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对各种深海海底海洋沉积物中的活原核生物进行量化，这些沉积物中含有地球上大多数原核生物。古生菌、细菌和选定的细菌群的丰度将使用基于核酸的定量分子技术实时聚合酶链式反应(Q-PCR)和催化报告者沉积-荧光原位杂交(CARD-FISH)结合总细胞计数来确定。Q-PCR还将用于对可能丰富的16S rDNA簇、JS1候选群和氯杆菌的细菌进行量化。此外，硫酸盐还原菌、产甲烷古生菌和自养微生物将使用功能基因cterAB、apra、mcra和cbbL进行量化。多样性将通过变性梯度凝胶电泳(DGGE)分离和片段测序来评估。沉积物中DMA和完整极脂的保存以及加热对微生物群落的影响将分别与K.U.Hinrichs和R.J.Parkes合作在培养实验中进行研究。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营养和碳氢化合物降解微生物。

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.