Abstract
Taxonomy of the genus Cordulegaster Leach in Brewster, 1815 in Greece is not completely understood. The taxonomic status of the subspecies C. helladica buchholzi (Lohmann, 1993), C. helladica kastalia (Lohmann, 1993), and C. heros pelionensis Theischinger, 1979 was still unclear. We applied a molecular genetic approach using sequences of mitochondrial and nuclear DNA fragments—cytochrome c oxidase I (COI) and Internal Transcribed Spacer 1 (ITS1). This approach revealed that specimens presently assigned to C. heros pelionensis should be considered as conspecific to the nominate subspecific taxon making C. heros a monotypic species. Two major monophyletic lines were found within the Greek representatives of the species grouped around C. bidentata Selys, 1843: the clade of the European endemic C. bidentata and the clade composed of three species: C. helladica (Lohmann, 1993), C. buchholzi (stat. nov., raised to species level), and C. insignis Schneider, 1845. Cordulegaster helladica is restricted to the Peloponnese. Cordulegaster buchholzi is not restricted to the Cyclades as previously thought, but widespread from the Cyclades over the island Euboea to south-east mainland Greece reaching in the west near Mount Parnassos, where it hybridize with C. bidentata. Hybridization between C. bidentata and C. buchholzi was detected at the Castalian Spring, where in ancient times the Oracle of Delphi was located, and some kilometres east of the Castalian Spring. These hybrids had been formerly named C. helladica kastalia. In the case of C. insignis montandoni St. Quentin, 1971 we have investigated specimens some kilometres away from the type locality in Romania, which all revealed hybrids between C. bidentata and C. insignis. However, we do not know if specimens phenotypically looking like C. insignis from further west in the SE Balkans represent isolated population of C. insignis within the range of C. bidentata or belong to a broader hybrid zone between C. bidentata and C. insignis.
References
Barido-Sottani, J., Bošková, V., du Plessis, L., Kühnert, D., Magnus, C., Mitov, V., Müller, N.F., Pečerska, J., Rasmussen, D.A., Zhang, C., Drummond, A.J., Heath, T.A., Pybus, O.G., Vaughan, T.G. & Stadler, T. (2018) Taming the BEAST—A community teaching material resource for BEAST 2. Systematic Biology, 67, 170–174. https://doi.org/10.1093/sysbio/syx060
Boudot, J.-P. (2001) Les Cordulegaster du Paléarctique occidental: identification et répartition (Odonata, Anisoptera, Cordulegastridae). Martinia, 17, 1–34.
Boudot, J.-P. (2010a) Cordulegaster helladica. The IUCN Red List of Threatened Species, 2010, e.T59706A11981390. https://doi.org/10.2305/IUCN.UK.2010-3.RLTS.T59706A11981390.en
Boudot, J.-P. (2010b) Cordulegaster helladica ssp. kastalia. The IUCN Red List of Threatened Species, 2010, e.T63227A12640908 https://doi.org/10.2305/IUCN.UK.2010-1.RLTS.T63227A12640908.en
Boudot, J.-P. & Holuša, O. (2015) Cordulegaster helladica (Lohmann, 1993). In: Boudot, J.-P. & Kalkman, V. (Eds.), Atlas of the European dragonflies and damselflies. KNNV publishing, Zeist, pp. 215–216.
Boudot, J.-P. & Jacquemin, G. (1995) Revision of Cordulegaster boltonii (Donovan, 1807) in southwestern Europe and northern Africa, with description of C. b. iberica ssp. from Spain (Anisoptera: Cordulegastridae). Odonatologica, 24, 149–173.
Boudot, J.-P. & Kalkman, V. (2014) Chapter 5. Odonata (dragonflies and damselflies). In: Smith, K.G., Barrios, V., Darwall, W.R.T. & Numa, C. (Eds.), The Status and Distribution of Freshwater Biodiversity in the Eastern Mediterranean, IUCN, Gland and Cambridge, pp. 57–69.
Boudot, J.-P, Marinov, M. & Kutsarov, Y. (2015) Cordulegaster insignis Schnder, 1845. In: Boudot, J.-P. & Kalkman, V. (Eds.), Atlas of the European dragonflies and damselflies. KNNV publishing, Zeist, pp. 218–219.
Corso, A. (2019) Morphological variability of Cordulegaster trinacriae in Italy (Odonata: Cordulegastridae). Odonatologica, 48, 167–174.
Darriba, D., Taboada, G.L., Doallo, R. & Posada, D. (2012) jModelTest 2: more models, new heuristics and parallel computing. Nature Methods, 9, 772. https://doi.org/10.1038/nmeth.2109
Drummond, A.J. & Rambaut, A. (2007) BEAST: Bayesian evolutionary analysis by sampling trees. BMC Evolutionary Biology, 7, 1–8. https://doi.org/10.1186/1471-2148-7-214
Drummond, A.J., Suchard, M.A., Xie, D. & Rambaut, A. (2012) Bayesische Phylogenetik mit BEAUti und dem BEAST 1.7. Molekularbiologie und Evolution, 22, 1185–1192. https://doi.org/10.1093/molbev/mss075
Earth Observation System (2021) Available from: https://eos.com/blog/natural-disasters-2021/ (accessed 29 December 2021)
Excoffier, L. & Lischer, H.E.L. (2010) Arlequin suite ver 3.5: A new series of programs to perform population genetics analyses under Linux and Windows. Molecular Ecology Resources, 10, 564–567. https://doi.org/10.1111/j.1755-0998.2010.02847.x
Flouri, T., Jiao, X., Rannala, B. & Yang, Z. (2018) Species Tree Inference with BPP using Genomic Sequences and the Multispecies Coalescent. Molecular Biology and Evolution, 35, 2585–2593. https://doi.org/10.1093/molbev/msy147
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, 294–299.
Froufe, E., Roufe, E., Ferreira, S., Boudot, J.-P., Alves, P.C. & Harris, D.J. (2014) Molecular phylogeny of the Western Palaearctic Cordulegaster taxa (Odonata Anisoptera: Cordulegastridae). Biological Journal of the Linnean Society, 111, 49–57. https://doi.org/10.1111/bij.12190
Fraser, F.C. (1929) A revision of the Fissilabioidea (Cordulegasteridae, Petaliidae and Petaluridae) (Order Odonata). Part 1. Cordulegasteridae. Memoirs of the Indian Museum, 9, 69–167, pls. IX–XII.
Galimberti, A., Assandri, G., Maggioni, D., Ramazzotti, F., Baroni, D., Bazzi, G., Chiandetti, I., Corso, A., Ferri, V., Galuppi, M., Ilahiane, L., La Porta, G., Laddaga, L., Landi, F., Ramellini, S., Santinelli, R., Soldato, G., Surdo, S. & Casiraghi, M. (2021) DNA barcoding and eDNA-based biomonitoring of Italian Odonata. Molecular Ecology Resources, 21, 183–200. https://doi.org/10.1111/1755-0998.13235
Ge, Y., Xia, C., Wang, J., Zhang, X., Ma., X. & Zhou, Q. (2021) The efficacy of DNA barcoding in the classification, genetic differentiation, and biodiversity assessment of benthic macroinvertebrates. Ecology and Evolution, 11 (10), 5669–5681. https://doi.org/10.1002/ece3.7470
Geiger, M., Koblmüller, S., Assandri, G., Chovanec, A., Ekrem, T., Fischer, I., Galimberti, A., Grabowski, M., Haring, E., Hausmann, A., Hendrich, L., Koch, S., Mamos, T., Rothe, U., Rulik, B., Rewicz, T., Sittenthaler, M., Stur, E., Tonczyk, G., Zangl, L. & Moriniere, J. (2021) Coverage and quality of DNA barcode references for Central and Northern European Odonata. Peer Journal, 9, e11192. https://doi.org/10.7717/peerj.11192
Hebert, P.D.N., Cywinska, A., Ball, S.L. & de Waard, J.R. (2003a) Biological identifications through DNA barcodes. Proceedings of the Royal Society - Biological Sciences, Series B, 270, 313–321. https://doi.org/10.1098/rspb.2002.2218
Hebert, P.D.N., Ratnasingham S. & de Waard J.R. (2003b). Barcoding animal cytochrome c subunit 1 divergences among closely species. Proceedings of the Royal Society London, B (Suppl.), 270, 96–99. https://doi.org/10.1098/rsbl.2003.0025
Katoh, K., Rozewicki, J. & Yamada, K.D. (2019) MAFFT online service: multiple sequence alignment, interactive sequence choice and visualization. Brief Bioform, 20, 1160–1166. https://doi.org/10.1093/bib/bbx108
Koroiva, R., Pepinelli, M., Rodrigues, M.E., Roque. Fd. O., Lorenz-Lemke, A.P. & Kvist, S. (2017) DNA barcoding of odonates from the Upper Plata basin: Database creation and genetic diversity estimation. PLoS ONE, 12, e0182283. https://doi.org/10.1371/journal.pone.0182283
Kumar, S., Stecher, G., Li, M., Knyaz, C, & Tamura, K. (2018) MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms. Molecular Biology and Evolution, 35, 1547–1549. https://doi.org/10.1093/molbev/msy096
Leigh, J. & Bryant, D. (2015) POPART: full-feature software for haplotype network construction. Methods in Ecology and Evolution, 6, 1110–1116. https://doi.org/10.1111/2041-210X.12410
Lohmann, H. (1993) Revision der Cordulegastridae. 2. Beschreibung neuer Arten in den Gattungen Cordulegaster, Anotogaster, Neallogaster und Sonjagaster (Anisoptera). Odonatologica, 22, 273–294.
Lopau, W. (2010) Studien zur Libellenfauna Griechenland IV. Libellula, Supplement 10, 1–260.
Marinov, M., Grebe, B. & Kutsarov, Y. (2007) Cordulegaster insignis (Schneider, 1845) in Bulgaria with notes on its biology and ecology. In: Tyagi, B.K. (Ed.). Odonata. Biology of Dragonflies, Science Publishers, Madurai, India, pp. 51–61.
Morton, K.J. (1916) Some Palaearctic species of Cordulegaster. The Transactions of the Entomological Society of London, 273–290, pls. 34–37. https://doi.org/10.1111/j.1365-2311.1916.tb02535.x
Pimenta, A.L.A., Pinto, A.P. & Takiya, D.M. (2019) Integrative taxonomy and phylogeny of the damselfly genus Forcepsioneura Lencioni, 1999 (Odonata: Coenagrionidae: Protoneurinae) with description of two new species from the Brazilian Atlantic Forest. Arthropod Systematics & Phylogenie, 77, 1–19.
Rach, J., Desalle, R., Sarkar, I.N., Schierwater, B. & Hadrys, H. (2008) Character based DNA barcoding allows discrimination of genera, species and populations in Odonata. Proceedings of the Royal Society B: Biological Sciences, 275, 237–247. https://doi.org/10.1098/rspb.2007.1290
Rambout, A. (2021) GitHub. Available online: https://github.com/rambaut/figtree (accessed October 2021)
Ronquist, F., Teslenko, M., van der Mark, P., Ayres, D.L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M.A. & Huelsenbeck, J.P. (2012) MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology, 61, 539–542. https://doi.org/10.1093/sysbio/sys029
Schneider, T., Vierstraete, A., Müller, O., van Pelt, G.J., Casper, M., Ikemeyer, D., Snegovaya, N. & Dumont, H.J. (2021) Taxonomic Revision of eastern part of Western Palaearctic Cordulegaster using molecular phylogeny and morphology, with the description of two new species (Odonata: Anisoptera: Cordulegastridae). Diversity, 13, 667. https://doi.org/10.3390/d13120667
Selys Longchamps (de), E. (1850) Revue des Odonates ou Libellules d’Europe. Mémoires de la Société Royale des Sciences de Liège, 6, 1–408.
Solano, E., Hardersen, S., Audisio, P., Amorosi, V., Senczuk, G. & Antonini, G. (2018) Asymmetric hybridization in Cordulegaster (Odonata: Cordulegastridae): Secondary postglacial contact and the possible role of mechanical constraints. Ecology and Evolution, 8, 9657–9671. https://doi.org/10.1002/ece3.4368
St. Quentin, D. (1952) Der Rassenkreis Cordulegaster boltonii (Donovan) (Odonata). Entomologisches Nachrichtenblatt Österreichischer und Schweizer Entomologen, 4, 73–75.
St. Quentin, D. (1971) Zum Vorkommen von Cordulegaster insignis Schneider in Rumänien. Studii Communicari, Sibiu, 16, 205–208.
Swofford, D.L. (2003) PAUP*. Phylogenetic Analysis Using Parsimony (*and Other Methods). Version 4. Sinauer Associates, Sunderland, Massachusetts. [program]
Theischinger, G. (1979) Cordulegaster heros sp. nov. und Cordulegaster heros pelionensis ssp. nov., zwei neue Taxa des Cordulegaster boltonii (Donovan)-Komplexes aus Europa (Ansioptera: Cordulegastridae). Odonatologica, 8, 23–38.
Van Pelt, G.J. (1999) On dragonflies from Greece in the RMNH collection, Leiden, The Netherlands. Libellula Supplement, 2, 77–90.
Van Pelt, G.J. (2020) Cordulegaster. In: Dijkstra, K.-D., Schröter, A. & Lewington, R. (Eds.) Field guide to the dragonflies of Britain and Europe. Bloomsbury Wildlife, London, pp. 220-231.