Quantification of Living Microorganisms and of Novel Abundant Prokaryotic Groups in different Deeply Buried Marine Sediments

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The main objective of the project is the quantification of living prokaryotes in various deep sub-seafloor marine sediments, which contain the majority of all prokaryotic cells on Earth. The abundance of Archaea, Bacteria and selected bacterial groups will be determined using the nucleic-acid based quantitative molecular techniques real-time PCR (Q-PCR) and Catalyzed Reporter Deposition - Fluorescence In Situ Hybridisation (CARD - FISH) in combination with total cell counting. Q-PCR will also be used to quantify Bacteria from the presumably abundant 16S rDNA clusters JS1 candidate group and Chloroflexi. Furthermore, sulfate reducing bacteria, methanogenic archaea and autotrophic microorganisms will be quantified using the functional genes cterAB, aprA, mcrA, and cbbL The diversity will be assessed by separation via Denaturing Gradient Gel Electrophoresis (DGGE) and sequencing of excised bands. The preservation of DMA and Intact Polar Lipids in sediments as well as the effect of heating on the microbial community will be studied in incubation experiments in cooperation with K.-U. Hinrichs and R. J. Parkes, respectively. To complement the molecular studies, we also will enrich, isolate and describe novel anaerobic methanotrophic and hydrocarbon-degrading microorganisms.