Rapid development of 56 novel microsatellite markers for the benthic freshwater bug Aphelocheirus aestivalis using Illum
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SHORT COMMUNICATION
Rapid development of 56 novel microsatellite markers for the benthic freshwater bug Aphelocheirus aestivalis using Illumina paired‑end sequencing data and M13‑tailed primers Agnieszka Kaczmarczyk‑Ziemba1 Received: 11 September 2020 / Accepted: 3 November 2020 © The Author(s) 2020
Abstract The freshwater true bug Aphelocheirus aestivalis (Aphelocheiridae) is widely distributed in Europe but occurs rather locally and often in isolated populations. Moreover, it is threatened with extinction in parts of its range. Unfortunately, little is known about the genetic diversity and population structure due to the lack of molecular tools for this species. Thus, to overcome the limitations, a whole-genome sequencing has been performed to identify polymorphic microsatellite markers for A. aestivalis. The whole-genome sequencing has been performed with the Illumina MiSeq platform. Obtained paired-end reads were processed and overlapped into 2,378,426 sequences, and the subset of 267 sequences containing microsatellite motifs were then used for in silico primer designing. Finally, 56 microsatellite markers were determined and 34 of them were polymorphic. Analyses performed in two samples (collected from Drawa and Gowienica rivers, respectively) showed that the number of alleles per locus ranged from 2 to 21, and the observed and expected heterozygosity varied from 0 to 0.933 and 0.064 to 0.931, respectively. The microsatellite markers developed in the present study provide new suitable tools available for the scientific community to study A. aestivalis population dynamics. The assessment of its genetic diversity and population structure will provide important data, that can be used in population management and conservation efforts, elucidating the broad- and fine-scale population genetic structure of A. aestivalis. Keywords Aphelocheiridae · Freshwater true bug · High-throughput sequencing · Microsatellites · Illumina
Introduction The benthic freshwater true bug Aphelocheirus aestivalis (Fabricius 1794) and two endemic congeneric species from the Iberian Peninsula, A. murcius (Nieser and Millán 1989) and A. occidentalis (Nieser and Millán 1989) [1] are the only representatives of the family Aphelocheiridae in Europe. A. aestivalis is distributed throughout Europe [2], where lives in the middle and upper reaches of streams and rivers with β-mesosaprobic waters [3, 4]. The individuals of A. aestivalis occurring in Europe are predominantly micropterous [2]. Due to the fact that only a very few macropterous specimens capable of flight have been collected in the past [5], it has * Agnieszka Kaczmarczyk‑Ziemba agnieszka.kaczmarczyk‑[email protected] 1
Department of Genetics and Biosystematics, Faculty of Biology, University of Gdansk, Wita Stwosza 59, 80‑308 Gdansk, Poland
been assumed that A. aestivalis disperses mainly through the watercourses [6]. The remarkable downstream drift of nymphs and adults was observed e.g. in the Danube [6] and the Rhein rivers [5]. Although A. aestivalis is relatively toleran
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