Skip to main content
Log in

Is the copulation of silver-washed fritillary inverted? Function of the female genital projection (Lepidoptera: Nymphalidae: Argynnini)

  • Published:
Organisms Diversity & Evolution Aims and scope Submit manuscript


Females of the silver-washed fritillary butterfly, Argynnis paphia (Linnaeus), and its relatives are known to have a unique genital projection called the cornucopia. Previous observations showed that the cornucopia partly penetrates the male genitalia during copulation, but its detailed structures and functions are unknown to date. Our observations using light microscopy, confocal laser scanning microscopy, scanning electron microscopy, and microcomputed tomography suggested that penetration by the female cornucopia occurs passively, and it is hooked and pulled by the male towards the male genitalia using sharp teeth on the uncus. This hooking and pulling causes wounds on the dorsal region of the cornucopia. Artificial amputation of a cornucopia confirmed that it is indispensable for spermatophore transfer from males and functions to break off the male phallic spines (cornuti). The broken spines may be used by females to help the digestion of spermatophores in the female’s corpus bursae. Females with larger body sizes had more broken spines in the corpus bursae, indicating that secure holding of a large female during copulation is challenging for males.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Data availability

The statistical data generated or analyzed during this study are included in the online supplementary files. Scanned µCT images (zip file) are available from Figshare at 10.6084/m9.figshare.21992420.


  • Anzaldo, S., Domobroskie, J., & Brown, J. W. (2014). Morphological variation, taxonomic distribution, and phylogenetic significance of cornuti in Tortricinae (Lepidoptera: Tortricidae). Proceedings of the Entomological Society of Washington, 116, 1–31.

    Article  Google Scholar 

  • Arnold, R. A., & Fischer, R. L. (1977). Operational mechanisms of copulation and oviposition in Speyeria (Lepidoptera: Nymphalidae). Annals of the entomological Society of America, 70, 455–468.

    Article  Google Scholar 

  • Bartoń, K. (2022). MuMIn: multi-model inference. R package ver., 1(47), 1.

    Google Scholar 

  • Bayard, A. (1944). Observations sur l’accouplement de Dryas paphia (L.) [Lep, Nymphalidae]. Bulletin de la Société entomologique de France, 42, 92–95.

    Article  Google Scholar 

  • Burnham, K. P., & Anderson, D. R. (2002). Model selection and multimodel inference: a practical information-theoretic approach (2nd ed.). Springer-Verlag.

    Google Scholar 

  • Camacho-Garcia, J., Baixeras, J., & Cordero, C. (2018). Males of the tortricid moth Amorbia cuneana (Walsingham, 1879) shed their genital spines inside the female during copulation. Zoologischer Anzeiger, 277, 197–202.

    Article  Google Scholar 

  • Cheng, Z., & Yoshizawa, K. (2019). Functional morphology of Trichadenotecnum male and female genitalia analyzed using µCT (Insecta: Psocodea: Psocomorpha). Journal of Morphology, 280, 555–567.

    Article  PubMed  Google Scholar 

  • Cordero, C. (2005). The evolutionary origin of signa in female Lepidoptera: Natural and sexual selection hypotheses. Journal of Theoretical Biology, 232, 443–449.

  • Cordero, C. (2010). On the function of cornuti, sclerotized structures of the endophallus of Lepidoptera. Genetica, 138, 27–35.

    Article  PubMed  Google Scholar 

  • Cordero, C., & Miller, J. S. (2012). On the evolution and function of caltrop cornuti in Lepidoptera – potentially damaging male genital structures transferred to females duringause copulatory wound copulation. Journal of Natural history, 46, 701–715.

    Article  Google Scholar 

  • Crawley, M. J. (2012). The R book. Wiley & Sons.

    Book  Google Scholar 

  • Dos Passos, C. F., & Grey, L. P. (1945). A genitalic survey of Argynninae (Lepidoptera, Nymphalidae). American Museum of Natural History, 1296, 1–29.

    Google Scholar 

  • Eberhard, W. G. (1985). Sexual selection and animal genitalia. Harvard University Press.

    Book  Google Scholar 

  • Hannemann, H. J. (1954a). Zur funktionellen anatomie des männlichen kopulationsapparates von Argynis paphia (L.). Zoologischer Anzeiger, 152, 266–274.

  • Hannemann, H. J. (1954b). Zur muskelfunktion der weiblichen genitalsegmente von Argynnis paphia (L.) (Lep.). Zoologischer Anzeiger, 153, 149–154.

  • Jolivet, P. (2008). Inverted Copulation. Encyclopedia of Entomology, 55, 2041–2044.

    Google Scholar 

  • Kamimura, Y., & Mitsumoto, H. (2011). Comparative copulation anatomy of the Drosophila melanogaster species complex (Diptera: Drosophilidae). Entomological Science., 14, 399–410.

    Article  Google Scholar 

  • Klier, E. (1956). Zur Konstructionsmorphologie des männlichen Geschlechtsapparates der Psocopteren. Zoologische Jahbücher (Anatomie)., 75, 207–286.

    Google Scholar 

  • Klots, A. B. (1970). Lepidoptera. In S. L. Tuxen (Ed.), Taxonomist’s glossary of genitalia in insects (pp. 115–130). Munksgaard.

    Google Scholar 

  • Lienhard, C. (2007). Descritption of a new African genus and a new tribe of Speleketorinae (Pscodea: ‘Psocoptera’: Prionoglarididae). Revue Suisse de Zoologie, 114, 441–469.

    Article  Google Scholar 

  • Lienhard, C., Do Carmo, T. O., & Ferreira, R. L. (2010). A new genus of Sensitibillini from Brazilian caves (Psocodea: ‘Psocoptera’: Prionoglarididae). Revue Suisse de Zoologie, 117, 611–635.

    Article  Google Scholar 

  • Michels, J., & Gorb, S. N. (2011). Detailed three-dimensional visualization of resilin in the exoskeleton of arthropods using confocal laser scanning microscopy. Journals of Microscopy., 245, 1–16.

    Google Scholar 

  • Michels, J., Gorb, S. N., & Reinhardt, K. (2015). Reduction of female copulatory damage by resilin represents evidence for tolerance in sexual conflict. Journal of the Royal Society Interface, 12, 20141107.

    Article  PubMed  PubMed Central  Google Scholar 

  • Ockenfels, P., Boppré, M., Fischer, O. W. & Schultz, S. (1998). Chemical communication in the silver-washed fritillary, Argynnis paphia (Lepidoptera, Nymphalidae). Poster from 15th Annual Meeting of the International Society of Chemical Ecology. Available via

  • R Core Team. (2022). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria.

  • Sánchez, V., & Cordero, C. (2014). Sexual coevolution of spermatophore envelopes and female genital traits in butterflies: Evidence of male coercion? PeerJ, 2, e247.

  • Simonsen, T. J. (2006a). Fritillary phylogeny, classification, and larval host plants: reconstructed mainly on the basis of male and female genitalic morphology (Lepidoptera: Nymphalidae: Argynnini). Biological Journal of the Linnean Society, 89, 627–673.

  • Simonsen, T. J. (2006b). Glands, muscles and genitalia. Morphological and phylogenetic implications of histological characters in the male genitalia of Fritillary butterflies (Lepidoptera: Nymphalidae: Argynnini). Zoologica Scripta, 35, 231–241.

  • Simonsen, T. J. (2008). Phylogeny of the cactus-feeding phycitines and their relatives (Lepidoptera, Pyralidae) based on adult morphology: Evaluation of adult character systems in phyctine systematics and evidence for a single origin of Cactaceae-feeding larvae. Insect Systematics & Evolution, 39, 303–325.

    Article  Google Scholar 

  • Sloan, N. S., & Simmons, L. W. (2019). The evolution of female genitalia. Journal of Evolutionary Biology, 32, 882–899.

    Article  PubMed  Google Scholar 

  • Uesugi, K., Hoshino, M., Takeuchi, A., Suzuki, Y., & Yagi, N. (2012). Development of fast and high throughput tomography using CMOS image detector at SPring-8. Developments in X-Ray tomography VII, I, 850601.

    Google Scholar 

  • Uesugi, K., Hoshino, M., & Takeuchi, A. (2017). Introducing high efficiency image detector to X-ray imaging tomography. Journal of Physics: Conference Series, 849, 012051.

    Google Scholar 

  • Wedell, N. (2005). Female receptivity in butterflies and moths. Journal of Experimental Biology, 208, 3433–3440.

    Article  PubMed  Google Scholar 

  • Yoshizawa, K., Ferreira, R. L., Kamimura, Y., & Lienhard, C. (2014). Female penis, male vagina, and their correlated evolution in a cave insect. Current Biology, 24, 1006–1010.

    Article  CAS  PubMed  Google Scholar 

  • Yoshizawa, K., Ferreira, R. L., Lienhard, C., & Kamimura, Y. (2019). Why did a female penis evolve in a small group of cave insects. BioEssays, 41, 1900005.

    Article  Google Scholar 

  • Yushkevich, P. A., Piven, J., Hazlett, H. C., Smith, R. G., Ho, S., Gee, J. C., & Gerig, G. (2006). User-guided 3D active contour segmentation of anatomical structures: Significantly improved efficiency and reliability. NeuroImage, 31, 1116–1128.

    Article  PubMed  Google Scholar 

Download references


We thank Toyohei Saigusa and Takashi Adachi‑Yamada for valuable advice about rearing and collection of the butterflies, and Akihisa Takeuchi and Masayuki Uesugi for their support at SPring-8. The research at SPring-8 was approved through project number 2022A1201 to KY. We also thank Thomas J. Simonsen and an anonymous reviewer for their constructive comments. This study was partly supported by JSPS grant numbers 15H04409 and 19H03278 to KY and 15K07133 to YK. This study was conducted to fulfill the master's degree of TM, who thanks Shin-ichi Akimoto and Masahiro Ohara for their continuous guidance.

Author information

Authors and Affiliations



YK and KY conceived the project. TM designed the experiments, collected specimens, analyzed morphology, and performed experiments. KY performed µCT analysis. YK performed SEM imaging and statistical analyses. All authors wrote and approved the final manuscript.

Corresponding author

Correspondence to Kazunori Yoshizawa.

Ethics declarations

Competing interest

All authors approved the final version of the manuscript. The authors declare no competing interests.

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 file1 (PDF 119 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Matoba, T., Kamimura, Y. & Yoshizawa, K. Is the copulation of silver-washed fritillary inverted? Function of the female genital projection (Lepidoptera: Nymphalidae: Argynnini). Org Divers Evol 23, 871–879 (2023).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: