Geobiology of saline fluids from the deep biosphere obtained from the KTB pump test

拟议项目的主要目标是确定在受控环境生物膜模型系统中形成的微生物生物膜的特征，这些生物膜模型系统用来自大陆深层生物圈的KTB抽水试验的流体进行接种和维护。为此，指定了两个受控环境生物膜模型系统来研究生物膜的形成。系统A基于改良的罗宾斯装置(MRD)，允许测试不同的底物，在不同的曝光时间后，可以独立地取出优惠券，以检查生物膜或测试不同介质对生物膜生长的影响。不同类型岩石的薄片将被用作生物膜形成的天然基质。对于假定的嗜热古生菌和从深层地下流体中获得的细菌的生物膜形成，指定了一个改进的可加热恒化器(系统B)。将使用荧光素测定细胞总数和荧光原位杂交(FISH)来研究从这两个系统获得的生物膜的总体系统发育结构、空间安排和生物膜相关细菌的细胞活性。对标志性脂质生物标志物的分析还提供了对所附社区的丰度、分布和营养状况的定量测量。在进行微生物研究的同时，还将进行生物化学分析，特别是对流体中特定有机化合物(如脂类)的d13C、d15N、2H同位素组成进行分析，从而能够阐明深层生物圈内的微生物驱动过程。

The major goal of the proposed project is the characterization of microbial biofilms formed in controlled environment biofilm model systems inoculated and maintained with fluids from the KTB pumping test originated from the deep continental biosphere. For this purpose, two controlled environment biofilm model systems are designated for the investigation of biofilm formation. System A is based on a modified Robbins Devices (MRD), that allows testing of different substrata, and coupons can be removed independently after various exposure times for examination of the biofilm or testing the effects of different media on biofilm growth in situ. Thin sections of different types of rock will be used as natural substrata for biofilm formation. For biofilm formation of putative thermophilic Archaea and Bacteria obtained from the deep subsurface fluids, a modified heatable chemostat (System B) is designated. The biofilms obtained from both systems will be investigated with regard to their overall phylogenetic structure, spatial arrangement and the cellular activity of the biofilm associated bacteria using fluorochromes for total cell count determination and fluorescent in situ hybridization (FISH). Analysis of the signature lipid biomarkers additonally provides quantitative measures of abundance, distribution and the nutritional status of the attached communities. Along with the microbiological investigations, biochemical analysis, especially for d13C, d15N, 2H isotopic compositions of specific organic compounds in the fluids (e.g. lipids) will be performed that could enable to elucidate microbiological driven processes within the deep biosphere.