Molecular markers in Megalobulimus sanctipauli (Ihering & Pilsbry, 1900)

BELTRAMINO, A.A.; VOGLER, R.E.; NÚÑEZ, V.; GUTIÉRREZ GREGORIC, D.E.; RUMI, A.

Rio de Janeiro

Congreso; XI International Congress on Medical and Applied Malacology; 2012

The terrestrial mollusk Megalobulimus sanctipauli (Ihering & Pilsbry, 1900) was registered for the first time in the city of Botucatú, São Paulo, Brazil (type locality), and its distribution area comprises southern Brazil, Paraguay and northeastern Argentina (Misiones and Corrientes Provinces). Original description of M. sanctipauli was based on shell features. More than a century later, shell data are still used for specific diagnosis, given that anatomical, morphometric, genetic and ecological data are still lacking. In this study, two molecular markers were characterized for the first time for the species. Samples of M. sanctipauli were collected from Puerto Iguazú city, Misiones, Argentina (n=5) in 2011. DNA was extracted by using standard CTAB protocol. Universal PCR primers were partial regions from the cytochrome b (Cb) and the cytochrome oxidase subunit I (COI) genes of the mtDNA. Both amplified genes were confirmed through sequencing. After editing, sequences of Cb and COI were 361 bp and 655 bp in length, respectively. One haplotype for Cb and two haplotypes for COI were found among the five individuals. Comparison of COI gene with other available DNA sequences for terrestrial gastropods demonstrates accuracy for species specific identification, the same as for Cb. To our knowledge, this is also the first time than Cb is obtained for a Megalobulimus species. Both markers could be considered as diagnostic tools which can offer valuable support to the traditional taxonomic methods, and partial sequence of both genes are expected to contribute further studies to elucidate phylogenetic relationships of M. sanctipauli.