Abstract
Hymenopteran egg parasitoids of the genus Trichogramma are among the tiniest insects on Earth (< 0.5 mm in size, 8 µg) and are important biological control agents of moth pests. The existence of sex pheromones has been demonstrated in Trichogramma species, but none has yet been fully characterized. Here, we report absolute configuration, improved synthesis and behavioral activity of the sex pheromone of T. turkestanica, making it one of the smallest insects with a fully characterized sex pheromone. Enantioselective synthesis and chiral gas chromatography were used to determine the absolute configuration of the previously identified, stereochemically complex dienol (2E,4E,6S,8S,10S)-4,6,8,10-tetramethyltrideca-2,4-dien-1-ol and to newly establish the same stereochemistry for the corresponding diene (2E,4E,6S,8S,10S)-4,6,8,10-tetramethyltrideca-2,4-diene. In a 3:1 mixture of dienol and diene applied to two solvent-washed wasps (dummies), these compounds triggered attraction, arrestment, and courtship (“casting”) in males at doses, as little as 670 attograms per dummy. This pheromone could be studied to monitor T. turkestanica field populations, with the aim to improve the efficiency of biocontrol strategies.
Data availability
All data, including raw data underlying the figures, can be found in the Supplementary Information.
References
Li, Y. L. in Biological Control with Egg Parasitoids (eds Wajnberg, E. & Hassan, S. A.) 37–53 (CAB International, 1994).
Schöller, M., Prozell, S., Suma, P. & Russo, A. in Recent Advances in Stored Product Protection (eds Athanassiou, C. G. & Arthur, F. H.) 183–209 (Springer, 2018).
Cherif, A., Mansour, R. & Grissa-Lebdi, K. The egg parasitoids Trichogramma: From laboratory mass rearing to biological control of lepidopteran pests. Biocontrol Sci. Technol. 31, 661–693 (2021).
Stouthamer, R., Hu, J., van Kan, F. J. P. M., Platner, G. R. & Pinto, J. D. The utility of internally transcribed spacer 2 DNA sequences of the nuclear ribosomal gene for distinguishing sibling species of Trichogramma. BioControl 43, 421–440 (1999).
Greenberg, L. O., Huigens, M. E., Groot, A. T., Cusumano, A. & Fatouros, N. E. Finding an egg in a haystack: Variation in chemical cue use by egg parasitoids of herbivorous insects. Curr. Opin. Insect Sci. 55, 101002, 1–8 (2023).
Ruther, J. in Chemical Ecology of Insect Parasitoids (eds E. Wajnberg & S. Colazza) Ch. 6, pp. 112–143 (Wiley-Blackwell, 2013).
Hase, A. Beiträge zur Lebensgeschichte der Schlupfwespe Trichogramma evanescens Westwood. Arbeiten Biol. Reichsanst Land- Forstw. 14, 171–224 (1925).
Al-Jalely, B. H. & Xu, W. Olfactory sensilla and olfactory genes in the parasitoid wasp Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae). Insects 12, 998, (2021).
Pintureau, B. & Toonders, T. Quelques résultats concernant l’étude de l’attraction des mâles par les femelles vierges chez Trichogramma maidis (Hym. Trichogrammatidae). Bull. Mensuel Soc. Linnéenne Lyon. 52, 81–87 (1983).
Pompanon, F., de Schepper, B., Mourer, Y., Fouillet, P. & Bouletreau, M. Evidence for a substrate-borne sex pheromone in the parasitoid wasp Trichogramma brassicae. J. Chem. Ecol. 23, 1349–1360 (1997).
Al Dabel, F. Etude des Comportements et des Molécules Impliqués dans la Rencontre des Sexes chez les Parasitoïdes du genre Trichogramma spp. PhD thesis (Université de Paris-Sud, 2004).
Wang, D. et al. Mate choice and host discrimination behavior of the parasitoid Trichogramma chilonis. Bull. Entomol. Res. 106, 530–537 (2016).
Silva, I. M. M. S. Identification and Evaluation of Trichogramma Parasitoids for Biological Pest Control PhD thesis thesis, (Wageningen University, 1999).
van Beek, T. A., Silva, I. M. M. S., Posthumus, M. A. & Melo, R. Partial elucidation of Trichogramma putative sex pheromone at trace levels by solid-phase microextraction and gas chromatography-mass spectrometry studies. J. Chromatogr. A. 1067, 311–321 (2005).
Tröger, A. et al. Structure elucidation of female-specific volatiles released by the parasitoid wasp Trichogramma turkestanica (Hymenoptera: Trichogrammatidae). Beilstein J. Org. Chem. 10, 767–773 (2014).
Geerdink, D., Buter, J., van Beek, T. A. & Minnaard, A. J. Asymmetric total synthesis of a putative sex pheromone component from the parasitoid wasp Trichogramma turkestanica. Beilstein J. Org. Chem. 10, 761–766 (2014).
Francke, W. & Schulz, S. in Comprehensive Natural Products II – Chemistry and Biology (eds L. Mander, H.-W. Lui, & K. Mori) Vol. 4, pp. 153–223 (Elsevier, 2010).
Ando, T. & Yamakawa, R. Chiral methyl-branched pheromones. Nat. Prod. Rep. 32, 1007–1041 (2015).
Cosse, A. A. et al. Female-produced sex pheromone of Tetrastichus planipennisi, a parasitoid introduced for biological control of the invasive emerald ash borer, Agrilus planipennis. J. Chem. Ecol. 46, 508–519 (2020).
Schmidt, Y. et al. Enantioselective total synthesis of the unnatural and the natural stereoisomers of vittatalactone. J. Org. Chem. 75, 4424–4433 (2010).
Lin, Z. et al. Asymmetric total synthesis and structural revision of DAT2, an antigenic glycolipid from Mycobacterium tuberculosis. Angew. Chem. Int. Ed. 63, e202318582 (2024).
Souza, J. P. A., Bandeira, P. T., Bergmann, J. & Zarbin, P. H. G. Recent advances in the synthesis of insect pheromones: An overview from 2013 to 2022. Nat. Prod. Rep. 40, 866–889 (2023).
Morozova, V., Moriya, K., Karaghiosoff, K. & Knochel, P. Preparation of optically enriched secondary alkyllithium and alkylcopper reagents—Synthesis of (–)-lardolure and siphonarienal. Angew. Chem. Int. Ed. 57, 5516–5519 (2018).
Ruther, J., Homann, M. & Steidle, J. L. M. Female-derived sex pheromone mediates courtship behaviour in the parasitoid Lariophagus distinguendus. Entomol. Exp. Appl. 96, 265–274 (2000).
Kennedy, J. S. Zigzagging and casting as a programmed response to wind-borne odor – A review. Physiol. Entomol. 8, 109–120 (1983).
Steiner, S., Hermann, N. & Ruther, J. Characterization of a female-produced courtship pheromone in the parasitoid Nasonia vitripennis. J. Chem. Ecol. 32, 1687–1702 (2006).
Ruther, J., Döring, M. & Steiner, S. Cuticular hydrocarbons as contact sex pheromone in the parasitoid Dibrachys cavus. Entomol. Exp. Appl. 140, 59–68 (2011).
Würf, J., Pokorny, T., Wittbrodt, J., Millar, J. G. & Ruther, J. Cuticular hydrocarbons as contact sex pheromone in the parasitoid wasp Urolepis rufipes. Front. Ecol. Evol. 8, 180, (2020).
Jungwirth, S., Ruther, J. & Pokorny, T. Similar is not the same – mate recognition in a parasitoid wasp. Front. Ecol. Evol. 9, 64667, (2021).
Kühbandner, S., Sperling, S., Mori, K. & Ruther, J. Deciphering the signature of cuticular lipids with contact sex pheromone function in a parasitic wasp. J. Exp. Biol. 215, 2471–2478 (2012).
Hrabar, M. et al. (7E,11E)-3,5,9,11-Tetramethyltridecadienal: Sex pheromone of the Strepsipteran Xenos peckii. J. Chem. Ecol. 41, 732–739 (2015).
Lagoutte, R. et al. Total synthesis, proof of absolute configuration, and biosynthetic origin of Stylopsal, the first isolated sex pheromone of strepsiptera. Chem. Eur. J. 19, 8515–8524 (2013).
Park, S. C., Wi, A. J. & Kim, H. S. Response of Matsucoccus thunbergianae males to synthetic sex pheromone and its utilization for monitoring the spread of infestation. J. Chem. Ecol. 20, 2185–2196 (1994).
Yang, C. Y., Mori, K., Kim, J. & Kwon, K. B. Identification and field bioassays of the sex pheromone of Eurytoma maslovskii (Hymenoptera: Eurytomidae). Sci. Rep. 10, 10281, 1–8 (2020).
Kaissling, K. E. in Olfactory Concepts of Insect Control – Alternative to Insecticides (eds Picimbon, J. F.) Vol. 2, pp. 1–27 (Springer International Publishing, 2019).
Fatouros, N. E. & Huigens, M. E. Phoresy in the field: Natural occurrence of Trichogramma egg parasitoids on butterflies and moths. Biocontrol 57, 493–502 (2012).
Hammer, Ø., Harper, D. A. & Ryan, P. D. PAST: Paleontological statistics software package for education and data analysis. Palaeontol. Electron. 4, 9 (2001).
Faul, F., Erdfelder, E., Lang, A. G. & Buchner, A. G*Power 3: A flexible statistical power analysis program for the social, behavioral, and biomedical sciences. Behav. Res. Meth. 39, 175–191 (2007).
Holm, S. A simple sequentially rejective multiple test procedure. Scand. J. Stat. 6, 65–70 (1979).
Acknowledgements
We wish to thank Frank Claassen and Elbert van der Klift for technical support and Birgit Lautenschläger for producing the SEM images.
Funding
No funding reported.
Author information
Authors and Affiliations
Contributions
T.A.v.B, J.J.J.A.v.L., W.F., A.J.M and J.R. designed research; T.A.v.B, J.P.K., A.D., H.M.S., M.H., K.C. and H.B. performed research; T.A.v.B, J.P.K., A.D., M.H., K.C., H.B., W.F., A.J.M and J.R. analyzed data; T.A.v.B., A.J.M., and J.R. wrote the paper with input from all other authors.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Reporting summary
Further information on research design is available in the Nature Research Reporting Summary linked to this article.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Supplementary Material 1 (download XLSX )
Supplementary Material 2
Supplementary Material 3 (download DOCX )
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
Reprints and permissions
About this article
Cite this article
van Beek, T.A., Kaniraj, J.P., Dornbusch, A. et al. Absolute configuration, improved synthesis and femtogram-level behavioral activity of the sex pheromone of the minute parasitoid wasp Trichogramma turkestanica.
Sci Rep (2026). https://doi.org/10.1038/s41598-026-46414-z
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-46414-z
Supplementary Material 2
Source: Ecology - nature.com
