Multiplex species-specific PCR identification of native and non-native oysters (Crassostrea) in Brazil: a useful tool for application in oyster culture and stock management |
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Authors: | Mauro André Damasceno Melo Adam Rick Bessa da Silva Colin Robert Beasley Claudia Helena Tagliaro |
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Institution: | 1. Laboratório de Conserva??o e Biologia Evolutiva, Instituto de Estudos Costeiros, Universidade Federal do Pará, Campus de Bragan?a, Alameda Leandro Ribeiro s/n, Bragan?a, Pará, CEP 68600-000, Brazil 2. Instituto Federal do Pará, Campus de Bragan?a, rua da Escola Agrícola s/n, Bragan?a, Pará, CEP 68600-000, Brazil 3. Laboratório de Moluscos, Instituto de Estudos Costeiros, Universidade Federal do Pará, Campus de Bragan?a, Alameda Leandro Ribeiro s/n, Bragan?a, Pará, CEP 68600-000, Brazil
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Abstract: | In an effort to develop molecular tools for oyster identification, this study reveals the usefulness of a multiplex polymerase chain reaction (PCR) for the reliable, rapid and low-cost identification of the four oyster species found along the Brazilian coast: Crassostrea gasar, Crassostrea rhizophorae, Crassostrea gigas and Crassostrea sp. Canela originally found at Pará, Brazil. In order to perform a simultaneous identification of these species, we used a set of five primers developed and adapted from the cytochrome oxidase subunit I (COI) and internal transcribed spacer 1 (ITS1) fragments, respectively. Amplification was successful in all four species and PCR products were visualized in agarose gel. A single reaction was capable of distinguishing between the species: C. gigas, with two fragments (236 bp for COI and 718 bp for ITS1); C. gasar, with one fragment (718 bp for ITS1); Crassostrea sp., with one fragment (621 bp for ITS1) and C. rhizophorae with two fragments (377 bp for COI and 718 bp for ITS1). This molecular approach provides a simple and rapid identification of the oyster species from the Brazilian coast, thus increasing the efficient and quality of oyster culture programs by reducing the risk of wrong species identification. |
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