Provenance and age of bacteria-like structures on mid-Palaeozoic plant fossils

Structures, termed microbioids, comforming to bacteria in size and shape (e.g. rods, spheres, chains and clusters of spheres) have been observed by field emission scanning electron microscopy (FE-SEM) on coalified Silurian and Lower Devonian spores, sporangia, cuticles and coprolites. Some were sectioned for transmission electron microscopy. The elemental composition of both microbioids and ‘substrates’ was investigated using a X-ray microanalysis system. These analyses combined with comparative studies on recent bacteria and cyanobacteria were undertaken to evaluate the biogenicity, nature and age of the microbioids. Spheres with a Si signature (0.03–0.5 μm diameter) and assumed composed of silica are interpreted as artefacts produced abiotically during the extraction procedures. A similar origin is proposed for hollow spheres that are composed of CaF2. These occur singly, in short chains simulating filaments, and in clusters. Considerable differences in size (0.2–2.0 μm diameter) and appearance relate to local variation in the chemical environment during extraction. Spheres (0.2–1.5 μm diameter), that lack a mineral signature, with a framboidal surface ornament and occur within sporangia are identified as by-products of spore development. A biotic origin is also postulated for C-containing rod-shaped structures (>3.1 μm long, <1.4 μm wide), some with collapsed surfaces, although comparisons with living bacteria indicate recent contamination. More elongate rod-shaped microbioids (<8.6 μm long, 1.2 μm wide) have been identified as detrital rutile crystals (TiO2). Minute naviculate structures (<2.2 μm long) resembling diatoms are of unknown origin but are probably composed of thorium hydroxide. Unmineralized filaments of cyanobacterial morphology are recent contaminants. Some of the sporangia and spore masses are partially covered by associations of fragmented sheets, interconnecting strands, rods and spheres that are interpreted as dehydrated biofilms. Being unmineralized they are probably also of recent origin, although they might have survived wild-fire along with the charcoalified mesofossils. Many of the structures illustrated here were initially identified casually as bacteria on the small fossils extracted for biodiversity studies using well-tried, conventional, palaeobotanical techniques. Our subsequent more detailed analyses have shown how such processes can produce artefacts that are morphological analogues of mineralized bacteria, leave residues that mimic bacterial shapes and, despite some efforts such as storage in dilute HCl to eliminate living bacteria, introduce contamination. They reinforce previous concerns that verification of the biogenicity and syngenicity of bacterial-like objects in ancient Earth and extra-terrestrial rocks should not only rely on size and morphological look-alikes, but must encompass a thorough understanding of fossilization processes and extraction techniques plus, ideally, other measures of biogenicity (e.g. biomarkers) and syngenicity

Analysis of DGGE profiles to explore the relationship between prokaryotic community composition and biogeochemical processes in deep subseafloor sediments from the Peru Margin

The aim of this work was to relate depth profiles of prokaryotic community composition with geochemical processes in the deep subseafloor biosphere at two shallow-water sites on the Peru Margin in the Pacific Ocean (ODP Leg 201, sites 1228 and 1229). Principal component analysis of denaturing gradient gel electrophoresis banding patterns of deep-sediment Bacteria, Archaea, Euryarchaeota and the novel candidate division JS1, followed by multiple regression, showed strong relationships with prokaryotic activity and geochemistry (R2=55–100%). Further correlation analysis, at one site, between the principal components from the community composition profiles for Bacteria and 12 other variables quantitatively confirmed their relationship with activity and geochemistry, which had previously only been implied. Comparison with previously published cell counts enumerated by fluorescent in situ hybridization with rRNA-targeted probes confirmed that these denaturing gradient gel electrophoresis profiles described an active prokaryotic communit

Diversity of prokaryotes and methanogenesis in deep subsurface sediments from the Nankai Trough, Ocean Drilling Program Leg 190

Diversity of Bacteria and Archaea was studied in deep marine sediments by PCR amplification and sequence analysis of 16S rRNA and methyl co-enzyme M reductase (mcrA) genes. Samples analysed were from Ocean Drilling Program (ODP) Leg 190 deep subsurface sediments at three sites spanning the Nankai Trough in the Pacific Ocean off Shikoku Island, Japan. DNA was amplified, from three depths at site 1173 (4.15, 98.29 and 193.29 mbsf; metres below the sea floor), and phylogenetic analysis of clone libraries showed a wide variety of uncultured Bacteria and Archaea. Sequences of Bacteria were dominated by an uncultured and deeply branching ‘deep sediment group’ (53% of sequences). Archaeal 16S rRNA gene sequences were mainly within the uncultured clades of the Crenarchaeota. There was good agreement between sequences obtained independently by cloning and by denaturing gradient gel electrophoresis. These sequences were similar to others retrieved from marine sediment and other anoxic habitats, and so probably represent important indigenous bacteria. The mcrA gene analysis suggested limited methanogen diversity with only three gene clusters identified within the Methanosarcinales and Methanobacteriales. The cultivated members of the Methanobacteriales and some of the Methanosarcinales can use CO2 and H2 for methanogenesis. These substrates also gave the highest rates in 14C-radiotracer estimates of methanogenic activity, with rates comparable to those from other deep marine sediments. Thus, this research demonstrates the importance of the ‘deep sediment group’ of uncultured Bacteria and links limited diversity of methanogens to the dominance of CO2/H2 based methanogenesis in deep sub-seafloor sediment

Culturable prokaryotic diversity of deep, gas hydrate sediments: First use of a continuous high-pressure, anaerobic, enrichment and isolation system for subseafloor sediments (DeepIsoBUG)

Deep subseafloor sediments may contain depressurization-sensitive, anaerobic, piezophilic prokaryotes. To test this we developed the DeepIsoBUG system, which when coupled with the HYACINTH pressure-retaining drilling and core storage system and the PRESS core cutting and processing system, enables deep sediments to be handled without depressurization (up to 25 MPa) and anaerobic prokaryotic enrichments and isolation to be conducted up to 100 MPa. Here, we describe the system and its first use with subsurface gas hydrate sediments from the Indian Continental Shelf, Cascadia Margin and Gulf of Mexico. Generally, highest cell concentrations in enrichments occurred close to in situ pressures (14 MPa) in a variety of media, although growth continued up to at least 80 MPa. Predominant sequences in enrichments were Carnobacterium, Clostridium, Marinilactibacillus and Pseudomonas, plus Acetobacterium and Bacteroidetes in Indian samples, largely independent of media and pressures. Related 16S rRNA gene sequences for all of these Bacteria have been detected in deep, subsurface environments, although isolated strains were piezotolerant, being able to grow at atmospheric pressure. Only the Clostridium and Acetobacterium were obligate anaerobes. No Archaea were enriched. It may be that these sediment samples were not deep enough (total depth 1126–1527 m) to obtain obligate piezophiles

Archaeal community diversity and abundance changes along a natural salinity gradient in estuarine sediments

Archaea are widespread in marine sediments, but their occurrence and relationship with natural salinity gradients in estuarine sediments is not well understood. This study investigated the abundance and diversity of Archaea in sediments at three sites (Brightlingsea [BR], Alresford [AR] and Hythe [HY]) along the Colne Estuary, using quantitative real-time PCR (qPCR) of 16S rRNA genes, DNA hybridisation, Archaea 16S rRNA and mcrA gene phylogenetic analyses. Total archaeal 16S rRNA abundance in sediments were higher in the low-salinity brackish sediments from HY (2–8 × 107 16S rRNA gene copies cm−3) than the high-salinity marine sites from BR and AR (2 × 104–2 × 107 and 4 × 106–2 × 107 16S rRNA gene copies cm−3, respectively), although as a proportion of the total prokaryotes Archaea were higher at BR than at AR or HY. Phylogenetic analysis showed that members of the ‘Bathyarchaeota’ (MCG), Thaumarchaeota and methanogenic Euryarchaeota were the dominant groups of Archaea. The composition of Thaumarchaeota varied with salinity, as only ‘marine’ group I.1a was present in marine sediments (BR). Methanogen 16S rRNA genes from low-salinity sediments at HY were dominated by acetotrophic Methanosaeta and putatively hydrogentrophic Methanomicrobiales, whereas the marine site (BR) was dominated by mcrA genes belonging to methylotrophic Methanococcoides, versatile Methanosarcina and methanotrophic ANME-2a. Overall, the results indicate that salinity and associated factors play a role in controlling diversity and distribution of Archaea in estuarine sediments

Deep sub-seafloor prokaryotes stimulated at interfaces over geological time

The sub-seafloor biosphere is the largest prokaryotic habitat on Earth1 but also a habitat with the lowest metabolic rates2. Modelled activity rates are very low, indicating that most prokaryotes may be inactive or have extraordinarily slow metabolism2. Here we present results from two Pacific Ocean sites, margin and open ocean, both of which have deep, subsurface stimulation of prokaryotic processes associated with geochemical and/or sedimentary interfaces. At 90m depth in the margin site, stimulation was such that prokaryote numbers were higher (about 13-fold) and activity rates higher than or similar to near-surface values. Analysis of high-molecular-mass DNA confirmed the presence of viable prokaryotes and showed changes in biodiversity with depth that were coupled to geochemistry, including a marked community change at the 90-m interface. At the open ocean site, increases in numbers of prokaryotes at depth were more restricted but also corresponded to increased activity; however, this time they were associated with repeating layers of diatomrich sediments (about 9Myr old). These results show that deep sedimentary prokaryotes can have high activity, have changing diversity associated with interfaces and are active over geological timescales

Subsurface microbiology and biogeochemistry of a deep, cold-water carbonate mound from the Porcupine Seabight (IODP Expedition 307)

The Porcupine Seabight Challenger Mound is the first carbonate mound to be drilled (∼270 m) and analyzed in detail microbiologically and biogeochemically. Two mound sites and a non-mound Reference site were analyzed with a range of molecular techniques [catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH), quantitative PCR (16S rRNA and functional genes, dsrA and mcrA), and 16S rRNA gene PCR-DGGE] to assess prokaryotic diversity, and this was compared with the distribution of total and culturable cell counts, radiotracer activity measurements and geochemistry. There was a significant and active prokaryotic community both within and beneath the carbonate mound. Although total cell numbers at certain depths were lower than the global average for other subseafloor sediments and prokaryotic activities were relatively low (iron and sulfate reduction, acetate oxidation, methanogenesis) they were significantly enhanced compared with the Reference site. In addition, there was some stimulation of prokaryotic activity in the deepest sediments (Miocene, > 10 Ma) including potential for anaerobic oxidation of methane activity below the mound base. Both Bacteria and Archaea were present, with neither dominant, and these were related to sequences commonly found in other subseafloor sediments. With an estimate of some 1600 mounds in the Porcupine Basin alone, carbonate mounds may represent a significant prokaryotic subseafloor habitat

Regulation of anaerobic methane oxidation in sediments of the Black Sea

Anaerobic oxidation of methane (AOM) and sulfate reduction (SRR) were investigated in sediments of the western Black Sea, where upward methane transport is controlled by diffusion. To understand the regulation and dynamics of methane production and oxidation in the Black Sea, rates of methanogenesis, AOM, and SRR were determined using radiotracers in combination with pore water chemistry and stable isotopes. In the Danube Canyon and the Dnjepr palaeo-delta AOM did not consume methane effectively and upwards diffusing methane created an extended sulfate-methane transition zone (SMTZ) that spread over more than 2.5 m and was located in brackish and limnic sediment. Measurable AOM rates occurred mainly in the lower part of the SMTZ, sometimes even at depths where sulfate seemed to be unavailable. The inefficiency of methane oxidation appears to be linked to the paleoceanographic history of the sediment, since in all cores methane was completely oxidized at the transition from the formerly oxic brackish clays to marine anoxic sediments. The upward tailing of methane was less pronounced in a core from the deep sea in the area of the Dnjepr Canyon, the only station with a SMTZ close to the marine deposits. Sub-surface sulfate reduction rates were mostly extremely low, and in the SMTZ were even lower than AOM rates. Rates of bicarbonate-based methanogenesis were below detection limit in two of the cores, but δ13C values of methane indicate a biogenic origin. The most δ13C- depleted isotopic signal of methane was found in the SMTZ of the core from the deep sea, most likely as a result of carbon recycling between AOM and methanogenesis

Research and Design of a Routing Protocol in Large-Scale Wireless Sensor Networks

无线传感器网络，作为全球未来十大技术之一，集成了传感器技术、嵌入式计算技术、分布式信息处理和自组织网技术，可实时感知、采集、处理、传输网络分布区域内的各种信息数据，在军事国防、生物医疗、环境监测、抢险救灾、防恐反恐、危险区域远程控制等领域具有十分广阔的应用前景。 本文研究分析了无线传感器网络的已有路由协议，并针对大规模的无线传感器网络设计了一种树状路由协议，它根据节点地址信息来形成路由，从而简化了复杂繁冗的路由表查找和维护，节省了不必要的开销，提高了路由效率，实现了快速有效的数据传输。 为支持此路由协议本文提出了一种自适应动态地址分配算——ADAR（AdaptiveDynamicAddre...As one of the ten high technologies in the future, wireless sensor network, which is the integration of micro-sensors, embedded computing, modern network and Ad Hoc technologies, can apperceive, collect, process and transmit various information data within the region. It can be used in military defense, biomedical, environmental monitoring, disaster relief, counter-terrorism, remote control of haz...学位：工学硕士院系专业：信息科学与技术学院通信工程系_通信与信息系统学号：2332007115216