Attracting a mate is critical in all species that sexually reproduce. Most animals, particularly insects, do this using chemical compounds called pheromones which can be sensed by potential mates. But how these vast range of different compounds encode and convey the information needed to secure a partner is not fully understood, and the genes that drive this complex communication mechanism are largely unknown.

To address this knowledge gap, Sun et al. studied the parasitic wasp Nasonia vitripennis. Like other insects, female N. vitripennis contain a wide range of chemical compounds on their cuticle, the outer waxy layer coating their surface. Sun et al. set out to find exactly which of these compounds, known as cuticular hydrocarbons, are involved in sexual communication.

They did this by simultaneously inactivating two related genes that they hypothesized to be responsible for synthesizing and maintaining chemical compounds on the cuticle of insects. The genetic modification altered the pattern of chemicals on the surface of the female wasps by specifically up- and down-regulating compounds with similar branching structures. The mutant females were also much less sexually attractive to male wasps.

These findings suggest that the chemical pattern identified by Sun et al. is responsible for communicating and maintaining sexual attractiveness in N. vitripennis female wasps. This is a significant stepping stone towards unravelling how sexual attractiveness can be encoded in complex mixtures of pheromones.

The results also have important implications for agriculture, as this parasitic wasp species is routinely used to exterminate particular fly populations that cause agricultural damage. The work by Sun et al. provides new insights into how these wasps sexually communicate, which may help scientists improve their rearing conditions and sustain them over multiple generations. This could contribute to a wider application of this more sustainable, eco-friendly alternative to destructive agricultural pesticides.

Tightly coordinated chemical signaling has repeatedly been shown to be of fundamental importance for successful reproduction in a wide range of animal species (Blomquist and Ginzel, 2021; Wyatt, 2014). Particularly insects have exploited this type of signaling as their primary mode of communication (Buellesbach et al., 2018a; Greenfield, 2002). Nevertheless, exactly how specific information such as mating status or attractiveness is encoded in the myriad of signaling molecules documented to be involved in sexual communication remains poorly understood (Stökl and Steiger, 2017; Allison and Carde, 2016). Cuticular hydrocarbons (CHCs), major components on the epicuticle of insects, are capable of chemically encoding and conveying a wide variety of biologically relevant information (Blomquist and Ginzel, 2021; Blomquist and Bagnères, 2010). Most prominently, CHCs have been shown to play pivotal roles in sexual communication as the main cues to attract and elicit courtship from conspecific mates (Würf et al., 2020; Finck et al., 2016b) to enable discrimination of con- from heterospecific mating partners (Shahandeh et al., 2018; Xue et al., 2018) and to signal receptivity and mating status (Berson and Simmons, 2019; Simmons, 2015).

Despite such diversified CHC-encoded signals and mediated behaviors, our knowledge on exactly how CHCs encode biologically relevant information has remained surprisingly scarce. This is particularly problematic in studies considering CHC profiles in their entirety as the main signaling entities (Berson and Simmons, 2019; Buellesbach et al., 2018b). The exact compounds or their combinations actually encoding the relevant information within CHC profiles remain largely elusive, except for a few case studies mainly involving the dipteran model organism Drosophila melanogaster, where single unsaturated CHC compounds appear to be the main mediators in sexual communication (Marcillac and Ferveur, 2004; Grillet et al., 2006). In most other cases, chemical information appears to be encoded in a much more complex manner, involving several CHC compounds in different quantitative combinations, with no deeper understanding on the actual coding patterns conveying specific information (Würf et al., 2020; Buellesbach et al., 2013).

In addition to our limited understanding on how CHC profiles encode information, our knowledge of the genetic basis of CHC biosynthesis and its impact on sexual signaling has remained comparably restricted and biased toward the Drosophila model system as well (Blomquist and Bagnères, 2010; Holze et al., 2021). In short, the CHC biosynthetic pathway consists of the elongation of fatty-acyl-Coenzyme A units to produce very long-chain fatty acids that are subsequently converted to CHCs (Blomquist and Bagnères, 2010; Nelson and Blomquist, 1995). An important early switch in CHC biosynthesis is either the incorporation of malonyl-CoA or methyl-malonyl-CoA, eventually leading to the production of straight-chain (n-) or methyl-branched (MB-) alkanes, respectively (Holze et al., 2021; Nelson and Blomquist, 1995, Figure 1). It has been hypothesized that these processes are mediated by two types of fatty-acyl-synthases (FAS), microsomal for methylmalonyl-CoA and cytosolic for malonyl-CoA (Gu et al., 1997; Juárez et al., 1992). To produce olefins (mono- and poly-unsaturated CHCs), desaturases introduce double bonds into straight-chain CHC precursors (Blomquist and Bagnères, 2010; Holze et al., 2021). In Drosophila, a couple of genes have been identified that mainly affect the biosynthesis and ratios of unsaturated CHC compounds that function in sexual signaling. For instance, two desaturases (Desat1 and DesatF) and one elongase (eloF) are involved in female diene production and consequently in their functionality as main sex pheromonal compounds (Chertemps et al., 2007; Chertemps et al., 2006). Furthermore, in the Australian congeneric species D. serrata, the male-specific D. melanogaster orthologue FASN2 has been shown to affect the biosynthesis of three MB-CHCs, among them the additional female mating stimulant 2-Me-C26 (Wicker-Thomas et al., 2015; Chung et al., 2014). Apart from these case studies limited to Drosophila, we know very little about the genetic basis linking CHC biosynthesis and sexual signaling in other insects (Holze et al., 2021).

Figure 1

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The parasitoid jewel wasp Nasonia vitripennis (Hymenoptera: Pteromalidae) has emerged as a suitable model organism to combine studies on functional genetics as well as chemical communication systems in Hymenoptera (Buellesbach et al., 2022; Niehuis et al., 2013). Female CHCs serve as sexual cues capable of eliciting male courtship and copulation behavior in N. vitripennis (Buellesbach et al., 2018b; Steiner et al., 2006). The CHC profiles of N. vitripennis females exhibit a high complexity, consisting of a mixture of n-alkanes, n-alkenes, and MB-alkanes in various quantities. Specifically, the latter fraction, which makes up more than 85% of the whole profile, displays a rich diversity in methyl-branch numbers, chain lengths, and respective relative abundances, hinting at a considerable potential for encoding differential information (Buellesbach et al., 2013; Steiner et al., 2006). However, the entire female CHC profile has long been regarded as encoding their sexual attractiveness, with no single compound, compound classes, or particular patterns being identifiable as the main conveyers of the sexual signaling function (Steiner et al., 2006; Buellesbach, 2018). Moreover, despite recent advances in unraveling the genetic architecture of CHC biosynthesis and variation in the Nasonia genus (Buellesbach et al., 2022), the effects of individual genes on CHC profiles and, more importantly, on the encoded sexual signaling function, could not be determined as of yet.

In this study, we characterize the phenotypic effects of two fatty acid synthase genes whose knockdown impacts the variation of several structurally related CHC components concordant with female sexual attractiveness. CHC profiles of knockdown females primarily showed significant up- and downregulations of MB-alkane compounds with correlated branching patterns. At the same time, these knockdown females elicited significantly less courtship and copulation attempts from conspecific males. These constitute the first hymenopteran genes with a demonstrated function in governing the variation of primarily MB-alkanes as well as communication, hinting at a chemical coding pattern for sexual attractiveness mostly conveyed by this CHC compound class.

In our study, we shed light on how sexual attractiveness can be encoded by differentially branched CHCs and unravel its genetic architecture to be based on two highly similar fatty acid synthase genes. Knocking down these two genes in N. vitripennis females leads to a consistent pattern of primarily up- and downregulated MB-alkanes with specific branching patterns. This dramatic shift is accompanied by a significant reduction of courtship and copulation behavior toward knockdown females by conspecific males which we demonstrate to be mainly determined by the altered MB-alkane fraction. This advances our understanding of how genetic information is translated into chemical information and brings us a step closer in decoding biologically relevant information from complex chemical profiles.

Most conspicuously in females, quantities of CHCs with their first methyl groups at the 7th and 9th (and higher) C-atom positions are dramatically downregulated, whereas the ones with their first methyl groups at the respective 3rd and 5th positions appear mainly upregulated, in most cases by several orders of magnitude (Figure 2 and Supplementary file 3). Intriguingly, overall CHC quantities do not differ between knockdown and control individuals (Figure 2—figure supplement 3A), suggesting a very specific regulatory function for fas5/fas6 in governing these opposing and potentially compensatory branching patterns. Concordantly, this emphasizes the pivotal role of both these particular genes and the wild type MB-alkane branching patterns in encoding and maintaining the attractiveness of female N. vitripennis CHC profiles. Moreover, there appear to be a number of sex-specific differences in the knockdown effects on the different CHC compound classes, most notably apparent in significant upregulations of both total and n-alkane CHC quantities in knockdown males as opposed to females and partially differentially affected MB-alkane quantities (compare Figure 2 to Figure 2—figure supplement 4). Curiously, GFP dsRNAi appears to have a generally upregulating effect as opposed to WT controls despite for n-alkene quantities in males, which has not been reported before. However, a recent study hints at potential off-target effects of GFP dsRNAi on a small subset of N. vitripennis genes mainly involved in microtubule and sperm development (Rougeot et al., 2021). Interestingly, this appears to be also the case for the expressions of fas1 and fas2 gene transcripts, which are significantly upregulated in the GFP dsRNAi controls, again exclusively in males (Figure 2—figure supplement 1B). Though no off-target effects on CHC profile biosynthesis and variation have been reported so far for GFP dsRNAi, we cannot exclude this possibility, particularly since a trend toward higher CHC quantities in GFP dsRNAi controls as opposed to WT controls is also discernible in some cases for the females (Figure 2E–G). Therefore, as Rougeot et al. already hinted at , we strongly suggest alternative non-target controls in future studies on the genetics of CHC biosynthesis and variation. Concerning the partially different effects of fas5 on CHC quantities in females and males, as no functionality in chemical communication could be attributed to the latters’ CHC profiles so far, these could not be investigated any further. However, opposing sex-specific effects of CHC biosynthesis genes on the quantity of different compound classes appear to be a rather common occurrence, particularly in the insect model system D. melanogaster (Holze et al., 2021). Based mainly on research on the latter, CHC-based sexual signaling mechanisms have long been assumed to be comparatively simple, mainly mediated by two doubly unsaturated dienes in females and a mono-unsaturated n-alkene in males (Marcillac and Ferveur, 2004; Grillet et al., 2006). However, experimental evidence has accumulated that other CHC compounds can also complement sexual signaling in Drosophila in various ways (Chung et al., 2014; Everaerts et al., 2010). When regarding CHC profiles in their entirety as opposed to single compounds, direct causal links between sexual attractiveness and CHC profiles properties have rarely been experimentally demonstrated and have most often defied clear patterns (Würf et al., 2020; Xue et al., 2018). In the genus Nasonia and other related parasitoid wasp species, it has so far been assumed that sexual attractiveness is a trait attributed to their entire CHC profile as either present or absent depending on the studied species and also potentially reinforced by other factors such as polar cuticular compounds (Buellesbach et al., 2013; Mair et al., 2017). Our study clearly shows that CHC-mediated sexual attractiveness is primarily conveyed through a relatively complex chemical pattern with a comparatively simple genetic basis.

Interestingly, knockdown of fas5 also upregulated n-alkene quantities (Figure 2—figure supplement 3C), which have recently been shown to have a repellent effect on N. vitripennis males, preventing them from engaging in homosexual courtship behavior (Wang et al., 2022). However, our CHC compound class separation greatly reduced the increased proportion of n-alkenes in fas5 knockdown females to levels almost equivalent to those found in wild type females (0.94% and 0.91 %, respectively, in the extract of fas5 and WT MB fractions, compare Figure 4—source data 1 to Supplementary file 3). This renders the contribution of n-alkenes to the sharp reduction in sexual attractiveness in fas5 knockdown females unlikely and strongly suggests that the female sexual signaling function is mainly mediated by MB-alkanes (Figure 4). We argue that this CHC compound class indeed possesses the highest potential for encoding a wide variety of chemical information through the myriad of possible positions and numbers of methyl branches. In fact, a couple of studies have already hinted at MB-alkanes as the main carriers for chemical information in insect CHC profiles, providing evidence for the involvement of MB-alkanes in chemical communication processes (Spikes et al., 2010; Holman et al., 2016). This might be of particular importance in Hymenoptera, an insect order with CHC profiles largely dominated by MB-alkanes (Kather and Martin, 2015; Martin and Drijfhout, 2009) and in which both theoretical considerations and empirical evidence have accumulated for the increased complexity and high sophistication of their chemical communication systems (Saad et al., 2018; Obiero et al., 2021). Contrary to this, it has been argued that olefins (mainly n-alkenes and dienes) have a higher potential for encoding chemical information than MB-alkanes (Chung and Carroll, 2015). However, this view might have been biased and mainly informed by findings from Drosophila, where unsaturated compounds appear to be, in fact, the main mediators of sexual communication (Marcillac and Ferveur, 2004; Ferveur, 2005). In direct comparison, MB-alkanes constitute the dominant fraction in Nasonia CHC profiles (>85%) (Buellesbach et al., 2022) as opposed to Drosophila profiles (16–24%) (Dembeck et al., 2015). Since the split between Hymenoptera and other holometabolous insects including Diptera has been estimated to have occurred 327 mya (Misof et al., 2014), fundamental shifts in basic properties of both surface profile compositions and chemical signaling functionalities might be expected. Therefore, the promising role of MB-alkanes in conveying chemical information should be investigated more prominently in future studies. Through a wider evolutionary lens, it will be interesting to investigate how the present findings compare to other CHC-based communication systems in the vastly diverse insect order Hymenoptera. Notably, whereas in solitary Hymenoptera CHCs can function as contact sex pheromones, they are prominent and fundamental nestmate and caste recognition cues in eusocial Hymenoptera (Buellesbach et al., 2018b; Vereecken et al., 2007; Leonhardt et al., 2016). However, the main encoding mechanisms as well as the underlying genetic basis still remain largely elusive in most taxa, and our study constitutes an important stepping stone for investigating potential similarities in these important communication modalities in a larger evolutionary context. Moreover, since CHC profiles can be highly species-specific, their potential involvement in species recognition and assortative mating warrants further investigation to elucidate the exact underlying chemical and genetic differentiation mechanisms (Buellesbach et al., 2018b; Wurdack et al., 2015; Weiss et al., 2015).

Concerning gene orthology, fas5 has been annotated as a homolog to FASN3 in D. melanogaster (Buellesbach et al., 2022; Lammers et al., 2019). Interestingly, knockdowns of FASN3 alone do not induce any compound changes in D. melanogaster CHC profiles but increase the flies’ sensitivity to desiccation (Wicker-Thomas et al., 2015). A FASN3 ortholog expressed in the kissing bug Rhodinius prolixus also contributes to desiccation resistance, but simultaneously downregulates MB- while upregulating straight-chain alkanes (Moriconi et al., 2019). Similarly, in the migratory locust Locusta migratoria, silencing two FAS genes decreased insect survival under desiccation stress while altering the amounts of both MB- and straight-chain alkanes (Yang et al., 2020). In contrast to these studies, we were not able confirm any functionality in desiccation resistance for fas5 in both N. vitripennis males and females (Figure 5), nor did the previous studies report any impact on CHC-based chemical signaling. There are two additional FAS genes characterized in D. melanogaster: FASN1, which is responsible for overall CHC production with no specific effects on any particular compound classes (Wicker-Thomas et al., 2015) and FASN2, which exclusively regulates MB-alkane production in males (Chung et al., 2014). Although not a direct homolog, the main impact on MB-alkane variations of FASN2 in D. melanogaster is comparable to that of fas5 in N. vitripennis, with the exception that we were able to document effects on this compound class for both sexes (compare Figure 2 and Figure 2—figure supplement 4). Such diversified functionalities of the FAS genes characterized so far indicate their high versatility as early mediators in the CHC biosynthesis pathway (Figure 1). FAS genes have been implied as instrumental for generating the huge diversity of different CHC profiles across insects, with high evolutionary turnover rates and differences in the specific functional recruitments of FAS gene family members (Finck et al., 2016a; Moriconi et al., 2019). However, FAS genes are far from restricted to impact CHC biosynthesis alone and have been documented to be involved in a wide variety of other physiological processes, ranging from lipogenesis to diapause induction (Tan et al., 2017; Lammers et al., 2019; Moriconi et al., 2019). Therefore, specifically predicting functionalities of FAS genes and unambiguously associating them with CHC biosynthesis and variation has been notoriously difficult (Holze et al., 2021). Our study suggests a very specific affinity of fas5 for particular methyl-branching patterns, potentially originating in an enzymatic preference for methyl-malonyl-CoA predecessors with pre-existing branching patterns (Blomquist and Bagnères, 2010; Nelson and Blomquist, 1995). To clarify this, future studies should determine whether the fas5 gene product constitutes a soluble cytosolic or membrane-bound microsomal FAS enzyme, the latter of which has been postulated to be specific for the incorporation of methyl-malonyl-CoA subunits (Gu et al., 1997; Juárez et al., 1992, Figure 1).

The previously uncharacterized fas gene fas6 not only shows high sequence similarity to fas5,but is also its physical neighbor (Figure 2—figure supplement 2). Moreover, both of these genes show similarly high expression patterns in wild type N. vitripennis wasps (Figure 2H and Figure 2—figure supplement 4I). This basically implies that these two genes originated from a tandem gene duplication event and therefore constitute paralogs of the D. melanogaster FASN3. Hence, we cannot state at this point whether either one of these two genes alone or both are responsible for the observed phenotypic changes. Interestingly, this mirrors a similar finding in N. vitripennis males concerning the biosynthesis of a lactone functioning in a long-range pheromonal blend attractive to virgin females (Niehuis et al., 2013). The production of this compound was unambiguously attributed to three short-chain dehydrogenase/reductase genes with high sequence similarity, which could also not be targeted independently by dsRNAi knockdowns. This demonstrates the difficulty to narrow down specific genes responsible for the biosynthesis of pheromonally active compounds when their sequences show particularly high degrees of similarity and simultaneously high expression patterns.

In conclusion, our study demonstrates the considerable impact of two highly similar fatty acid synthase genes on female sexual attractiveness in a parasitoid wasp, thereby immediately suggesting how this trait can be encoded through specific patterns of MB-alkanes. To the best of our knowledge, these are the first identified hymenopteran genes with a specific effect on MB-alkane ratios that simultaneously impact sexual attractiveness. Transcending the demonstrated impact on sexual signaling and elicited mating behavior, the present findings also substantially advance our general knowledge on the so far little investigated genetic underpinnings of MB-alkane variation and production. This particular compound class dominates the surface profiles of many insects, most notably in the ecologically and economically important insect order Hymenoptera, and harbors a considerable potential for encoding chemical information, inviting a stronger emphasis on these compounds in future studies on chemical signaling and its behavioral impact.

The datasets generated or analyzed during this study are available at the figshare data repository under https://doi.org/10.6084/m9.figshare.20411958.

1. 1. Sun W
   2. Lange MI
   3. Gadau J
   4. Buellesbach J

   (2023) figshare

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

   https://doi.org/10.6084/m9.figshare.20411958

1. Book

   1. Allison JD
   2. Carde RT

   (2016) Pheromone Communication in Moths: Evolution, Behavior, and Application.

   Univ of California Press.

   https://doi.org/10.1525/9780520964433

   • Google Scholar
2. 1. Bello JE
   2. McElfresh JS
   3. Millar JG

   (2015) Isolation and determination of absolute configurations of insect-produced methyl-branched hydrocarbons

   PNAS 112:1077–1082.

   https://doi.org/10.1073/pnas.1417605112

   • PubMed
   • Google Scholar
3. 1. Benjamini Y
   2. Yekutieli D

   (2001) The control of the false discovery rate in multiple testing under dependency

   The Annals of Statistics 29:1165–1188.

   https://doi.org/10.1214/aos/1013699998

   • Google Scholar
4. 1. Berson JD
   2. Simmons LW

   (2019) Female cuticular hydrocarbons can signal indirect fecundity benefits in an insect

   Evolution; International Journal of Organic Evolution 73:982–989.

   https://doi.org/10.1111/evo.13720

   • PubMed
   • Google Scholar
5. Book

   1. Blomquist GJ
   2. Bagnères AG

   (2010) Insect hydrocarbons

   In: Blomquist GJ, editors. Insect Hydrocarbons: Biology, Biochemistry, and Chemical Ecology.. Cambridge University Press. pp. 221–252.

   https://doi.org/10.1017/CBO9780511711909

   • Google Scholar
6. 1. Blomquist GJ
   2. Ginzel MD

   (2021) Chemical ecology, biochemistry, and molecular biology of insect hydrocarbons

   Annual Review of Entomology 66:45–60.

   https://doi.org/10.1146/annurev-ento-031620-071754

   • PubMed
   • Google Scholar
7. 1. Buellesbach J
   2. Gadau J
   3. Beukeboom LW
   4. Echinger F
   5. Raychoudhury R
   6. Werren JH
   7. Schmitt T

   (2013) Cuticular hydrocarbon divergence in the jewel wasp Nasonia: evolutionary shifts in chemical communication channels?

   Journal of Evolutionary Biology 26:2467–2478.

   https://doi.org/10.1111/jeb.12242

   • PubMed
   • Google Scholar
8. 1. Buellesbach J
   2. Greim C
   3. Schmitt T

   (2014) Asymmetric interspecific mating behavior reflects incomplete prezygotic isolation in the jewel wasp genus Nasonia

   Ethology: Formerly Zeitschrift Fur Tierpsychologie 120:834–843.

   https://doi.org/10.1111/eth.12250

   • Google Scholar
9. Book

   1. Buellesbach J

   (2018)

   Pheromone communication systems in the Nasonia species complex

   In: Jenkins OP, editors. Advances in Animal Science and Zoology.. Nova Science Publishers, Inc. pp. 205–219.

   • Google Scholar
10. Book

    1. Buellesbach J
    2. Cash E
    3. Schmitt T

    (2018a)

    Communication, insects

    In: Skinner M, editors. Encyclopedia of Reproduction. Elsevier Science Publishing Co Inc. pp. 78–83.

    • Google Scholar
11. 1. Buellesbach J
    2. Vetter SG
    3. Schmitt T

    (2018b) Differences in the reliance on cuticular hydrocarbons as sexual signaling and species discrimination cues in parasitoid wasps

    Frontiers in Zoology 15:22.

    https://doi.org/10.1186/s12983-018-0263-z

    • PubMed
    • Google Scholar
12. 1. Buellesbach J
    2. Holze H
    3. Schrader L
    4. Liebig J
    5. Schmitt T
    6. Gadau J
    7. Niehuis O

    (2022) Genetic and genomic architecture of species-specific cuticular hydrocarbon variation in parasitoid wasps

    Proceedings. Biological Sciences 289:20220336.

    https://doi.org/10.1098/rspb.2022.0336

    • PubMed
    • Google Scholar
13. 1. Chertemps T
    2. Duportets L
    3. Labeur C
    4. Ueyama M
    5. Wicker-Thomas C

    (2006) A female-specific desaturase gene responsible for diene hydrocarbon biosynthesis and courtship behaviour in Drosophila melanogaster

    Insect Molecular Biology 15:465–473.

    https://doi.org/10.1111/j.1365-2583.2006.00658.x

    • PubMed
    • Google Scholar
14. 1. Chertemps T
    2. Duportets L
    3. Labeur C
    4. Ueda R
    5. Takahashi K
    6. Saigo K
    7. Wicker-Thomas C

    (2007) A female-biased expressed elongase involved in long-chain hydrocarbon biosynthesis and courtship behavior in Drosophila melanogaster

    PNAS 104:4273–4278.

    https://doi.org/10.1073/pnas.0608142104

    • PubMed
    • Google Scholar
15. 1. Chung H
    2. Loehlin DW
    3. Dufour HD
    4. Vaccarro K
    5. Millar JG
    6. Carroll SB

    (2014) A single gene affects both ecological divergence and mate choice in Drosophila

    Science 343:1148–1151.

    https://doi.org/10.1126/science.1249998

    • PubMed
    • Google Scholar
16. 1. Chung H
    2. Carroll SB

    (2015) Wax, sex and the origin of species: Dual roles of insect cuticular hydrocarbons in adaptation and mating

    BioEssays 37:822–830.

    https://doi.org/10.1002/bies.201500014

    • PubMed
    • Google Scholar
17. 1. Davies NJ
    2. Tauber E

    (2015) WaspAtlas: a Nasonia vitripennis gene database and analysis platform

    Database 2015:bav103.

    https://doi.org/10.1093/database/bav103

    • PubMed
    • Google Scholar
18. 1. Dembeck LM
    2. Böröczky K
    3. Huang W
    4. Schal C
    5. Anholt RRH
    6. Mackay TFC

    (2015) Genetic architecture of natural variation in cuticular hydrocarbon composition in Drosophila melanogaster

    eLife 4:e09861.

    https://doi.org/10.7554/eLife.09861

    • PubMed
    • Google Scholar
19. 1. Everaerts C
    2. Farine JP
    3. Cobb M
    4. Ferveur JF

    (2010) Drosophila cuticular hydrocarbons revisited: mating status alters cuticular profiles

    PLOS ONE 5:e9607.

    https://doi.org/10.1371/journal.pone.0009607

    • PubMed
    • Google Scholar
20. 1. Ferveur JF

    (2005) Cuticular hydrocarbons: their evolution and roles in Drosophila pheromonal communication

    Behavior Genetics 35:279–295.

    https://doi.org/10.1007/s10519-005-3220-5

    • PubMed
    • Google Scholar
21. 1. Finck J
    2. Berdan EL
    3. Mayer F
    4. Ronacher B
    5. Geiselhardt S

    (2016a) Divergence of cuticular hydrocarbons in two sympatric grasshopper species and the evolution of fatty acid synthases and elongases across insects

    Scientific Reports 6:33695.

    https://doi.org/10.1038/srep33695

    • PubMed
    • Google Scholar
22. 1. Finck J
    2. Kuntze J
    3. Ronacher B

    (2016b) Chemical cues from females trigger male courtship behaviour in grasshoppers

    Journal of Comparative Physiology. A, Neuroethology, Sensory, Neural, and Behavioral Physiology 202:337–345.

    https://doi.org/10.1007/s00359-016-1081-4

    • PubMed
    • Google Scholar
23. Book

    1. Greenfield MD

    (2002)

    Signalers and Receivers: Mechanisms and Evolution of Arthropod Communication

    Oxford University Press.

    • Google Scholar
24. 1. Grillet M
    2. Dartevelle L
    3. Ferveur JF

    (2006) A Drosophila male pheromone affects female sexual receptivity

    Proceedings. Biological Sciences 273:315–323.

    https://doi.org/10.1098/rspb.2005.3332

    • PubMed
    • Google Scholar
25. 1. Gu P
    2. Welch WH
    3. Guo L
    4. Schegg KM
    5. Blomquist GJ

    (1997) Characterization of a novel microsomal fatty acid synthetase (FAS) compared to a cytosolic FAS in the housefly, Musca domestica

    Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology 118:447–456.

    https://doi.org/10.1016/s0305-0491(97)00112-0

    • PubMed
    • Google Scholar
26. 1. Holman L
    2. Hanley B
    3. Millar JG

    (2016) Highly specific responses to queen pheromone in three Lasius ant species

    Behavioral Ecology and Sociobiology 70:387–392.

    https://doi.org/10.1007/s00265-016-2058-6

    • Google Scholar
27. 1. Holze H
    2. Schrader L
    3. Buellesbach J

    (2021) Advances in deciphering the genetic basis of insect cuticular hydrocarbon biosynthesis and variation

    Heredity 126:219–234.

    https://doi.org/10.1038/s41437-020-00380-y

    • PubMed
    • Google Scholar
28. 1. Juárez P
    2. Chase J
    3. Blomquist GJ

    (1992) A microsomal fatty acid synthetase from the integument of Blattella germanica synthesizes methyl-branched fatty acids, precursors to hydrocarbon and contact sex pheromone

    Archives of Biochemistry and Biophysics 293:333–341.

    https://doi.org/10.1016/0003-9861(92)90403-j

    • PubMed
    • Google Scholar
29. 1. Kather R
    2. Martin SJ

    (2015) Evolution of cuticular hydrocarbons in the hymenoptera: A meta-analysis

    Journal of Chemical Ecology 41:871–883.

    https://doi.org/10.1007/s10886-015-0631-5

    • PubMed
    • Google Scholar
30. 1. Lammers M
    2. Kraaijeveld K
    3. Mariën J
    4. Ellers J

    (2019) Gene expression changes associated with the evolutionary loss of a metabolic trait: lack of lipogenesis in parasitoids

    BMC Genomics 20:309.

    https://doi.org/10.1186/s12864-019-5673-6

    • PubMed
    • Google Scholar
31. 1. Leonhardt SD
    2. Menzel F
    3. Nehring V
    4. Schmitt T

    (2016) Ecology and evolution of communication in social insects

    Cell 164:1277–1287.

    https://doi.org/10.1016/j.cell.2016.01.035

    • PubMed
    • Google Scholar
32. 1. Lynch JA
    2. Desplan C

    (2006) A method for parental RNA interference in the wasp Nasonia vitripennis

    Nature Protocols 1:486–494.

    https://doi.org/10.1038/nprot.2006.70

    • PubMed
    • Google Scholar
33. 1. Mair MM
    2. Kmezic V
    3. Huber S
    4. Pannebakker BA
    5. Ruther J

    (2017) The chemical basis of mate recognition in two parasitoid wasp species of the genus Nasonia

    Entomologia Experimentalis et Applicata 164:1–15.

    https://doi.org/10.1111/eea.12589

    • Google Scholar
34. 1. Marcillac F
    2. Ferveur JF

    (2004) A set of female pheromones affects reproduction before, during and after mating in Drosophila

    The Journal of Experimental Biology 207:3927–3933.

    https://doi.org/10.1242/jeb.01236

    • PubMed
    • Google Scholar
35. 1. Martin S
    2. Drijfhout F

    (2009) A review of ant cuticular hydrocarbons

    Journal of Chemical Ecology 35:1151–1161.

    https://doi.org/10.1007/s10886-009-9695-4

    • PubMed
    • Google Scholar
36. 1. Misof B
    2. Liu S
    3. Meusemann K
    4. Peters RS
    5. Donath A
    6. Mayer C
    7. Frandsen PB
    8. Ware J
    9. Flouri T
    10. Beutel RG
    11. Niehuis O
    12. Petersen M
    13. Izquierdo-Carrasco F
    14. Wappler T
    15. Rust J
    16. Aberer AJ
    17. Aspöck U
    18. Aspöck H
    19. Bartel D
    20. Blanke A
    21. Berger S
    22. Böhm A
    23. Buckley TR
    24. Calcott B
    25. Chen J
    26. Friedrich F
    27. Fukui M
    28. Fujita M
    29. Greve C
    30. Grobe P
    31. Gu S
    32. Huang Y
    33. Jermiin LS
    34. Kawahara AY
    35. Krogmann L
    36. Kubiak M
    37. Lanfear R
    38. Letsch H
    39. Li Y
    40. Li Z
    41. Li J
    42. Lu H
    43. Machida R
    44. Mashimo Y
    45. Kapli P
    46. McKenna DD
    47. Meng G
    48. Nakagaki Y
    49. Navarrete-Heredia JL
    50. Ott M
    51. Ou Y
    52. Pass G
    53. Podsiadlowski L
    54. Pohl H
    55. von Reumont BM
    56. Schütte K
    57. Sekiya K
    58. Shimizu S
    59. Slipinski A
    60. Stamatakis A
    61. Song W
    62. Su X
    63. Szucsich NU
    64. Tan M
    65. Tan X
    66. Tang M
    67. Tang J
    68. Timelthaler G
    69. Tomizuka S
    70. Trautwein M
    71. Tong X
    72. Uchifune T
    73. Walzl MG
    74. Wiegmann BM
    75. Wilbrandt J
    76. Wipfler B
    77. Wong TKF
    78. Wu Q
    79. Wu G
    80. Xie Y
    81. Yang S
    82. Yang Q
    83. Yeates DK
    84. Yoshizawa K
    85. Zhang Q
    86. Zhang R
    87. Zhang W
    88. Zhang Y
    89. Zhao J
    90. Zhou C
    91. Zhou L
    92. Ziesmann T
    93. Zou S
    94. Li Y
    95. Xu X
    96. Zhang Y
    97. Yang H
    98. Wang J
    99. Wang J
    100. Kjer KM
    101. Zhou X

    (2014) Phylogenomics resolves the timing and pattern of insect evolution

    Science 346:763–767.

    https://doi.org/10.1126/science.1257570

    • PubMed
    • Google Scholar
37. 1. Moriconi DE
    2. Dulbecco AB
    3. Juárez MP
    4. Calderón-Fernández GM

    (2019) A fatty acid synthase gene (FASN3) from the integument tissue of Rhodnius prolixus contributes to cuticle water loss regulation

    Insect Molecular Biology 28:850–861.

    https://doi.org/10.1111/imb.12600

    • PubMed
    • Google Scholar
38. Book

    1. Nelson DR
    2. Blomquist GJ

    (1995)

    Insect waxes

    In: Hamilton RJ, editors. Waxes: Chemistry, Molecular Biology and Functions.. Oily Press. pp. 1–90.

    • Google Scholar
39. 1. Niehuis O
    2. Buellesbach J
    3. Gibson JD
    4. Pothmann D
    5. Hanner C
    6. Mutti NS
    7. Judson AK
    8. Gadau J
    9. Ruther J
    10. Schmitt T

    (2013) Behavioural and genetic analyses of Nasonia shed light on the evolution of sex pheromones

    Nature 494:345–348.

    https://doi.org/10.1038/nature11838

    • PubMed
    • Google Scholar
40. 1. Obiero GF
    2. Pauli T
    3. Geuverink E
    4. Veenendaal R
    5. Niehuis O
    6. Große-Wilde E

    (2021) Chemoreceptor diversity in apoid wasps and its reduction during the evolution of the pollen-collecting lifestyle of bees (hymenoptera: Apoidea)

    Genome Biology and Evolution 13:evaa269.

    https://doi.org/10.1093/gbe/evaa269

    • PubMed
    • Google Scholar
41. 1. Rao X
    2. Huang X
    3. Zhou Z
    4. Lin X

    (2013)

    An improvement of the 2ˆ(-delta delta CT) method for quantitative real-time polymerase chain reaction data analysis

    Biostatistics, Bioinformatics and Biomathematics 3:71–85.

    • PubMed
    • Google Scholar
42. 1. Rougeot J
    2. Wang Y
    3. Verhulst EC
    4. Nevels M

    (2021) Effect of using green fluorescent protein double-stranded RNA as non-target negative control in Nasonia vitripennis RNA interference assays

    Experimental Results 2:67.

    https://doi.org/10.1017/exp.2020.67

    • Google Scholar
43. 1. Saad R
    2. Cohanim AB
    3. Kosloff M
    4. Privman E

    (2018) Neofunctionalization in ligand binding sites of ant olfactory receptors

    Genome Biology and Evolution 10:2490–2500.

    https://doi.org/10.1093/gbe/evy131

    • PubMed
    • Google Scholar
44. 1. Shahandeh MP
    2. Pischedda A
    3. Turner TL

    (2018) Male mate choice via cuticular hydrocarbon pheromones drives reproductive isolation between Drosophila species

    Evolution; International Journal of Organic Evolution 72:123–135.

    https://doi.org/10.1111/evo.13389

    • PubMed
    • Google Scholar
45. 1. Simmons LW

    (2015) Sexual signalling by females: do unmated females increase their signalling effort?

    Biology Letters 11:20150298.

    https://doi.org/10.1098/rsbl.2015.0298

    • PubMed
    • Google Scholar
46. 1. Spikes AE
    2. Paschen MA
    3. Millar JG
    4. Moreira JA
    5. Hamel PB
    6. Schiff NM
    7. Ginzel MD

    (2010) First contact pheromone identified for a longhorned beetle (Coleoptera: Cerambycidae) in the subfamily Prioninae

    Journal of Chemical Ecology 36:943–954.

    https://doi.org/10.1007/s10886-010-9837-8

    • PubMed
    • Google Scholar
47. 1. Steiner S
    2. Hermann N
    3. Ruther J

    (2006) Characterization of a female-produced courtship pheromone in the parasitoid Nasonia vitripennis

    Journal of Chemical Ecology 32:1687–1702.

    https://doi.org/10.1007/s10886-006-9102-3

    • PubMed
    • Google Scholar
48. 1. Stökl J
    2. Steiger S

    (2017) Evolutionary origin of insect pheromones

    Current Opinion in Insect Science 24:36–42.

    https://doi.org/10.1016/j.cois.2017.09.004

    • PubMed
    • Google Scholar
49. 1. Tan QQ
    2. Liu W
    3. Zhu F
    4. Lei CL
    5. Wang XP

    (2017) Fatty acid synthase 2 contributes to diapause preparation in a beetle by regulating lipid accumulation and stress tolerance genes expression

    Scientific Reports 7:40509.

    https://doi.org/10.1038/srep40509

    • PubMed
    • Google Scholar
50. Book

    1. Therneau TM
    2. Grambsch PM
    3. Therneau TM
    4. Grambsch PM

    (2000) Modeling Survival Data: Extending the Cox Model.

    Springer.

    https://doi.org/10.1007/978-1-4757-3294-8

    • Google Scholar
51. 1. Vereecken NJ
    2. Mant J
    3. Schiestl FP

    (2007) Population differentiation in female sex pheromone and male preferences in a solitary bee

    Behavioral Ecology and Sociobiology 61:811–821.

    https://doi.org/10.1007/s00265-006-0312-z

    • Google Scholar
52. 1. Wang Y
    2. Sun W
    3. Fleischmann S
    4. Millar JG
    5. Ruther J
    6. Verhulst EC

    (2022) Silencing Doublesex expression triggers three-level pheromonal feminization in Nasonia vitripennis males

    Proceedings of the Royal Society of London, Series B: Biological Sciences 1:20212002.

    https://doi.org/10.1101/2021.05.21.445141

    • Google Scholar
53. 1. Weiss I
    2. Hofferberth J
    3. Ruther J
    4. Stökl J

    (2015) Varying importance of cuticular hydrocarbons and iridoids in the species-specific mate recognition pheromones of three closely related Leptopilina species

    Frontiers in Ecology and Evolution 3:00019.

    https://doi.org/10.3389/fevo.2015.00019

    • Google Scholar
54. 1. Wicker-Thomas C
    2. Garrido D
    3. Bontonou G
    4. Napal L
    5. Mazuras N
    6. Denis B
    7. Rubin T
    8. Parvy JP
    9. Montagne J

    (2015) Flexible origin of hydrocarbon/pheromone precursors in Drosophila melanogaster

    Journal of Lipid Research 56:2094–2101.

    https://doi.org/10.1194/jlr.M060368

    • PubMed
    • Google Scholar
55. 1. Wurdack M
    2. Herbertz S
    3. Dowling D
    4. Kroiss J
    5. Strohm E
    6. Baur H
    7. Niehuis O
    8. Schmitt T

    (2015) Striking cuticular hydrocarbon dimorphism in the mason wasp Odynerus spinipes and its possible evolutionary cause (Hymenoptera: Chrysididae, Vespidae)

    Proceedings. Biological Sciences 282:20151777.

    https://doi.org/10.1098/rspb.2015.1777

    • PubMed
    • Google Scholar
56. 1. Würf J
    2. Pokorny T
    3. Wittbrodt J
    4. Millar JG
    5. Ruther J

    (2020) Cuticular hydrocarbons as contact sex pheromone in the parasitoid wasp urolepis rufipes

    Frontiers in Ecology and Evolution 8:180.

    https://doi.org/10.3389/fevo.2020.00180

    • Google Scholar
57. Book

    1. Wyatt TD

    (2014) Pheromones and Animal Behaviour: Communication by Smell and Taste.

    Cambridge University Press.

    https://doi.org/10.1017/CBO9781139030748

    • Google Scholar
58. 1. Xue HJ
    2. Segraves KA
    3. Wei J
    4. Zhang B
    5. Nie RE
    6. Li WZ
    7. Yang XK

    (2018) Chemically mediated sexual signals restrict hybrid speciation in a flea beetle

    Behavioral Ecology 29:1462–1471.

    https://doi.org/10.1093/beheco/ary105

    • Google Scholar
59. 1. Yang Y
    2. Zhao X
    3. Niu N
    4. Zhao Y
    5. Liu W
    6. Moussian B
    7. Zhang J

    (2020) Two fatty acid synthase genes from the integument contribute to cuticular hydrocarbon biosynthesis and cuticle permeability in Locusta migratoria

    Insect Molecular Biology 29:555–568.

    https://doi.org/10.1111/imb.12665

    • PubMed
    • Google Scholar

Author details

1. Weizhao Sun

   Institute for Evolution & Biodiversity, University of Münster, Hüfferstr, Münster, Germany

   Contribution

   Data curation, Formal analysis, Validation, Investigation, Visualization, Methodology, Writing – original draft, Writing – review and editing

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0003-2781-1228

2. Michelle Ina Lange

   Institute for Evolution & Biodiversity, University of Münster, Hüfferstr, Münster, Germany

   Contribution

   Investigation

   Competing interests

   No competing interests declared

3. Jürgen Gadau

   Institute for Evolution & Biodiversity, University of Münster, Hüfferstr, Münster, Germany

   Contribution

   Resources, Supervision, Project administration, Writing – review and editing

   Competing interests

   No competing interests declared

4. Jan Buellesbach

   Institute for Evolution & Biodiversity, University of Münster, Hüfferstr, Münster, Germany

   Contribution

   Conceptualization, Resources, Formal analysis, Supervision, Funding acquisition, Validation, Visualization, Writing – original draft, Project administration, Writing – review and editing

   For correspondence

   buellesb@uni-muenster.de

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0001-8493-692X

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