Wspólnotowy Serwis Informacyjny Badan i Rozwoju - CORDIS

DGGE, RFLP, sequencing, biomarker and biogeochemical data of seep and sediment samples from Kvitebjorn, Tommeliten, Holene

Overview of the result:
Six different water samples, taken from above gas seeps and above a pockmark grope at Tommeliten, Kvitebjørn and Holene, have been analysed for microbial diversity. Primers against both bacteria and Archaea have been used. Some samples have been drawn at varying water depth at the same location.

Corresponding sediment samples have been taken close to the seep and at the pockmark grope.

PCR, DGGE, cloning and sequencing has been done. Restriction enzyme pattern (RFLP) has been run with HeaIII and RsaI.

BLAST analyses have been performed and closest match and phylogenetic dendrogram constructed.

Description of the result:
A direct measurement on seep water samples by flow cytometer reveals a particle distribution of very small size particles, e.g. 200 – 600 nm. Different staining techniques show that most of the organisms are dead when received on shore.

The DGGE run gives different pattern for the different locations and also a variation for the depth of sampling. There is no resemblance to traditional sea water run as a control.

The amplified ribosomal DNA restriction analysis of the sediments samples show a distinct differentiation between the various sites. Only ANDRA-type 10 is found in all three locations.

Corresponding results were found for the seep water samples. ANDRA-types, 8 and 11 are common for Kvitebjørn and Western Slope. The others are distinct and also different from what was found in the sediment samples.

Grouping of clones of Archaea within RFLP types based on restriction enzyme mapping of M13PCR products with HaeIII and RsaI, showed RFLP-type 1 to be common for Tommeliten, Kvitebjørn and Holene. Three other types were distinct to each location.

Based on the RFLP/ANDRA mapping and sequence analysis, the distribution of bacterial groups within the community from the different seep samples could be evaluated.

A phylogenetic dendrogram is generated and closest match to gene bank affiliations looked upon for better understanding of possible origin and function in the actual habitat.

Current status and use:
Sequence analyses of specific clones are used for DNA probe synthesis. These will be used as specific markers in analysis of samples from other locations and other type of samples associated with oil exploration.

Key innovations:
- Unknown gene sequences indicated presence of new species.
- Differentiation between different locations.
- Differentiation between water versus sediment samples.

Users interested in the result:
- Oil exploration industry.
- Scientists interested in constructing new DNA probes/markers.

Expected benefits:
- Use in oil exploration and extended understanding of oil related/ oil associated microorganisms.
- Publication planned together with MPI.

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Reported by

Statoil Research Centre
Arkitekt Ebbels Vei 10, Rotboll
7005 Trondheim
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