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Int. J. Mol. Sci. 2012, 13(8), 9893-9899; doi:10.3390/ijms13089893
Article

Development of Microsatellite Markers Using Pyrosequencing in Galium trifidum (Rubiaceae), a Rare Species in Central Europe

1,* , 2
, 1
, 2
, 3
 and 4
1 Department of Botany and Nature Protection, University of Warmia and Mazury in Olsztyn, Plac Łódzki 1, Olsztyn 10-727, Poland 2 Department of Genetics, University of Silesia, Jagiellońska 28, Katowice 40-032, Poland 3 Department of Genetics, University of Warmia and Mazury in Olsztyn, Oczapowskiego 1A, Olsztyn 10-719, Poland 4 Regional Directorate of Environmental Protection in Olsztyn, Dworcowa 60, Olsztyn 10-437, Poland
* Author to whom correspondence should be addressed.
Received: 9 July 2012 / Revised: 23 July 2012 / Accepted: 25 July 2012 / Published: 8 August 2012
(This article belongs to the Section Bioinorganic Chemistry)
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Abstract

We identify a large number of microsatellites from Galium trfidum, a plant species considered rare and endangered in Central and Western Europe. Using a combination of a total enriched genomic library and small-scale 454 pyrosequencing, we determined 9755 contigs with a length of 100 to 6192 bp. Within this dataset, we identified 153 SSR motifs in 144 contigs. Here, we tested 14 microsatellite loci in 2 populations of G. trifidum. The number of alleles and expected heterozygosity were 1–8 (mean 3.2) and 0.00–0.876 (0.549 on average), respectively. The markers described in this study will be useful for evaluating genetic diversity within and between populations, and gene flow between G. trifidum populations. These markers could also be applied to investigate the biological aspects of G. trifidum, such as the population dynamics and clonal structure, and to develop effective conservation programs for the Central European populations of this species.
Keywords: SSR markers; Galium trifidum; 454 pyrosequencing SSR markers; Galium trifidum; 454 pyrosequencing
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Szczecińska, M.; Kwaśniewski, M.; Sawicki, J.; Chwiałkowska, K.; Szandar, K.; Pisarek, W. Development of Microsatellite Markers Using Pyrosequencing in Galium trifidum (Rubiaceae), a Rare Species in Central Europe. Int. J. Mol. Sci. 2012, 13, 9893-9899.

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