Genetic variability of the endangered fish lake minnow, Eupallasella percnurus (Pall.) in populations newly established by translocations in Poland
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1
University of Warmia and Mazury in Olsztyn, Poland
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2
Stanisław Sakowicz Inland Fisheries Institute, Poland
Introduction
In Poland cyprinid lake minnow, Eupallasella percnurus (Pall.), is a threatened with extinction native fish species. It was covered with strict protection with the requirement of active protection in 2004. The first protective undertakings took place in 2004-2009. All of them were aimed at the initiation of new lake minnow populations by single or repeated translocations of juvenile fish at the age of 0+. Juveniles, offspring of maternal fish originated from a local population, were obtained using standard procedures of reproduction and rearing (Kamiński et al. 2004, Wolnicki et al. 2004). The fish were translocated to 12 small water bodies where lake minnow did not occur earlier. Out of all the attempts five proved to be successful so far. The aim of the present work was to compare the basic parameters of the genetic variability of 5 new lake minnow populations, 4-8 years after the final fish translocation, with the respective data for 13 populations existing over several decades.
Materials and methods
Adult fish from 18 lake minnow populations from throughout the species' entire Polish range were caught in baited traps. From each population, 48 fish were taken randomly. Material for genetic analyses were small (20 mm2) fragments of the fishes’ fins. The samples were preserved by air drying (Kaczmarczyk and Wolnicki 2016).
Genomic DNA was extracted from fin tissues using Genomic Mini AX Tissue SPIN DNA Extraction and Purification Kit (A&A Biotechnology Poland). The extraction procedure followed manufacturer’s recommendation.
The assessment of genetic variation was based on 13 polymorphic microsatellites. They were Z9878, 10362, 13419, Ca3, 4, 12, and Eupe 1, 2, 4, 5, 6, 9.
The forward primer of each primer pair was 5’ end labelled with fluorescent dyes (6FAM, VIC, NED, and PET). The composition of the PCR mixture and thermal profiles of the PCR reaction followed the method by Kaczmarczyk and Wolnicki (2016).
The lengths of amplified DNA fragments were determined using an Applied Biosystems 3130 Genetic Analyser against GS500LIZ size standard. Fragment size and allele determination was performed by using GeneMapper 3.0 software (Applied Biosystems), following the manufacturer’s recommendations.
In each population the genetic diversity was evaluated, based on the observed heterozygosity (Ho) and expected heterozygosity (He) at given locus and across loci. An average number of alleles across a loci (AN) was determined. All calculations were computed by MSA software (Dieringer and Schlötterer 2003).
The Exact Hardy Weinberg (H-W) test (Guo and Thompson 1992) was used to test a deviation from H-W equilibrium. Those calculations were performed by Arlequin 3.0 software (Excoffier et al. 2005).
The likely occurrence of a bottleneck or founder effect, and their influence on within-population genetic variability was based on the Garza-Williamson M index (Garza and Williamson 2001) including Excoffier’s adjustment (Excoffier et al. 2005).
The values of indicators of the genetic variation (Ho, He, AN), and M value of Garza-Williamson index were compared between the group of new lake minnow populations and the populations existing over decades. The significance of the differences between these groups were tested by using Kruskal-Wallis test.
Results
The values of all indicators of genetic variation (Ho, He, and AN) were higher in group of new populations than in old (Table 1). The differences were significant at p=0.007 (Ho), p= 0.004 (He) and p=0.009 (AN). The populations remained at H-W equilibrium; deviations occurred only at several loci in some populations.
The M value of G-W index was higher in group of new populations than in old and the differences were significant at p=0.031.
Acknowledgements
The present studies were carried out within the project No. 2014/15/B/NZ9/05240 granted by the National Science Centre (Poland) for years 2015-2019.
References
Dieringer D., Schlötterer C. 2003. Microsatellite analyzer (MSA): a platform independent analysis tool for large microsatellite data sets. Mol. Ecol. Not. 3: 167-169.
Excoffier L., Laval G., Schneider S. 2005. Arlequin ver. 3.0: An integrated software packane for population genetics data analysis. Evol. Bioinform. On. 1: 47-50.
Garza J.C., Williamson E.G. 2001. Detection of reduction in population size using data from microsatellite loci. Mol. Ecol. 10: 305-318.
Guo S.W., Thompson E.A. 1992. Performing the exact test of Hardy-Weinberg proportion for multiple alleles. Biometrics, 48: 361-372.
Kaczmarczyk D., Wolnicki J. 2016. Genetic diversity of the endangered cyprinid fish lake minnow Eupallasella percnurus in Poland and its implications for conservation. PLoS One 12: 1-16.
Kamiński R., Kusznierz J., Myszkowski L., Wolnicki J. 2004. The first attempt to artificially reproduce the endangered cyprinid lake minnow Eupallasella perenurus (Pallas). Aquacult. Int. 12: 3-10.
Wolnicki J., Kamiński R., Korwin-Kossakowski M., Kusznierz J., Myszkowski L. 2004. The influence of water temperature on laboratory-reared lake minnow Eupallasella perenurus (Pallas) larvae and juveniles. Arch. Pol. Fish. 12: 61-69.
Keywords:
lake minnow,
Active Protection,
initiation of new populations,
fish translocations,
Genetic Variation
Conference:
XVI European Congress of Ichthyology, Lausanne, Switzerland, 2 Sep - 6 Sep, 2019.
Presentation Type:
Poster
Topic:
THREATS AND CONSERVATION
Citation:
Kaczmarczyk
D,
Wolnicki
J,
Kamiński
R,
Sikorska
J and
Radtke
G
(2019). Genetic variability of the endangered fish lake minnow, Eupallasella percnurus (Pall.) in populations newly established by translocations in Poland.
Front. Mar. Sci.
Conference Abstract:
XVI European Congress of Ichthyology.
doi: 10.3389/conf.fmars.2019.07.00163
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Received:
29 Jul 2019;
Published Online:
14 Aug 2019.
*
Correspondence:
Mx. Dariusz Kaczmarczyk, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland, d.kaczmarczyk@uwm.edu.pl