A simple method for species identification of the ghost crabs using PCR-RFLP

Abstract

Ghost crabs (genus Ocypode spp.) found on sandy beaches are susceptible with the loss of an ecotone, making them useful as indicators of anthropogenic alteration, such as seawall construction. On the sandy beaches of Sendai Bay on the Pacific coast of the Tohoku region of northern Japan, three Ocypode species have been sympatrically observed in recent years: O. stimpsoni, O. ceratophthalmus, and O. sinensis. However, these three Ocypode species only occur sympatrically from late summer to autumn, immediately after new recruitment. Since they are all juveniles at that period, it is difficult to identify them by morphological characteristics. Thus, we have developed a method for identifying these three Ocypode species. By using two restriction enzymes of AciI and BfaI, we can perform polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis on the 12S-rRNA region of the mitochondrial DNA. As a result, our PCR-RFLP analysis is able to identify all three Ocypode species. Therefore, this method will be a helpful tool for understanding population dynamics and evaluating the sandy beach ecosystem in cases where the three Ocypode species occur sympatrically but identification by morphological features is difficult.