Microbial diversity in surface sediments of the Xisha Trough, the South China Sea

Microbial communities were obtained from the surface sediments of the Xisha Trough using the culture-independent technique. The characteristics of the 16S rDNA gene amplified from the sediments indicated that archaeal clones could be grouped into Euryarchaeota and Crenarchaeota, respectively. Two archaeal groups, Marine Crenarchaeotic GroupI and Terrestrial Miscellaneous Euryarchaeotal Group, were the most dominant archaeal 16S rDNA gene components in the sediments. The remaining components were related to the members of Marine Benthic Group B, Marine Benthic Group A, Marine Benthic Group D, Novel Euryarchaeotic Group and C3. The bacterial clones exhibited greater diversity than the archaeal clones with the 16S rDNA gene sequences from the members of Proteobacteria, Planctomycetes, Actinobacteria, Firmicutes, Chloroflexi, Acidobacteria, candidate division OP8, Bacterioidetes/Chlorobi and Verrucomicrobia. Most of these lineages represented uncultured microorganisms. The result suggests that a vast amount of microbial resource in the surface sediments of the South China Sea has not been known.     

南海西沙海槽表层沉积物微生物多样性

利用非培养的分子技术研究了西沙海槽表层沉积物中的微生物群落.沉积物中扩增的古菌16S rDNA 序列分属两个大类:泉古生菌(Crenarchaeota)和广古生菌(Euryarchaeota).以Marine Crenarchaeotic GroupⅠ (古菌16S rDNA文库的49.2%)和Terrestrial Miscellaneous Euryarchaeotal Group (16.9%)为主要类群;其余为Marine Benthic Group B (9.7%)、 Marine Benthic Group A (4%)、 Marine Benthic Group D (1.6%)、Novel Euryarchaeotic Group (0.8%)和 C3(0.8%).细菌克隆子多样性明显高于古菌,16S rDNA序列分别来自变形杆菌(Proteobacteria)(细菌16S rDNA文库的30.5%)、浮霉菌(Planctomycetes)(20.3%)、放线菌(Actinobacteria)(14.4%)、厚壁菌(Firmicutes)(15.3%)、屈桡杆菌(Chloroflexi)(8.5%)、酸杆菌(Acidobacteria)(3.4%)、candidate division OP8 (2.5%)、拟杆菌/绿菌(Bacterioidetes/Chlorobi)(1.7%)和疣微菌(Verrucomicrobia)(1.7%).变形杆菌为优势类群(包括Alpha-和Delta-Proteobacteria亚群).多数克隆子为未培养细菌和古菌.结果表明南海表层沉积物中蕴含大量未知的微生物资源.     

Archaeal communities associated with shallow to deep subseafloor sediments of the New Caledonia Basin

The distribution of the archaeal communities in deep subseafloor sediments [0–36 m below the seafloor (mbsf)] from the New Caledonia and Fairway Basins was investigated using DNA- and RNA-derived 16S rRNA clone libraries, functional genes and denaturing gradient gel electrophoresis (DGGE). A new method, Co-Migration DGGE (CM-DGGE), was developed to access selectively the active archaeal diversity. Prokaryotic cell abundances at the open-ocean sites were on average ∼3.5 times lower than at a site under terrestrial influence. The sediment surface archaeal community (0–1.5 mbsf) was characterized by active Marine Group 1 (MG-1) Archaea that co-occurred with ammonia monooxygenase gene ( amoA ) sequences affiliated to a group of uncultured sedimentary Crenarchaeota . However, the anoxic subsurface methane-poor sediments (below 1.5 mbsf) were dominated by less active archaeal communities, such as the Thermoplasmatales , Marine Benthic Group D and other lineages probably involved in the methane cycle ( Methanosarcinales , ANME-2 and DSAG/MBG-B). Moreover, the archaeal diversity of some sediment layers was restricted to only one lineage (Uncultured Euryarchaeota , DHVE6, MBG-B, MG-1 and SAGMEG). Sequences forming two clusters within the Thermococcales order were also present in these cold subseafloor sediments, suggesting that these uncultured putative thermophilic archaeal communities might have originated from a different environment. This study shows a transition between surface and subsurface sediment archaeal communities.     

南海南部陆坡表层沉积物细菌和古菌多样性

从南海南部陆坡表层沉积物中扩增了细菌和古菌16S rDNA序列,并对克隆子文库进行系统发育分析.细菌序列以变形杆菌(Proteobacteria)居多,其次是浮霉菌(Planctomycete)、酸杆菌(Acidobacteria)和candidate division OP10,另外还有少量铁还原杆菌(Deferrobacteres)、candidate division OP3、OP11、OP8、TM6、疣微菌(Verrucomicrobia)和螺旋体(Spirochaetes).古菌序列分别来自泉古生菌(Crenarchaeota)和广古生菌(Euryarchaeota),以Marine Benthic Group B(MBGB)、MarineCrenarchaeotic Group Ⅰ(MGⅠ)、Marine Benthic Group D(MBGD)和South African Gold Mine Euryarchaeotic Group(SAGMEG)为主.少量序列为C3、甲烷杆菌(Methanobacteriales)和Novel Euryarchaeotic Group(NEG).结果表明海底表层沉积物中有丰富多样的微生物群落.     

Community Structure of Archaea from Deep-Sea Sediments of the South China Sea

Using the archaeal 16S rRNA gene, we determined the community structures of archaea of subseafloor sediments (～9-11 m below seafloor) from two geographically distant cores (MD05-2896, south, water depth 1,657 m; MD05-2902, north, water depth 3,697 m) in the South China Sea. Euryarchaeota accounted for 61.4% of total archaeal clone libraries at MD05-2896 and 56.2% at MD05-2902. At both locations, the Euryarchaeota-related sequences were dominated by Marine Benthic Group D, Terrestrial Miscellaneous Eryarchaeotal Group, and South African GoldMine Euryarchaeotal Group; the Crenarchaeota-related sequences were dominated by Marine Benthic Group B, Marine Group I, pSL12, and C3. The community structure showed no significant difference with depth at each location, suggesting the lack of stratification of archaeal populations in the deep-sea marine sediments in the South China Sea. On the other hand, the community structure is significantly different between the two sites, which may be related to geographical difference in the South China Sea.     

Active Archaeal Communities at Cold Seep Sediments Populated by Siboglinidae Tubeworms from the Storegga Slide

Siboglinid tubeworms in cold seep sediments can locally modify the geochemical gradients of electron acceptors and donors, hence creating potential microhabitats for prokaryotic populations. The archaeal communities associated with sediments populated by Oligobrachia haakonmosbiensis and Sclerolinum contortum Siboglinid tubeworms in the Storegga Slide were examined in this study. Vertical distribution of archaeal communities was investigated using denaturing gradient gel electrophoresis based on 16S rRNA genes. The active fraction of the archaeal community was assessed by using reverse-transcribed rRNA. Archaeal communities associated with sediments colonized by tubeworms were affiliated with uncultivated archaeal lineages of the Crenarchaeota and Euryarchaeota. The composition of the active archaeal populations changed with depth indicating a reorganization of microbial communities. 16S rRNA gene libraries were dominated by sequences affiliated to the Rice Cluster V which are unusual in marine sediment samples. Moreover, this study provides the first evidence of living Crenarchaeota of the Rice Cluster V in cold seep sediments. Furthermore, the Storegga Slide sediments harbored a high diversity of other minor groups of uncultivated lineages including Terrestrial Miscellaneous Euryarchaeotal Group, Marine Benthic Group (MBG)-D, MBG-E, Deep-Sea Hydrothermal Vent Euryarchaeotal Group, Lake Dagow Sediment, Val Kotinen Lake clade III, and Sippenauer Moor 1. Thus, we hypothesize that the vertical geochemical imprint created by the tubeworms could support broad active archaeal populations in the Siboglinidae-populated Storegga Slide sediments.     

Diversity of Archaea in Bottom Sediments of the Discharge Areas With Oil- and Gas-Bearing Fluids in Lake Baikal

Using massively parallel sequencing (the Roche 454 platform) we have studied the diversity of archaeal 16S rRNA gene sequences in oxic and anoxic sediments at six sites in Lake Baikal with oil- and gas-bearing fluids discharge. Archaeal communities appeared to be represented mainly by five phyla: Euryarchaeota, Crenarchaeota, Thaumarchaeota, Bathyarchaeota (miscellaneous Crenarchaeotic group), and Woesearchaeota (deep sea hydrothermal vent group 6). Among them we detected sequences of methanogens of the orders Methanomicrobiales, Methanosarsinales, Methanococcales, as well as representatives of the following uncultured archaeal lineages: Group C3, Marine Benthic Group D, and Terrestrial Miscellaneous Group. We have also identified sequences of ammonia-oxidizing archaea of the phyla Crenarchaeota and Thaumarchaeota. Phylogenetic analysis showed the presence ANME-2d-related sequences. However, the analysis of mcrA genes libraries has not revealed typical representatives of ANME groups. Comparison of amplicon libraries 16S rRNA gene fragments from different samples proved the widespread presence of previously detected Baikal archaeal lineages, which are members of the phylum Crenarchaeota and Thaumarchaeota (formerly Group C3 of Crenarchaeota).     

Phylogenetic diversity of planktonic archaea in the estuarine region of East China Sea

To examine the diversity and structure of archaeal communities in the Yangtze River estuarine region of East China Sea (ECS), the 16S rRNA gene clone libraries of two typical sites were constructed with the archaea-specific primers. In total, 71 clones randomly selected were screened by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis and 21 clones with unique RFLP pattern were sequenced. All the sequences are clustered into the two groups of Marine Group I (MGI) and Marine Group II (MGII) which are affiliated with the phyla Crenarchaeota and Euryarchaeota, respectively. MGI clones dominate both libraries with 20 MGI sequences obtained. The majority of sequences are closely related to uncultured marine archaea except for two sequences of which the nearest neighbor is a newly identified isolate of nitrifying marine archaeon Nitrosopumilus maritimus (98% identity). The results indicate that ECS coastal waters are inhabited by archaeal community with low dominance and high diversity corresponding to the complex estuarine environments, suggesting that archaea may perform an important role in the estuarine ecosystem.     

Stratification of archaea in the deep sediments of a freshwater meromictic lake: vertical shift from methanogenic to uncultured archaeal lineages

As for lineages of known methanogens, several lineages of uncultured archaea were recurrently retrieved in freshwater sediments. However, knowledge is missing about how these lineages might be affected and structured according to depth. In the present study, the vertical changes of archaeal communities were characterized in the deep sediment of the freshwater meromictic Lake Pavin. For that purpose, an integrated molecular approach was performed to gain information on the structure, composition, abundance and vertical stratification of archaeal communities thriving in anoxic freshwater sediments along a gradient of sediments encompassing 130 years of sedimentation. Huge changes occurred in the structure and composition of archaeal assemblages along the sediment core. Methanogenic taxa (i.e. Methanosaeta and Methanomicrobiales) were progressively replaced by uncultured archaeal lineages (i.e. Marine Benthic Group-D (MBG-D) and Miscellaneous Crenarchaeal Group (MCG)) which are suspected to be involved in the methane cycle.     

Vertical structure of archaeal communities and the distribution of ammonia monooxygenase A gene variants in two meromictic High Arctic lakes

The distribution of archaeal amoA and 16S rRNA genes was evaluated in two marine-derived, meromictic lakes in the Canadian High Arctic: Lake A and Lake C1 on the northern coast of Ellesmere Island. The amoA gene was recorded in both lakes, with highest copy numbers in the oxycline. Sequence analysis showed that amoA from the two lakes shared 94% similarity, indicating at least two phylogenetically distinct clusters. Clone libraries of archaeal 16S rRNA genes from Lake A revealed strong vertical differences in archaeal community diversity and composition down the water column. The oxic layer was dominated by one group of Euryarchaeota affiliated to the Lake Dagow Sediment (LDS) cluster. This group was absent from the oxycline, which had an extremely low archaeal diversity of two phylotypes. Both belonged to the Crenarchaeota Marine Group I (MGI), the marine group that has been linked to archaeal amoA ; however, there was a low ratio of amoA to MGI copy numbers, suggesting that many MGI Archaea did not carry the amoA gene. The anoxic zone contained representatives of the RC-V (Rice Cluster-V) and LDS clusters of Euryarchaeota. These results show the strong vertical differentiation of archaeal communities in polar meromictic lakes, and they suggest archaeal nitrification within the oxycline of these highly stratified waters.     

High archaeal richness in the water column of a freshwater sulfurous karstic lake along an interannual study

We surveyed the archaeal assemblage in a stratified sulfurous lake (Lake Vilar, Banyoles, Spain) over 5 consecutive years to detect potential seasonal and interannual trends in the free-living planktonic Archaea composition. The combination of different primer pairs and nested PCR steps revealed an unexpectedly rich archaeal community. Overall, 140 samples were analyzed, yielding 169 different 16S rRNA gene sequences spread over 14 Crenarchaeota (109 sequences) and six Euryarchaeota phylogenetic clusters. Most of the Crenarchaeota (98% of the total crenarchaeotal sequences) affiliated within the Miscellaneous Crenarchaeota Group (MCG) and were related to both marine and freshwater phylotypes. Euryarchaeota mainly grouped within the Deep Hydrothermal Vent Euryarchaeota (DHVE) cluster (80% of the euryarchaeotal sequences) and the remaining 20% distributed into three less abundant taxa, most of them composed of soil and sediment clones. The largest fraction of phylotypes from the two archaeal kingdoms (79% of the Crenarchaeota and 54% of the Euryarchaeota) was retrieved from the anoxic hypolimnion, indicating that these cold and sulfide-rich waters constitute an unexplored source of archaeal richness. The taxon rank-frequency distribution showed two abundant taxa (MCG and DHVE) that persisted in the water column through seasons, plus several rare ones that were only detected occasionally. Differences in richness distribution and seasonality were observed, but no clear correlations were obtained when multivariate statistical analyses were carried out.     

中国南海北部陆坡沉积物古菌多样性及丰度分析

【目的】海洋沉积物中的古菌在全球生物地球化学循环中充当重要的角色,深入了解沉积物中古菌群落的结构及功能特征是探究海洋沉积物中古菌参与生物地球化学循环和生态学功能的基础。【方法】采用高通量测序技术,分别对南海北部陆坡不同海域(东部,西部和神狐海域的7个站位)沉积物中古菌16SrRNA基因进行Illumina Mi Seq测序。【结果】中国南海北部陆坡沉积物中古菌的主要门类是Bathyarchaeota、Thermoplasmata、Woesearchaeota(DHVEG-6)、Thaumarchaeota(Marine Group I)、Lokiarchaeota和Marine Hydrothermal Vent Group(MHVG),还存在少量的AK8、Marine Benthic Group A和Terrestrial Hot Spring Crenarchaeota Group(THSCG)等。在潜在水合物区沉积物中还发现了甲烷代谢相关古菌(Anaerobic methanotrophic archaea,ANME)类群,主要为ANME-1、ANME-2ab和ANME-2c等。甲烷代谢古菌的分布特征也从甲烷代谢保守功能基因mcr A(Methyl coenzyme-Mreductase A)的扩增中得到了验证。利用定量PCR对南海沉积物中的细菌、古菌的16SrRNA基因和mcrA基因进行了定量,发现细菌16SrRNA基因拷贝数为10~5-10~7 copies/g(湿重),古菌16SrRNA基因拷贝数为10~5-10~6 copies/g(湿重),潜在水合物区mcrA基因拷贝数为10~3-10~5 copies/g(湿重)。【结论】揭示了中国南海北部陆坡沉积物中具有丰富的微生物资源,其中古菌种类多样且丰度较高,同时发现冷泉特征古菌群落,为深入认识和理解南海沉积物中微生物丰度和古菌多样性,以及解析古菌地球化学功能奠定基础。     

海南东寨港红树林不同植被土壤微生物群落结构比较

【目的】比较不同植被下红树林土壤细菌和古菌的多样性及群落结构,认识红树林土壤微生物资源多样性。【方法】直接提取红树林土壤总DNA,采用细菌通用引物27F/1492R和古菌通用引物Arch21F/Arch958R进行PCR扩增,构建细菌和古菌16S rRNA基因文库,对海南东寨港自然保护区秋茄林、无瓣海桑林和无红树林裸滩土壤的细菌和古菌多样性和群落结构进行分析和比较。【结果】3种土壤样品的细菌类群包括变形细菌门(Proteobacteria)等16个类群,其中变形细菌门(Proteobacteria)与绿屈挠菌门(Chloroflexi)是优势类群;古菌包括6个嗜泉古菌界(Crenarchaeota)类群和7个广域古菌界(Euryarchaeota)类群,分别以Marine Benthic Group C、Marine Benthic Group D为优势类群。多样性指数(H’)和物种丰富度指数(Schao1)表明,本地种秋茄林下土壤细菌和古菌的多样性指数最高,外来种无瓣海桑显著低于秋茄林,甚至明显低于相邻无红树林裸滩沉积物;不同植被下土壤细菌和古菌群落结构存在显著差异,秋茄林土壤微生物群落结构和无红树林裸滩沉积物更相似。【结论】红树林土壤微生物类群丰富,不同植被下土壤细菌和古菌多样性和群落结构存在显著差异。     

The phylogeny of endolithic microbes associated with marine basalts

We examined the phylogenetic diversity of microbial communities associated with marine basalts, using over 300 publicly available 16S rDNA sequences and new sequence data from basalt enrichment cultures. Phylogenetic analysis provided support for 11 monophyletic clades originating from ocean crust (sediment, basalt and gabbro). Seven of the ocean crust clades (OCC) are bacterial, while the remaining four OCC are in the Marine Group I (MGI) Crenarchaeota. Most of the OCC were found at diverse geographic sites, suggesting that these microorganisms have cosmopolitan distributions. One OCC in the Crenarchaeota consisted of sequences derived entirely from basalts. The remaining OCC were found in both basalts and sediments. The MGI Crenarchaeota were observed in all studies where archaeal diversity was evaluated. These results demonstrate that basalts are occupied by cosmopolitan clades of microorganisms that are also found in marine sediments but are distinct from microorganisms found in other marine habitats, and that one OCC in the ubiquitous MGI Crenarchaeota clade may be an ecotype specifically adapted to basalt.     

Bacterial and Archaeal 16S rRNA Genes in Late Pleistocene to Holocene Muddy Sediments from the Kanto Plain of Japan

Microbial communities in ancient marine sediments composed of clay and silt obtained from the terrestrial subsurface were phylogenetically analyzed based on their 16S rRNA gene sequences. Chloroflexi and Miscellaneous Crenarchaeotic Group were predominant in bacterial and archaeal clone libraries, respectively. Of 44 operational taxonomic units (OTUs) that had close relatives in the database, 30 were close to sequences obtained from marine environments. Some sequences belonged to the candidate groups JS1, ANME-I, and Marine Benthic Group-C, which are typically found in marine sediments. Low chloride concentrations in the sediments suggest that these marine-affiliated sequences may not reflect currently active microbial communities. Our results indicate the existence of long-term preserved DNA or descendants of ancient oceanic microbial components in subsurface muddy sediments in a temperate region, which may reflect indigenous population of paleoenvironments.     

南海北部陆坡神狐海域HS-PC500 岩心微生物多样性

[目的]本文研究南海北部陆坡神狐HS-PC500重力活塞岩心沉积物中微生物多样性.[方法]使用吖啶橙染色法计数沉积物中微生物丰度;提取沉积物微生物总DNA,使用特异性引物扩增古菌及细菌16SrRNA基因序列;对克隆文库进行系统发育分析.[结果]系统发育分析显示表层PC500-l(0-5 cm below sea floor,bsf)古菌以C3为主要类群,占该层总序列的25.6%;中层PC500-6(350-355 cm bsf)和底层PC500-11(790-795 cm bsf)古菌以Marine Benthic Group(MBG)-B为主要类群,分别占该层总序列的48.1%和38.9%.另有部分克隆序列属于MBG-A、Miscellaneous Crenarchaeotic Group(MCG)、Thermoprotei、NGC、Halobacteriales、MBG-E、South African Gold Mine Euryarchaeotic Group(SAGMEG).表层细菌以变形菌(Proteobacteria)为主要类群,占该层文库的38.3%.中层和底层细菌以绿弯菌(Choloflexi)和JS1为主要类群,分别占该层文库的28.1%、29.2%和39%、24.7%.另有部分克隆序列属于硝化螺旋菌(Nitrospirae)、放线菌(Actinobacteria)、酸杆菌(Acidobacteria)、OP8、螺旋体菌(Spirochaetes)、TM6、脱铁杆菌(Deferribacteres)、浮霉菌(Plantomycete).[结论]结果显示,HS-PC500岩心微生物丰度与甲烷浓度变化相吻合;微生物丰度较低可能与较低的总有机碳量有关;微生物多样性较高,并且随深度的增加群落结构变化明显;岩心中有关硫酸盐还原的微生物类群占优势,说明微生物的硫代谢在该海区沉积物的物质循环过程中占有重要地位.     

Identification of novel Archaea in bacterioplankton of a boreal forest lake by phylogenetic analysis and fluorescent in situ hybridization(1)

We report here on novel groups of Archaea in the bacterioplankton of a small boreal forest lake studied by the culture-independent analysis of the 16S rRNA genes amplified directly from lake water in combination with fluorescent in situ hybridization (FISH). Polymerase chain reaction products were cloned and 28 of the 160 Archaea clones with around 900-bp-long 16S rRNA gene inserts, were sequenced. Phylogenetic analysis, including 642 Archaea sequences, confirmed that none of the freshwater clones were closely affiliated with known cultured Archaea. Twelve Archaea sequences from lake Valkea Kotinen (VAL) belonged to Group I of uncultivated Crenarchaeota and affiliated with environmental sequences from freshwater sediments, rice roots and soil as well as with sequences from an anaerobic digestor. Eight of the Crenarchaeota VAL clones formed a tight cluster. Sixteen sequences belonged to Euryarchaeota. Four of these formed a cluster together with environmental sequences from freshwater sediments and peat bogs within the order Methanomicrobiales. Five were affiliated with sequences from marine sediments situated close to marine Group II and three formed a novel cluster VAL III distantly related to the order Thermoplasmales. The remaining four clones formed a distinct clade within a phylogenetic radiation characterized by members of the orders Methanosarcinales and Methanomicrobiales on the same branch as rice cluster I, detected recently on rice roots and in anoxic bulk soil of flooded rice microcosms. FISH with specifically designed rRNA-targeted oligonucleotide probes revealed the presence of Methanomicrobiales in the studied lake. These observations indicate a new ecological niche for many novel 'non-extreme' environmental Archaea in the pelagic water of a boreal forest lake.     

Large-Scale Distribution and Activity of Prokaryotes in Deep-Sea Surface Sediments of the Mediterranean Sea and the Adjacent Atlantic Ocean

The deep-sea represents a substantial portion of the biosphere and has a major influence on carbon cycling and global biogeochemistry. Benthic deep-sea prokaryotes have crucial roles in this ecosystem, with their recycling of organic matter from the photic zone. Despite this, little is known about the large-scale distribution of prokaryotes in the surface deep-sea sediments. To assess the influence of environmental and trophic variables on the large-scale distribution of prokaryotes, we investigated the prokaryotic assemblage composition (Bacteria to Archaea and Euryarchaeota to Crenarchaeota ratio) and activity in the surface deep-sea sediments of the Mediterranean Sea and the adjacent North Atlantic Ocean. Prokaryotic abundance and biomass did not vary significantly across the Mediterranean Sea; however, there were depth-related trends in all areas. The abundance of prokaryotes was positively correlated with the sedimentary concentration of protein, an indicator of the quality and bioavailability of organic matter. Moving eastwards, the Bacteria contribution to the total prokaryotes decreased, which appears to be linked to the more oligotrophic conditions of the Eastern Mediterranean basins. Despite the increased importance of Archaea, the contributions of Crenarchaeota Marine Group I to the total pool was relatively constant across the investigated stations, with the exception of Matapan-Vavilov Deep, in which Euryarchaeota Marine Group II dominated. Overall, our data suggest that deeper areas of the Mediterranean Sea share more similar communities with each other than with shallower sites. Freshness and quality of sedimentary organic matter were identified through Generalized Additive Model analysis as the major factors for describing the variation in the prokaryotic community structure and activity in the surface deep-sea sediments. Longitude was also important in explaining the observed variability, which suggests that the overlying water masses might have a critical role in shaping the benthic communities.     

Archaea in the Gulf of Aqaba

Using a polyphasic approach, we examined the presence of Archaea in the Gulf of Aqaba, a warm marine ecosystem, isolated from major ocean currents and subject to pronounced seasonal changes in hydrography. Catalyzed reported deposition FISH analyses showed that Archaea make up to >20% of the prokaryotic community in the Gulf. A spatial separation between the two major phyla of Archaea was observed during summer stratification. Euryarchaeota were found exclusively in the upper 200 m, whereas Crenarchaeota were present in greater numbers in layers below the summer thermocline. 16S rRNA gene-based denaturing gradient gel electrophoresis confirmed this depth partitioning and revealed further diversity of Crenarchaeota and Euryarchaeota populations along depth profiles. Phylogenetic analysis showed pelagic Crenarchaeota and Euryarchaeota to differ from coral-associated Archaea from the Gulf, forming distinct clusters within the Marine Archaea Groups I and II. Endsequencing of fosmid libraries of environmental DNA provided a tentative identification of some members of the archaeal community and their role in the microbial community of the Gulf. Incorporation studies of radiolabeled leucine and bicarbonate in the presence of different inhibitors suggest that the archaeal community participates in autotrophic CO₂ uptake and contributes little to the heterotrophic activity.     

Archaeal diversity associated with in situ samplers deployed on hydrothermal vents on the East Pacific Rise (13 degrees N)

To evaluate possible compositional changes in archaeal communities at a deep-sea hydrothermal vent field scale, we examined five different samples obtained after deploying in situ collectors for different times on three spatially separated venting sulphide structures on the East Pacific Rise (13 degrees N). Direct cell counts and whole-cell hybridizations with fluorescently labelled 16S rRNA-based oligonucleotide probes revealed that the relative abundance of archaeal populations represented from 14 to 33% of the prokaryotic community. 16S rDNA sequence analysis of the archaeal clone libraries indicated that a large percentage of clones were closely related to known archaeal isolates recovered from similar habitats. Among the 24 different phylotypes identified, Thermococcales-related sequences were dominant in all the libraries that also included representative genera of orders Methanopyrales, Methanococcales, Archaeoglobales and Desulfurococcales. The presence of most of these phylogenetic groups was confirmed in enrichment cultures performed at temperatures from 60 to 90 degrees C. Additional sequences with no known cultivated relatives grouped with the Marine group I Crenarchaeota, Korarchaeota and Deep-sea Hydrothermal Vent Euryarchaeota (DHVE) within which a novel lineage was identified. Furthermore, the archaeal community composition was distinct from vent to vent within the same vent field and varied within short time scales. This study provides new insights into microbial diversity and distribution at deep-sea hydrothermal vents.