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Restriction fragment length polymorphism of mitochondrial DNA and phylogenetic relationships among five species of Indian freshwater turtles

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Abstract

DNA-based identification of species for phylogenetic analysis as well as forensic identification is widely being carried out with the help of polymerase chain reaction (PCR). In this study, a successful effort has been made to identify 5 species of Indian freshwater turtles, including 3 hard-shell turtles (Geoemydidae), i.e.Kachuga dhongoka, K. kachuga andGeoclemys hamiltoni, and 2 species of soft-shell turtles (Trionychidae), i.e.Aspideretes gangeticus andLissemys punctata punctata, by using a well-optimized PCR-RFLP method. The analysis of nucleotide sequence variations in the PCR-amplified mitochondrialcyt-b genes (encoding cytochromeb) from the 5 species revealed its usefulness in the taxonomic differentiation of these species. On the basis ofcyt-b sequence data and the PCR-RFLP pattern, a phylogeny was developed to resolve the genetic relationships between these species, living in the same habitat type. In comparison, the PCR-RFLP of mitochondrial 16S rDNA genes appeared less decisive in analysing phylogenetic relationships or even in species differentiation. Further, the molecular method (PCR-RFLP) developed here is simple, rapid, reliable and reproducible; hence it can be routinely applied for species identification, essential for conservation and management of endangered chelonian species.

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  1. School of Studies Zoology, Jiwaji University, 474 011, Gwalior, India

    Manoj S. Rohilla & Pramod K. Tiwari

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  1. Manoj S. Rohilla
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  2. Pramod K. Tiwari
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Correspondence to Pramod K. Tiwari.

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Rohilla, M.S., Tiwari, P.K. Restriction fragment length polymorphism of mitochondrial DNA and phylogenetic relationships among five species of Indian freshwater turtles. J Appl Genet 49, 167–182 (2008). https://doi.org/10.1007/BF03195610

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