Have a personal or library account? Click to login
New multiplex PCR assays for estimating genetic diversity in European grayling (Thymallus thymallus) by polymorphism of microsatellite DNA Cover

New multiplex PCR assays for estimating genetic diversity in European grayling (Thymallus thymallus) by polymorphism of microsatellite DNA

Open Access
|Jan 2026

References

  1. Akimov, I. 2009. The Red Data Book of Ukraine. Animals. Global Consulting, Kyiv, 1-486.
  2. Brown, S.S., Chen, Y-W., Wang, M., Clipson, A., Ochoa, E., Du, M-Q. (2017). PrimerPooler: automated primer pooling to prepare library for targeted sequencing. Biology Methods and Protocols. Oxford University Press. 2(1).
  3. Brzuzan, P., Jurczyk, Ł. (2003). Analysis of mtDNA sequences of European grayling, Thymallus thymallus, from southwestern Poland. Archives of Polish Fisheries, 11, 301-306.
  4. Butler, J.M., Ruitberg, C.M., Vallone, P.M. (2001). Capillary electrophoresis as a tool for optimization of multiplex PCR reactions. Fresenius Journal of Analytical Chemistry, 369(3), 200-205.
  5. Dieringer, D., Schlötterer, C. (2003). Microsatellite analyser (MSA): a platform independent analysis tool for large microsatellite data sets. Molecular Ecology Notes, 3(1), 167-169.
  6. Duftner, N., Koblmüller, S., Weiss, S., Medgyesy, N., Sturmbauer, C. (2005). The impact of stocking on the genetic structure of European grayling Thymallus thymallus, Salmonidae) in two alpine rivers. Hydrobiologia, 542(1), 121-129.
  7. Edwards, M.C., Gibbs, R.A. (1994). Multiplex PCR: advantages, development, and applications. PCR Methods and Applications, 3(4), S65-S75.
  8. Excoffier, L., Lischer, H.E. (2010). Arlequin suite ver 3.5: a new series of programs to perform population genetics analyses under Linux and Windows. Molecular Ecology Resources, 10(3), 564-567.
  9. Fishback, A.G., Danzmann, R.G., Sakamoto, T., Ferguson, M.M. (1999). Optimization of semi-automated microsatellite multiplex polymerase chain reaction systems for rainbow trout (Oncorhynchus mykiss). Aquaculture, 172(3-4), 247-254.
  10. Ford, M. (2024). Thymallus thymallus. The IUCN Red List of Threatened Species 2024: e.T266178365A135093349. https://dx.doi.org/10.2305/IUCN.UK.2024-2.RLTS.T266178365A135093349.en.
  11. Garza, J.C., Williamson, E.G. (2001). Detection of reduction in population size using data from microsatellite loci. Molecular Ecology,10(2), 305-318.
  12. Haddeland, P.J., Junge, C., Serbezov, D., Vrllestad, L.A. (2015). Genetic parentage analysis confirms a polygynandrous breeding system in the European grayling (Thymallus thymallus). PloS one, 10(3), e0122032.
  13. He, L., Kierzek, R., SantaLucia, Jr J., Walter, A.E., Turner, D.H. (1991). Nearest-neighbor parameters for G.U mismatches. Biochemistry, 30, 11124-11132.
  14. HELCOM, (2013). HELCOM Red List of Baltic Sea species in danger of becoming extinct. Balt. Sea Environment Proceedings, 140, 1-72.
  15. Henegariu, O., Heerema, N.A., Dlouhy, S.R., Vance, G.H., Vogt, P.H. (1997). Multiplex PCR: critical parameters and step-by-step protocol. Biotechniques, 23(3), 504-511.
  16. Junge, C., Primmer, C.R., Vrllestad, L.A., Leder, E.H. (2010). Isolation and characterization of 19 new microsatellites for European grayling, Thymallus thymallus (Linnaeus, 1758), and their cross-amplification in four other salmonid species. Conservation Genetics Resources, 2(Suppl 1), 219-223.
  17. Jurczyk, £. (2006). The genetic characteristics of selected grayling populations (Thymallus thymallus Linnaeus, 1758) based on the molecular markers. PhD thesis, University of Warmia and Mazury (in Polish).
  18. Kaczmarczyk, D., Wolnicki, J. (2020). Genassemblage 2.0 software facilitates conservation of genetic variation of captively propagated species. Scientific Reports, 10(1), 17871.
  19. Kaczmarczyk, D., Wolnicki, J. (2022). Management of genetic variation in the gamete bank of the endangered lake minnow Eupallasella percnurus, using Genassemblage 2.2 software. Animals, 12(23), 3329.
  20. Kim, J.E., Withler, R.E., Ritland, C., Cheng, K.M. (2004). Genetic variation within and between domesticated chinook salmon, Oncorhynchus tshawytscha, strains and their progenitor populations. Environmental Biology of Fishes, 69(1), 371-378
  21. Koskinen, M.T., Primmer, C.R. (2001). High throughput analysis of 17 microsatellite loci in grayling (Thymallus spp. Salmonidae). Conservation Genetics, 2(2), 173-177.
  22. Koskinen, M.T., Sundell, P., Piironen, J., Primmer, C.R. (2002). Genetic assessment of spatiotemporal evolutionary relationships and stocking effects in grayling (Thymallus thymallus, Salmonidae). Ecology Letters, 5(2), 193-205.
  23. Lerceteau-Köhler, E., Weiss, S. (2006). Development of a multiplex PCR microsatellite assay in brown trout Salmo trutta, and its potential application for the genus. Aquaculture, 258, 641-645.
  24. Lyach, R., Remr, J. (2019). The effect of a large-scale fishing restriction on angling harvest: a case study of grayling Thymallus thymallus in the Czech Republic. Aquatic Living Resources, 32, 11.
  25. Mallet, J.P., Lamouroux, N., Sagnes, P., Persat, H. (2000). Habitat preferences of European grayling in a medium size stream, the Ain River, France. Journal of Fish Biology, 56(6), 1312-1326.
  26. Marić, S., Razpet, A., Nikolić, V., Simonović, P. (2011). Genetic differentiation of European grayling (Thymallus thymallus) populations in Serbia, based on mitochondrial and nuclear DNA analyses. Genetics Selection Evolution, 43(1), 2, http://www.gsejournal.org/content/43/1/2
  27. Marsh, J.E., Cove, R.J., Britton, J.R., Wellard, R.G., House, A., Gregory, S.D. (2021). Medium-term environmental changes influence age-specific survival estimates in a salmonid population. Freshwater Biology, 66(8), 1530-1545.
  28. Mruk, A., Kucheruk, A., Bielikova, O., Èiampor Jr, F. (2024). Current state of the European grayling (Thymallus thymallus L.) in Ukraine and conservation strategy for population support. A review. Journal for Nature Conservation, 81, 126676.
  29. Nei, M. (1987). Molecular Evolutionary Genetics. Columbia University Pres, New York, USA.
  30. Nielsen, J.L., Sage G.K. 92001). Microsatellite analyses of the trout of northwest Mexico. Genetica, 111, 269-27.
  31. Olsen, J.B., Bentzen, P., Seeb, J.E. (1998). Characterization of seven microsatellite loci derived from pink salmon. Molecular Ecology, 7, 1087-1089.
  32. Pastor, T., Garza, J. C., Allen, P., Amos, W., Aguilar, A. (2004). Low genetic variability in the highly endangered Mediterranean monk seal. Journal of Heredity, 95(4), 291-300.
  33. Persat, H. (1996). Threatened populations and conservation of the European grayling, Thymallus thymallus (L., 1758). In: Conservation of endangered fishes of Europe (Ed.) Kirchhofer A, Hefti D., Birkhäuser Verlag, Basel: 233-247.
  34. Rai, A. J., Udar, N., Saad, R., Fleisher, M. (2009). A multiplex assay for detecting genetic variations in CYP2C9, VKORC1, and GGCX involved in warfarin metabolism. Clinical Chemistry, 55(4), 823-826.
  35. Rich, J., Cove, R.J., Taylor, R.G. (2018). European grayling conservation, ecology & management: a practical conservation guide for the United Kingdom. Grayling Research Trust, UK.
  36. Shrimpton, J.M., Clarke, A.D. (2012). Genetic analysis of Arctic grayling population structure in the Williston Water-shed: samples from the Finlay River. Fish and Wildlife Compensation Program – Peace Region Report No. 354.
  37. Stamford, M.D., Taylor, E.B. (2005). Population subdivision and genetic signatures of demographic changes in Arctic grayling (Thymallus arcticus) from an impounded water-shed. Canadian Journal of Fisheries and Aquatic Sciences, 62(11), 2548-2559.
  38. Sušnik S., Snoj A., Dovč P. (1999). Microsatellites in grayling (Thymallus thymallus): comparison of two geographically remote populations from the Danubian and Adriatic river basin in Slovenia. Molecular Ecology, 8: 1756-1758.
  39. Sušnik, S., Berrebi, P., Dovč, P., Hansen, M.M., Snoj, A. (2004). Genetic introgression between wild and stocked salmonids and the prospects for using molecular markers in population rehabilitation: the case of the Adriatic grayling (Thymallus thymallus L. 1785). Heredity, 93(3), 273-282.
  40. Swatdipong, A., Vasemägi, A., Koskinen, M.T., Piironen, J., Primmer, C.R. (2009). Unanticipated population structure of European grayling in its northern distribution: implications for conservation prioritization. Frontiers in Zoology, 6(1), 6, doi:10.1186/1742-9994-6-6
  41. Weiss, S.J., Gonçalves, D.V., Secci-Petretto, G., Englmaier, G.K., Gomes-Dos-Santos, A., Denys, G.P., ... & Froufe, E. (2021). Global systematic diversity, range distributions, conservation and taxonomic assessments of graylings (Teleostei: Salmonidae; Thymallus spp.). Organisms Diversity & Evolution, 21(1), 25-42.
  42. Zemanová, B., Hájková, P., Hájek, B., Martínková, N., Mikulíček, P., Zima, J., Bryja, J. (2015). Extremely low genetic variation in endangered Tatra chamois and evidence for hybridization with an introduced Alpine population. Conservation Genetics, 16(3), 729-741.
DOI: https://doi.org/10.2478/aopf-2025-0014 | Journal eISSN: 2545-059X | Journal ISSN: 2545-0255
Language: English
Page range: 153 - 165
Submitted on: Nov 8, 2024
|
Accepted on: Nov 4, 2025
|
Published on: Jan 18, 2026
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year

© 2026 Dariusz Kaczmarczyk, Anna Nitkiewicz, Martyna Gadomska, published by Stanisław Sakowicz Inland Fisheries Institute
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.