Denaturing Gel Electrophoresis Band Research Articles

An investigation of the physiology and potential role of components of the deep ocean bacterial community (of the NE Atlantic) by enrichments carried out under minimal environmental change

Samples of deep-ocean water (3170m) taken from the Rockall Trough (North-East Atlantic) were incubated for one-month at atmospheric and in-situ pressure (31MPa), at 4°C and in the absence and presence of added nutrients. Prokaryotic abundance (direct cell counts) increased by at least 28-fold in enrichments without added nutrients. However, the magnitude of increase in abundance was less for incubations carried out at in-situ pressure (131–181-fold) than those incubations at surface pressure (163–1714-fold increase in abundance). Changes in the prokaryotic community profile as a result of one-month incubation were measured by means of Denaturing Gel Electrophoresis (DGGE) of extracted 16S rDNA. The profiles of post-incubation samples incubated at in-situ pressure were separated from all other profiles as were those of unpressurised samples with added nutrients. The behaviour (fitness) of individual community members (Operational Taxonomic Units: OTUs) was determined on the basis of change in relative DGGE band intensities between pre- and post-incubation samples. Of twenty-one OTUs examined, six were fitter when incubated in the presence of added nutrients and at in-situ pressure and one of these was advantaged when grown in the absence of added nutrients and at in-situ pressure. These represented autochthonous and active members of the deep-ocean prokaryotic community. In contrast, seven OTUs were disadvantaged when grown under in-situ pressure and were indicative surface-derived allochtonous microorganisms. A further two OTUs came to dominance in incubations with added nutrients (pressurised and unpressurised) and similar to the previous category were probably surface-derived microorganisms. A single OTU showed characteristics of piezophilic and oliogrophic behaviour and four OTUs were disadvantaged under all incubation conditions examined. The twenty-one DGGE bands were sequenced and the bacterial communities were dominated by Gamma proteobactria and to a lesser extent members of the Cytophaga–Flavobacterium–Bacteroides and δ groups of prokaryotes.