Abstract
The stellate sturgeon, Acipenser stellatus, is a critically endangered fish species. Knowledge on its genetic diversity and population structure is urgently needed to enable the identification of management units in order to prevent extinction. Therefore, 18 species-specific, polymorphic microsatellite loci have been isolated using GS-FLX Titanium pyrosequencing, arranged into 6 multiplex PCR sets, and characterized in 52 individuals (20 farmed and 32 wild). The total number of alleles per locus varied between 3 and 36 with an average of 8.44. The wild individuals were more diverse with an average number of 8.17 alleles per locus than the farmed ones with 3.28 alleles per locus. Observed heterozygosities ranged from 0.050 to 0.950 in the farmed and from 0.094 to 0.969 in the wild individuals. Significant deviations from Hardy-Weinberg equilibrium were found at 3 loci of the farmed and 5 loci of the wild individuals. The two sturgeon groups were significantly differentiated (F ST = 0.118). The high sensitivity and discriminatory power of the 18 loci were proven by a very low overall probability of identity for siblings (PIsib = 8.73 × 10−6) and a high accuracy of self-classification (98%). Thus, these newly developed markers represent a valuable genetic toolbox to identify management units for species conservation and sustainable fisheries.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Barmintseva AE, Mugue NS (2013) The use of microsatellite loci for identification of sturgeon species (Acipenseridae) and hybrid forms. Russ J Genet 49:950–961
Berg LS (1935) Summer and winter races in anadromous fish species (Sommer und Winterrassen bei den anadromen Fischen). Archiv für Naturgeschichte 4:376–403
Cornuet JM, Piry S, Luikart G, Estoup A, Solignac M (1999) New methods employing multilocus genotypes to select or exclude populations as origins of individuals. Genetics 153:1989–2000
DeHaan PW, Libants SV, Elliott RF, Scribner KT (2006) Genetic population structure of remnant Lake sturgeon populations in the Upper Great Lakes Basin. T Am Fish Soc 135:1478–1492
Goudet J (2002) FSTAT, a program to estimate and test gene diversities and fixation indices (version 2.9.3.2). (http://www2.unil.ch/popgen/softwares/fstat.htm)
Kaplinski L, Andreson R, Puurand T, Remm M (2005) MultiPLX: automatic grouping and evaluation of PCR primers. Bioinformatics 21:1701–1702
Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948
Liu ZJ, Cordes JF (2004) DNA marker technologies and their applications in aquaculture genetics. Aquaculture 238:1–37
Ludwig A, Belfiore NM, Pitra C, Svirsky V, Jeneckens I (2001) Genome duplication events and functional reduction of ploidy levels in sturgeon (Acipenser, Huso and Scaphirhynchus). Genetics 158:1203–1215
Malausa T, Gilles A, Meglécz E, Blanquart H, Duthoy S, Costedoat C, Dubut V, Pech N, Castagnone-Sereno P, Délye C, Feau N, Frey P, Gauthier P, Guillemaud T, Hazard L, Le Corre V, Lung-Escarmant B, Malé P-J, Ferreira S, Martin J-F (2011) High-throughput microsatellite isolation through 454 GS-FLX Titanium pyrosequencing of enriched DNA libraries. Mol Ecol Resour 11:638–644
Meglécz E, Costedoat C, Dubut V, Gilles A, Malausa T, Pech N, Martin J-F (2010) QDD: a user-friendly program to select microsatellite markers and design primers from large sequencing projects. Bioinformatics 26:403–404
O’Leary SJ, Dunton KJ, King TL, Frisk MG, Chapman DD (2014) Genetic diversity and effective size of Atlantic sturgeon, Acipenser oxyrhinchus oxyrhinchus river spawning populations estimated from the microsatellite genotypes of marine-captured juveniles. Conserv Genet 15:1173–1181
Qiwei W (2010) Acipenser stellatus. The IUCN Red List of Threatened Species 2010: e.T229A13040387. https://doi.org/10.2305/IUCN.UK.2010-1.RLTS.T229A13040387.en. Downloaded on 02 January 2017
Rajkov J, Shao Z, Berrebi P (2014) Evolution of ploidy and functional diploidization in sturgeons: microsatellite analysis in 10 sturgeon species. J Hered 105:521–531
Rousset F (2008) GENEPOP’007: a complete re-implementation of the GENEPOP software for Windows and Linux. Mol Ecol Res 8:103–106
Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol 132:365–386
Van Oosterhout C, Hutchinson WF, Wills DPM, Shipley P (2004) MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data. Mol Ecol Notes 4:535–538
Waits LP, Luikart G, Taberlet P (2001) Estimating the probability of identity among genotypes in natural populations: cautions and guidelines. Mol Ecol 10:249–256
Wilberg MJ, Dreher BP (2004) GENECAP: a program for analysis of multilocus genotype data for non-invasive sampling and capture-recapture population estimation. Mol Ecol Notes 4:783–785
Acknowledgments
We thank Mr. Peter Groß, fish farm “Rhönforelle,” Gersfeld, Germany, for generously providing access to the offspring of the ex situ stocks for sampling. Marian Paraschiv, Ștefan Honț, and Marian Iani of the DDNI Sturgeon Research Group collected most of the tissue samples from the Danube River in Romania.
Funding
This project was sponsored by the COFASP (Cooperation in Fisheries, Aquaculture and Seafood Processing) ERA-NET partners, which has received funding from the European Union’s Seventh Framework Programme for Research, Technological Development and Demonstration under grant agreement no. 321553. Financial support was provided by the German Federal Ministry of Food and Agriculture (BMEL) through the Federal Office for Agriculture and Food (BLE), grant number 2814ERA01G, and the Romanian Executive Agency for Higher Education, Research, Development and Innovation Funding (UEFISCDI), contract number 26/2015.
Data availability
DNA sequences of the 18 suitable microsatellite loci were submitted to GenBank (accession numbers KY496652-KY496669).
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Table S1
(DOCX 19 kb)
Rights and permissions
About this article
Cite this article
Kohlmann, K., Kersten, P., Geßner, J. et al. Isolation and characterization of 18 polymorphic microsatellite markers for the critically endangered stellate sturgeon, Acipenser stellatus . Eur J Wildl Res 63, 75 (2017). https://doi.org/10.1007/s10344-017-1132-3
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s10344-017-1132-3