Isolation and characterization of 13 microsatellite DNA loci for the odorous frog Odorrana margaretae and O. graminea (A
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TECHNICAL NOTE
Isolation and characterization of 13 microsatellite DNA loci for the odorous frog Odorrana margaretae and O. graminea (Anura: Ranidae) Haiyan Gao • Liang Qiao • Weizhao Yang Jinzhong Fu
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Received: 29 March 2013 / Accepted: 6 May 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract Thirteen microsatellite DNA markers were developed from a DNA library of Odorrana margaretae. To characterize these loci, one population from O. margaretae (N = 24) and another from O. graminea (N = 42) were genotyped. All loci were variable, with the number of alleles ranging from 1 to 14 within populations. The average observed and expected heterozygosity per locus ranged from 0.00 to 0.83 and from 0.00 to 0.87, respectively. These markers are readily cross-amplified between the two species and are likely amplifiable in other species of the genus Odorrana. Odorous frogs have a very high level of cryptic species diversity, and we expect that these markers will offer useful tools in species delineation as well as for population genetic study in this ecologically important group. Keywords Odorous frogs Microsatellite DNA Species delineation Odorrana margaretae Odorrana graminea
The odorous frogs (genus Odorrana) are large high-gradient stream dwellers and are primarily distributed in tropical and subtropical Asia (Frost 2013). They form a major component of the forest ecosystem; however, the
H. Gao L. Qiao W. Yang J. Fu Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China H. Gao Graduate University of Chinese Academy of Sciences, Beijing 100049, China J. Fu (&) Department of Integrative Biology, University of Guelph, Guelph, ON N1G 2W1, Canada e-mail: [email protected]
species diversity of the group is likely underestimated to a great extent. For example, using a combination of morphometric and molecular data, Bain et al. (2003) discovered six new cryptic species from the northern Vietnamese populations, which were previously considered as part of a widespread species, O. livida. Stuart et al. (2006) further suggested that there is a high level of species diversity among odorous frogs and more cryptic species remain to be discovered. The lack of accurate estimates of local biodiversity significantly hinders the progress of prioritizing the conservation effort. Additionally, a northward distribution range shift has been observed in several Odorrana species (Fei et al. 2009), possibly as consequence of global warming. These frogs may serve as good environmental indicator species. Microsatellite DNA markers are highly variable and are particularly useful to identify newly established populations and their sources. Microsatellite DNA based methods have also been developed for species delineation (e.g. Noble et al. 2010). Here, we report 13 microsatellite DNA markers that are designed for two common odorous frogs, O. margaretae and O. graminea. A microsatellite-enriched library was constructed with the fast isolation by AFLP of sequences containing repeats (FIASCO) protoco
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