs) in Drosophila serrata. Despite strong directional sexual selection on CHCs, genetic variance differed between high and low fitness individuals and was greater among the low fitness males for seven of eight CHCs. Univariate tests of a difference in genetic variance were non-significant but have low power. My results implicate stabilizing selection, arising through pleiotropy, in generating a genetic limit to the evolution of CHCs in this species.

Directional selection is prevalent in nature yet phenotypes tend to remain relatively constant, suggesting a limit to trait evolution. The genetic basis of evolutionary limits in unmanipulated populations, however, is generally not known. Given widespread pleiotropy, opposing selection on a focal trait may arise from the effects of the underlying alleles on other fitness components, generating net stabilizing selection on trait genetic variance and thus limiting evolution. Here, I look for the signature of stabilizing selection for a suite of cuticular hydrocarbons (CHCs) in Drosophila serrata. Despite strong directional sexual selection on CHCs, genetic variance differed between high and low fitness individuals and was greater among the low fitness males for seven of eight CHCs. Univariate tests of a difference in genetic variance were non-significant but have low power. My results implicate stabilizing selection, arising through pleiotropy, in generating a genetic limit to the evolution of CHCs in this species.