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

该项目的主要目标是量化各种深海海底下海洋沉积物中的活原核生物，这些沉积物含有地球上大部分原核细胞。将使用基于核酸的定量分子技术实时PCR（Q-PCR）和催化报告分子沉积-荧光原位杂交（CARD-FISH）结合总细胞计数来确定细菌、细菌和选定细菌群的丰度。Q-PCR还将用于定量来自可能丰富的16 S rDNA簇JS 1候选组和绿杆菌的细菌。此外，将使用功能基因cterAB、aprA、mcrA和cbbL对硫酸盐还原菌、产甲烷古菌和自养微生物进行定量。将通过变性梯度凝胶电泳（DGGE）分离并对切下的条带进行测序来评估多样性。DMA和完整的极性脂质在沉积物中的保存以及加热对微生物群落的影响将在与K.联合Hinrichs和R. J.帕克斯，分别。为了补充分子研究，我们还将丰富，分离和描述新的厌氧甲烷氧化和烃降解微生物。

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.