Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

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


Sex pheromones play a pivotal role in the communication of many sexually reproducing organisms1. Accordingly, speciation is often accompanied by pheromone diversification enabling proper mate finding and recognition2. Current theory implies that chemical signals are under stabilizing selection by the receivers who thereby maintain the integrity of the signals3. How the tremendous diversity of sex pheromones seen today evolved is poorly understood4,5. Here we unravel the genetics of a newly evolved pheromone phenotype in wasps and present results from behavioural experiments indicating how the evolution of a new pheromone component occurred in an established sender–receiver system. We show that male Nasonia vitripennis evolved an additional pheromone compound differing only in its stereochemistry from a pre-existing one. Comparative behavioural studies show that conspecific females responded neutrally to the new pheromone phenotype when it evolved. Genetic mapping and gene knockdown show that a cluster of three closely linked genes accounts for the ability to produce this new pheromone phenotype. Our data suggest that new pheromone compounds can persist in a sender’s population, without being selected against by the receiver and without the receiver having a pre-existing preference for the new pheromone phenotype, by initially remaining unperceived. Our results thus contribute valuable new insights into the evolutionary mechanisms underlying the diversification of sex pheromones. Furthermore, they indicate that the genetic basis of new pheromone compounds can be simple, allowing them to persist long enough in a population for receivers to evolve chemosensory adaptations for their exploitation.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Prices vary by article type



Prices may be subject to local taxes which are calculated during checkout

Figure 1: Evolution of sex pheromone diversity and behavioural response in Nasonia parasitoid wasps.
Figure 2: Genetics of sex pheromone differences between N. vitripennis and N. giraulti males.
Figure 3: Phylogeny and dsRNA-mediated knockdown of candidate genes.

Accession codes

Primary accessions


Data deposits

The sequences reported in this article are deposited in GenBank under accession numbers FN429934FN429952, FN430419 and HE962018HE962021.


  1. Wyatt, T. D. Pheromones and Animal Behaviour Ch. 3 37–73 (Cambridge Univ. Press, 2003)

    Book  Google Scholar 

  2. Smadja, C. & Butlin, R. K. On the scent of speciation: the chemosensory system and its role in premating isolation. Heredity 102, 77–97 (2009)

    Article  CAS  Google Scholar 

  3. Butlin, R. K. & Trickett, A. J. in Insect Pheromone Research: New Directions (eds Cardé, R. T. & Minks, A. K. ) 548–562 (Chapman & Hall, 1997)

    Book  Google Scholar 

  4. Symonds, M. R. E. & Elgar, M. A. The evolution of pheromone diversity. Trends Ecol. Evol. 23, 220–228 (2008)

    Article  Google Scholar 

  5. Steiger, S., Schmitt, T. & Schaefer, H. M. The origin and dynamic evolution of chemical information transfer. Proc. R. Soc. B 278, 970–979 (2011)

    Article  Google Scholar 

  6. Roelofs, W. L. & Rooney, A. P. Molecular genetics and evolution of pheromone biosynthesis in Lepidoptera. Proc. Natl Acad. Sci. USA 100, 9179–9184 (2003)

    Article  CAS  ADS  Google Scholar 

  7. Ferveur, J.-F. Cuticular hydrocarbons: their evolution and roles in Drosophila pheromonal communication. Behav. Genet. 35, 279–295 (2005)

    Article  Google Scholar 

  8. Xue, B., Rooney, A. P., Kajikawa, M., Okada, N. & Roelofs, W. L. Novel sex pheromone desaturases in the genomes of corn borers generated through gene duplication and retroposon fusion. Proc. Natl Acad. Sci. USA 104, 4467–4472 (2007)

    Article  CAS  ADS  Google Scholar 

  9. Shirangi, T. R., Dufour, H. D., Williams, T. M. & Carroll, S. B. Rapid evolution of sex pheromone-producing enzyme expression in Drosophila . PLoS Biol. 7, e1000168 (2009)

    Article  Google Scholar 

  10. Lassance, J.-M. et al. Allelic variation in a fatty-acyl reductase gene causes divergence in moth sex pheromones. Nature 466, 486–489 (2010)

    Article  CAS  ADS  Google Scholar 

  11. Liénard, M. A., Hagström, A. K., Lassance, J.-M. & Löfstedt, C. Evolution of multicomponent pheromone signals in small ermine moths involves a single fatty-acyl reductase gene. Proc. Natl Acad. Sci. USA 107, 10955–10960 (2010)

    Article  ADS  Google Scholar 

  12. Albre, J. et al. Sex pheromone evolution is associated with differential regulation of the same desaturase gene in two genera of leafroller moths. PLoS Genet. 8, e1002489 (2012)

    Article  CAS  Google Scholar 

  13. Werren, J. H. & Loehlin, D. W. The parasitoid wasp Nasonia: an emerging model system with haploid male genetics. Cold Spring Harb. Protocols 2009, pdb.emo134 (2009)

    Google Scholar 

  14. Werren, J. H. et al. Functional and evolutionary insights from the genomes of three parasitoid Nasonia species. Science 327, 343–348 (2010)

    Article  CAS  Google Scholar 

  15. Ruther, J., Stahl, L. M., Steiner, S., Garbe, L. A. & Tolasch, T. A male sex pheromone in a parasitic wasp and control of the behavioral response by the female’s mating status. J. Exp. Biol. 210, 2163–2169 (2007)

    Article  CAS  Google Scholar 

  16. Ruther, J., Steiner, S. & Garbe, L.-A. 4-Methylquinazoline is a minor component of the male sex pheromone in Nasonia vitripennis . J. Chem. Ecol. 34, 99–102 (2008)

    Article  CAS  Google Scholar 

  17. Abdel-latief, M., Garbe, L. A., Koch, M. & Ruther, J. An epoxide hydrolase involved in the biosynthes is of an insect sex attractant and its use to localize the production site. Proc. Natl Acad. Sci. USA 105, 8914–8919 (2008)

    Article  CAS  ADS  Google Scholar 

  18. Steiner, S. & Ruther, J. Mechanism and behavioral context of male sex pheromone release in Nasonia vitripennis . J. Chem. Ecol. 35, 416–421 (2009)

    Article  CAS  Google Scholar 

  19. Munoz-Torres, M. C. et al. Hymenoptera Genome Database: integrated community resources for insect species of the order Hymenoptera. Nucleic Acids Res. 39, D658–D662 (2011)

    Article  CAS  Google Scholar 

  20. Niehuis, O. et al. Recombination and its impact on the genome of the haplodiploid parasitoid wasp Nasonia . PLoS ONE 5, e8597 (2010)

    Article  ADS  Google Scholar 

  21. Punta, M. et al. The Pfam protein families database. Nucleic Acids Res. 40, D290–D301 (2012)

    Article  CAS  Google Scholar 

  22. Tanner, M. E. Understanding nature’s strategies for enzyme-catalyzed racemization and epimerization. Acc. Chem. Res. 35, 237–246 (2002)

    Article  CAS  Google Scholar 

  23. Kavanagh, K. L., Jörnvall, H., Persson, B. & Oppermann, U. Medium- and short-chain dehydrogenase/reductase gene and protein families: the SDR superfamily: functional and structural diversity within a family of metabolic and regulatory enzymes. Cell. Mol. Life Sci. 65, 3895–3906 (2008)

    Article  CAS  Google Scholar 

  24. Cho, H., Oliveira, M. A. & Tai, H.-H. Critical residues for the coenzyme specificity of NAD+-dependent 15-hydroxyprostaglandin dehydrogenase. Arch. Biochem. Biophys. 419, 139–146 (2003)

    Article  CAS  Google Scholar 

  25. Niehuis, O., Judson, A. K. & Gadau, J. Cytonuclear genic incompatibilities cause increased mortality in male F2 hybrids of Nasonia giraulti and N. vitripennis . Genetics 178, 413–426 (2008)

    Article  CAS  Google Scholar 

  26. Broman, K. W., Wu, H., Sen, S. & Churchill, G. A. R/qtl: QTL mapping in experimental crosses. Bioinformatics 19, 889–890 (2003)

    Article  CAS  Google Scholar 

  27. Niehuis, O., Büllesbach, J., Judson, A. K., Schmitt, T. & Gadau, J. Genetics of cuticular hydrocarbon differences between males of the parasitoid wasps Nasonia giraulti and Nasonia vitripennis . Heredity 107, 61–70 (2011)

    Article  CAS  Google Scholar 

  28. Lynch, J. A. & Desplan, C. A method for parental RNA interference in the wasp Nasonia vitripennis . Nature Protocols 1, 486–494 (2006)

    Article  CAS  Google Scholar 

  29. Tamura, K. et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. 28, 2731–2739 (2011)

    Article  CAS  Google Scholar 

Download references


We thank J. H. Werren for providing laboratory strains of Nasonia and Trichomalopsis and D. Wheeler for discussion on the gene expression data. G. Amdam and J. Liebig allowed us to use their research facilities for conducting the gene expression studies, knockdown experiments and gas chromatography analyses. We thank D. D. McKenna and R. S. Peters for comments on an earlier draft of this paper. O.N. acknowledges the Alexander von Humboldt foundation for a Feodor Lynen postdoctoral research stipend. J.B. and T.S. were supported by the Excellence Initiative of the German Research Foundation (GSC-4, Spemann Graduate School). Parts of this research were supported by the German Research Foundation (DFG) grant RU 717/10-1 to J.R.

Author information

Authors and Affiliations



Authors J.B., J.D.G., J.R. and O.N. contributed equally to this work. T.S. initiated the study. J.B., J.G., J.D.G., J.R., O.N. and T.S. conceived the experiments. D.P. conducted the bioassays. C.H., J.B., J.R. and T.S. conducted the chemical analyses. A.K.J., C.H., J.B., J.D.G., J.G., N.S.M. and O.N. performed the QTL analyses and knockdown experiments. J.D.G. and O.N. performed the transcriptional analyses. J.G., J.R., O.N. and T.S. provided material and resources. O.N. was responsible for the comparative sequence analysis and took the lead in writing the manuscript. J.R., O.N. and T.S. were the main contributors to the writing of the manuscript.

Corresponding author

Correspondence to Oliver Niehuis.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Methods (Species and strains, Comparative chemical analysis, Preparation of enantiopure HDL stereoisomers, Behavioural bioassays, QTL and fine mapping, Introgression experiments, Gene knockdown experiments, Gene expression analysis, Rapid Amplification of cDNA ends (RACE), Comparative nucleotide and amino acid sequence analysis), Supplementary Data (Introgression experiments, Possible functions of NV10124, NV10126, NV10130 and NV30591, Annotation of putative SDR-coding genes in candidate region, Comparative nucleotide and amino acid sequence analysis), Supplementary References, Supplementary Tables 1–5 and Supplementary Figures 1–5. (PDF 452 kb)

Supplementary Data

This file contains additional Supplementary Data showing the detailed annotation of the putative SDR-coding candidate genes NV10127, NV10128 and NV10129. The annotations are provided in a file as plain text in generic feature format (.gff). (TXT 3 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Niehuis, O., Buellesbach, J., Gibson, J. et al. Behavioural and genetic analyses of Nasonia shed light on the evolution of sex pheromones. Nature 494, 345–348 (2013).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing