A fast and accurate molecular method for the detection of larvae of the golden mussel Limnoperna fortunei (Mollusca: Mytilidae) in plankton samples.

摘要

The golden mussel (Limnoperna fortunei) is native to continental China, but is emerging as an important invasive species, and has caused severe damage to different water bodies and artificial water supply systems. Efficient management of L. fortunei in its introduced range requires the ability to predict sites most likely to experience colonization, permitting targeting of control measures, which can be achieved by monitoring the presence of larvae in plankton samples. This paper describes the design of a set of primers that can be used to detect larvae of the golden mussel. This approach is based on the amplification of a DNA fragment of the species of interest and its digestion using restriction enzymes to generate species-specific banding profiles. A fragment of the cytochrome oxidase subunit 1 gene of L. fortunei was sequenced. Unique regions were identified by comparing the L. fortunei sequence with a sample of 4 mollusc species. Sequences of Crassostrea gigas, Corbicula fluminea and Modiolus brasiliensis were obtained. Sensitivity analyses were carried out with increasingly smaller amounts of template DNA to determine the extent of detectability of the method. Specificity tests were conducted by testing the designed primers against samples of the four taxa used for their development as well as a crustacean (Ucides cordatus) and six additional molluscs (Modiolus brasiliensis, Perna perna, Colisella sp., Littorinidae sp., Brachidontes sp. and Thais sp.). Results indicate that the primer sets provided in the study can become an important tool for testing for the presence of larvae of L. fortunei in its introduced range. This method is already being used to monitor the presence of the golden mussel in the Rio Iguacu basin..