Molecular evidence for the taxonomic resurrection of Stenurella sennii nom. res. within the phylogeographic framework of S. melanura (Coleoptera, Cerambycidae)

S. Melnyk; V. Malaniy; Y.-Y. Kasicki; O. Piddubnyi; O. Pozniak; L. Felbaba-Klushyna; V. Shparyk; O. Zinenko; A. Zamoroka

doi:10.15421/012602

S. Melnyk Vasyl Stefanyk Carpathian National University
V. Malaniy Vasyl Stefanyk Carpathian National University
Y.-Y. Kasicki Vasyl Stefanyk Carpathian National University
O. Piddubnyi Vasyl Stefanyk Carpathian National University
O. Pozniak Vasyl Stefanyk Carpathian National University
L. Felbaba-Klushyna Uzhhorod National University
V. Shparyk Vasyl Stefanyk Carpathian National University
O. Zinenko V. N. Karazin Kharkiv National University
A. Zamoroka Vasyl Stefanyk Carpathian National University

Keywords: biodiversity, saproxylic beetles, cryptic species, DNA barcoding, COI, phenotypic polymorphism, hybridization, speciation, genetic divergence, integrative taxonomy.

Abstract

We utilized DNA barcoding to analyze 241 mitochondrial COI sequences of Stenurella melanura across its European range, establishing a framework for understanding its intraspecific genetic variation with four genetically distinct haplogroups ( Sm1–Sm4 ). The two most widespread lineages, Sm1 and Sm2 , show a strong association with dorsal pube s cence coloration, particularly of the elytra: Sm1 is dominated by light-pubescent individuals, whereas Sm2 predominantly comprises dark-pubescent forms. Instances of discordance between haplogroup and pubescence coloration are best e x plained by ongoing hybridization, with geographically structured distributions across Europe, reflecting complex postgl a cial biogeography. Phylogenetic analysis revealed two highly divergent sequences forming a distinct clade , separated from S. melanura by a mean genetic distance of 0.077 substitutions per site. Morphological examination confirmed their co n formity with the original description of Stenurella sennii nom. res. , validating the taxonomic restoration of this neglected species. As S. sennii nom. res. is a cryptic sibling species morphologically indistinguishable from light-pubescent Sm1 haplogroup of S. melanura , traditional identification fails to capture this diversity. Our pan-European phylogeographic framework resolves this confusion, demonstrating that molecular diagnostics are essential for accurate species delimitation and for maintaining the integrity of taxonomic inventories.

References

Çakmak, Y. E., Soydabaş-Ayoub, H. K., & Uçkan, F. (2020). A preliminary phylogenetic analysis of ribbed pine borer (Rhagium inquisitor) based on mitochondrial COI sequences. Journal of Asia-Pacific Entomology, 23(3), 809–815.

Crowley, L. M., Poloni, R., Broad, G. R., Boyes, D., & Darwin Tree of Life Consortium (2024). The genome sequence of a longhorn beetle, Stenurella melanura (Linnaeus, 1758). Wellcome Open Research, 9, 414.

Gascuel, O. (1997). BIONJ: An improved version of the NJ algorithm based on a simple model of sequence data. Molecular Biology and Evolution, 14(7), 685–695.

Gouy, M., Tannier, E., Comte, N., & Parsons, D. P. (2021). SeaView version 5: A multiplatform software for multiple sequence alignment, molecular phylogenetic analyses, and tree reconciliation. Methods in Molecular Biology, 2231, 241–260.

Guindon, S., Dufayard, J.-F., Lefort, V., Anisimova, M., Hordijk, W., & Gascuel, O. (2010). New algorithms and methods to estimate maximum-likelihood phylogenies: Assessing the performance of PhyML 3.0. Systematic Biology, 59(3), 307–321.

Korlević, P., McAlister, E., Mayho, M., Makunin, A., Flicek, P., & Lawniczak, M. K. N. (2021). A minimally morphologically destructive approach for DNA retrieval and whole-genome shotgun sequencing of pinned historic dipteran vector species. Genome Biology and Evolution, 13(10), evab226.

Löbl, I., & Smetana, A. (Eds.). (2010). Catalogue of Palaearctic Coleoptera. Vol. 6: Chrysomeloidea. Stenstrup, Apollo Books.

Makunin, A., Korlević, P., Park, N., Goodwin, S., Waterhouse, R. M., von Wyschetzki, K., Jacob, C. G., Davies, R., Kwiatkowski, D., St. Laurent, B., Ayala, D., & Lawniczak, M. K. N. (2022). A targeted amplicon sequencing panel to simultaneously identify mosquito species and Plasmodium presence across the entire Anopheles genus. Molecular Ecology Resources, 22(1), 28–44.

Özdikmen, H., Mercan, N., & Cihan, N. (2012). A new species of the genus Stenurella Villiers, 1974 from Turkey (Coleoptera: Cerambycidae: Lepturinae). Munis Entomology and Zoology, 7(1), 18–21.

Rapuzzi, P., & Sama, G. (2009). Descriptions of new Cerambycidae from Greece, Turkey, Northern Syria and Southern Morocco (Coleoptera: Cerambycidae). Munis Entomology and Zoology, 4(1), 181–192.

Rapuzzi, P., Konvička, O., & Vít, D. (2011). The longhorn beetle Stenurella sennii (Coleoptera: Cerambycidae) in the Czech Republic and Croatia. Klapalekiana, 47, 237–238.

Sama, G. (2002). Atlas of the Cerambycidae of Europe and the Mediterranean Area. British Isles and Continental Europe from France (excl. Corsica) to Scandinavia and Urals. Vol. 1. Zlín, Nakladatelství Kabourek.

Sama, G., & Rapuzzi, P. (2011). Una nuova checklist dei Cerambycidae d’Italia (Insecta: Coleoptera: Cerambycidae). Quaderno di Studi e Notizie di Storia Naturale della Romagna, 32, 121–164.

Semaniuk, D. V., & Zamoroka, A. M. (2020). Preliminary phylogenetic analysis of Lepturini (Insecta: Coleoptera: Cerambycidae). In: Proceedings of the XVI International Scientific Conference for Students and PhD Students “Youth and Progress of Biology”. Lviv. P. 121.

Sire, L., Gey, D., Debruyne, R., Noblecourt, T., Soldati, F., Barnouin, T., Parmain, G., Bouget, C., Lopez-Vaamonde, C., & Rougerie, R. (2019). The challenge of DNA barcoding saproxylic beetles in natural history collections: Exploring the potential of parallel multiplex sequencing with Illumina MiSeq. Frontiers in Ecology and Evolution, 7, 495.

Srivathsan, A., Feng, V., Suárez, D., Emerson, B., & Meier, R. (2024). ONTbarcoder 2.0: Rapid species discovery and identification with real-time barcoding facilitated by Oxford Nanopore R10.4. Cladistics, 40(2), 192–203.

Srivathsan, A., Lee, L., Katoh, K., Hartop, E., Kutty, S. N., Wong, J., Yeo, D., & Meier, R. (2021). ONTbarcoder and MinION barcodes aid biodiversity discovery and identification by everyone, for everyone. BMC Biology, 19(1), 217.

Touroult, J., Cima, V., Bouyon, H., Hanot, C., Horellou, A., & Brustel, H. (2019). Longicornes de France – Atlas préliminaire (Coleoptera: Cerambycidae & Vesperidae) [Longhorn beetles of France – preliminary atlas (Coleoptera: Cerambycidae & Vesperidae)]. Paris, Supplément au Bulletin d’ACOREP-France (in French).

Villiers, A. (1974). Une nouvelle nomenclature des Lepturines de France (Coleoptera: Cerambycidae) [A new nomenclature of Lepturins of France (Coleoptera: Cerambycidae)]. L’Entomologiste, 30, 207–217 (in French).

Vitali, F. (2018). Atlas of the insects of the Grand Duchy of Luxembourg: Coleoptera, Cerambycidae. Ferrantia, 79. Musée National d’Histoire Naturelle, Luxembourg.

Zamoroka A. M., Ruicănescu A., & Manci C.-O. (2024) East and West of the Carpathian Arc: Evidence of postglacial ecological and morphological divergence of Phytoecia tigrina metapopulations (Coleoptera, Cerambycidae). Biosystems Diversity, 32(1), 12–29.

Zamoroka, A. (2023). New additions to the fauna of the longhorn beetles in Ukraine with new records of rare, poorly known and invasive species. Baltic Journal of Coleopterology, 23(2), 159–188.

Zamoroka, A. (2025). When morphology meets molecules: Barcoding confirms an ancient separation of Phytoecia tigrina subspecies (Coleoptera: Cerambycidae). Zoodiversity, 59(5), 477–492.

Zamoroka, A. M. (2022). The longhorn beetles (Coleoptera: Cerambycidae) of Ukraine: Results of two centuries of research. Biosystems Diversity, 30(1), 46–74.

Zamoroka, A. M., Panin, R. Y., Kapelukh, Y. I., & Podobivskiy, S. S. (2012). The catalogue of the longhorn beetles (Coleoptera: Cerambycidae) of Western Podillya, Ukraine. Munis Entomology and Zoology, 7(2), 1145–1177.

Zamoroka, A. M., Semaniuk, D. V., Shparyk, V. Yu., & Mykytyn, T. V. (2019). Taxonomic position of Anastrangalia reyi and A. sequensi (Coleoptera: Cerambycidae) based on molecular and morphological data. Vestnik Zoologii, 53(3), 209–226.

Zamoroka, A. M., Trócoli, S., Shparyk, V. Y., & Semaniuk, D. V. (2022). Polyphyly of the genus Stenurella (Coleoptera: Cerambycidae): Consensus of morphological and molecular data. Biosystems Diversity, 30(2), 119–136.

Zamoroka, A., & Zinenko, O. (2025). Molecular cross-validation of intraspecific structure of Rutpela maculata (Insecta: Coleoptera: Cerambycidae). Journal of Vasyl Stefanyk Precarpathian National University. Biology, 12, 46–59.