Development and characterization of 27 new microsatellite markers for the Indian bullfrog <i>Hoplobatrachus tigerinus</i> and its congeneric species

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  • Sultana Nasrin
    Institute for Amphibian Biology, Graduate School of Science, Hiroshima University
  • Igawa Takeshi
    Institute for Amphibian Biology, Graduate School of Science, Hiroshima University Division of Development Science, Graduate School of International Development and Cooperation, Hiroshima University
  • Nozawa Masafumi
    Center for Information Biology, National Institute of Genetics Department of Genetics, The Graduate University for Advanced Studies (Sokendai)
  • Islam Mohammed Mafizul
    Institute for Amphibian Biology, Graduate School of Science, Hiroshima University
  • Hasan Mahmudul
    Institute for Amphibian Biology, Graduate School of Science, Hiroshima University
  • Alam Mohammad Shafiqul
    Department of Genetics and Fish Breeding, Faculty of Fisheries, Bangabandhu Sheikh Mujibur Rahman Agricultural University
  • Khan Md. Mukhlesur Rahman
    Department of Fisheries Biology and Genetics, Faculty of Fisheries, Bangladesh Agricultural University
  • Sumida Masayuki
    Institute for Amphibian Biology, Graduate School of Science, Hiroshima University

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  • Development and characterization of 27 new microsatellite markers for the Indian bullfrog Hoplobatrachus tigerinus and its congeneric species

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Abstract

The Indian bullfrog Hoplobatrachus tigerinus and its four congeneric species are common frog species and distributed throughout South Asia. Due to recent human activity, they are facing a changing environment and reduction in natural population size. For effective conservation and molecular ecological studies, we therefore isolated and characterized microsatellite loci for these frogs. We obtained genomic data using an Ion Torrent PGM sequencer and designed 54 primer sets for candidate loci. By screening for polymorphic loci in individuals of H. tigerinus and its congeneric species, we isolated 27 loci as highly polymorphic microsatellite loci. Eight of these loci were commonly applicable for all species except H. chinensis. Within two populations of H. tigerinus, the total number of alleles per locus and expected heterozygosity ranged from 2 to 18 and 0.271 to 0.938, respectively. No significant linkage disequilibrium was observed across all loci, and five showed a significant deviation from Hardy-Weinberg equilibrium in some populations after Bonferroni correction. Consequently, our findings suggest that these novel markers will be applicable for conservation genetic studies across varying scales from inter-population to inter-individual.

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