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Phylogenetic Insights into Sitona Germar, 1817 (Coleoptera: Curculionidae) through DNA Barcoding of the COI Gene

Year 2025, Volume: 8 Issue: 2, 114 - 121, 29.12.2025
https://doi.org/10.46876/ja.1782989

Abstract

Sitona Germar is a large genus in the family Curculionidae, comprising approximately 100 species. However, recent revisions of subgenera or species groups classifications are lacking, making it difficult to compare newly discovered species with previously described ones. In this study, we performed a molecular phylogenetic analysis based on the COI (cytochrome c oxidase subunit I) gene to assess the validity of the traditional groupings within the genus Sitona. A total of 23 species were analyzed, 14 of them were collected in the eastern and southern regions of Türkiye: S. callosus Gyllenhal, S. concavirostris Hochhuth, S. cylindricollis Fåhraeus, S. fairmairei Allard, S. hispidulus (Fabricius), Sitona humeralis Stephens, S. lateralis Gyllenhal, S. lineatus (Linnaeus), S. lividipes Fåhraeus, S. longulus Gyllenhal, S. macularius (Marsham), S. obsoletus (Gmelin), S. puncticollis Stephens and S. sulcifrons Thunberg. In addition, nine species were incorporated from GenBank records: S. ambiguus Gyllenhal, S. brucki Allard, S. discoideus Gyllenhal, S. languidus Gyllenhal, S. striatellus Gyllenhal, S. suturalis Stephens and S. waterhousei Walton. Two outgroup taxa, Coelositona latipennis (Gyllenhal) and C. limosus (Rossi), were also included in the analysis. Phylogenetic reconstruction was performed using the Maximum Parsimony method with 1000 bootstrap replicates, implemented with Geneious Prime (2024.0.7) and the PAUP 4.0a plug-in. The resulting phylogram grouped the analyzed species into four distinct clusters, which was inconsistent with Reitter’s previous classification of Sitona. The traditional morphological characters used in the taxonomy of the genus are briefly reviewed, and a new species grouping framework is proposed.

Project Number

120O352

References

  • Aeschlimann, J. P. (1980). The Sitona (Coleoptera, Curculionidae) species occurring on Medicago and their natural enemies in the Mediterranean region. Entomophaga, 25, 139–153. https://doi.org/10.1007/bf02374316
  • Alonso-Zarazaga, M. A. (2002). Lista preliminar de los Coleoptera Curculionoidea del área ibero-balear, con descripción de Melicius gen. nov. y nuevas citas. Boletín de la S.E.A., 31, 9–33.
  • Alonso-Zarazaga, M. A., Barrios, H., Borovec, R., Bouchard, P., Caldara, R., Colonnelli, E., Gültekin, L., Hlaváč, P., Korotyaev, B., Lyal, C. H. C., Machado, A., Meregalli, M., Pierotti, H., Ren, L., Sánchez-Ruiz, M., Sforzi, A., Silfverberg, H., Skuhrovec, J., Trýzna, M., … Yunakov, N. N. (2023). Cooperative catalogue of Palaearctic Coleoptera Curculionoidea (2nd ed., Vol. 14). Zaragoza, Spain: Sociedad Entomológica Aragonesa S.E.A.
  • Boroumand, H. (1975). Key to the groups and species of the genus Sitona (Coleoptera: Curculionidae) in Iran. Journal of the Entomological Society of Iran, 2(2), 67–68 (in English), 101–110 (in Farsi).
  • Bright, D. E. (1994). Revision of the genus Sitona (Coleoptera: Curculionidae) of North America. Annals of the Entomological Society of America, 87(3), 277–306. https://doi.org/10.1093/aesa/87.3.277
  • Cantot, P. (2001). Influence de quelques Papilionacées sur la ponte et le développement larvaire de Sitona lineatus (L.) (Coleoptera, Curculionidae). Bulletin de la Société Entomologique de France, 106, 441–447.
  • Coissac, E., Hollingsworth, P. M., Lavergne, S., & Taberlet, P. (2016). From barcodes to genomes: Extending the concept of DNA barcoding. Molecular Ecology, 25(7), 1423–1428. https://doi.org/10.1111/mec.13549
  • Crozier, R. H., & Crozier, Y. C. (1993). The mitochondrial genome of the honeybee Apis mellifera: Complete sequence and genome organization. Genetics, 133, 97–117. https://doi.org/10.1093/genetics/133.1.97
  • Dowton, M., & Austin, A. D. (1995). Increased genetic diversity in mitochondrial genes correlated with the evolution of parasitism in the Hymenoptera. Journal of Molecular Evolution, 41(6), 958–965. https://doi.org/10.1007/bf00173176
  • Folmer, O., Black, M., Hoeh, W., Lutz, R., & Vrijenhoek, R. (1994). DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology, 3(5), 294–299.
  • Gözüaçık, C. (2023). Diversity of Sitona species (Coleoptera, Curculionidae) in cultivated plants belonging to the Fabaceae family. Applied Ecology and Environmental Research, 21(4), 3275–3284. https://doi.org/10.15666/aeer/2104_32753284
  • Hoffmann, A. (1950). Faune de France 52 Coléoptères Curculionides (Première partie). Paris: Lechevalier.
  • Hollingsworth, M. L., Clark, A. A., Forrest, L. L., Richardson, J., Pennington, R. T., Long, D. G., Cowan, R. S., Chase, M. W., Gaudeul, M., & Hollingsworth, P. M. (2009). Selecting barcoding loci for plants: Evaluation of seven candidate loci with species-level sampling in three divergent groups of land plants. Molecular Ecology Resources, 9(2), 439–457. https://doi.org/10.1111/j.1755-0998.2008.02439.x
  • Hollingsworth, P. M. (2011). Refining the DNA barcode for land plants. Proceedings of the National Academy of Sciences, 108(49), 19451–19452. https://doi.org/10.1073/pnas.1116812108
  • Hustache, A. (1946). Coléoptères nouveaux du Maroc et de l‘Algérie. Bulletin de la Société des Sciences Naturelles du Maroc, 24 (1944), 44–81.
  • Murray, P. J. (1996). Influence of food source on feeding, longevity and fecundity of Sitona flavescens (Coleoptera: Curculionidae), a major pest of white clover in the U.K. In J. Frame (Ed.), Recent Research and Development on White Clover in Europe (REU Technical Series 42). FAO, Estonia.
  • Porta, A. (1932). Fauna Coleopterorum Italica (Vol. V). Piacenza: Stabilimento Tipografico Piacentino.
  • Reitter, E. (1903). Bestimmungs-Tabellen der europäischen Coleopteren. Curculionidae. 9 Theil: Genus Sitona Germ, und Mesagroicus Schönh. aus der palaearctischen Fauna. Paskau: Edmund Reitter.
  • Scherf, H., 1964. Die Entwicklungsstadien der Mitteleuropäischen Curculioniden (Morphologie, Bionomie, Ökologie). Abhandlungen des Senckenbergischen Naturforschenden Gesellshaft 506, Verlag Waldemar Kramer, Frankfurt.
  • Schütte, A., & Stüben, P. E., & Sprick, P. (2013). The Molecular Weevil Identification Project. Part I. A contribution to integrative taxonomy and phylogenetic systematics. Snudebiller: Studies on Taxonomy, Biology and Ecology of Curculionoidea, 14, 77 pp.
  • Schütte, A., Stüben, P. E., & Astrin, J. J. (2023). Molecular Weevil Identification Project: A thoroughly curated barcode release of 1300 Western Palearctic weevil species (Coleoptera, Curculionoidea). Biodiversity Data Journal, 11, 1–42. https://doi.org/10.3897/BDJ.11.e96438
  • Stierlin, G. (1885). Bestimmungs-Tabellen europäischer Rüsselkäfer II, Brachyderidae (Fortsetzung). Mitteilungen der Schweizerischen Entomologischen Gesellschaft, 7(3), 99–146.
  • Stüben, P. (2022). Weevils of Macaronesia: Canary Islands, Madeira, Azores (Coleoptera: Curculionoidea). Mönchengladbach: Curculio Institute.
  • Stüben, P. E., & Bayer, C. (2015). New nomenclatural and taxonomic acts, and comments (2015). Snudebiller, 16, 1–8.
  • Stüben, P. E., Schütte, A., Bayer, C., & Astrin, J. J. (2015). The Molecular Weevil Identification Project (Coleoptera: Curculionoidea), Part II. Towards an integrative taxonomy. Snudebiller, 16, 237–294.
  • Swofford, D. L. (2003). PAUP. Phylogenetic Analysis Using Parsimony (and Other Methods) (Version 4). Sunderland, MA: Sinauer Associates.
  • Syrett, P. (1992). Feeding range of Sitona regensteinensis Hbst. (Coleoptera: Curculionidae), a potential agent for biological control of Cytisus scoparius (L.) Link (broom) in New Zealand. Biocontrol Science and Technology, 2, 261–265. https://doi.org/10.1080/09583159209355240
  • Velázquez De Castro, A., 1997. Estudio Morfológico y Taxonómico del Género Sitona Gerfllar,1817 (Coleoptera, Curculionidae). Tesis Doctoral, Universidad Complutense de Madrid Facultad de Ciencias Biológicas, Madrid.
  • Velázquez De Castro, A. J., Alonso-Zarazaga, M. A., & Outerelo, R. (2007). Systematics of Sitonini (Coleoptera: Curculionidae: Entiminae), with a hypothesis on the evolution of feeding habits. Systematic Entomology, 32, 312–331. https://doi.org/10.1111/j.1365-3113.2006.00368.x
  • Whitfield, J. B., & Cameron, S. A. (1998). Hierarchical analysis of variation in the mitochondrial 16S rRNA gene among Hymenoptera. Molecular Biology and Evolution, 15(12), 1728–1743. https://doi.org/10.1093/oxfordjournals.molbev.a025899

Phylogenetic Insights into Sitona Germar, 1817 (Coleoptera: Curculionidae) through DNA Barcoding of the COI Gene

Year 2025, Volume: 8 Issue: 2, 114 - 121, 29.12.2025
https://doi.org/10.46876/ja.1782989

Abstract

Sitona Germar is a large genus in the family Curculionidae, comprising approximately 100 species. However, recent revisions of subgenera or species groups classifications are lacking, making it difficult to compare newly discovered species with previously described ones. In this study, we performed a molecular phylogenetic analysis based on the COI (cytochrome c oxidase subunit I) gene to assess the validity of the traditional groupings within the genus Sitona. A total of 23 species were analyzed, 14 of them were collected in the eastern and southern regions of Türkiye: S. callosus Gyllenhal, S. concavirostris Hochhuth, S. cylindricollis Fåhraeus, S. fairmairei Allard, S. hispidulus (Fabricius), Sitona humeralis Stephens, S. lateralis Gyllenhal, S. lineatus (Linnaeus), S. lividipes Fåhraeus, S. longulus Gyllenhal, S. macularius (Marsham), S. obsoletus (Gmelin), S. puncticollis Stephens and S. sulcifrons Thunberg. In addition, nine species were incorporated from GenBank records: S. ambiguus Gyllenhal, S. brucki Allard, S. discoideus Gyllenhal, S. languidus Gyllenhal, S. striatellus Gyllenhal, S. suturalis Stephens and S. waterhousei Walton. Two outgroup taxa, Coelositona latipennis (Gyllenhal) and C. limosus (Rossi), were also included in the analysis. Phylogenetic reconstruction was performed using the Maximum Parsimony method with 1000 bootstrap replicates, implemented with Geneious Prime (2024.0.7) and the PAUP 4.0a plug-in. The resulting phylogram grouped the analyzed species into four distinct clusters, which was inconsistent with Reitter’s previous classification of Sitona. The traditional morphological characters used in the taxonomy of the genus are briefly reviewed, and a new species grouping framework is proposed.

Ethical Statement

There's no need

Supporting Institution

TUBITAK

Project Number

120O352

Thanks

This study was financially supported by The Scientific and Technological Research Council of Turkey with project number 120O352.

References

  • Aeschlimann, J. P. (1980). The Sitona (Coleoptera, Curculionidae) species occurring on Medicago and their natural enemies in the Mediterranean region. Entomophaga, 25, 139–153. https://doi.org/10.1007/bf02374316
  • Alonso-Zarazaga, M. A. (2002). Lista preliminar de los Coleoptera Curculionoidea del área ibero-balear, con descripción de Melicius gen. nov. y nuevas citas. Boletín de la S.E.A., 31, 9–33.
  • Alonso-Zarazaga, M. A., Barrios, H., Borovec, R., Bouchard, P., Caldara, R., Colonnelli, E., Gültekin, L., Hlaváč, P., Korotyaev, B., Lyal, C. H. C., Machado, A., Meregalli, M., Pierotti, H., Ren, L., Sánchez-Ruiz, M., Sforzi, A., Silfverberg, H., Skuhrovec, J., Trýzna, M., … Yunakov, N. N. (2023). Cooperative catalogue of Palaearctic Coleoptera Curculionoidea (2nd ed., Vol. 14). Zaragoza, Spain: Sociedad Entomológica Aragonesa S.E.A.
  • Boroumand, H. (1975). Key to the groups and species of the genus Sitona (Coleoptera: Curculionidae) in Iran. Journal of the Entomological Society of Iran, 2(2), 67–68 (in English), 101–110 (in Farsi).
  • Bright, D. E. (1994). Revision of the genus Sitona (Coleoptera: Curculionidae) of North America. Annals of the Entomological Society of America, 87(3), 277–306. https://doi.org/10.1093/aesa/87.3.277
  • Cantot, P. (2001). Influence de quelques Papilionacées sur la ponte et le développement larvaire de Sitona lineatus (L.) (Coleoptera, Curculionidae). Bulletin de la Société Entomologique de France, 106, 441–447.
  • Coissac, E., Hollingsworth, P. M., Lavergne, S., & Taberlet, P. (2016). From barcodes to genomes: Extending the concept of DNA barcoding. Molecular Ecology, 25(7), 1423–1428. https://doi.org/10.1111/mec.13549
  • Crozier, R. H., & Crozier, Y. C. (1993). The mitochondrial genome of the honeybee Apis mellifera: Complete sequence and genome organization. Genetics, 133, 97–117. https://doi.org/10.1093/genetics/133.1.97
  • Dowton, M., & Austin, A. D. (1995). Increased genetic diversity in mitochondrial genes correlated with the evolution of parasitism in the Hymenoptera. Journal of Molecular Evolution, 41(6), 958–965. https://doi.org/10.1007/bf00173176
  • Folmer, O., Black, M., Hoeh, W., Lutz, R., & Vrijenhoek, R. (1994). DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology, 3(5), 294–299.
  • Gözüaçık, C. (2023). Diversity of Sitona species (Coleoptera, Curculionidae) in cultivated plants belonging to the Fabaceae family. Applied Ecology and Environmental Research, 21(4), 3275–3284. https://doi.org/10.15666/aeer/2104_32753284
  • Hoffmann, A. (1950). Faune de France 52 Coléoptères Curculionides (Première partie). Paris: Lechevalier.
  • Hollingsworth, M. L., Clark, A. A., Forrest, L. L., Richardson, J., Pennington, R. T., Long, D. G., Cowan, R. S., Chase, M. W., Gaudeul, M., & Hollingsworth, P. M. (2009). Selecting barcoding loci for plants: Evaluation of seven candidate loci with species-level sampling in three divergent groups of land plants. Molecular Ecology Resources, 9(2), 439–457. https://doi.org/10.1111/j.1755-0998.2008.02439.x
  • Hollingsworth, P. M. (2011). Refining the DNA barcode for land plants. Proceedings of the National Academy of Sciences, 108(49), 19451–19452. https://doi.org/10.1073/pnas.1116812108
  • Hustache, A. (1946). Coléoptères nouveaux du Maroc et de l‘Algérie. Bulletin de la Société des Sciences Naturelles du Maroc, 24 (1944), 44–81.
  • Murray, P. J. (1996). Influence of food source on feeding, longevity and fecundity of Sitona flavescens (Coleoptera: Curculionidae), a major pest of white clover in the U.K. In J. Frame (Ed.), Recent Research and Development on White Clover in Europe (REU Technical Series 42). FAO, Estonia.
  • Porta, A. (1932). Fauna Coleopterorum Italica (Vol. V). Piacenza: Stabilimento Tipografico Piacentino.
  • Reitter, E. (1903). Bestimmungs-Tabellen der europäischen Coleopteren. Curculionidae. 9 Theil: Genus Sitona Germ, und Mesagroicus Schönh. aus der palaearctischen Fauna. Paskau: Edmund Reitter.
  • Scherf, H., 1964. Die Entwicklungsstadien der Mitteleuropäischen Curculioniden (Morphologie, Bionomie, Ökologie). Abhandlungen des Senckenbergischen Naturforschenden Gesellshaft 506, Verlag Waldemar Kramer, Frankfurt.
  • Schütte, A., & Stüben, P. E., & Sprick, P. (2013). The Molecular Weevil Identification Project. Part I. A contribution to integrative taxonomy and phylogenetic systematics. Snudebiller: Studies on Taxonomy, Biology and Ecology of Curculionoidea, 14, 77 pp.
  • Schütte, A., Stüben, P. E., & Astrin, J. J. (2023). Molecular Weevil Identification Project: A thoroughly curated barcode release of 1300 Western Palearctic weevil species (Coleoptera, Curculionoidea). Biodiversity Data Journal, 11, 1–42. https://doi.org/10.3897/BDJ.11.e96438
  • Stierlin, G. (1885). Bestimmungs-Tabellen europäischer Rüsselkäfer II, Brachyderidae (Fortsetzung). Mitteilungen der Schweizerischen Entomologischen Gesellschaft, 7(3), 99–146.
  • Stüben, P. (2022). Weevils of Macaronesia: Canary Islands, Madeira, Azores (Coleoptera: Curculionoidea). Mönchengladbach: Curculio Institute.
  • Stüben, P. E., & Bayer, C. (2015). New nomenclatural and taxonomic acts, and comments (2015). Snudebiller, 16, 1–8.
  • Stüben, P. E., Schütte, A., Bayer, C., & Astrin, J. J. (2015). The Molecular Weevil Identification Project (Coleoptera: Curculionoidea), Part II. Towards an integrative taxonomy. Snudebiller, 16, 237–294.
  • Swofford, D. L. (2003). PAUP. Phylogenetic Analysis Using Parsimony (and Other Methods) (Version 4). Sunderland, MA: Sinauer Associates.
  • Syrett, P. (1992). Feeding range of Sitona regensteinensis Hbst. (Coleoptera: Curculionidae), a potential agent for biological control of Cytisus scoparius (L.) Link (broom) in New Zealand. Biocontrol Science and Technology, 2, 261–265. https://doi.org/10.1080/09583159209355240
  • Velázquez De Castro, A., 1997. Estudio Morfológico y Taxonómico del Género Sitona Gerfllar,1817 (Coleoptera, Curculionidae). Tesis Doctoral, Universidad Complutense de Madrid Facultad de Ciencias Biológicas, Madrid.
  • Velázquez De Castro, A. J., Alonso-Zarazaga, M. A., & Outerelo, R. (2007). Systematics of Sitonini (Coleoptera: Curculionidae: Entiminae), with a hypothesis on the evolution of feeding habits. Systematic Entomology, 32, 312–331. https://doi.org/10.1111/j.1365-3113.2006.00368.x
  • Whitfield, J. B., & Cameron, S. A. (1998). Hierarchical analysis of variation in the mitochondrial 16S rRNA gene among Hymenoptera. Molecular Biology and Evolution, 15(12), 1728–1743. https://doi.org/10.1093/oxfordjournals.molbev.a025899
There are 30 citations in total.

Details

Primary Language English
Subjects Forest Entomology and Forest Protection
Journal Section Research Article
Authors

Celalettin Gözüaçık 0000-0002-6543-7663

Kaan Hürkan 0000-0001-5330-7442

Antonio J. Velázquez De Castro 0000-0002-5357-4126

Project Number 120O352
Submission Date September 12, 2025
Acceptance Date November 2, 2025
Early Pub Date November 21, 2025
Publication Date December 29, 2025
Published in Issue Year 2025 Volume: 8 Issue: 2

Cite

APA Gözüaçık, C., Hürkan, K., & Velázquez De Castro, A. J. (2025). Phylogenetic Insights into Sitona Germar, 1817 (Coleoptera: Curculionidae) through DNA Barcoding of the COI Gene. Journal of Agriculture, 8(2), 114-121. https://doi.org/10.46876/ja.1782989