Development and characterization of thirty novel microsatellite markers for the critically endangered Myanmar Roofed Tur
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TECHNICAL NOTE
Development and characterization of thirty novel microsatellite markers for the critically endangered Myanmar Roofed Turtle, Batagur trivittata, and cross-amplification in the Painted River Terrapin, B. borneoensis, and the Southern River Terrapin, B. affinis, using paired-end Illumina shotgun sequencing Cara N. Love • Cris Hagen • Brian D. Horne Kenneth L. Jones • Stacey L. Lance
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Received: 20 September 2012 / Accepted: 29 October 2012 Ó Springer Science+Business Media Dordrecht 2012
Abstract We isolated and characterized 30 microsatellite loci from the critically endangered Myanmar Roofed Turtle, Batagur trivittata. Loci were screened in 9 B. trivittata samples and in the congeners the Painted River Terrapin, Batagur borneoensis, with 22 of 30 amplifying, and the Southern River Terrapin, B. affinis, with 15 of 30 amplifying. In the B. trivittata samples, the number of alleles per locus ranged from 3 to 10 and the probability of identity values ranged from 0.031 to 0.354. These new loci will provide tools for captive management and reintroduction programs for B. trivittata and the other five species of Batagur, particularly B. affinis in Cambodia. Keywords Batagur Illumina Microsatellite PAL_FINDER PCR primers SSR
The genus Batagur (comprising 6 species) is one of two of the most critically endangered turtle genera in the world (Turtle Conservation Coalition 2011). Within the genus, the Myanmar Roofed Turtle, B. trivittata, is considered one of the most endangered vertebrates. Undetected by scientists since the 1930s, B. trivittata was feared extinct until 3 C. N. Love C. Hagen S. L. Lance (&) Savannah River Ecology Laboratory, University of Georgia, Drawer E, Aiken, SC 29802, USA e-mail: [email protected] B. D. Horne Wildlife Conservation Society, 2300 Southern Blvd Bronx, New York, NY 10460, USA K. L. Jones Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, Denver, CO 80045, USA
individuals (2 females and 1 male) were found in 2002 in a temple pond near the city of Mandalay in central Myanmar. Since that time, surveys have revealed two small wild populations with \10 nesting females remaining in the wild. By protecting nests in the field over a 10-year period as well as modest captive breeding, more than 500 juvenile B. trivittata are being head-started in captivity. Similarly, the Southern River Terrapin, B. affinis, was thought to be extinct within Cambodia, until a small population was found in the Sre Ambel River of Southern Cambodia. This population of \10 nesting females is isolated from declining populations in Malaysia and perhaps extirpated from Thailand, Myanmar, and Vietnam; and represents important genetic variation within a species, which may number \1,000 in the wild. A nest protection program has resulted in over 150 B. affinis from the Sre Ambel River now being head-started. Yet, when managing small populations, it is crucial to maximize genetic diversity of both the founder animals in captive assurance colonies a
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