Isolation by 454-sequencing and characterization of polymorphic microsatellite markers in the tetraploid perennial herb
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MICROSATELLITE LETTERS
Isolation by 454-sequencing and characterization of polymorphic microsatellite markers in the tetraploid perennial herb Campanula rotundifolia Jan Plue • Katrien Vandepitte • Olivier Honnay Sara A. O. Cousins
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Received: 16 January 2015 / Accepted: 14 February 2015 Ó Springer Science+Business Media Dordrecht 2015
Abstract Campanula rotundifolia is a perennial polyploid herb with a wide circumpolar distribution growing in a range of grassland, heathland and dune communities. The species is showing signs of strong decline in most parts of its range due to habitat degradation. Eight polymorphic microsatellites were developed using a 454-sequencing approach and subsequently characterized in 323 tetraploid individuals from 15 populations in a fragmented landscape in central Sweden. Between 7 and 26 alleles were observed per locus with observed and expected heterozygosity ranging between 0.71–0.98 and 0.71–0.93, respectively. C. rotundifolia may serve as a model species for studying the relative importance of life-history traits in genetic diversity responses to environmental changes. Keywords Environmental changes Microsatellites Polyploidy Traits Campanula rotundifolia L., common harebell, is an evergreen, perennial and self-incompatible herb, capable of limited clonal growth. Its wide ecological amplitude allows growth in grassland, heath and dune communities across its circumpolar boreo-temperate distribution (Stevens et al. 2012). C. rotundifolia has a complex cytotype distribution,
Electronic supplementary material The online version of this article (doi:10.1007/s12686-015-0443-5) contains supplementary material, which is available to authorized users. J. Plue (&) S. A. O. Cousins Department of Physical Geography, Stockholm University, Stockholm, Sweden e-mail: [email protected] K. Vandepitte O. Honnay Department of Biology, KU Leuven, Heverlee, Belgium
dominated by diploid, tetraploid and hexaploid forms, with central Swedish populations being strictly tetraploid (Bo¨cher 1966). C. rotundifolia forms remnant populations in response to habitat deterioration (Lindborg et al. 2005) and possesses a persistent seed bank (Plue and Cousins 2013), allowing for quick recovery following habitat restoration (Lindborg et al. 2005). The species is in strong decline across Europe (Stevens et al. 2012), with the species equally being listed as vulnerable and endangered in New York and Ohio (USA), respectively (http://plants. usda.gov). C. rotundifolia is an interesting model species to study the relative importance of its various life-history traits in mediating the genetic responses to environmental changes. For the detailed species ecology, see Stevens et al. (2012). Fifteen fresh leaf samples from 15 populations were sent to ecogenics GmbH to produce an enriched microsatellite library and test for polymorphism. Size-selected fragments from genomic DNA were enriched for SSR content by using magnetic streptavidin beads and biotin-labeled CT and GT repeat oligonucleotides. The SSR-enric
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