基于多种培养基和不同培养温度的马里亚纳海沟异养细菌多样性分析

马里亚纳海沟具有低温、高压、永久黑暗以及营养匮乏等深海环境特征,其中的细菌多样性对深海环境具有极为重要的作用。为研究马里亚纳海沟海水中异养细菌的物种多样性,采用多种培养基、不同培养温度同步筛选,单菌落16S rRNA基因序列鉴定,邻近法系统发育树构建分析等方法,对25个海水样品进行异养细菌多样性分析。共获得细菌531株,对其中371株进行16S rRNA基因鉴定,共分布41属,97种。经系统进化分析,异养菌株分布于4门:厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)、放线菌门(Actinobacteria)和拟杆菌门(Bacteroidetes)。优势菌群为Proteobacteria,占鉴定菌株数量的58%,其中γ-Proteobacteria占总菌数的52%。此外,还发现4株潜在新物种。     

Life inhabits rocks: clues to rock erosion from electron microscopy of pisolite at a UNESCO heritage site in Brazil

Rock erosion is attracting increasing attention from scientists worldwide. The area encompassing the Saint John Baptist Church, Saint John Village, XVII century ruins in Rio Grande do Sul at the UNESCO World Heritage Site is considered a Brazilian treasure. However, the risk of damage to this site from rock erosion has recently increased tremendously. Generally, the rocky construction such as fence, wall and tomb stone, seems strong but is actually extremely sensitive to erosion caused by lichens, fungi, molds and bacteria. Because of biological erosion and massive exposure, the fresh rock is dominated by clays and microorganisms. Water-adsorbing clays and microorganisms influence the mechanisms of the rock erosion. In this study, the formation of bio-clay-minerals in porous structure of pisolite was demonstrated using electron microscopy. Bacterial clay mineralization can deform the rock structure and even produce organic materials. Biological activity could easily corrode rocky constructions around the Saint John Baptist Church site. The rocks are pisolitic laterites possibly formed in Tertiary over the Kretaceous Parana flood Basalts. Samples inhabited by lichens and fungi were collected from a collapsed wall in the ancient church. The zonal reddish-brown pisolites are 4 mm in diameter in a matrix of clays associated with porous and empty spaces. Elemental distribution maps from X-ray fluorescence microscopy show iron-rich spherules of pisolite, whereas the matrix is composed of Al, Si, Mn, and Sr; thus producing goethite and kaolinite. Transmission electron microscopic observation showed that various types of bacteria inhabit the spherule and are associated with clay minerals and graphite. STEM elemental analysis confirmed the bio-clay-mineralization with Al, Si, S, and Fe, around bacterial cells. The results presented here will improve our understanding of nm-scale bio-mineralization and bio-erosion in lateritic rocks. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Micro-FTIR spectroscopic signatures of Bacterial lipids in Proterozoic microfossils

Precambrian microbial fossils show carbonaceous cellular structure, which often resemble in shape and size cyanobacteria and other prokaryotes. Morphological taxonomy of these minute, simple, and more or less degraded fossils is, however, often not enough to determine their precise phylogenetic positions. Here we report the results of micro-FTIR spectroscopic analyses of well-preserved microfossils in 850 Ma and 1900 Ma stromatolites, together with those of 8 species of extant prokaryotes and 5 of eukaryotes for comparison. These Proterozoic fossils have low CH₃/CH₂ absorbance ratios (R_3/2 < 0.5) of aliphatic CH moieties, suggesting selective preservation of long, straight, aliphatic carbon chains probably derived from bacterial membrane lipids. All the observed R_3/2 values of coccoids, filaments and amorphous organic matter resemble lipid fractions of extant Bacteria including cyanobacteria, but not Archaea. The results indicate that Proterozoic microfossils belong to Bacteria, which is consistent with the cyanobacterial origin inferred from morphology. Moreover, the R_3/2 value of fossilized cell would reflect chemical composition of its precursor membrane lipid, thus could be a useful new tracer for distinguishing Archaea, Bacteria and possibly Eucarya for fossilized and extant microorganisms.     

Metabolic patterning of biosilicification

Here we show a discernibly unique biosilicification pattern for live, metabolically active Synechococcus cyanobacterial cell surfaces compared to dead Synechococcus cells under identical experimental conditions. The live cell treatments showed signs of cell division and the growth of fimbriae indicating metabolic activity during the 5-day silicification experiment. Live treatment cells were also recultivable after the experiments confirming their continued viability. The metabolically active live cyanobacteria treatment bound twice the amount of colloidal SiO₂ and held it more tightly compared to the dead cell treatment. Further, biosilicification of the live, actively metabolizing bacteria was unipolar, leaving the core surface largely unencrusted. In contrast, biosilicification of the dead cells was heterogeneous, occurring across the entire cell surface with no observable localized pattern. The directed biosilicification localization of live cell surfaces is likely a bacterial strategy to protect the cell functionality against the potentially inhibitory effects of mineral encrustation. Localization of silica biominerals to the polar end of the cell is also consistent with reported bacteria regulated cell polarity, which, under the experimental pH of 3, would enable localized differential attraction between the charged colloidal silica (+) particles and the bacterial cell polar surface (−). Our results show a novel metabolically-linked distinct colloidal SiO₂ biomineralization fingerprint, suggesting a putative biomineralization signature.     

Benthic foraminifera from the deep-water Niger delta (Gulf of Guinea): Assessing present-day and past activity of hydrate pockmarks

We present ecological and isotopic (δ¹⁸O and δ¹³C) data on benthic foraminifera sampled from 4 deep-sea stations in a pockmark field from the deep-water Niger delta (Gulf of Guinea, Equatorial Atlantic Ocean). In addition, a series of sedimentological and (bio)geochemical data are shown to back up foraminiferal observations. All stations are located within 1.2 km of each other, so prevailing oceanographic conditions can be assumed to be similar at each site. Two of the sites (GMMC-01 and GMMC-02) are located in a pockmark (named “pockmark A”) where current methane seepages were recorded by ROV observations. A third station (GMMC-03) is located in the topographic depression interpreted as a collapsed pockmark (named “pockmark B”). The fourth site (GMMC-04) is a reference station, without evidence of past or present seepages. Our observations show that degraded organic matter with low bio-availability is present at all stations with a preferential burial of organic compounds in topographic depressions (GMMC-03 station). Authigenic aragonite is abundant in surface sediments at stations GMMC-01 and -02. Its precipitation is likely related to high rates of methane oxidation during past seep events in episodically active pockmark A. In contrast, the absence of anaerobic methanotrophic Archaea (ANME) during the sampling period (November 2011) suggests that only moderate sulphide and methane oxidation take place close to the sediment–water interface. Compared to the reference site GMMC-04, living foraminifera at the collapsed and episodically active pockmarks show minor changes in terms of diversity, standing stocks and faunal composition. However, the δ¹³C signal of living and dead (but well-preserved) foraminiferal species (Ceratobulimina contraria, Melonis barleeanus, Uvigerina peregrina) is depleted in the episodically active pockmark A compared to the other stations. Overgrowth of authigenic carbonate on altered foraminifera generates an important shift to lower δ¹³C values. Dead faunas carry a complex time-averaged message, integrating taphonomic gains and losses related to the temporal variability of gas emission. They reveal major faunal differences that may be useful to detect gas hydrate seepages in different pockmark stages.     

Bacterial wax esters in recent fluvial sediments

We investigated the molecular inventory of River Danube sediments and identified wax esters with 28–36 carbons, with C₃₀, C₃₁ and C₃₂ homologs the most abundant. They consist of various combinations of n-, iso- and anteiso-aliphatic acid and alcohol moieties, with different isomer distributions for the esters with odd and even carbon numbers. The short chain length (C₂₈–C₃₆), high proportion of methyl moieties and presence of monounsaturated esters together suggest a bacterial origin for the wax esters. The concentration in surface sediments varied from 0 to 741 μg/kg, being significantly enriched at locations with high nutrient concentration and high primary productivity. Cluster analysis of denaturing gradient gel electrophoresis (DGGE) band patterns revealed different bacterial communities in surface sediments from the stream and surface sediments from the reservoir. The concentration in a 70 cm core of rapidly deposited sediments decreased significantly with depth, following first order kinetics, suggesting a loss of the esters within ca. 3 years following burial and hence a low probability for preservation in the geological record. Wax esters in rivers have rarely been investigated; our results suggest that they represent a bacterial response to enhanced primary production, triggered by higher nutrient supply.     

Sublimation extraction coupled with gas chromatography-mass spectrometry: A new technique for future in situ analyses of purines and pyrimidines on Mars

We have developed a sublimation technique coupled with chemical derivatization and gas chromatography mass spectrometry (GC-MS) to detect nucleobases and other volatile organic compounds derived from bacteria in Mars analog materials. To demonstrate this technique, a sample of serpentine inoculated with Escherichia coli (E. coli) cells was heated to 500 °C for several seconds under Martian ambient pressure. The sublimate was collected on a cold finger, then derivatized and analyzed by GC-MS. We found that adenine, cytosine, thymine and uracil were the most abundant molecules detected in the sublimed E. coli extract by GC-MS. In addition, nucleobases were also detected in sublimed extracts of a deep-sea sediment sample, seawater, and soil collected from the Atacama Desert in Chile after heating the samples under the same conditions. Our results indicate that nucleobases can be easily isolated directly from natural samples using sublimation and then detected by GC-MS after chemical derivatization. The sublimation-based extraction technique is one approach that should be considered for use by future in situ instruments designed to detect organic compounds relevant to life in the Martian regolith.     

Carbonate and silicate biomineralization in a hypersaline microbial mat (Mesaieed sabkha,Qatar): Roles of bacteria,extracellular polymeric substances and viruses

In a modern peritidal microbial mat from Qatar, both biomediated carbonates and Mg‐rich clay minerals (palygorskite) were identified. The mat, ca 5 cm thick, shows a clear lamination reflecting different microbial communities. The initial precipitates within the top millimetres of the mat are composed of Ca–Mg–Si–Al–S amorphous nanoparticles (few tens of nanometres) that replace the ultrastructure of extracellular polymeric substances. The extracellular polymeric substances are enriched in the same cations and act as a substrate for mineral nucleation. Successively, crystallites of palygorskite fibres associated with carbonate nanocrystals develop, commonly surrounding bacterial bodies. Micron‐sized crystals of low‐Mg calcite are the most common precipitates, together with subordinate aragonite, very high‐Mg calcite/dolomite and ankerite. Pyrite nanocrystals and framboids are present in the deeper layers of the mat. Calcite crystallites form conical structures, circular to triangular/hexagonal in cross‐section, evolving to crystals with rhombohedral terminations; some crystallite bundles develop into dumb‐bell and stellate forms. Spheroidal organo‐mineral structures are also common within the mat. Nanospheres, a few tens of nanometres in diameter, occur attached to coccoid bacteria and within their cells; these are interpreted as permineralized viruses and could be significant as nuclei for crystallite‐crystal precipitation. Microspheres, 1 to 10 μm in diameter, result from intracellular permineralization within bacteria or the mineralization of the bacteria themselves. Carbonates and clay minerals are commonly aggregated to form peloids, tens of microns in size, surrounded by residual organic matter. Magnesium silicate and carbonate precipitation are likely to have been driven by pH – saturation index – redox changes within the mat, related to microenvironmental chemical changes induced by the microbes – extracellular polymeric substances – viruses and their degradation.     

Factors influencing benthic bacterial abundance, biomass, and activity on the northern continental margin and deep basin of the Gulf of Mexico

As part of a larger project on the deep benthos of the Gulf of Mexico, an extensive data set on benthic bacterial abundance (n>750), supplemented with cell-size and rate measurements, was acquired from 51 sites across a depth range of 212–3732 m on the northern continental slope and deep basin during the years 2000, 2001, and 2002. Bacterial abundance, determined by epifluorescence microscopy, was examined region-wide as a function of spatial and temporal variables, while subsets of the data were examined for sediment-based chemical or mineralogical correlates according to the availability of collaborative data sets. In the latter case, depth of oxygen penetration helped to explain bacterial depth profiles into the sediment, but only porewater DOC correlated significantly (inversely) with bacterial abundance (p<0.05, n=24). Other (positive) correlations were detected with TOC, C/N ratios, and % sand when the analysis was restricted to data from the easternmost stations (p<0.05, n=9–12). Region-wide, neither surface bacterial abundance (3.30–16.8×10⁸ bacteria cm⁻³ in 0–1 cm and 4–5 cm strata) nor depth-integrated abundance (4.84–17.5×10¹³ bacteria m⁻², 0–15 cm) could be explained by water depth, station location, sampling year, or vertical POC flux. In contrast, depth-integrated bacterial biomass, derived from measured cell sizes of 0.027–0.072 μm³, declined significantly with station depth (p<0.001, n=56). Steeper declines in biomass were observed for the cross-slope transects (when unusual topographic sites and abyssal stations were excluded). The importance of resource changes with depth was supported by the positive relationship observed between bacterial biomass and vertical POC flux, derived from measures of overlying productivity, a relationship that remained significant when depth was held constant (partial correlation analysis, p<0.05, df=50). Whole-sediment incubation experiments under simulated in situ conditions, using ³H-thymidine or ¹⁴C-amino acids, yielded low production rates (5–75 μg C m⁻² d⁻¹) and higher respiration rates (76–242 μg C m⁻² d⁻¹), with kinetics suggestive of resource limitation at abyssal depths. Compared to similarly examined deep regions of the open ocean, the semi-enclosed Gulf of Mexico (like the Arabian Sea) harbors in its abyssal sediments a greater biomass of bacteria per unit of vertically delivered POC, likely reflecting the greater input of laterally advected, often unreactive, material from its margins.     

Lipid biomarkers and bacterial lipase activities as indicators of organic matter and bacterial dynamics in contrasted regimes at the DYFAMED site, NW Mediterranean

This study investigated the relationships between dissolved organic matter (DOM) composition and bacterial dynamics on short time scale during spring mesotrophic (March 2003) and summer oligotrophic (June 2003) regimes, in a 0–500 m depth water column with almost no advection, at the DYFAMED site, NW Mediterranean. DOM was characterized by analyzing dissolved organic carbon (DOC), colored dissolved organic matter (CDOM) and lipid class biotracers. Bacterial dynamic was assessed through the measurement of in situ bacterial lipase activity, abundance, production and bacterial community structure. We made the assumption that by coupling the ambient concentration of hydrolysable acyl-lipids with the measurement of their in situ bacterial hydrolysis rates (i.e. the free fatty acids release rate) would provide new insights about bacterial response to change in environmental conditions. The seasonal transition from spring to summer was accompanied by a significant accumulation of excess DOC (+5 μM) (ANOVA, p<0.05, n=8) in the upper layer (0–50 m). In this layer, the free fatty acids release rate to the bacterial carbon demand (BCD) ratio increased from 0.6±0.3 in March to 1.3±1.0 in June (ANOVA, p<0.05, n=8) showing that more uncoupling between the hydrolysis of the acyl-lipids and the BCD occurred during the evolution of the season, and that free fatty acids contributed to the excess DOC. The increase of lipolysis index and CDOM absorbance (from 0.24±0.17 to 0.39±0.13 and from 0.076±0.039 to 0.144±0.068; ANOVA, p<0.05, n=8, respectively), and the higher contribution of triglycerides, wax esters and phospholipids (from <5% to 12–31%) to the lipid pool reflected the change in the DOM quality. In addition to a strong increase of bacterial lipase activity per cell (51.4±29.4–418.3±290.6 Ag C cell⁻¹ h⁻¹), a significant percentage of ribotypes (39%) was different between spring and summer in the deep chlorophyll maximum (DCM) layer in particular, suggesting a shift in the bacterial community structure due to the different trophic conditions. At both seasons, in the chlorophyll layers, diel variations of DOM and bacterial parameters reflected a better bioavailability and/or DOM utilization by bacteria at night (the ratio of free fatty acids release rate to bacterial carbon demand decreased), most likely related to the zooplankton trophic behaviour. In mesotrophic conditions, such day/night pattern was driving changes in the bacterial community structure. In more oligotrophic period, diel variations in bacterial community structure were depth dependent in relation to the strong summer stratification.