Development and characterization of microsatellite markers for the seagrass Halodule wrightii
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TECHNICAL NOTE
Development and characterization of microsatellite markers for the seagrass Halodule wrightii Patrick Larkin • Tabitha Schonacher Michael Barrett • Mindy Paturzzio
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Received: 30 November 2011 / Accepted: 1 December 2011 / Published online: 18 December 2011 Ó Springer Science+Business Media B.V. 2011
Abstract Halodule wrightii is a seagrass widely distributed in tropical areas, particularly in the western Atlantic. We have developed eight microsatellite markers from H. wrightii for use in genetic studies. The number of alleles ranged from 2 to 12 and the observed and expected heterozygosity ranged from 0.080 to 0.88, and from 0.077 to 0.86, respectively. No locus pair showed significant linkage disequilibrium or deviation from Hardy–Weinberg proportions after Bonferroni correction (P \ 0.05). These markers should prove useful for elucidating genetic structure, phylogenetic relationships, and ecological processes in this foundational coastal species. Keywords Halodule wrightii Microsatellites Seagrass Genetic variation
Seagrasses are aquatic angiosperms, monocots that colonized the marine environment some 100 million years ago (den Hartog 1970). They are especially noted for the number of ecosystem ‘‘services’’ they provide (Costanza et al. 1997), including primary production of organic nutrients, stabilization of near-shore environments and formation of habitat and nursery grounds for a number of species (Larkum et al. 2006). Like many ecosystems, seagrass-based communities face increasing fragmentation and degradation from anthropogenic sources (Duarte 2002; Orth et al. 2006). Conservation of seagrass ecosystems will
P. Larkin (&) T. Schonacher M. Barrett M. Paturzzio Department of Physical and Environmental Sciences, Texas A&M University-Corpus Christi, Corpus Christi, TX 78412, USA e-mail: [email protected]
require a multi-pronged approach that should incorporate genetic principles into any management plan. Halodule wrightii (synonymous with Halodule beaudettei) is a dioecious, generally perennial seagrass common throughout the Caribbean and Gulf of Mexico (Phillips and Menez 1988). It is the dominant seagrass species found in the Laguna Madre of Texas and Tamaulipas, the only hypersaline lagoon on the North American continent and the largest hypersaline system in the world (Tunnell and Judd 2002; Onuf 2007). Here we describe the development of 8 polymorphic microsatellite markers in H. wrightii. Genomic DNA was extracted from rhizome tissue collected near Corpus Christi, TX, USA using the DNeasy Plant Maxi Kit (Qiagen) according to the manufacturer’s instructions. DNA library construction, enrichment and screening were performed by Genetic Identification Services, Inc. as described previously (Jones et al. 2002). Genomic DNA was partially restricted with a cocktail of seven blunt-end cutting enzymes (RsaI, HaeIII, BsrB1, PvuII, StuI, ScaI, EcoRV). Fragments in the size range of 300–750 bp were adapted and subjected to magnetic bead capture (CPG), using biotinylated captur
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