Isolation and characterization of novel microsatellite markers in endangered mahseer, Tor putitora (Hamilton 1822) (Fami
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TECHNICAL NOTE
Isolation and characterization of novel microsatellite markers in endangered mahseer, Tor putitora (Hamilton 1822) (Family: Cyprinidae, Pisces) Prabhati Kumari Sahoo • Rohit Kumar • Jyoti Sati • Rakesh Matura • Ashoktaru Barat
Received: 20 February 2013 / Accepted: 8 March 2013 / Published online: 14 March 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract A set of 12 microsatellites was developed for a freshwater fish golden mahseer, Tor putitora. Golden mahseer is an endangered and endemic in mid-hill regions of South-Asian countries. Constructing a partial genomic library, we obtained 44 clones containing putative microsatellite motifs and 12 microsatellite with enough flanking sequence for primer designing. The number of alleles per locus ranged from 3 to 8 (mean 5.41), with a mean expected and observed heterozygosity of 0.68 and 0.66, respectively. Most of them were in Hardy–Weinberg equilibrium except TPM20a. There was no significant evidence for linkage disequilibrium between any pair of loci. These microsatellites were highly polymorphic (mean PIC 0.68) and will be useful for genetic variability and conservation genetics studies. Keywords Microsatellite markers Partial genomic library Conservation genetics Tor putitora
Tor putitora also known as Golden Mahseer belongs to the family Cyprinidae and is distributed through mid-hill regions of South-Asian countries (Mohindra et al. 2007; Naeem et al. 2011; Hussain 2012). But this species is declining in several water bodies and presently ranked as endangered (IUCN 2012). Identification and selection of better brood stocks using modern genetic tools such as microsatellite markers can speed up the species recovery process with the knowledge of stock structure and genetic
P. K. Sahoo R. Kumar J. Sati R. Matura A. Barat (&) Molecular Genetics Laboratory, Directorate of Coldwater Fisheries Research, Indian Council of Agricultural Research, Bhimtal-263136, Nainital, Uttarakhand, India e-mail: [email protected]
variability of the species. The microsatellite is a powerful co-dominant genetic marker that has been applied to the classification and systematics (Amado et al. 2011), parentage identification (Norris et al. 2000), germplasm conservation (Fopp-Bayat and Ciereszko 2012) etc. Identification of polymorphic microsatellites markers is prerequisite to generate genetic data for proper management in fisheries (Ferguson et al. 1995). Hence, we developed and characterized few highly polymorphic microsatellite loci of T. putitora by constructing a partial genomic library. Fin tissues were collected by non-invasive technique (Wasko et al. 2003) followed by proper treatment of organism before they were reverted to their habitat. Total genomic DNA was extracted from fin tissues using standard protocol of Sambrook and Russell (2001). After the qualitative assay and quantification, 10 lg of genomic DNA was digested with Mbo I (Fermentas, USA) and separated on a 1.2 % agarose gel. With the aid of U 9 174 DNA/HinfI marker (Fermenta
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