Skip to main content

Advertisement

Springer Nature Link
Log in

DNA barcoding provides new insights on the distribution, systematics and conservation of the freshwater genus Pelasgus (Cypriniformes: Cyprinidae) in Greece

  • Primary Research Paper
  • Published:
Hydrobiologia Aims and scope Submit manuscript

Abstract

The freshwater ichthyofauna of Greece is characterized by high diversity and endemism and faces various conservation challenges. Despite significant efforts, mainly with traditional assessment methods, the precise taxonomic status and geographic range of some of the currently acknowledged fish species remain insufficiently explored. Here, the results of the first comprehensive DNA barcoding survey of the genus Pelasgus are presented, covering its entire known range in Greece. Two hundred and forty COI sequences (retrieved from collected samples, museum specimens and the GenBank) were analysed to determine the number of putative species and delineate their current distribution. Data support the existence of seven haplogroups, corresponding to the currently valid Pelasgus species. Analysis of museum samples of the critically endangered species P. epiroticus from its type locality (now extinct there) uncovers its presence in contemporary samples from a neighbouring aquatic system (Zaravina Lake) and from aquatic systems well outside its native range (possibly as a result of past translocations) for the first time. A subgroup of P. epiroticus haplotypes was also identified in North-western Greece in regions previously believed to host P. thesproticus or P. stymphalicus. Based on the above, hotspots for further Pelasgus taxonomic investigations are identified and conservation actions are discussed.

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

Access this article

Log in via an institution

Subscribe and save

Springer+
from $39.99 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

References

  • Anisimova, M., M. Gil, J. F. Dufayard, C. Dessimoz & O. Gascuel, 2011. Survey of branch support methods demonstrates accuracy, power, and robustness of fast likelihood-based approximation schemes. Systematic Biology 60: 685–699.

    PubMed  PubMed Central  Google Scholar 

  • Apostolidis, A. P., M. T. Stoumboudi, E. Kalogianni, G. Cote & L. Bernatchez, 2011. Genetic divergence among native trout Salmo trutta populations from southern Balkans based on mitochondrial DNA and microsatellite variation. Journal of Fish Biology 79: 1950–1960.

    CAS  PubMed  Google Scholar 

  • April, J., R. Mayden, R. Hanner & L. Bernatchez, 2011. Genetic calibration of species diversity among North America’s freshwater fishes. Proceedings of the National Academy of Sciences of USA 108: 10602–10607.

  • Bandelt, H. J., P. Forster, & A. Röhl, 1999. Median-joining networks for inferring intraspecific phylogenies. Molecular Biology and Evolution 16: 37–48.

    CAS  PubMed  Google Scholar 

  • Barbieri, R., S. Zogaris, E. Kalogianni, M. T. Stoumboudi, Y. Chatzinikolaou, S. Giakoumi, Y. Kapakos, D. Kommatas, N. Koutsikos, V. Tachos, L. Vardakas & A. Economou, 2015. Freshwater Fishes and Lampreys of Greece. Monographs on Marine Sciences, Vol 8 Hellenic Centre for Marine Research, Athens.

    Google Scholar 

  • Buj, I., R. Šanda, S. Zogaris, J. Freyhof, M. F. Geiger & J. Vukić, 2019. Cryptic diversity in Telestes pleurobipunctatus (Actinopterygii; Leuciscidae) as a consequence of historical biogeography in the Ionian Freshwater Ecoregion (Greece, Albania). Hydrobiologia 835: 147–163.

    CAS  Google Scholar 

  • Chase, M. W., N. Salamin, M. Wilkinson, J. M. Dunwell, R. P. Kesanakurthi, N. Haidar & V. Savolainen, 2005. Land plants and DNA barcodes: short-term and long-term goals. Philosophical Transactions of the Royal Society B: Biological Sciences 360: 1889–1895.

    CAS  Google Scholar 

  • Crivelli, A.J., 2006. Pelasgus epiroticus. The IUCN Red List of Threatened Species 2006: e.T61256A12454831.

    Google Scholar 

  • Economou, A. N., R. Barbieri, C. Daoulas, T. Psarras, M. T. Stoumboudi, I. Bertahas, S. Giakoumi & A. Patsias, 1999. Endangered Freshwater Fish of Western Greece and Peloponnese: Distribution, Abundance, Threats and Recommended Conservation Measures. PENED, Technical Report. Anavyssos Attiki: Hellenic Centre for Marine Research, Institute of Inland Waters; 1999 (in Greek).

    Google Scholar 

  • Economou, A. N., S. Giakoumi, L. Vardakas, R. Barbieri, M. Stoumboudi & S. Zogaris, 2007. The freshwater ichthyofauna of Greece – an update based on a hydrographic basin survey. Mediterranean Marine Science 8: 91–166.

    Google Scholar 

  • Funk, D. J., & K. E. Omland, 2003. Species-level paraphyly and polyphyly: frequency, causes, and consequences, with insights from animal mitochondrial DNA. Annual Review of Ecology, Evolution, and Systematics 34: 397–423.

    Google Scholar 

  • Geiger, M. F., F. Herder, M. T. Monaghan, V. Almada, R. Barbieri, M. Bariche, P. Berrebi, J. Bohlen, M. Casal-Lopez, G.B. Delmastro, G.P.J. Denys, A. Dettai, I. Doadrio, E. Kalogianni, H. Kärst, M. Kottelat, M. Kovačić, M. Laporte, M. Lorenzoni, Z. Marčič, M. Özuluğ, A. Perdices, S. Perea, H. Persat, S. Porcelotti, C. Puzzi, J. Robalo, R. Šanda, M. Schneider, V. Šlechtová, M. Stoumboudi, S. Walter & J. Freyhof, 2014. Spatial heterogeneity in the Mediterranean biodiversity hotspot affects barcoding accuracy of its freshwater fishes. Molecular Ecology Resources 14: 1210–1221.

    CAS  PubMed  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  • Hajibabaei, M., G. A. C. Singer, P. D. N. Hebert & D. A. Hickey, 2007. DNA barcoding: how it complements taxonomy, molecular phylogenetics and population genetics. Trends in Genetics 23: 167–172.

    CAS  PubMed  Google Scholar 

  • IUCN, 2019. The IUCN Red List of Threatened Species. [available on internet at http://www.iucnredlist.org]. Accessed 21 March 2019.

  • Ivanova, N. V., T. S. Zemlak, R. H. Hanner & P. D. N. Hebert, 2007. Universal primer cocktails for fish DNA barcoding. Molecular Ecology Notes 7: 544–548.

    CAS  Google Scholar 

  • Kimura M., 1980. A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution 16:111-120.

    CAS  PubMed  Google Scholar 

  • Kottelat, M. & R. Barbieri, 2004. Pseudophoxinus laconicus, a new species of minnow from Peloponnese, Greece, with comments on the West Balkan Pseudophoxinus species (Teleostei: Cyprinidae). Ichthyological Exploration of Freshwaters 15: 147–160.

    Google Scholar 

  • Kottelat, M. & J. Freyhof, 2007a. Handbook of European Freshwater Fishes. Berlin: Kottelat, Cornol and Freyhof.

    Google Scholar 

  • Kottelat, M. & J. Freyhof, 2007b. Pelasgus, a new genus name for the Balkan species of Pseudophoxinus (Teleostei: Cyprinidae). Ichthyological Exploration of Freshwaters 18: 103–108.

    Google Scholar 

  • Kumar S., G. Stecher & K. Tamura, 2016. MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33:1870-1874.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lefort, V., J.E. Longueville & O. Gascuel, 2017. SMS: Smart Model Selection in PhyML. Molecular Biology and Evolution 34: 2422–2424.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Leonardos, I., I. Paschos & M. Prassa, 2005. Threatened fishes of the world: Phoxinellus epiroticus (Steindachner 1895) (Cyprinidae). Environmental Biology of Fishes 72: 250.

    Google Scholar 

  • Leonardos, I. D., I. Kagalou, M. Tsoumani & P. S. Economidis, 2008. Fish fauna in a Protected Greek lake: biodiversity, introduced fish species over a 80-year period and their impacts on the ecosystem. Ecology of Freshwater Fish 17: 165–173.

    Google Scholar 

  • Malaquias, M. A. E., L. T. Ohnheiser, T. R. Oskars, & E. Willassen, 2017. Diversity and systematics of philinid snails (Gastropoda: Cephalaspidea) in West Africa with remarks on the biogeography of the region. Zoological Journal of the Linnean Society 180: 1–35.

    Google Scholar 

  • Masters, B. C., V. Fan & H. A. Ross, 2010. Species delimitation plugin manual. https://assets.geneious.com/plugins/3rdparty/Species_Delimitation_Plugin_Manual.pdf

  • Masters, B. C., V. Fan & H.A. Ross, 2011. Species delimitation – a Geneious Plugin for the exploration of species boundaries. Molecular Ecology Resources 11: 154–157.

    PubMed  Google Scholar 

  • Messing, J., 1983. New M13 vectors for cloning. Methods in Enzymology 101: 20–78.

    CAS  PubMed  Google Scholar 

  • Miller, S. A., D. D. Dykes & H. F. Polesky, 1988. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Research 16: 1215.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Nei, M. & S. Kumar, 2000. Molecular Evolution and Phylogenetics. Oxford University Press, New York.

    Google Scholar 

  • Perdikaris, C., C. Nathanailides, E. Gouva, C. Karipoglou & I. Paschos, 2005. Collapse of Epirus minnow, Phoxinellus epiroticus (Steindachner, 1895) fisheries in Lake Pamvotis, Greece. Ichthyological Exploration of Freshwaters 16: 171–174.

    Google Scholar 

  • Perea, S., J. Vukić, R. Šanda & I. Doadrio, 2016. Ancient mitochondrial capture as factor promoting mitonuclear discordance in freshwater fishes: a case study in the genus Squalius (Actinopterygii, Cyprinidae) in Greece. PLoS ONE 11: e0166292.

    PubMed  PubMed Central  Google Scholar 

  • Puillandre, N., A. Lambert, S. Brouillet & G. Achaz, 2012a. ABGD, Automatic Barcode Gap Discovery for primary species delimitation. Molecular Ecology 21: 1864–1877.

    CAS  PubMed  Google Scholar 

  • Puillandre, N., M. V. Modica, Y. Zhang, L. Sirovich, M. C. Boisselier, C. Cruaud, M. Holford & S. Samadi, 2012b. Large-scale species delimitation method for hyperdiverse groups. Molecular Ecology 21: 2671–2691.

    CAS  PubMed  Google Scholar 

  • Rodrigo A. G., F. Bertels, J. Heled, R. Noder, H. Shearman & P. Tsai, 2008. The perils of plenty: what are we going to do with all these genes? Philosophical Transactions of the Royal Society London. Series B, Biological Sciences 363: 3893–3902.

    Google Scholar 

  • Rosenberg N. A., 2007. Statistical tests for taxonomic distinctiveness from observations of monophyly. Evolution 61: 317–323.

    PubMed  Google Scholar 

  • Satler, J. D., B. C. Carstens & M. Hedin, 2013. Multilocus species delimitation in a complex of morphologically conserved trapdoor spiders (Mygalomorphae, Antrodiaetidae, Aliatypus). Systematic Biology 62: 805–823.

    PubMed  Google Scholar 

  • Skoulikidis, N. T., A. N. Economou, K. C. Gritzalis & S. Zogaris, 2009. Rivers of the Balkans. In Tockner, K, C. T. Robinson & U. Uehlinger (eds), Rivers of Europe. Academic/Elsevier, San Diego.https://doi.org/10.1016/B978-0-12-369449-2.00011-4

    Chapter  Google Scholar 

  • Tamura, K., G. Stecher, D. Peterson, A. Filipski & S. Kumar, 2013. MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution 30: 2725–2729.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Vardakas, L., E. Kalogianni, C. Papadaki, T. Vavalidis, A. Mentzafou, D. Koutsoubas & T. N. Skoulikidis, 2017. Defining critical habitat conditions for the conservation of three endemic and endangered cyprinids in a Mediterranean intermittent river before the onset of drought. Aquatic Conservation: Marine and Freshwater Ecosystems 27: 1270–1280.

    Google Scholar 

  • Vogiatzi, E., E. Kalogianni, B. Zimmerman, S. Giakoumi, R. Barbieri, P. Paschou, A. Magoulas, D. Tsaparis, N. Poulakakis & C. S. Tsigenopoulos, 2014. Reduced genetic variation and strong genetic population structure in the freshwater killifish Valencia letourneuxi (Valenciidae) based on nuclear and mitochondrial markers. Biological Journal of the Linnean Society 111: 334–349.

    Google Scholar 

  • Wandeler, P., P.E.A. Hoeck & L.F. Keller, 2007. Back to the future: museum specimens in population genetics. Trends in Ecology and Evolution 22: 634–642.

    PubMed  Google Scholar 

  • Ward, R. D., T. S. Zemlak, B. H. Innes, P. R. Last & P. D. N. Hebert, 2005. DNA barcoding Australia’s fish species. Philosophical Transactions of the Royal Society B: Biological Sciences 360: 1847–1857.

    CAS  Google Scholar 

  • Xia, X., 2013. DAMBE5: a comprehensive software package for data analysis in molecular biology and evolution. Molecular Biology and Evolution 30: 1720–1728.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Zalidis, G. & A. Mantzavelas, 1996. Inventory of Greek wetlands as natural resources. Wetlands 16: 548–556.

    Google Scholar 

  • Zardoya, R., P. S. Economidis & I. Doadrio, 1999. Phylogenetic relationships of Greek Cyprinidae: molecular evidence for at least two origins of the Greek cyprinid fauna. Molecular Phylogenetics and Evolution 13: 122–131.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

Field work to collect genetic material was conducted mainly within the framework of the Programme “Monitoring of ecological water quality of rivers, coastal and transitional waters of Greece” (Article 8 Directive 2000/60/ΕU), (2012–2015), funded by the European Regional Development Fund (ESPA). For field work and collection of specimens, a permit was secured by HCMR from the Ministry of Environment and Energy of Greece. We are grateful to Mrs. Margaret Eleftheriou for comments and linguistic editing of the manuscript. We also thank the two anonymous reviewers who improved the final version of this manuscript with their constructive criticism.

Author information

Authors and Affiliations

  1. Institute of Marine Biology, Biotechnology and Aquaculture, Hellenic Centre for Marine Research, Heraklion, Greece

    Dimitris Tsaparis & Costas S. Tsigenopoulos

  2. Department of Ichthyology and Aquatic Environment, School of Agricultural Sciences, University of Thessaly, Vólos, Greece

    Ioannis Konstantinidis

  3. First Zoological Department, Vienna Museum of Natural History, Vienna, Austria

    Anja Palandacic

  4. Institute of Marine Biological Resources and Inland Waters, Hellenic Centre for Marine Research, Anavissos, Greece

    Eleni Kalogianni, Maria Th. Stoumboudi, Roberta Barbieri, Leonidas Vardakas & Nicholas Koutsikos

Authors
  1. Dimitris Tsaparis
    View author publications

    Search author on:PubMed Google Scholar

  2. Ioannis Konstantinidis
    View author publications

    Search author on:PubMed Google Scholar

  3. Anja Palandacic
    View author publications

    Search author on:PubMed Google Scholar

  4. Eleni Kalogianni
    View author publications

    Search author on:PubMed Google Scholar

  5. Maria Th. Stoumboudi
    View author publications

    Search author on:PubMed Google Scholar

  6. Roberta Barbieri
    View author publications

    Search author on:PubMed Google Scholar

  7. Leonidas Vardakas
    View author publications

    Search author on:PubMed Google Scholar

  8. Nicholas Koutsikos
    View author publications

    Search author on:PubMed Google Scholar

  9. Costas S. Tsigenopoulos
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Dimitris Tsaparis.

Additional information

Handling editor: Christian Sturmbauer

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.

(XLSX 22 kb)

(DOCX 2365 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tsaparis, D., Konstantinidis, I., Palandacic, A. et al. DNA barcoding provides new insights on the distribution, systematics and conservation of the freshwater genus Pelasgus (Cypriniformes: Cyprinidae) in Greece. Hydrobiologia 848, 1163–1176 (2021). https://doi.org/10.1007/s10750-021-04526-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10750-021-04526-9

Keywords

Profiles

  1. Leonidas Vardakas View author profile