Mutualistic ants and parasitoid communities associated with a facultative myrmecophilous lycaenid,Arhopala japonica,and the effects of ant attendance on the avoidance of parasitism

Herbivorous insects have evolved various defensive strategies to avoid their primary enemies, parasitoids. Many species of Lycaenidae (Lepidoptera) have food‐for‐protection mutualism with ants in their larval stages, where larvae produce nectar for ants and in return ants exclude parasitoids as well as predators. Myrmecophilous relationships are divided into two categories, obligate and facultative, by degrees of myrmecophily. Although parasitoids attacking obligate lycaenids always encounter lycaenid‐specific ant species, parasitoids that use facultative lycaenids are likely to encounter diverse ant species showing various defense systems. However, we know little about the parasitoid community of facultative lycaenid larvae. In this study, we investigated the mutualistic ant and parasitoid communities of a facultative myrmecophilous species, Arhopala japonica, in seven localities in Japan. The present field observation newly recorded four ant species attending A. japonica larvae, and combined with the previous data, the number of attending ant species reached 16, which is nearly the maximum number of reported attending ant species among myrmecophilous lycaenids. However, the present study revealed that almost all parasitized A. japonica larvae were attacked by a single braconid species, Cotesia sp. near inducta. We also assessed the efficiency of facultative ant defense against the parasitoid in the laboratory and revealed that oviposition by Cotesia sp. near inducta females was almost completely hindered when A. japonica larvae were attended by ants. This suggests that the dominant parasitoid does not have effective traits to overcome defensive behavior of ants and that the female wasps oviposit mainly in A. japonica larvae without intensive attendance.     

Cuticular hydrocarbon profiles of codling moth larvae,Cydia pomonella (Lepidoptera: Tortricidae), reflect those of their host plant species

The cuticular hydrocarbons (CHCs) of codling moth larvae collected in the field from their host plant species, apple and walnut, were analyzed and compared with the CHCs of fruits from these two phylogenetically distant hosts. The CHC profiles of the larvae consisted solely of n‐alkanes (C₂₃–C₃₁) and differed quantitatively between host populations. Amounts of the CHCs from the walnut‐collected larvae were shifted towards longer‐chain alkanes compared to those from apple‐collected larvae. A similar shift was observed for the CHC profiles of walnut and apple fruits. Analysis of the CHCs of larvae reared on artificial diet, in comparison with hydrocarbons from the diet, confirmed that larval CHCs scarcely reflect hydrocarbons from the food source. This finding indicates that the larval hydrocarbons must be biosynthesized to a large degree by the insect, rather than being gained directly from its diet. Hence, codling moth populations from apple and walnut each synthesize their own CHC profiles, which largely resemble those of their respective host plant, yielding a potential tool of chemical camouflage from certain natural antagonists of the larvae. The findings of the present study, together with recent molecular population analyses, provides evidence for a process that might ultimately lead to sympatric speciation of this herbivore species. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 376–384.     

The influence of ants on patterns of colonization and establishment within a set of coexisting lycaenid butterflies in a south-east Asian tropical rain forest

In Peninsular Malaysia ten species of lycaenid butterflies use leaf flushes or inflorescences of the legume tree Saraca thaipingensis as larval hostplant. Resource partitioning among these species is regulated by a complex mixture of patterns of interaction with ants. Females of obligately myrmecophilous species lay their eggs exclusively on trees colonized by their specific host ants. On trees colonized by weaver ants, only specialist mutualists adapted to these territorial ants are able to survive, while larvae of other species are killed. The formicine ant Cladomyrma petalae, which inhabits hollow twigs of the myrmecophytic hostplant, likewise precludes oviposition by female butterflies. Lycaenid larvae confronted with this ant species never survive, but one concealed feeding species (Jamides caeruleus) escapes removal due to the cryptic life-habits of the larvae. Two facultative myrmecophiles associate in a mutualistic way with a wide and largely overlapping range of ant genera which forage at the extrafloral nectaries of leaf flushes. One species (Cheritra freja) is not myrmecophilous, but is tolerated by all but the most territorial ants. Ant-dependent hostplant selection and egg-clustering characterize the obligate mutualists, whereas facultative myrmecophiles and the non-myrmecophile distribute their eggs singly over appropriate hostplants. Signals mediating caterpillar-ant communication are highly specialized in one obligate myrmecophile (Drupadia theda), but rather unspecific in four other species tested. Altogether our observations indicate that colonization and establishment of lycaenid butterflies on S. thaipingensis trees are governed by specializations as well as opportunistic use of resources (ants and hostplant parts). Therefore, the diversity of this species assemblage is maintained by deterministic as well as stochastic factors.     

Chemical convergence between a guild of facultative myrmecophilous caterpillars and host plants

1. Ants exert strong selective pressure on herbivorous insects, although some caterpillars can live in symbiosis with them using chemical defensive strategies.
2. We investigated the adaptive resemblance of cuticular hydrocarbons (CHCs) in multitrophic systems involving a guild of facultative myrmecophilous caterpillar species (Lepidoptera: Lycaenidae), tending ants (Hymenoptera: Formicidae), and host plants from three families. We hypothesised that the CHCs of the caterpillars would resemble those of their host plants (chemical camouflage).
3. We analysed CHCs using gas chromatography/mass spectrometry. Morisita's similarity index (SI) was used to compare CHC profiles of caterpillar species with different types of ant associations (commensal or mutualistic), ants, and host plants.
4. We found strong convergence between caterpillars' CHCs and plants, especially for commensal species that do not provide secretion rewards for ants. Moreover, we found unexpected chemical convergence among mutualistic (trophobiotic) caterpillar species that offer caloric reward secretions to ants.
5. These results show that the studied caterpillars acquire CHCs through their diet and that they vary according to host plant species and type of ant association (commensalism or mutualism). This ‘chemical camouflage’ of myrmecophilous caterpillars may have arisen as a defensive strategy allowing coexistence with ants on plants, whereas ‘chemical conspicuousness’ may have evolved in the context of honest signalling between mutualistic partners.
6. We suggest the existence of chemical mimicry among myrmecophilous species, especially between mutualistic caterpillars. Cuticular chemical mixtures can play a key adaptive role in decreasing ant attacks and increasing caterpillar survival in multimodal sensory systems.

     

Evolution of influence: signaling in a lycaenid-ant interaction

Some phytophagous insects gain defense from natural enemies by associating with otherwise potentially harmful top predators. Many lycaenid butterfly caterpillars are involved in such interactions with ants: larvae provide carbohydrate rewards from the dorsal nectary organ (DNO) to associated ants in return for protection from natural enemies. The stability of these interactions involves signals that identify the lycaenid caterpillar as a mutualist. However, larvae of some lycaenid species, such as Lycaena xanthoides, are found in close association with ants but do not possess the reward producing DNO. Evaluating the relationship in a phylogenetic framework, we show that the association between L. xanthoides and ants likely evolved from a non-ant-associated ancestor. Behavioral trials also show that L. xanthoides larvae are capable of influencing ant behavior to increase ant tending when faced with a simulated predator attack, without providing DNO-derived rewards to ant associates. These results demonstrate that the DNO is not necessary to maintain associations between lycaenid larvae and ants. Third-party interactions may affect the evolution of mutualisms and consideration of underlying evolutionary history is necessary to understand contemporary species associations.     

Behavioral strategy of a lycaenid (Lepidoptera) caterpillar against aggressive ants in a Brazilian savanna

Myrmecophily is widespread in lycaenid butterflies, in which ants receive food resources and, in turn, protect caterpillars against natural enemies. This interaction ranges from obligate myrmecophily, in which immatures are invariably associated with ants and are dependent on ants for survival, and facultative myrmecophily, in which larvae are not dependent on ants for survival, but the presence of the latter may increase larvae survival. Lycaenids also include non-myrmecophilous butterflies, which do not have positive associations with ants and have developed strategies to avoid being attacked or preyed upon by them. In this study, we examined the relationship between the lycaenid Michaelus ira and two ant species associated with Distictella elongata (Bignoniaceae). This plant has extrafloral nectaries and is patrolled by Camponotus crassus and Ectatomma tuberculatum. Morphological analyses revealed that M. ira larvae have ant organs, such as dorsal nectary organs and perforated cupolas, structures associated with myrmecophily. We performed larval exposure experiments in the field, predicting that, in the absence of myrmecophily, the butterfly larva would present strategies to avoid ant attack. Results showed that larvae were attacked by both ant species. To escape ant molestation, larvae lived and fed inside silk-sealed D. elongata flower buds. We concluded that the M. ira bud-sheltering behavior was a defensive strategy against these ant species, while the dorsal nectary organs were apparently nonfunctional. Nonetheless, myrmecophily, in general, cannot be excluded in M. ira since relationships with other ant species may exist.     

The aphid-tending ant Lasius fuji exhibits reduced aggression toward aphids marked with ant cuticular hydrocarbons

Some aphid species are attended by ants, which protect aphids against enemies, but ants sometimes prey on the aphids they are attending depending on the resource conditions. A previous study indicated that the ant Lasius niger preys less on the aphid individuals that experienced ant attendance than on those that did not. This observation leads to the hypothesis that ants transfer some substances to the aphids they attend and selectively prey on the aphids without the substances. In this study, we focus on cuticular hydrocarbons (CHCs), which are used by ants as nestmate recognition substances, and test whether ants discriminate the aphids on the basis of CHCs. We confirmed that the ant Lasius fuji preyed less on the aphids that were attended by their nestmates than those that were not attended. Glass dummies treated with CHCs from attended aphids were attacked less by ants than those treated with CHCs from non-attended aphids. The CHC profiles of ant attended aphids resembled those of the ants, suggesting that ants’ CHCs are transferred to the aphids’ body surface through ant attendance. These results support the hypothesis that ants “mark” their attended aphids with their CHCs and the CHCs reduce ant predation intensity.     

Seasonal changes in cuticular hydrocarbons in response to polyphenism in the host-alternating aphid Prociphilus oriens

In most terrestrial arthropods, cuticular hydrocarbons (CHCs) function to assist in desiccation tolerance and chemical communications. However, few studies have clarified whether CHC profiles change among developmental stages or among different morphs in non-social insects. In the present study, we evaluated how CHC profiles change in accordance with polyphenism in the host-alternating aphid Prociphilus oriens, which exhibits a complex life cycle and five distinct morphs. These morphs are sexual or asexual and adapt to different host plants. We found that all generations of P. oriens shared high proportions of n-alkanes, but its composition varied among morphs. Three morphs that are attended by ants were characterized by relatively high proportions of n-C25 to n-C27, whereas two morphs that are not attended by ants had higher proportions of longer-chain n-alkanes, such as n-C27 and n-C29. The CHC profiles of sexual females were largely different from those of males. Considering that sexual females of Prociphilus spp. lack organs that secrete sex pheromones (scent plaques), the CHCs of sexual females are likely to function as a sex attractant. High proportions of methyl-branched alkanes were detected in the long and flocculent waxy substances of autumnal migrants. These methyl-branched alkanes are considered a cue to recognize conspecifics. We concluded that the functions and components of CHCs differ among morphs, and that those of sexual females differ from those of males and asexual generations because of their function in sexual communication.     

Of crazy-ants and Curetinae: are Curetis butterflies tended by ants?

The larvae of the lycaenid subfamily Curetinae have never been reported to be associated with ants. Observations on Curetis regula Evans from Brunei are presented which show that this species may be tended by ants both as larvae and adults. The observations are discussed in relation to a recent review on lycaenid/ant associations, u is suggested that the Curetinae will be found to be associated with ants when more species have been reared, on evidence of the larval tentacle organs and apparent ‘pore cupolas’, both of which are ant adaptations. More studies are needed on Curetis biology and larval morphology to resolve the relationships of this enigmatic genus within the Lycaenidae.     

A single lycaenid caterpillar gets an ant‐constructed shelter and uninterrupted ant attendance

Ant‐lycaenid associations range from mutualism to parasitism and the caterpillars of some species of lycaenids are reported to enter ant nests for shelter, diapause, or pupation. The present study aimed to examine the nature of the association between Euchrysops cnejus (Fabricius) (Lepidoptera: Lycaenidae) and Camponotus compressus (Fabricius) (Hymenoptera: Formicidae) worker ants on the extrafloral nectary‐bearing cowpea plant, Vigna unguiculata (L.) Walp. (Fabaceae). The abundance patterns of the ants and the lycaenid caterpillars together with the spatial patrolling patterns of the ants on the plants revealed that ant abundance increased with the occurrence of the lycaenid caterpillars and the ants preferred the lycaenids over the extrafloral nectar. Camponotus compressus worker ants constructed a shelter at the cowpea plant base after interaction with one or more lycaenid caterpillar(s) and tended the caterpillars and pupae till the emergence of the butterfly. The ant‐constructed shelters (ACSs) inhabited by the minor caste workers (13 ± 1.3 ants per ACS), were utilized by the caterpillars to undergo pupation. The ants confined their activities predominantly to tending the pod‐feeding caterpillars and the solitary pupa within each ACS. It appears that the behavior of the tending worker ants is modulated by the lycaenid vulnerable stages.     

The distribution and density of a lycaenid butterfly in relation to Lasius ants

Summary Larvae and pupae of lycaenid butteflies are often associated with ants: this is usually a mutualism in which ants guard the lycaenids from natural enemies, and the lycaenid larvae and pupae provide sugars and amino acids for the ants. A possible consequence of the interaction is spatially correlated ant and lycaenid distributions, but the phenomenon is poorly documented. We examined the lycaenid Plebejus argus, which is tended by Lasius ants. Within habitat patches, P. argus eggs, larvae and pupae were all spatially associated with Lasius. On a larger scale, the densities of butterflies in different habitat patches and populations, and whether the butterfly was present or not, were correlated with Lasius ant densities. The association of P. argus with Lasius ants is consistent among populations, and occurs at several spatial scales. Other aspects of the ecology of P. argus are more variable.     

Ants benefit from attending facultatively myrmecophilous Lycaenidae caterpillars: evidence from a survival study

Workers of three ant species (Lasius niger, Lasius flavus, Myrmica rubra) were caged in the laboratory together with caterpillars and pupae of five species of lycaenid butterflies. Mortality of ants was 3–5 times higher when the ants were confined with larvae lacking a dorsal nectar organ (Lycaena phlaeas, Lycaena tityrus) rather than with caterpillars which possess a nectar gland (Aricia agestis, Polyommatus bellargus, P. icarus). For all five species, ant survival was always lower at the pupal stage (where a nectar organ is always absent) than at the caterpillar stage and was largely equivalent for the butterfly species tested. The experimental data confirm earlier estimates that ants can derive nutritive benefits from tending facultatively myrmecophilous lycaenid caterpillars, even though these caterpillars produce nectarlike secretions at low rates.     

Specificity of an ant-lycaenid interaction

Summary Many lycaenid butterflies are believed to be mutualists of ants — the butterfly larvae secrete sugars and amino acids as rewards for the ants, and the ants protect the larvae from predation or parasitism. We examined the specificity of the relationship between the lycaenid Plebejus argus and ants in the genus Lasius. Eggs were not attractive to Lasius ants until the emerging larvae had broken through the chorion. First instar larvae were palpated and picked up by Lasius workers and taken to the nest. First instars were mostly ignored by Myrmica sabuleti ants and they were rarely detected by Formica fusca. Older larvae were more attractive to Lasius than to the other ant genera. Pupae were very attractive to Lasius, moderately so to Myrmica, and were ignored by Formica fusca. Teneral adults were palpated by Lasius, but were attacked by Myrmica and Formica workers. We conclude that P. argus is a specialist associate of Lasius ants. Two populations of Plebejus argus were compared: one is naturally associated with Lasius niger, and the other with Lasius alienus. In reciprocal trials, larvae were slightly more attractive to their natural host ant species. Since test larvae were reared on a single host plant species in captivity, this differentiation probably has a genetic basis.     

The setae of parasitic Liphyra brassolis butterfly larvae form a flexible armour for resisting attack by their ant hosts (Lycaenidae: Lepidoptera)

Caterpillars of the lycaenid butterfly, Liphyra brassolis, live inside the nests of arboreal weaver ants, Oecophylla smaragdina, and eat their brood. Observations of mature larvae suggest that they are impervious to relentless ant molestation, yet they lack sclerotized cuticular plates. We document a novel form of integumental defence that imparts protection from ant attack whilst maintaining the flexibility necessary to walk with a hydraulic skeleton. Analysis of the trunk integument and cuticular structures of early and late instars of L. brassolis using light microscopy, scanning electron microscopy, and histology revealed three new setae types (disc, clavate, and lanceolate), as well as three new cuticular structures (pored sockets, cuticular pores, and cuticular domes). The unique cuticle is covered with lanceolate setae, which act as endocuticular struts, and overlapping scale‐like sockets, which form a hard, flexible integument. The imperfect armour of the early‐instar larvae suggests that abundant, putatively secretory pores are likely to be homologous to pore cupola organs (PCOs) found in other lycaenid larvae and thus may exude semiochemicals to allay ant aggression. The importance of these pores presumably wanes as structural (setal) cuticular defenses are reinforced in later instars, when adult ants have been observed attacking caterpillars to no avail. The caterpillar's antennae are unusual and seem to be involved in manipulating ant larvae into the caterpillar's mouth. Behavioural observations indicate that the dexterity of these structures is associated with eating ants (myrmecophagy).     

Two ways to be a myrmecophilous butterfly: natural history and comparative immature‐stage morphology of two species of Theope (Lepidoptera: Riodinidae)

Symbiotic interactions between butterfly larvae and ants, termed myrmecophily, require a range of behavioural and morphological adaptations (ant‐organs). Here, using light and scanning electron microscopy, we describe the complete life cycle of two species of Theope (Lepidoptera: Riodinidae) that have contrasting ways of life. Theope thestias larvae are facultatively tended by several ant species, whereas Theope pieridoides have obligate symbiotic interactions with Azteca ants that inhabit a myrmecophytic tree. Morphological differences associated with their different degrees of intimacy with tending ants are visible from hatching. In T. thestias, the untended first‐instar larva has elongated bifurcated setae and a few tiny perforated cupola organs (PCOs), whereas in T. pieridoides, the ant‐tended first instar has short dendritic setae, larger and more numerous PCOs, and functional tentacle nectary organs (TNOs). Throughout ontogeny, T. pieridoides always shows more conspicuous ant‐organs than T. thestias, with the exception of balloon setae, which are larger and more numerous in T. thestias. In addition, mature T. pieridoides larvae have an anterior set of ant‐organs, including a new type, here described and termed anterior glandular openings (AGOs). Based on the behavioural responses of ants in contact with these structures, a new interpretation for the mechanism whereby Theope larvae can manipulate the behaviour of their tending ants is proposed. Until now, three ecological syndromes can be defined among Theope species: (1) oligophagous larvae with facultative myrmecophily; (2) monophagous larvae with obligate myrmecophily; and (3) polyphagous larvae with obligate myrmecophily. These results suggest that differences in the degree of specificity in the ant–plant interactions may have an important role in the evolution of host‐plant use in Theope. © 2013 The Linnean Society of London     

Chemical disguise as particular caste of host ants in the ant inquiline parasite Niphanda fusca (Lepidoptera: Lycaenidae)

The exploitation of parental care is common in avian and insect 'cuckoos' and these species engage in a coevolutionary arms race. Caterpillars of the lycaenid butterfly Niphanda fusca develop as parasites inside the nests of host ants (Camponotus japonicus) where they grow by feeding on the worker trophallaxis. We hypothesized that N. fusca caterpillars chemically mimic host larvae, or some particular castes of the host ant, so that the caterpillars are accepted and cared for by the host workers. Behaviourally, it was observed that the host workers enthusiastically tended glass dummies coated with the cuticular chemicals of larvae or males and those of N. fusca caterpillars living together. Cuticular chemical analyses revealed that N. fusca caterpillars grown in a host ant nest acquired a colony-specific blend of cuticular hydrocarbons (CHCs). Furthermore, the CHC profiles of the N. fusca caterpillars were particularly close to those of the males rather than those of the host larvae and the others. We suggest that N. fusca caterpillars exploit worker care by matching their cuticular profile to that of the host males, since the males are fed by trophallaxis with workers in their natal nests for approximately ten months.     

Convergence of chemical mimicry in a guild of aphid predators

Abstract.  1. A variety of insects prey on honeydew-producing Homoptera and many do so even in the presence of ants that tend, and endeavour to protect, these trophobionts from natural enemies. Few studies have explored the semiochemical mechanisms by which these predators circumvent attack by otherwise aggressive ants.
2. Ants use specific mixtures of cuticular hydrocarbons (CHCs) as recognition labels, but this simple mechanism is frequently circumvented by nest parasites that engage in 'chemical mimicry' of their host ants by producing or acquiring a critical suite of these CHCs.
3. Analysis of the CHCs from the North American woolly alder aphid, Prociphilus tessellatus (Homoptera: Aphididae), their tending ants, and aphid predators from three insect orders, Feniseca tarquinius (Lepidoptera: Lycaenidae), Chrysopa slossonae (Neuroptera: Chrysopidae), and Syrphus ribesii (Diptera: Syrphidae), showed that while the CHC profile of each predatory species was distinct, each was chemically more similar to the aphids than to either tending ant species. Further, the CHCs of each predator species were a subset of the compounds found in the aphids' profile.
4. These results implicate CHCs as a recognition cue used by ants to discriminate trophobionts from potential prey and a probable mechanism by which trophobiont predators circumvent detection by aphids and their tending ants.
5. Although several features of the aphids' CHC profile are shared among the chemically mimetic taxa, variation in the precision of mimicry among the members of this predatory guild demonstrates that a chemical mimic need not replicate every feature of its model.     

Effect of grazing intensity on the interaction between Shijimiaeoides divinus asonis (Matsumura, 1929) and its attendant ants

This study was carried out in the natural herbaceous grassland of Mt. Aso, which had been almost completely grazed, and which is subjected to routine grassland burning every spring (February or March) to conserve Shijimiaeoides divinus asonis (Matsumura, 1929). We clarify that ants protect the larvae of this butterfly and evaluate the effects of grazing intensity on the attendant ant population. The results obtained are summarized as follows: (i) Five species of attendant ant were identified, with the dominant species being Formica japonica (Motschulsky, 1866) and Camponotus japonicus (Mayr, 1866). (ii) The number of attendant ants decreased at night time; however, only one or two ants attended the larva until the following morning. (iii) Ten species of insect excluding ants and three species of spiders that approached the larva were recorded on the larval host‐plant. Formica japonica and C. japonicus fought off most newcomers of other insects including the natural enemies of these butterfly larvae. (iv) The number of ants in this butterfly habitat under regular grazing intensity was significantly higher than during low grazing intensity and non‐grazing periods. (v) A positive correlation was found between the number of attendant ants and the number of butterfly larvae on the host‐plant. We concluded that the interaction between this butterfly and attendant ants is one of facultative mutualism because the attendant ants protect the butterfly larva. Therefore, the numbers of this butterfly species may decrease if the number of attendant ants decreases due to the cessation of pasturage.     

The role of cuticular hydrocarbons as chemical cues for nestmate recognition in the invasive Argentine ant (<Emphasis Type="Italic">Linepithema humile</Emphasis>)

Argentine ants (Linepithema humile) in their native South American range, like most other ant species, form spatially restricted colonies that display high levels of aggression toward other such colonies. In their introduced range, Argentine ants are unicolonial and form massive supercolonies composed of numerous nests among which territorial boundaries are absent. Here we examine the role of cuticular hydrocarbons (CHCs) in nestmate recognition of this highly damaging invasive ant using three supercolonies from its introduced range. We conducted behavioral assays to test the response of Argentine ants to workers treated with colonymate or non-colonymate CHCs. Additionally, we quantified the amount of hydrocarbons transferred to individual ants and performed gas chromatography-mass spectrometry (GC/MS) to qualitatively characterize our manipulation of CHC profiles. The GC/MS data revealed marked differences in the hydrocarbon profiles across supercolonies and indicated that our treatment effectively masked the original chemical profile of the treated ants with the profile belonging to the foreign individuals. We found that individual workers treated with foreign CHCs were aggressively rejected by their colonymates and this behavior appears to be concentration-dependent: larger quantities of foreign CHCs triggered higher levels of aggression. Moreover, this response was not simply due to an increase in the amount of CHCs applied to the cuticle since treatment with high concentrations of nestmate CHCs did not trigger aggression.The results of this study bolster the findings of previous studies on social insects that have implicated CHCs as nestmate recognition cues and provide insight into the mechanisms of nestmate recognition in the invasive Argentine ant. Received 6 February 2007; revised 31 May and 27 July 2007; accepted 16 August 2007.     

The taste of origin in a lady beetle: do males discriminate between females based on cuticular hydrocarbons?

The Asian lady beetle Harmonia axyridis originates from Asia and has established invasive populations worldwide. Recent population genetic studies trace their invasion routes and demonstrate that bottlenecks in population size have reduced their genetic diversity. Consequently, phenotypical differences are highlighted between native and invasive populations. Among phenotypical traits, cuticular hydrocarbons (CHCs) might reflect the geographical origin of a lady beetle, especially because of their genetic basis. The present study investigates whether (i) the CHC profiles qualitatively and quantitatively differ between females of H. axyridis from native and invasive populations and (ii) males discriminate females from native and invasive populations using CHC profiles. CHCs are solvent‐extracted before being quantified and identified by gas chromatography and mass spectrometry. In total, 17 CHCs are detected from female elytra, including six alkanes, three polyunsaturated and eight monounsaturated alkenes. The total quantity of CHCs differs among the populations, with lady beetles from Tai'an (China) displaying higher CHC concentrations than lady beetles from Gembloux (Belgium) and from Beijing (China) populations. Multivariate analyses detect differences in CHC qualitative profiles, with females from Tai'an being different from the two other populations. Finally, behavioural assays show that females originating from the native Tai'an population are less preferred by males, whereas females from the invasive population are mounted more often. The behavioural assays suggest that CHCs are not involved in discrimination of mating partners based on their origin.