Multiplex PCR-based genotyping of mitochondrial DNA from two species of the genus Batillaria (B. multiformis and B. flectosiphonata) from Amami-Oshima, Japan

Abstract

Snails of the genus Batillaria are common on most muddy tidal flats throughout Japan. To conserve tidal flat ecosystems, it is essential to understand the ecology of the Batillaria snails. A previous mitochondrial (mt) cytochrome c oxidase subunit I (COI) gene sequence analysis indicated that two Batillaria species, B. flectosiphonata and B. multiformis, occur together on the tidal flats on Amami-Oshima, Japan. However, it is difficult to distinguish them by their morphological traits. In this study, a simple multiplex PCR (mPCR)-based method was developed to identify the two species without the need for DNA sequencing. The method allows the discrimination of species by the electrophoretic detection of species-specific DNA fragments of the COI gene amplified by mPCR. This method will enhance the ecological study of these Batillaria snails on Amami-Oshima.