Molecular Discrimination of Perna (Mollusca: Bivalvia) Species Using the Polymerase Chain Reaction and Species-Specific Mitochondrial Primers
This work was prompted by the need to be able to identify the invasive mussel species, Perna viridis, in tropical Australian seas using techniques that do not rely solely on morphology. DNA-based molecular methods utilizing a polymerase chain reaction (PCR) approach were developed to distinguish unambiguously between the three species in the genus Perna. Target regions were portions of two mitochondrial genes, cox1 and nad4, and the intergenic spacer between these that occurs in at least two Perna species. Based on interspecific sequence comparisons of the nad4 gene, a conserved primer has been designed that can act as a forward primer in PCRs for any Perna species. Four reverse primers have also been designed, based on nad4 and intergenic spacer sequences, which yield species-specific products of different lengths when paired with the conserved forward primer. A further pair of primers has been designed that will amplify part of the cox1 gene of any Perna species, and possibly other molluscs, as a positive control to demonstrate that the PCR is working.
KeywordsInvasive species Perna canaliculus Perna perna Perna viridis polymerase chain reaction (PCR)
We thank the following for provision of samples: Patrick Baker (University of Florida, USA); Dr. Marcos di Donato (Universidad de Oriente, Venezuela); Sarah Jane Bownes (Rhodes University, South Africa); Wo King-Tai (Agriculture, Fisheries and Conservation Department, Hong Kong Special Administrative Region of the People's Republic of China). This project was funded by the Department of Agriculture, Fisheries and Forestry, Australia and managed through the Cooperative Research Centre for the Great Barrier Reef World Heritage Area.
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