Implementation of DNA Mitochondrial Analysis in Rhinoclemmys nasuta (Testudines: Geoemydidae)

Abstract

Rhinoclemmys nasuta (Testudines:Geoemydidae) is considered an almost endemic specie to Colombia and the most primitive species of Rhynoclemmys. However, it is classified data deficient by IUCN because the available information is not enough to make a direct or indirect assessment of its extinction risk. Here, we describe the implementation of the method to analyze the mitochondrial DNA control sequence (mtDNA) of R. nasuta in order to generate tools for future studies in systematics and population conservation. Genomic mtDNA was extracted by salting-out from blood samples from Isla Palma and Playa Chucheros (Bahía Málaga–Colombian Pacific Coast) and we used a pair of degenerate primers (reported for Chrysemys picta, Testudines:Emydidae) to perform amplification. Fragments of 800pb were obtained and the sequencing reaction was effective. A homology percentage above of 92 % was established between the obtained sequences and mtDNA sequences from Sacalia quadriocellata (Testudines:Geoemydidae),and Cuora aurocapitata (Testudines:Geoemydidae) reported in the GenBank. This result shows that the described method can be a useful tool for the study of R. nasuta populations in the Colombian Pacific region, achieving an effective sequencing of the mtDNA control region of this species.