Applying an easy molecular method to differentiate Pterois volitans from Pterois miles by RFLPs

Abstract

Numerous papers have reported that Pterois volitans and Pterois miles have invaded the Western Atlantic. Identification of Petrois sp. has been based in morphological differences. However, given their similarities, these are not enough to differentiate both species with certainty. Identifying these species by biological analysis faces difficulties because available methodologies are inaccurate and results unreliable. We developed a method to differentiate these two species using a PCR-RFLPs based protocol. The 476 samples of Pteroides sp. included in the study were collected in the Caribbean and the Gulf of Mexico. To classify them as P. miles or volitans, a DNA sample of each specimen was first analyzed by PCR reaction to amplify cytochrome b (CytB, 1160 bp) gene, followed by digestion of the PCR product using restriction enzymes. Specific banding patterns were visualized for each species by electrophoresis agarose gel. To confirm the presence of these enzyme sites, PCR amplifications of CytB gene were sequenced; resulting sequences were compared with GenBank public database. Our method was 100% effective in the identification of each Pteroids species, providing a quick, easy, and reliable differentiation method. It will help to know the territorial expansion and the basic biology of both species.