Molecular and morphological identification of settlement-stage vent mussel larvae, Bathymodiolus azoricus (Bivalvia: Mytilidae), preserved in situ at active vent fields on the Mid-Atlantic Ridge
Limnol. Oceanogr., 45(7), 2000, 1655-1661 | DOI: 10.4319/lo.2000.45.7.1655
ABSTRACT: This paper describes the first successful attempt to trap and identify the larvae of a deep-sea vent organism using a combination of sediment traps and molecular analysis. During the European Union-funded MARVEL cruise in August and September 1997, sediment traps containing a high-salt DNA preservative were deployed around active black smoker chimneys on the newly discovered Rainbow vent field in an attempt to collect larval stages of the dominant vent bivalve Bathymodiolus azoricus. A total of 2,055 shelled mussel larvae was collected within a 2-week period, all of which were of identical size and shell morphology that indicated that they were at the settlement, prodissoconch II stage. Scanning electron microscopy of the shell hinge indicated that they belonged to the family Mytilidae, but it required molecular analysis to confirm their species identity. Polymerase chain reaction (PCR) amplification of the ITS2 region was performed on a subset of the larvae, and the resulting PCR products were cut with diagnostic restriction endonucleases to allow comparison with a DNA database based on adult specimens. The DNA restriction patterns typifying the Rainbow larvae were identical to those of adult B. azoricus from the Menez Gwen, Lucky Strike, and Rainbow vent sites and were significantly different from those that typified Bathymodiolus thermophilus (Pacific) and Bathymodiolus puteoserpentis (Snake Pit vent field on the Mid-Atlantic Ridge), which clearly points to the Rainbow larvae having their origin on that part of the ridge close to the Azores. These findings point to the value of sediment traps as a way to study the temporal and spatial aspects of larval settlement in deep-sea hydrothermal vent environments.