Cultivation of the ubiquitous SAR11 marine bacterioplankton clade

The alpha-proteobacterial lineage that contains SAR11 and related ribosomal RNA gene clones was among the first groups of organisms to be identified when cultivation-independent approaches based on rRNA gene cloning and sequencing were applied to survey microbial diversity in natural ecosystems. This group accounts for 26% of all ribosomal RNA genes that have been identified in sea water and has been found in nearly every pelagic marine bacterioplankton community studied by these methods. The SAR11 clade represents a pervasive problem in microbiology: despite its ubiquity, it has defied cultivation efforts. Genetic evidence suggests that diverse uncultivated microbial taxa dominate most natural ecosystems, which has prompted widespread efforts to elucidate the geochemical activities of these organisms without the benefit of cultures for study. Here we report the isolation of representatives of the SAR11 clade. Eighteen cultures were initially obtained by means of high-throughput procedures for isolating cell cultures through the dilution of natural microbial communities into very low nutrient media. Eleven of these cultures have been successfully passaged and cryopreserved for future study. The volume of these cells, about 0.01 micro m(3), places them among the smallest free-living cells in culture.     

Novel pathway for assimilation of dimethylsulphoniopropionate widespread in marine bacteria

Dimethylsulphoniopropionate (DMSP) accounts for up to 10% of carbon fixed by marine phytoplankton in ocean surface waters, producing an estimated 11.7-103?Tmol S per year, most of which is processed by marine bacteria through the demethylation/demethiolation pathway. This pathway releases methanethiol (MeSH) instead of the climatically active gas dimethylsulphide (DMS) and enables marine microorganisms to assimilate the reduced sulphur. Despite recognition of this critical microbial transformation for over two decades, the biochemical pathway and enzymes responsible have remained unidentified. Here we show that three new enzymes related to fatty acid β-oxidation constitute the pathway that assimilates methylmercaptopropionate (MMPA), the first product of DMSP demethylation/demethiolation, and that two previously unknown coenzyme A (CoA) derivatives, 3-methylmercaptopropionyl-CoA (MMPA-CoA) and methylthioacryloyl-CoA (MTA-CoA), are formed as novel intermediates. A member of the marine roseobacters, Ruegeria pomeroyi DSS-3, requires the MMPA-CoA pathway for MMPA assimilation and MeSH production. This pathway and the ability to produce MeSH from MMPA are present in diverse bacteria, and the ubiquitous SAR11 clade bacterium Pelagibacter ubique possesses enzymes for at least the first two steps. Analysis of marine metagenomic data indicates that the pathway is widespread among bacterioplankton in the ocean surface waters, making it one of the most important known routes for acquisition of reduced carbon and sulphur by surface ocean heterotrophs.     

Proteorhodopsin in the ubiquitous marine bacterium SAR11

Proteorhodopsins are light-dependent proton pumps that are predicted to have an important role in the ecology of the oceans by supplying energy for microbial metabolism. Proteorhodopsin genes were first discovered through the cloning and sequencing of large genomic DNA fragments from seawater. They were later shown to be widely distributed, phylogenetically diverse, and active in the oceans. Proteorhodopsin genes have not been found in cultured bacteria, and on the basis of environmental sequence data, it has not yet been possible to reconstruct the genomes of uncultured bacterial strains that have proteorhodopsin genes. Although the metabolic effect of proteorhodopsins is uncertain, they are thought to function in cells for which the primary mode of metabolism is the heterotrophic assimilation of dissolved organic carbon. Here we report that SAR11 strain HTCC1062 ('Pelagibacter ubique'), the first cultivated member of the extraordinarily abundant SAR11 clade, expresses a proteorhodopsin gene when cultured in autoclaved seawater and in its natural environment, the ocean. The Pelagibacter proteorhodopsin functions as a light-dependent proton pump. The gene is expressed by cells grown in either diurnal light or in darkness, and there is no difference between the growth rates or cell yields of cultures grown in light or darkness.     

A newly discovered Roseobacter cluster in temperate and polar oceans

Bacterioplankton phylotypes of alpha-Proteobacteria have been detected in various marine regions, but systematic biogeographical studies of their global distribution are missing. Alpha-Proteobacteria comprise one of the largest fractions of heterotrophic marine bacteria and include two clades, SAR11 and Roseobacter, which account for 26 and 16% of 16S ribosomal RNA gene clones retrieved from marine bacterioplankton. The SAR11 clade attracted much interest because related 16S rRNA gene clones were among the first groups of marine bacteria to be identified by cultivation-independent approaches and appear to dominate subtropical surface bacterioplankton communities. Here we report on the global distribution of a newly discovered cluster affiliated to the Roseobacter clade, comprising only as-yet-uncultured phylotypes. Bacteria of this cluster occur from temperate to polar regions with highest abundance in the Southern Ocean, but not in tropical and subtropical regions. Between the south Atlantic subtropical front and Antarctica, we detected two distinct phylotypes, one north and one south of the polar front, indicating that two adjacent but different oceanic provinces allow the persistence of distinct but closely related phylotypes. These results suggest that the global distribution of major marine bacterioplankton components is related to oceanic water masses and controlled by their environmental and biogeochemical properties.     

SAR11 marine bacteria require exogenous reduced sulphur for growth

Sulphur is a universally required cell nutrient found in two amino acids and other small organic molecules. All aerobic marine bacteria are known to use assimilatory sulphate reduction to supply sulphur for biosynthesis, although many can assimilate sulphur from organic compounds that contain reduced sulphur atoms. An analysis of three complete 'Candidatus Pelagibacter ubique' genomes, and public ocean metagenomic data sets, suggested that members of the ubiquitous and abundant SAR11 alphaproteobacterial clade are deficient in assimilatory sulphate reduction genes. Here we show that SAR11 requires exogenous sources of reduced sulphur, such as methionine or 3-dimethylsulphoniopropionate (DMSP) for growth. Titrations of the algal osmolyte DMSP in seawater medium containing all other macronutrients in excess showed that 1.5 x 10(8) SAR11 cells are produced per nanomole of DMSP. Although it has been shown that other marine alphaproteobacteria use sulphur from DMSP in preference to sulphate, our results indicate that 'Cand. P. ubique' relies exclusively on reduced sulphur compounds that originate from other plankton.     

Endospore abundance, microbial growth and necromass turnover in deep sub-seafloor sediment

Two decades of scientific ocean drilling have demonstrated widespread microbial life in deep sub-seafloor sediment, and surprisingly high microbial-cell numbers. Despite the ubiquity of life in the deep biosphere, the large community sizes and the low energy fluxes in this vast buried ecosystem are not yet understood. It is not known whether organisms of the deep biosphere are specifically adapted to extremely low energy fluxes or whether most of the observed cells are in a dormant, spore-like state. Here we apply a new approach--the D:L-amino-acid model--to quantify the distributions and turnover times of living microbial biomass, endospores and microbial necromass, as well as to determine their role in the sub-seafloor carbon budget. The approach combines sensitive analyses of unique bacterial markers (muramic acid and D-amino acids) and the bacterial endospore marker, dipicolinic acid, with racemization dynamics of stereo-isomeric amino acids. Endospores are as abundant as vegetative cells and microbial activity is extremely low, leading to microbial biomass turnover times of hundreds to thousands of years. We infer from model calculations that biomass production is sustained by organic carbon deposited from the surface photosynthetic world millions of years ago and that microbial necromass is recycled over timescales of hundreds of thousands of years.     

A hydrogen-based subsurface microbial community dominated by methanogens.

The search for extraterrestrial life may be facilitated if ecosystems can be found on Earth that exist under conditions analogous to those present on other planets or moons. It has been proposed, on the basis of geochemical and thermodynamic considerations, that geologically derived hydrogen might support subsurface microbial communities on Mars and Europa in which methanogens form the base of the ecosystem. Here we describe a unique subsurface microbial community in which hydrogen-consuming, methane-producing Archaea far outnumber the Bacteria. More than 90% of the 16S ribosomal DNA sequences recovered from hydrothermal waters circulating through deeply buried igneous rocks in Idaho are related to hydrogen-using methanogenic microorganisms. Geochemical characterization indicates that geothermal hydrogen, not organic carbon, is the primary energy source for this methanogen-dominated microbial community. These results demonstrate that hydrogen-based methanogenic communities do occur in Earth's subsurface, providing an analogue for possible subsurface microbial ecosystems on other planets.     

Anaerobic benzene oxidation coupled to nitrate reduction in pure culture by two strains of Dechloromonas.

Benzene contamination is a significant problem. It is used in a wide range of manufacturing processes and is a primary component of petroleum-based fuels. Benzene is a hydrocarbon that is soluble, mobile, toxic and stable, especially in ground and surface waters. It is poorly biodegraded in the absence of oxygen. However, anaerobic benzene biodegradation has been documented under various conditions. Although benzene biomineralization has been demonstrated with nitrate, Fe(III), sulphate or CO2 as alternative electron acceptors, these studies were based on sediments or microbial enrichments. Until now there were no organisms in pure culture that degraded benzene anaerobically. Here we report two Dechloromonas strains, RCB and JJ, that can completely mineralize various mono-aromatic compounds including benzene to CO2 in the absence of O2 with nitrate as the electron acceptor. This is the first example, to our knowledge, of an organism of any type that can oxidize benzene anaerobically, and we demonstrate the potential applicability of these organisms to the treatment of contaminated environments.     

海水养殖循环系统不同功能区域微生物群落结构比较

为了探究海水养殖循环系统不同功能区域微生物群落结构特征,深入了解水质理化因子与微生物群落之间的相关性,通过16S rRNA扩增子高通量测序技术对系统不同功能区域水体微生物群落结构进行了分析。结果表明,变形菌门(Proteobacteria)和拟杆菌门(Bacteroidetes)为整个系统绝对优势菌门,不同功能区域优势属组成差异较大,但是弧菌(Vibrio)丰度都在1%以上。脉红螺养殖池和生物滤池水体α多样性最高,鲍鱼养殖池水体和自然海水多样性最低。NMDS分析结果显示生物滤池和养殖池进水口微生物群落结构相似,其他区域微生物群落结构有明显差异。生物滤池到养殖池NH₄⁺和NO₂^-去除率分别为8.77%和45.12%,NO₃^-质量浓度在整个系统内都处于较高水平(148.50~200.47mg/L)。环境因子与微生物群落结构没有显著相关性,温度T和NO₂^-对微生物群落结构的影响相对较大。研究结果为下一步循环水养殖系统设计的完善和日常管理提供了参考。     

有机紫外吸收剂在海洋中的时空分布及其生态风险评价

以Web of Science数据库收录的65篇英文文献为研究对象，综合分析近15年来世界各海域海水、沉积物和海洋生物中多种有机紫外吸收剂(Organic Ultraviolet Absorbers，OUVs)的赋存特征和时空分布，并结合已有的毒理学数据，采用风险商(Risk Quotient, RQ)法评估海水中OUVs对海洋生物的生态风险。结果表明，从空间分布上看，近岸海域的OUVs浓度水平普遍高于离岸海域，这可能是由于人类活动对近岸海域的影响较大。从时间分布上看，夏季由于抗紫外线产品使用量增加，导致OUVs的释放量高于其他季节。风险评估结果表明，在全球范围内，二苯酮-3 (BP-3)、甲氧基肉桂酸乙基己酯(EHMC)、奥克立林(OC)、辛基二甲基对氨基苯甲酸(OD-PABA)、阿伏苯宗(AVO)和水杨酸异辛酯(EHS)分别在37.5%、87.0%、100.0%、44.4%、60.0%和60.0%的海域对海洋生物具有中高风险，而4-甲基苄亚基樟脑(4-MBC)在所有海域对海洋生物均无风险。该研究旨在评估OUVs在世界海洋中的污染状态，以期更好地从源头控制和管理OUVs及其对海洋生态环境的影响。     

Volcanic carbon dioxide vents show ecosystem effects of ocean acidification

The atmospheric partial pressure of carbon dioxide (p(CO(2))) will almost certainly be double that of pre-industrial levels by 2100 and will be considerably higher than at any time during the past few million years. The oceans are a principal sink for anthropogenic CO(2) where it is estimated to have caused a 30% increase in the concentration of H(+) in ocean surface waters since the early 1900s and may lead to a drop in seawater pH of up to 0.5 units by 2100 (refs 2, 3). Our understanding of how increased ocean acidity may affect marine ecosystems is at present very limited as almost all studies have been in vitro, short-term, rapid perturbation experiments on isolated elements of the ecosystem. Here we show the effects of acidification on benthic ecosystems at shallow coastal sites where volcanic CO(2) vents lower the pH of the water column. Along gradients of normal pH (8.1-8.2) to lowered pH (mean 7.8-7.9, minimum 7.4-7.5), typical rocky shore communities with abundant calcareous organisms shifted to communities lacking scleractinian corals with significant reductions in sea urchin and coralline algal abundance. To our knowledge, this is the first ecosystem-scale validation of predictions that these important groups of organisms are susceptible to elevated amounts of p(CO(2)). Sea-grass production was highest in an area at mean pH 7.6 (1,827 (mu)atm p(CO(2))) where coralline algal biomass was significantly reduced and gastropod shells were dissolving due to periods of carbonate sub-saturation. The species populating the vent sites comprise a suite of organisms that are resilient to naturally high concentrations of p(CO(2)) and indicate that ocean acidification may benefit highly invasive non-native algal species. Our results provide the first in situ insights into how shallow water marine communities might change when susceptible organisms are removed owing to ocean acidification.     

Rapid removal of Chernobyl fallout from Mediterranean surface waters by biological activity

The sinking of particulate organic matter from the euphotic zone is an important pathway for the vertical transport of many elements and organic compounds in the sea. Many natural and artificial radionuclides in surface waters are readily adsorbed onto suspended particles and are presumably scavenged and removed to depth on time scales commensurate with both particle sinking rate and retention time of the radionuclide on the particle. Previously, abyssal benthic organisms from the northeast Pacific were found to contain short-lived fission products which entered the sea surface as fallout from nuclear testing. The presence of these radionuclides at great depth could not be explained by Stokesian settling of small fallout particles and it was hypothesized that zooplankton grazing in the surface layers packaged these particle-reactive radionuclides into large, relatively dense faecal pellets which rapidly sank to depth. We report here data from a time-series sediment trap experiment and concomitant zooplankton collections which show conclusively that Chernobyl radioactivity, in particular the rare earth nuclides 141Ce and 144Ce, entering the Mediterranean as a single pulse, was rapidly removed from surface waters and transported to 200 m in a few days primarily by zooplankton grazing.     

Unsuspected diversity among marine aerobic anoxygenic phototrophs

Aerobic, anoxygenic, phototrophic bacteria containing bacteriochlorophyll a (Bchla) require oxygen for both growth and Bchla synthesis. Recent reports suggest that these bacteria are widely distributed in marine plankton, and that they may account for up to 5% of surface ocean photosynthetic electron transport and 11% of the total microbial community. Known planktonic anoxygenic phototrophs belong to only a few restricted groups within the Proteobacteria alpha-subclass. Here we report genomic analyses of the photosynthetic gene content and operon organization in naturally occurring marine bacteria. These photosynthetic gene clusters included some that most closely resembled those of Proteobacteria from the beta-subclass, which have never before been observed in marine environments. Furthermore, these photosynthetic genes were broadly distributed in marine plankton, and actively expressed in neritic bacterioplankton assemblages, indicating that the newly identified phototrophs were photosynthetically competent. Our data demonstrate that planktonic bacterial assemblages are not simply composed of one uniform, widespread class of anoxygenic phototrophs, as previously proposed; rather, these assemblages contain multiple, distantly related, photosynthetically active bacterial groups, including some unrelated to known and cultivated types.     

江苏海岸带生物体的重金属水平与生态评价

通过对江苏海岸带几种主要生物的重金属含量测定,分析生物体的重金属富集效应和元素间的变化．分析结果表明,底栖生物的重金属含量高于浮游生物,生物肉体和壳体对外界金属元素的吸收具有选择性,一般壳体吸收更多的有害元素;对于不同的生物个体,吸收的主要方式和含量不同．根据有关海洋生物体污染物标准,认为江苏沿海的生物体已受到轻度污染．     

Novel major archaebacterial group from marine plankton.

Marine bacteria often dominate the plankton biomass and are responsible for much of the cycling of organic matter, but bacterial diversity is poorly understood because conventional identification methods (requiring culturing) miss about 99% of the organisms. Recent advances permit characterization of microbial communities by analysis of 16S ribosomal RNA gene sequences directly from biomass without the need to culture the organisms; such studies from surface ocean samples have found only eubacteria, not archaebacteria (or Archaea), which are profoundly different. Here we report 16S rRNA sequences obtained from Pacific Ocean bacterioplankton samples collected from depths of 100 m and 500 m. Among these we found sequences only distantly related to those of any organisms previously characterized by 16S rRNA sequences, with similarities to the nearest such relatives (extreme thermophiles) approximately the same as those between animals and plants. We suggest that these sequences are from a previously undescribed archaebacterial group that may have diverged from the ancestors of characterized organisms very early in evolution.     

Genetic diversity in Sargasso Sea bacterioplankton

Bacterioplankton are recognized as important agents of biogeochemical change in marine ecosystems, yet relatively little is known about the species that make up these communities. Uncertainties about the genetic structure and diversity of natural bacterioplankton populations stem from the traditional difficulties associated with microbial cultivation techniques. Discrepancies between direct counts and plate counts are typically several orders of magnitude, raising doubts as to whether cultivated marine bacteria are actually representative of dominant planktonic species. We have phylogenetically analysed clone libraries of eubacterial 16S ribosomal RNA genes amplified from natural populations of Sargasso Sea picoplankton by the polymerase chain reaction. The analysis indicates the presence of a novel microbial group, the SAR11 cluster, which appears to be a significant component of this oligotrophic bacterioplankton community. A second cluster of lineages related to the oxygenic phototrophs--cyanobacteria, prochlorophytes and chloroplasts--was also observed. However, none of the genes matched the small subunit rRNA sequences of cultivated marine cyanobacteria from similar habitats. The diversity of 16S rRNA genes observed within the clusters suggests that these bacterioplankton may be consortia of independent lineages sharing surprisingly distant common ancestors.     

Agricultural runoff fuels large phytoplankton blooms in vulnerable areas of the ocean

Biological productivity in most of the world's oceans is controlled by the supply of nutrients to surface waters. The relative balance between supply and removal of nutrients--including nitrogen, iron and phosphorus--determines which nutrient limits phytoplankton growth. Although nitrogen limits productivity in much of the ocean, large portions of the tropics and subtropics are defined by extreme nitrogen depletion. In these regions, microbial denitrification removes biologically available forms of nitrogen from the water column, producing substantial deficits relative to other nutrients. Here we demonstrate that nitrogen-deficient areas of the tropical and subtropical oceans are acutely vulnerable to nitrogen pollution. Despite naturally high nutrient concentrations and productivity, nitrogen-rich agricultural runoff fuels large (54-577 km2) phytoplankton blooms in the Gulf of California. Runoff exerts a strong and consistent influence on biological processes, in 80% of cases stimulating blooms within days of fertilization and irrigation of agricultural fields. We project that by the year 2050, 27-59% of all nitrogen fertilizer will be applied in developing regions located upstream of nitrogen-deficient marine ecosystems. Our findings highlight the present and future vulnerability of these ecosystems to agricultural runoff.     

SAR差分干涉测量技术在地震区域形变测量中的应用研究

合成孔径雷达(SAR)差分干涉测量是一种最新的大地形变测量遥感技术方法.利用欧洲空间局1996年4月15日获取的地球资源卫星ERS-1数据和4月16日获取的ERS-2数据,以及1997年12月2日获取的ERS-2数据,应用差分干涉测量技术对发生于1997年11月8日的西藏玛尼地震进行了提取区域形变场的应用研究.从得到的变化检测条纹图中可识别出地表破裂带,并可定量推算震中周围和两条断裂带附近的变形情况.分析结果表明:断裂为左行平移性质,断裂带南侧水平错动的最大变形量至少超过2.8 m,北侧水平错动的最大变形量至少超过1.75 m.差分干涉测量分析结果与地面调查及已有的相关研究资料符合得很好.     

Bacterial photosynthesis in surface waters of the open ocean

The oxidation of the global ocean by cyanobacterial oxygenic photosynthesis, about 2,100 Myr ago, is presumed to have limited anoxygenic bacterial photosynthesis to oceanic regions that are both anoxic and illuminated. The discovery of oxygen-requiring photosynthetic bacteria about 20 years ago changed this notion, indicating that anoxygenic bacterial photosynthesis could persist under oxidizing conditions. However, the distribution of aerobic photosynthetic bacteria in the world oceans, their photosynthetic competence and their relationship to oxygenic photoautotrophs on global scales are unknown. Here we report the first biophysical evidence demonstrating that aerobic bacterial photosynthesis is widespread in tropical surface waters of the eastern Pacific Ocean and in temperate coastal waters of the northwestern Atlantic. Our results indicate that these organisms account for 2-5% of the photosynthetic electron transport in the upper ocean.     

浅析合成孔径雷达遥感图像海岸带应用

海岸带自然资源丰富、变动频繁,是人类活动的重要区域。合成孔径雷达(SAR)以其全天时、全天候和高分辨率等特征,成为海岸带探测和动态监测的重要遥感器。论述了SAR探测地物的原理、海岸带地物的SAR图像解译标志和SAR图像处理方法。在此基础上,重点探讨了SAR图像在海岸带地质地貌探测、海岸分类、海岸带土地开发利用探测和海岸带动态变迁的监测等几方面的应用。