Isolation and characterization of twelve novel microsatellite markers for the sea-bulrush ( Scirpus mariqueter Tang et W
- PDF / 170,069 Bytes
- 2 Pages / 595.276 x 790.866 pts Page_size
- 15 Downloads / 180 Views
MICROSATELLITE LETTERS
Isolation and characterization of twelve novel microsatellite markers for the sea-bulrush (Scirpus mariqueter Tang et Wang, Cyperaceae) Xinji Li • Yin Zhou • Yuguo Wang Wenju Zhang
•
Received: 26 February 2014 / Accepted: 24 March 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract Twelve microsatellite markers were isolated and characterized for the sea-bulrush (Scirpus mariqueter Tang et Wang), a vanward species endemic to coastal areas of East China. The number of alleles per locus ranged from 2 to 8. Observed and expected heterozygosities ranged from 0.125 to 0.694 and from 0.120 to 0.704, respectively. All microsatellites showed relatively high polymorphism, indicating potential usage of these microsatellite markers in population genetic assessment and molecular ecology studies of this species. Keywords Wetland Conservation Scripus mariqueter Microsatellites Population genetics Sea-bulrush (Scirpus mariqueter Tang et Wang) is a clonal herbaceous plant distributed only in salt marshes of Yangtze Estuary and Hangzhou Bay, East China, where is an important stopover place of migratory birds from Australia to East Asia. As a vanward species, it used to form vast meadows in the frontiers of coastal areas, depositing silt and providing food for migratory birds (Zhu et al. 2007). However, its distribution during last decade has been severely shrunk and fragmentized due to human activities and biological invasion (Chen et al. 2004). More and more academic concern have been placed on ecological values and conservation of this species, while there are still limited microsatellite markers available for molecular study. Electronic supplementary material The online version of this article (doi:10.1007/s12686-014-0188-6) contains supplementary material, which is available to authorized users. X. Li Y. Zhou Y. Wang W. Zhang (&) Institute of Biodiversity Science, School of Life Sciences, Fudan University, Shanghai 200433, China e-mail: [email protected]
To evaluate genetic potential and population dynamics of the sea-bulrush, we developed twelve novel polymorphic microsatellite markers from this species. Isolation of microsatellites followed the enrichment protocol of Glenn and Schable (2005). Genomic DNA from 12 S. mariqueter individuals was pooled to construct a microsatellite library. Total DNA, extracted from silicadried leaves using the modified cetyltrimethyl ammonium bromide (CTAB) method, was digested with RsaI and XmnI (New England Biolabs, Beijing, China) and ligated to SuperSNX24 double-stranded adaptors by T4 DNA ligase (TaKaRa Biotechnology Co., Dalian, Dalian, China). Polymerase chain reactions (PCR) were conducted with the adaptors as priming sites. PCR products were hybridized with six 50 -biotinylated probes (CT)13, (CCG)6, (AGG)6, (AAG)6, (ATC)6 and (AGC)6 and then enriched using streptavidin-coated magnetic beads (Invitrogen Dynal AS, Oslo, Norway). Captured gDNA was amplified with SNX24 primers, ligated to pMD18-T plasmid (TaKaRa) and transformed into
Data Loading...
