Marine Benthic Group D Research Articles

An Ancient Respiratory System in the Widespread Sedimentary Archaea Thermoprofundales.

Thermoprofundales, formerly Marine Benthic Group D (MBG-D), is a ubiquitous archaeal lineage found in sedimentary environments worldwide. However, its taxonomic classification, metabolic pathways, and evolutionary history are largely unexplored because of its uncultivability and limited number of sequenced genomes. In this study, phylogenomic analysis and average amino acid identity values of a collection of 146 Thermoprofundales genomes revealed five Thermoprofundales subgroups (A–E) with distinct habitat preferences. Most of the microorganisms from Subgroups B and D were thermophiles inhabiting hydrothermal vents and hot spring sediments, whereas those from Subgroup E were adapted to surface environments where sunlight is available. H2 production may be featured in Thermoprofundales as evidenced by a gene cluster encoding the ancient membrane-bound hydrogenase (MBH) complex. Interestingly, a unique structure separating the MBH gene cluster into two modular units was observed exclusively in the genomes of Subgroup E, which included a peripheral arm encoding the [NiFe] hydrogenase domain and a membrane arm encoding the Na+/H+ antiporter domain. These two modular structures were confirmed to function independently by detecting the H2-evolving activity in vitro and salt tolerance to 0.2 M NaCl in vivo, respectively. The peripheral arm of Subgroup E resembles the proposed common ancestral respiratory complex of modern respiratory systems, which plays a key role in the early evolution of life. In addition, molecular dating analysis revealed that Thermoprofundales is an early emerging archaeal lineage among the extant MBH-containing microorganisms, indicating new insights into the evolution of this ubiquitous archaea lineage.

Discovery of the non-cosmopolitan lineages in Candidatus Thermoprofundales.

The recently proposed order Candidatus Thermoprofundales, currently containing only one family-level lineage Marine Benthic Group-D (MBG-D), is distributed in global subsurface ecosystems and ecologically important, but its diversity, evolution and metabolism remain largely unknown. Here we described two novel family-level specialized lineages in Ca. Thermoprofundales, JdFR-43 and HyVt, which are restricted to specific biotopes (primarily in marine hydrothermal vents and occasionally in oil reservoirs and hot springs) in contrast to the cosmopolitan lineage MBG-D. The comparative genomics revealed that the specialized lineages have streamlined genomes, higher GC contents, enriched genes associated with nucleotide biosynthesis, ribosome biogenesis and DNA repair and additional thermostable aminopeptidases, enabling them to adapt to high-temperature habitats such as marine hydrothermal vents, deep subsurface oil reservoirs and hot springs. On the contrary, the unique metabolic traits of the cosmopolitan MBG-D, motility, glycolysis, butanoate metabolism, secondary metabolites production and additional genes for specific peptides and carbohydrates degradation potentially enhance its response to environmental change. Substrate preference is found for most MAGs across all lineages with the ability to utilize both polysaccharides (chitin and starch) and proteinaceous substances, whereas JdFR-43 members from oil reservoirs can only utilize proteins. These results expand the diversity of Ca. Thermoprofundales significantly and further improve our understandings of the adaptations of Ca. Thermoprofundales to various environments.

Contrasting bacterial and archaeal distributions reflecting different geochemical processes in a sediment core from the Pearl River Estuary

Microbial community structure and metabolic activities have profound impacts on biogeochemical processes in marine sediments. Functional bacteria such as nitrate- and sulfate-reducing bacteria respond to redox gradients by coupling specific reactions amenable to relevant energy metabolisms. However, similar functional patterns have not been observed for sedimentary archaea (except for anaerobic methanotrophs and methanogens). We coupled taxonomic composition with comprehensive geochemical species to investigate the participation of distinct bacteria and archaea in sedimentary geochemical cycles in a sediment core (300 cm) from Pearl River Estuary (PRE). Geochemical properties (NO3−, dissolved Mn and Fe, SO42+, NH4+; dissolved inorganic carbon (DIC), δ13CDIC, dissolved organic carbon (DOC), total organic carbon (TOC), δ13CTOC, and fluorescent dissolved organic matter (FDOM)) exhibited strong depth variability of different trends. Bacterial 16S rRNA- and dsrB gene abundance decreased sharply with depth while archaeal and bathyarchaeotal 16S rRNA gene copies were relatively constant. This resulted in an increase in relative abundance of archaea from surface (11.6%) to bottom (42.8%). Network analysis showed that bacterial groups of Desulfobacterales, Syntrophobacterales and Gammaproteobacteria were significantly (P < 0.0001) associated with SO42− and dissolved Mn while archaeal groups of Bathyarchaeota, Group C3 and Marine Benthic Group D (MBGD) showed close positive correlations (P < 0.0001) with NH4+, δ13CTOC values and humic-like FDOM. Our study suggested that these bacterial groups dominated in redox processes relevant to sulfate or metal oxides, while the archaeal groups are more like to degrade recalcitrant organic compounds in anaerobic sediments.

Genomic and transcriptomic insights into the ecology and metabolism of benthic archaeal cosmopolitan, Thermoprofundales (MBG-D archaea)

Marine Benthic Group D (MBG-D) archaea, discovered by 16S rRNA gene survey decades ago, are ecologically important, yet understudied and uncultured sedimentary archaea. In this study, a comprehensive meta-analysis based on the 16S rRNA genes of MBG-D archaea showed that MBG-D archaea are one of the most frequently found archaeal lineages in global sediment with widespread distribution and high abundance, including 16 subgroups in total. Interestingly, some subgroups show significant segregations toward salinity and methane seeps. Co-occurrence analyses indicate significant non-random association of MBG-D archaea with Lokiarchaeota (in both saline and freshwater sediments) and Hadesarchaea, suggesting potential interactions among these archaeal groups. Meanwhile, based on four nearly complete metagenome-assembled genomes (MAGs) and corresponding metatranscriptomes reconstructed from mangrove and intertidal mudflat sediments, we provide insights on metabolic potentials and ecological functions of MBG-D archaea. MBG-D archaea appear to be capable of transporting and assimilating peptides and generating acetate and ethanol through fermentation. Metatranscriptomic analysis suggests high expression of genes for acetate and amino acid utilization and for peptidases, especially the M09B-type extracellular peptidase (collagenase) showing high expression levels in all four mangrove MAGs. Beyond heterotrophic central carbon metabolism, the MBG-D genomes include genes that might encode two autotrophic pathways: Wood–Ljundahl (WL) pathways using both H4MPT and H4folate as C1 carriers, and an incomplete dicarboxylate/4-hydroxybutyrate cycle with alternative bypasses from pyruvate to malate/oxaloacetate during dicarboxylation. These findings reveal MBG-D archaea as an important ubiquitous benthic sedimentary archaeal group with specific mixotrophic metabolisms, so we proposed the name Thermoprofundales as a new Order within the Class Thermoplasmata. Globally, Thermoprofundales and other benthic archaea might synergistically transform benthic organic matter, possibly playing a vital role in sedimentary carbon cycle.

Genomic reconstruction of multiple lineages of uncultured benthic archaea suggests distinct biogeochemical roles and ecological niches.

Genomic bins belonging to multiple archaeal lineages were recovered from distinct redox regimes in sediments of the White Oak River estuary. The reconstructed archaeal genomes were identified as belonging to the rice cluster subgroups III and V (RC-III, RC-V), the Marine Benthic Group D (MBG-D), and a newly described archaeal class, the Theionarchaea. The metabolic capabilities of these uncultured archaea were inferred and indicated a common capability for extracellular protein degradation, supplemented by other pathways. The multiple genomic bins within the MBG-D archaea shared a nearly complete reductive acetyl-CoA pathway suggesting acetogenic capabilities. In contrast, the RC-III metabolism appeared centered on the degradation of detrital proteins and production of H2, whereas the RC-V archaea lacked capabilities for protein degradation and uptake, and appeared to be specialized on carbohydrate fermentation. The Theionarchaea appeared as complex metabolic hybrids; encoding a complete tricarboxylic acid cycle permitting carbon (acetyl-CoA) oxidation, together with a complete reductive acetyl-CoA pathway and sulfur reduction by a sulfhydrogenase. The differentiated inferred capabilities of these uncultured archaeal lineages indicated lineage-specific linkages with the nitrogen, carbon and sulfur cycles. The predicted metabolisms of these archaea suggest preferences for distinct geochemical niches within the estuarine sedimentary environment.

Geochemical and microbial context of the gassy sediments in the Ría de Vigo (NW of Spain)

Multidisciplinary data coming from high-resolution seismic profiles, gravity core analysis (sedimentology and geochemistry), radiocarbon dating and metagenomics has been used to establish the stratigraphic framework as well as the geochemical and microbial context of the gassy sediments in the Ría de Vigo. Gas appears within the muddy sediments that compound the youngest Holocene seismic unit of the infilling of the ria and is mainly methane, generated by microbial decomposition in a rich organic matter depositional environment. Within the ria, the biogeochemical zonation is affected by the quality, quantity and age of the buried organic matter. The 16S ribosomal RNA gene amplicon clone libraries sequencing suggests that prokaryotic composition may be linked to the sediment type. Uncultured archaea common to the marine sub-sea floor of the Miscellaneous Crenarchaeotal Group (MCG) and Marine Benthic Group-D (MBGD) are dominant in these organic-rich estuarine and shallow marine sediments. The presence of Methanomicrobia and ANME in the sulphate-methane transition zone (SMTZ) from the estuarine gassy area indicates anaerobic oxidation of methane (AOM) in these sediments. Desulfobacteraceae are also homogeneously distributed along the sediment columns, even in the methanogenic zone, suggesting the existence of a cryptic sulphur cycle driven by iron under the SMTZ. The older sediments retrieved from the non-gassy outer area of the ria contain less organic matter and appear to represent mainly a sulphate-reducing environment where SMTZ is not yet reached at the bottom of the sediment core. Methanogens of various affiliations and traces of methane are detected in this sulphate-reducing environment, with an unusual increase in concentration towards the sediment surface. Microbial methanogenesis in the sulphate-reducing zone of surface sediments, a process recently described in organic-rich sediments able to supply methanogenic substrates could reconcile these observations.

Microbial Community Structure Associated with Biogeochemical Processes in the Sulfate–Methane Transition Zone (SMTZ) of Gas-hydrate-bearing Sediment of the Ulleung Basin, East Sea

ABSTRACTWe investigated the biogeochemical constituents, microbial communities and functional genes (mcr and dsr) associated with anaerobic methane oxidation and sulfate reduction, and metabolic activities by sulfate reduction in the sulfate–methane transition zone (SMTZ) of gas-hydrate-bearing sediment of the Ulleung Basin in the East Sea. Maxima in the sulfate reduction rate (12.6 nmol cm−3 d−1), CO concentration (83 μM), and gene abundances of dsrA (9.1 × 106 copies cm−3) and mcrA (11.6 × 106 copies cm−3) occurred in the SMTZ. The peaks of CO consistently found in the SMTZ suggested that CO is an intermediate metabolic product related to methane oxidation. Candidate division JS1, the predominant bacterial group that comprised 59.0–63.7% of the 16S rRNA gene sequences, was recognized as an important organic carbon oxidizer. Both Marine Benthic Group D (MBGD) and Marine Benthic Group B (MBGB), which constituted 40.8–52.9 and 10.3–43.9% of the 16S rRNA gene sequences, respectively, were the dominant archaeal groups. Analysis of functional gene diversity revealed that anaerobic methanotroph-1-related phylotypes appeared to be the major CH4 oxidizer, whereas Firmicutes-like group was a predominant sulfate reducer in the 0.8 mbsf in SMTZ with low SO42− concentration. Overall results indicated that JS1 and two archaeal groups (MBGB and MBGD) seem to play a significant role in carbon and elements cycles in the gas-hydrate-bearing subsurface sediment of the Ulleung Basin.

Microbial communities associated with benthic faunal assemblages at cold seep sediments of the Sonora Margin, Guaymas Basin

The Sonora Margin cold seeps present a seafloor mosaic pattern consisting of different faunal assemblages and microbial mats. To better understand if sedimentary microbial communities reflect this patchy distribution, all major habitats were investigated using four complementary approaches: 16S rRNA 454 pyrosequencing, quantitative polymerase chain reaction, fluorescence in situ hybridization and geochemistry analyses. This study reveals that sediments populated by different surface assemblages show distinct porewater geochemistry features and are associated with distinct microbial communities. In the sediments underlying the microbial mat and the surrounding macrofauna, microbial communities were dominated by anaerobic methane oxidizers (archaeal anaerobic methanotroph ANME) and sulfate-reducing Deltaproteobacteria. In contrast, sediment-associated microbial communities underlying the megafauna habitats (vesicomyids and siboglinids) were characterized by a lower biomass and important proportions of the Marine Benthic Group D (MBG-D), Chloroflexi as well as filamentous Gammaproteobacteria and Deltaproteobacteria. Together, geochemical and microbial surveys indicate that porewater methane concentrations play an important role in the microbial community structure and subsequently in the establishment of the surface colonizers. Furthermore, presence and activity of the surface colonizers influence the underlying microbial communities probably because of modification of energy source availabilities.

A long-term cultivation of an anaerobic methane-oxidizing microbial community from deep-sea methane-seep sediment using a continuous-flow bioreactor.

Anaerobic oxidation of methane (AOM) in marine sediments is an important global methane sink, but the physiological characteristics of AOM-associated microorganisms remain poorly understood. Here we report the cultivation of an AOM microbial community from deep-sea methane-seep sediment using a continuous-flow bioreactor with polyurethane sponges, called the down-flow hanging sponge (DHS) bioreactor. We anaerobically incubated deep-sea methane-seep sediment collected from the Nankai Trough, Japan, for 2,013 days in the bioreactor at 10°C. Following incubation, an active AOM activity was confirmed by a tracer experiment using 13C-labeled methane. Phylogenetic analyses demonstrated that phylogenetically diverse Archaea and Bacteria grew in the bioreactor. After 2,013 days of incubation, the predominant archaeal components were anaerobic methanotroph (ANME)-2a, Deep-Sea Archaeal Group, and Marine Benthic Group-D, and Gammaproteobacteria was the dominant bacterial lineage. Fluorescence in situ hybridization analysis showed that ANME-1 and -2a, and most ANME-2c cells occurred without close physical interaction with potential bacterial partners. Our data demonstrate that the DHS bioreactor system is a useful system for cultivating fastidious methane-seep-associated sedimentary microorganisms.

Microbial diversity in the hydrate-containing and -free surface sediments in the Shenhu area, South China Sea

Microbial diversity in the hydrate-containing (sites SH3B and SH7B) and -free (sites SH1B, SH5B, SH5C) sediments collected from the Shenhu area of the South China Sea (SCS) was investigated using 16S rRNA gene phylogenetic analysis. The phylogenetic results indicate difference in microbial communities between hydrate-containing and -free sediments. At the gas hydrate-containing sites, bacterial communities were dominated by Deltaproteobacteria (30.5%), and archaeal communities were dominated by Miscellaneous Crenarchaeotic Group (33.8%); In contrast, Planctomycetes was the major group (43.9%) in bacterial communities, while Marine Benthic Group-D (MBG-D) (32.4%) took up the largest proportion in the archaeal communities. Moreover, the microbial communities have characteristics different from those in other hydrate-related sediments around the world, indicating that the presence of hydrates can affect the microbial distribution. In addition, the microbial community composition in the studied sediments has its own uniqueness, which may result from co-effect of geochemical characteristics and presence/absence of hydrate.

Predominant archaea in marine sediments degrade detrital proteins

Half of the microbial cells in the Earth's oceans are found in sediments. Many of these cells are members of the Archaea, single-celled prokaryotes in a domain of life separate from Bacteria and Eukaryota. However, most of these archaea lack cultured representatives, leaving their physiologies and placement on the tree of life uncertain. Here we show that the uncultured miscellaneous crenarchaeotal group (MCG) and marine benthic group-D (MBG-D) are among the most numerous archaea in the marine sub-sea floor. Single-cell genomic sequencing of one cell of MCG and three cells of MBG-D indicated that they form new branches basal to the archaeal phyla Thaumarchaeota and Aigarchaeota, for MCG, and the order Thermoplasmatales, for MBG-D. All four cells encoded extracellular protein-degrading enzymes such as gingipain and clostripain that are known to be effective in environments chemically similar to marine sediments. Furthermore, we found these two types of peptidase to be abundant and active in marine sediments, indicating that uncultured archaea may have a previously undiscovered role in protein remineralization in anoxic marine sediments.

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.

Diversity of Achaea Communities in Deep Sea Sediments from the Sulu Sea

To study the archaeal diversity of deep-sea sediments in the tropical Western Pacific,we investigated the archaeal community structure in six sediments subsamples along the MD3059 sediment core collected during the IMAGES ⅪⅤ cruise with 16S rDNA analysis. A total of 543 archaeal 16S rDNA clones were examined and a total of 137 OTUs were obtained. Phylogenetic results showed that the archaeal diversity in the collected samples was very diverse,and the obtained OTUs were grouped into Crenarchaeota and Euryarchaeota,with the former dominated by Miscellaneous Crenarchaeotic Group ( MCG,54% of total archaeal clones) and the latter dominated by Marine Benthic Group D ( MBG-D) ,South African Gold Mine Euryarchaeotic Group ( SAGMEG) and Marine Benthic Group B ( MBG-B). These data have important implications for our understanding of archaeal community in deep-sea sediments of the tropical Western Pacific.

Archaeal and bacterial community structures in the anoxic sediment of Antarctic meromictic lake Nurume-Ike

Prokaryotic community structures in the anoxic sediment of the Antarctic meromictic Lake Nurume-Ike were revealed by sequence analysis of 16S rRNA gene clones. The archaeal clones obtained (205 total) consisted of only three phylotypes, and were dominantly affiliated with uncultured euryarchaeotes. Specifically, 93% of the clones were identified as marine benthic group-D archaeal phylotype. In contrast to the limited archaeal diversity, 53 phylotypes were detected within 312 bacterial clones. Major bacterial phylotypes were affiliated with α-Proteobacteria (20% of clones), d-Proteobacteria (9%), Planctmycetales (7%), and Cyanobacteria (7%). A small numbers of clones belonging to γ-Proteobacteria, Actinobacteria, Spirochaetes, Flavobacteria, and Verrucomicrobia were also found. A total of 53% of the bacterial clones, consisting of 13 phylotypes, could not be classified into any known group. These results indicated that the bacterial community of Lake Nurume-Ike sediment consisted of numerous phylogenetic groups and had a diversity comparable to the diversity of other Antarctic lakes communities previously reported. Interestingly, however, there were very few phylotypes shared between the communities of lakes Nurume-Ike and five other lakes located in the Vestfold Hills area. This is the first comprehensive study to analyze more than 500 16S rDNA clones for microbial community analysis of an Antarctic lake sediment sample, and the results significantly expand current views of bacterial diversity in Antarctic lakes.

Microbial Diversity in the Deep Marine Sediments from the Qiongdongnan Basin in South China Sea

The continental shelf and slope in the northern South China Sea is well known for its prospect of oil/gas/gas-hydrate resources. To study microbial communities and their roles in carbon cycling, a 4.9-m sediment core was collected from the Qiongdongnan Basin on the continental slope of the South China Sea during our cruise HY4-2005-5 in 2005. Geochemical, mineralogical, and molecular phylogenetic analyses were carried out. Sulfate concentration in pore water decreased with depth. Abundant authigenic carbonates and pyrite were observed in the sediments. The bacterial community was dominated by aerobic and facultative organisms. Bacterial clone sequences belonged to the Gamma-, Alpha-, Deltaproteobacteria and Firmicutes group, and they were related to Fe(III) and/or Mn(IV) reducers, sulfate reducers, aromatic hydrocarbon degraders, thiosulfate/sulfite oxidizers, and denitrifiers. Archaeal clone sequences exhibited greater overall diversity than the bacterial clones with most sequences related to Deep-Sea Archaeal Group (DSAG), Miscellaneous Crenarchaeotic Group (MCG), and Uncultured Euryarchaeotic Clusters (UECs). Archaeal sequences related to Methanosarcinales, South African Gold Mine Euryarchaeotic Group (SAGMEG), Marine Benthic Group-D (MBG-D) were also present. Most of these groups are commonly present in deep-sea sediments, particularly in methane/organic-rich or putative methane hydrate-bearing sediments.

Soil–Methanogen Interactions in Two Peatlands (Bog, Fen) in Central New York State

Rates of methanogenesis vary widely in peat soils, yet the reasons are poorly known. We examined rates of methanogenesis and methanogen diversity in relation to soil chemical and biological characteristics in 2 peatlands in New York State. One was an acidic (pH < 4.5) bog dominated by Sphagnum mosses and ericaceous shrubs, although deeper peat was derived from sedges. The other was a fen dominated by Carex lacustris sedges with near-neutral pH soil. At both sites, the most active rates of methanogenesis occurred in the top 20 cm of the peat profile, even when using a substrate-induced methanogenesis technique with added glucose that stimulated rates up to 2 μ mol g − 1 day −1 in the bog and 6 μ mol g −1 day −1 in the fen. Rates of anaerobic CO 2 production were greater in the bog (0–36 μ mol g −1 day −1 ) than in the fen (0–5 μ mol g −1 day −1 ), and added glucose induced greater rates in the sedge-derived peat from the bog than the fen. The peat soil was much more decomposed throughout the profile in the fen. Analysis of chemical elements in the peat profile revealed a striking anomaly: a very high concentration of Pb in surface peat of the bog, which might have constrained methanogenesis. Application of T-RFLP analysis to methanogens revealed dominance by a Methanomicrobiales E2 clade of H 2 /CO 2 users in the acidic peat soil of the bog, whereas deeper peat had a different Methanomicrobiales E1 clade, uncultured euryarchaeal rice cluster (RC)-I and RC-II groups, marine benthic group D (MBD) and a new cluster called subaqueous cluster (SC). In contrast, T-RFLP analysis of peat from the fen revealed co-dominance by Methanosaetaceae and Methanomicrobiales E1. The results showed complex relationships between rates of methanogenesis, methanogen populations and metabolic substrate availability with idiosyncratic interactions of trace chemical elements.