Development and characterization of polymorphic microsatellite loci in Ficus altissima
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MICROSATELLITE LETTERS
Development and characterization of polymorphic microsatellite loci in Ficus altissima Yi-Ping Ma • Yi-Fan Kong • Yawen Cao Yi-Su Shi
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Received: 23 September 2014 / Accepted: 27 September 2014 / Published online: 8 October 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract Fig trees are keystone resource species in tropical forests. In this study we developed microsatellite loci for Ficus altissima, a common dominant species in tropical China, using an enrichment approach. Nine polymorphic loci were developed, and their characteristics were analyzed in a population. Number of alleles per locus ranged from 2 to 6. The expected and observed heterozygosities were 0.108–0.740 and 0.111–0.800, respectively. These polymorphic microsatellites can be applied to further studies about genetic diversity and structure of F. altissima populations. Keywords Ficus altissima Genetic diversity Microsatellites Tropical rainforests support the richest biodiversity among terrestrial ecosystems, but suffering from rapid loss of biodiversity due to human disturbance. Fig trees are keystone resources in tropical forests, playing a critical role in biodiversity conservation in the tropics. Ficus altissima is a common dominant tree in tropical and subtropical China, but threatened by habitat alternations and transplanting for ornamental aims. Population genetic data can provide
Y.-P. Ma Y.-F. Kong School of Ecology and Environmental Sciences, East China Normal University, Shanghai 200241, China Y. Cao The Hun School of Princeton, Princeton, NJ 08540, USA Y.-S. Shi (&) School of Environmental and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang, Jiangsu 212003, China e-mail: [email protected]
important information in designing conservation strategies, and microsatellites are among the most widely used molecular markers. Polymorphic microsatellite markers had been developed in some Ficus species, e.g., F. pumila (Zhang et al. 2011). Amplification of existing primers from species of the same genus is a rapid and cheap approach to obtain microsatellite loci. However, null alleles have been frequently detected using cross-amplification of microsatellite markers from related species, and may lead to bias in population genetic studies. In this study, we developed polymorphic microsatellite loci using an enrichment approach (Liu et al. 2009). The total genomic DNA was extracted silica gel-dried leaves using the Plant Genomic DNA Kit (Tiangen, China). The microsatellites were developed following the protocol of Tong et al. (2012). Approximately 250 ng DNA was digested by the restriction enzyme Mse I, and then was linked to an Mse I-adapter pair (F: 50 -TACTCAGGACTCAT-30 , R: 50 -GACGATGAGTCCTGAG-30 ). After amplified with Mse I–N primer (50 -GATGAGTCCTGAG TAAN-30 ), the hybridization and enrichment were hybridized with 50 -biotinylated probe (AG)15, and then the streptavidin-coated magnetic beads (Promega, Madison, Wisconsin, USA) were used to capture the hybridized produc
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