Subseafloor Sediments Research Articles

Past and future evolution of the onshore-offshore groundwater system of a carbonate archipelago: The case of the Maltese Islands, central Mediterranean Sea

Offshore freshened groundwater (OFG) is groundwater with a salinity below that of seawater that is stored in sub-seafloor sediments and rocks. OFG has been proposed as an alternative solution to relieve water scarcity in coastal regions and to enhance oil recovery. Although OFG has been documented in most continental margins, we still have a poor understanding of the extent and flow characteristics of OFG systems, and their evolution through time. In view of the general absence of appropriate field data, paleohydrogeological models have been used. The majority of these models are based on 2D approaches, and they rarely consider the future evolution of OFG systems, especially in response to predicted climate change. Here we utilize recently acquired geological, geophysical and hydrogeological data from onshore and offshore the Maltese Islands, and employ 2D and 3D numerical models, to: (i) reconstruct the evolution of the onshore-offshore groundwater system during the last 188 ka, (ii) predict the evolution of the OFG system in response to climate-related changes. We show that the mechanisms emplacing OFG include a combination of active meteoric recharge at present as well as at sea-level lowstands. The Maltese onshore-offshore groundwater system is relatively dynamic, with 23% of groundwater being preserved in the last 18 ka. The control of geology is expressed by the more prevalent distribution of OFG north of the Great Fault, which is associated to the occurrence of low permeability units, and the asymmetry of the groundwater lens during the 18 ka lowstand. A 30% decrease in recharge predicted in the coming 100 years will diminish OFG extent by 38%, whereas sea-level rise will play a negligible role. At present the estimated volume of OFG is 1 km3, which could potentially provide an alternative supply of potable water to the Maltese Islands for 75 years. Exploitation of OFG with minimal salinization of onshore groundwater bodies would require locating pumping wells close to the coast.

Deep Subseafloor Biogeochemical Processes and Microbial Populations Potentially Associated with the 2011 Tohoku-oki Earthquake at the Japan Trench Accretionary Wedge (IODP Expedition 343).

Post-mega-earthquake geochemical and microbiological properties in subseafloor sediments of the Japan Trench accretionary wedge were investigated using core samples from Hole C0019E, which was drilled down to 851‍ ‍m below seafloor (mbsf) at a water depth of 6,890 m. Methane was abundant throughout accretionary prism sediments; however, its concentration decreased close to the plate boundary decollement. Methane isotope systematics indicated a biogenic origin. The content of mole-cular hydrogen (H2) was low throughout core samples, but markedly increased at specific depths that were close to potential faults predicted by logging-while-drilling ana-lyses. Based on isotopic systematics, H2 appeared to have been abundantly produced via a low-temperature interaction between pore water and the fresh surface of crushed rock induced by earthquakes. Subseafloor microbial cell density remained constant at approximately 105‍ ‍cells‍ ‍mL-1. Amplicon sequences revealed that predominant members at the phylum level were common throughout the units tested, which also included members frequently found in anoxic subseafloor sediments. Metabolic potential assays using radioactive isotopes as tracers revealed homoacetogenic activity in H2-enriched core samples collected near the fault. Furthermore, homoacetogenic bacteria, including Acetobacterium carbinolicum, were isolated from similar samples. Therefore, post-earthquake subseafloor microbial communities in the Japan Trench accretionary prism appear to be episodically dominated by homoacetogenic populations and potentially function due to the earthquake-induced low-temperature generation of H2. These post-earthquake microbial communities may eventually return to the steady-state communities dominated by oligotrophic heterotrophs and hydrogenotrophic and methylotrophic methanogens that are dependent on refractory organic matter in the sediment.

Grinding Beads Influence Microbial DNA Extraction from Organic-Rich Sub-Seafloor Sediment.

Sub-seafloor sediment is the largest microbial habitat on Earth. The study of microbes in sub-seafloor sediment is largely limited by the technical challenge of acquiring ambient microbial DNA because of sediment heterogeneity. Changes in the extraction method, even just by one step, can affect the extraction yields for complicated sediment samples. In this work, sub-seafloor sediment samples from the Baltic Sea with high organic carbon content were used to evaluate the influence of different grinding beads on DNA extraction. We found that the grinding beads can affect the DNA extraction from the organic-matter- and biosiliceous-clay-rich samples. A mixture of 0.5-mm and 0.1-mm grinding beads exhibited higher DNA yields and recovered more unique taxa than other bead combinations, such as Stenotrophomonas from Gammaproteobacteria and Leptotrichia from Fusobacteria; therefore, these beads are more suitable than the others for DNA extraction from the samples used in this study. This advantage might be magnified in samples with high biomass. On the contrary, the use of only small beads might lead to underestimation for certain Gram-positive strains. Overall, the discovery of abundant widespread deep biosphere clades in our samples indicated that our optimized DNA extraction method successfully recovered the in situ microbial community.

First description of archaeal communities in carbonate-rich seafloor and subseafloor sediments from the Southwestern Atlantic slope

First description of archaeal communities in carbonate-rich seafloor and subseafloor sediments from the Southwestern Atlantic slope

Dissolved Microbial Methane in the Deep Crystalline Crust Fluids–Current Knowledge and Future Prospects

Methane is a powerful greenhouse gas, of which most is produced by microorganisms in a process called methanogenesis. One environment where methanogenic microorganisms occur is the deep biosphere. The deep biosphere environment comprises a variety of ecosystem settings; marine habitats such as subseafloor sediments, rock pore volumes within subseafloor basalts, and terrestrial settings such as sedimentary rocks and crystalline bedrock fracture networks. Microbial methane formed in these environments influence the biological, chemical, and geological cycles of the upper crust, and may seep out of the deep into the atmosphere. This review focuses on the process of microbial methanogenesis and methane oxidation in the relatively underexplored deep crystalline-bedrock hosted subsurface, as several works in recent years have shown that microbial production and consumption occur in this energy-poor rock-fracture-hosted environment. These recent findings are summarized along with techniques to study the source and origins of methane in the terrestrial crust. Future prospects for exploration of these processes are proposed to combine geochemical and microbial techniques to determine whether microbial methanogenesis is a ubiquitous phenomenon in the crystalline crust across space and time. This will aid in determining whether microbial methane in the globally vast deep rock-hosted biosphere environment is a significant contributor to the global methane reservoir.

Geologic processes in marine “muddy” sediments

Marine muds, or “muddy” sediments, represent a broad range of sediments containing a mix of clay and silt size components, often with some smaller amount of sand sized and larger material. Sediments characterized by these broad definitions comprise most of the modern global seafloor, except for areas proximal to terrigenous inputs, such as continental shelves and glacial margins, which can receive and retain a higher proportion of coarser sediment. Beyond the coastal zone, the distribution of marine sediments is driven by a combination of geological, biological, and physical oceanographic processes that themselves are a response to variations in environmental conditions (e.g., global climate, nutrient availability, tectonics, terrestrial erosion). At the local scale, sediment source variation, seafloor bathymetry, primary transport and near-bed currents, resuspension and reworking, and bioturbation contribute to the short- and long-term geologic nature of marine muds. The structure, quantitative compositional descriptions, and physical properties of marine muds from several environments will be compared and the techniques used to analyze them will be presented. New methods that may facilitate an improved understanding of the role and importance of muddy seafloor and shallow sub-seafloor sediments in influencing the acoustic environment will be highlighted.

Structure and bioactivity of polysaccharide from a subseafloor strain of Schizophyllum commune 20R-7-F01

Fungal polysaccharide is a kind of biomacromolecule with multiple biological activities, which has a wide application prospect and may play an important role in organisms to cope with extreme environments. Herein, we reported an extracellular polysaccharide (EPS) produced by Schizophyllum commune 20R-7-F01 that was isolated from subseafloor sediments at ~2 km below the seafloor, obtained during expedition 337. The monosaccharide of EPS was glucose and its molecular weight was 608.8 kDa. Methylation and NMR analysis indicated that the backbone of the EPS was (1 → 3)-β-D-glucan with a side chain (1 → 6) β-D-glucan linking at every third residue. Bio-active assays revealed that the EPS had potent antioxidant activity and could promote RAW264.7 cells viability and phagocytosis. These results suggest that fungi derived from sediments below seafloor are important and new source of polysaccharides and may be involved in the adaptation of fungi to anoxic subseafloor extreme ecosystem.

Anaerobic biodegradation of polycyclic aromatic hydrocarbons (PAHs) by fungi isolated from anaerobic coal-associated sediments at 2.5km below the seafloor.

Fungi represent the dominant eukaryotic group in the deep biosphere and well-populated in the anaerobic coal-bearing sediments up to ∼2.5km below seafloor (kmbsf). But whether fungi are able to degrade and utilize coal to sustain growth in the anaerobic sub-seafloor environment remains unknown. Based on biodegradation investigation, we found that fungi isolated from sub-seafloor sediments at depths of ∼1.3-∼2.5 kmbsf showed a broad range of polycyclic aromatic hydrocarbons (PAHs) anaerobic degradation rates (3-25%). Among them, the white-rot fungus Schizophyllium commune 20R-7-F01 exhibited the highest degradation, 25%, 18% and 13%, of phenanthrene (Phe), pyrene (Pyr) and benzo[a]pyrene (BaP); respectively, after 10 days of anaerobic incubation. Phe was utilized well and about 40.4% was degraded by the fungus, after 20 days of anaerobic incubation. Moreover, the ability of fungi to degrade PAHs was positively correlated with the anaerobic growth of fungi, indicating that fungi can use PAHs as a sole carbon source under anoxic conditions. In addition, fungal degradation of PAHs was found to be related to the activity of carboxylases, but little or nothing to do with the activity of lignin modifying enzymes such as laccase (Lac), manganese peroxidase (MnP) and lignin peroxidase (LiP). These results suggest that sub-seafloor fungi possess a special mechanism to degrade and utilize PAHs as a carbon and energy source under anaerobic conditions. Furthermore, fungi living in sub-seafloor sediments may not only play an important role in carbon cycle in the anaerobic environments of the deep biosphere, but also be able to persist in deep sediment below seafloor for millions of years by using PAHs or related compounds as carbon and energy source. This anaerobic biodegradation ability could make these fungi suitable candidates for bioremediation of toxic pollutants such as PAHs from anoxic environments.

Genome-wide characterization of laccase gene family in Schizophyllum commune 20R-7-F01, isolated from deep sediment 2 km below the seafloor.

Laccases are ligninolytic enzymes that play a crucial role in various biological processes of filamentous fungi, including fruiting-body formation and lignin degradation. Lignin degradation is a complex process and its degradation in Schizophyllum commune is greatly affected by the availability of oxygen. Here, a total of six putative laccase genes (ScLAC) were identified from the S. commune 20R-7-F01 genome. These genes, which include three typical Cu-oxidase domains, can be classified into three groups based on phylogenetic analysis. ScLAC showed distinct intron-exon structures and conserved motifs, suggesting the conservation and diversity of ScLAC in gene structures. Additionally, the number and type of cis-acting elements, such as substrate utilization-, stress-, cell division- and transcription activation-related cis-elements, varied between ScLAC genes, suggesting that the transcription of laccase genes in S. commune 20R-7-F01 could be induced by different substrates, stresses, or other factors. The SNP analysis of resequencing data demonstrated that the ScLAC of S. commune inhabiting deep subseafloor sediments were significantly different from those of S. commune inhabiting terrestrial environments. Similarly, the large variation of conserved motifs number and arrangement of laccase between subseafloor and terrestrial strains indicated that ScLAC had a diverse structure. The expression of ScLAC5 and ScLAC6 genes was significantly up-regulated in lignin/lignite medium, suggesting that these two laccase genes might be involved in fungal utilization and degradation of lignite and lignin under anaerobic conditions. These findings might help in understanding the function of laccase in white-rot fungi and could provide a scientific basis for further exploring the relationship between the LAC family and anaerobic degradation of lignin by S. commune.

Sulfur isotope fractionation derived from reaction-transport modelling in the Eastern Equatorial Pacific

Microbial sulfate reduction in subseafloor sediments regulates a significant portion of the marine organic matter burial flux. Over secular timescales, sulfate reduction is the fundamental process connecting the biogeochemical cycles of sulfur, carbon, oxygen and phosphorus. Similar to carbon reduction, sulfate reduction is associated with a strong isotope fractionation process that allows us to track this process through time. It depends on a variety of factors and it has been argued previously that systematic differences between shallow and deep-sea environments might explain secular changes in the marine S-isotope ratio. However, observational data of in situ fractionation from deep-sea areas are scarce. Here we use a reaction-transport model to analyse the S-isotope fractionation during microbial sulfate reduction in the interstitial water of Ocean Drilling Program (ODP) Site 1226 (Leg 201). We find that the upper 100 m below seafloor are best explained with fractionation around −76‰, whereas below this depth the degree of fractionation drops to around −42‰. We propose that this shift is caused by changes in the ratio of the rate of microbial sulfate reduction relative to the rate of abiotic sulfide oxidation. Because large parts of deep oceans are characterized by exceedingly low sulfate reduction rates, this process may be widespread and possibly explain why pyrite S-isotope data suggest that the average S-isotope fractionation is around −50‰, rather than the theoretically predicted value below −70‰. Supplementary material: Supplementary figures and model results are available at https://doi.org/10.6084/m9.figshare.c.5976772 Thematic collection: This article is part of the Sulfur in the Earth system collection available at: https://www.lyellcollection.org/cc/sulfur-in-the-earth-system

Cultivation and biogeochemical analyses reveal insights into methanogenesis in deep subseafloor sediment at a biogenic gas hydrate site

Gas hydrates deposited in subseafloor sediments are considered to primarily consist of biogenic methane. However, little evidence for the occurrence of living methanogens in subseafloor sediments has been provided. This study investigated viable methanogen diversity, population, physiology and potential activity in hydrate-bearing sediments (1–307 m below the seafloor) from the eastern Nankai Trough. Radiotracer experiments, the quantification of coenzyme F430 and molecular sequencing analysis indicated the occurrence of potential methanogenic activity and living methanogens in the sediments and the predominance of hydrogenotrophic methanogens followed by methylotrophic methanogens. Ten isolates and nine representative culture clones of hydrogenotrophic, methylotrophic and acetoclastic methanogens were obtained from the batch incubation of sediments and accounted for 0.5–76% of the total methanogenic sequences directly recovered from each sediment. The hydrogenotrophic methanogen isolates of Methanocalculus and Methanoculleus that dominated the sediment methanogen communities produced methane at temperatures from 4 to 55 °C, with an abrupt decline in the methane production rate at temperatures above 40 °C, which is consistent with the depth profiles of potential methanogenic activity in the Nankai Trough sediments in this and previous studies. Our results reveal the previously overlooked phylogenetic and metabolic diversity of living methanogens, including methylotrophic methanogenesis.

Construction of Aerobic/Anaerobic-Substrate-Induced Gene Expression Procedure for Exploration of Metagenomes From Subseafloor Sediments.

Substrate-induced gene expression (SIGEX) is a high-throughput promoter-trap method. It is a function-based metagenomic screening tool that relies on transcriptional activation of a reporter gene green fluorescence protein (gfp) by a metagenomic DNA library upon induction with a substrate. However, its use is limited because of the relatively small size of metagenomic DNA libraries and incompatibility with screening metagenomes from anaerobic environments. In this study, these limitations of SIGEX were addressed by fine-tuning metagenome DNA library construction protocol and by using Evoglow, a green fluorescent protein that forms a chromophore even under anaerobic conditions. Two metagenomic libraries were constructed for subseafloor sediments offshore Shimokita Peninsula (Pacific Ocean) and offshore Joetsu (Japan Sea). The library construction protocol was improved by (a) eliminating short DNA fragments, (b) applying topoisomerase-based high-efficiency ligation, (c) optimizing insert DNA concentration, and (d) column-based DNA enrichment. This led to a successful construction of metagenome DNA libraries of approximately 6 Gbp for both samples. SIGEX screening using five aromatic compounds (benzoate, 3-chlorobenzoate, 3-hydroxybenzoate, phenol, and 2,4-dichlorophenol) under aerobic and anaerobic conditions revealed significant differences in the inducible clone ratios under these conditions. 3-Chlorobenzoate and 2,4-dichlorophenol led to a higher induction ratio than that for the other non-chlorinated aromatic compounds under both aerobic and anaerobic conditions. After the further screening of induced clones, a clone induced by 3-chlorobenzoate only under anaerobic conditions was isolated and characterized. The clone harbors a DNA insert that encodes putative open reading frames of unknown function. Previous aerobic SIGEX attempts succeeded in the isolation of gene fragments from anaerobes. This study demonstrated that some gene fragments require a strict in vivo reducing environment to function and may be potentially missed when screened by aerobic induction. The newly developed anaerobic SIGEX scheme will facilitate functional exploration of metagenomes from the anaerobic biosphere.

Not dead yet: Diatom resting spores can survive in nature for several millennia.

Understanding the adaptive capacities of species over long timescales lies in examining the revived recent and millennia-old resting spores buried in sediments. We show for the first time the revival, viability, and germination rate of resting spores of the diatom Chaetoceros deposited in sub-seafloor sediments from three ages (recent: 0 to 80 years; ancient: ~1250 (Medieval Climate Anomaly) and ~6600 (Holocene Thermal Maximum) calendar year before present. Recent and ancient Chaetoceros spores were revived to examine their viability and germination rate. Light and scanning electron microscopy and Sanger sequencing was done to identify the species. We show that ~6600 cal. year BP old Chaetoceros resting spores are still viable and that the vegetative reproduction in recent and ancient resting spores varies. The time taken to germinate is three hours to 2 to 3 days in both recent and ancient spores, but the germination rate of the spores decreased with increasing age. The germination rate of the recent spores was ~41% while that of the ancient spores were ~31% and ~12% for the ~1250 and ~6600 cal. year BP old resting spores, respectively. Based on the morphology of the germinated vegetative cells we identified the species as Chaetoceros muelleri var. subsalsum. Sanger sequences of nuclear and chloroplast markers identified the species as Chaetoceros muelleri. We identify a unique model system, Chaetoceros muelleri var. subsalsum and show that recent and ancient resting spores of the species buried in sediments in the Baltic Sea can be revived and used for long-term evolutionary studies.

First description of archaeal communities in carbonate-rich seafloor and subseafloor sediments from the Southwestern Atlantic slope

Deep-sea sediments comprise one of the largest habitats on Earth. The archaeal groups contribute to a large fraction of the deep-sea benthic biomass, playing a key role in biogeochemical cycles. However, their diversity in deep-sea benthic ecosystems remains poorly understood, mostly because only recently novel taxa have been proposed, thus remodeling the phylogenetic tree. Despite the dificulty in obtaining cultivated representatives, the metabolic capabilities of archaea have lately been described through metagenomic data, indicating that archaeal taxa are highly versatile. Here, we aimed to reveal the diversity of archaeal communities in surface (0 to 15 cm depth) and subsurface (200 cm depth) sediments from a carbonate-rich region in the Southwestern Atlantic upper slope. We performed 16S rRNA gene sequencing, and found that the archaeal composition in surface sediments was mainly dominated by ammonia-oxidizing archaea within Nitrososphaeria class. The distribution of Nitrososphaeria ASVs (amplicon sequence variants) indicates the presence of several species or ecotypes. Contrastingly, the subsurface sediment was dominated by uncultivated anaerobic and poorly known archaea, including representatives of all supergroups (Asgard, TACK, DPANN and Euryarchaeota). These archaea have been described as having potentially diverse metabolic capabilities, including autotrophic and heterotrophic pathways, such as acetogenesis, methylotrophy, and degradation of labile and recalcitrant organic compounds. This indicates an important role in the remineralization of organic matter in the SW (Southwest) Atlantic slope. They are likely enriched due to the transport and mixing of sediments by the IWBC (Intermediate Western Boundary Current) along the continental slope. However, further studies are needed to reveal the geochemical and oceanographic drivers of the archaeal distribution. This study provides the first description of the archaeal communities in carbonate-rich sediments in the SW Atlantic slope, and adds new biodiversity insights to this geological feature, which is considered a vulnerable marine ecosystem, thus helping for future conservation strategies.

Life Discovered in Super Nutrient Poor, Cretaceous Subseafloor Sediments

Life Discovered in Super Nutrient Poor, Cretaceous Subseafloor Sediments

Visualization of materials below shallow subseafloor sediment via Acoustic Mapping, an Example from Shallow Methane Hydrate-bearing Areas along the eastern margin of the Sea of Japan

Visualization of materials below shallow subseafloor sediment via Acoustic Mapping, an Example from Shallow Methane Hydrate-bearing Areas along the eastern margin of the Sea of Japan

Two Processes of Anglesite Formation and a Model of Secondary Supergene Enrichment of Bi and Ag in Seafloor Hydrothermal Sulfide Deposits

The in situ element concentrations and the sulfur (S), and lead (Pb) isotopic compositions in anglesite were investigated for samples from seafloor hydrothermal fields in the Okinawa Trough (OT), Western Pacific. The anglesite grains are of two kinds: (1) low Pb/high S primary hydrothermal anglesite (PHA), which is formed by mixing of fluid and seawater, and (2) high Pb/low S secondary supergene anglesite (SSA), which is the product of low-temperature (<100 °C) alteration of galena in the seawater environment. The Ag and Bi in the SSA go through a second enrichment process during the formation of high Pb/low S anglesite by galena alteration, indicating that the SSA and galena, which may be the major minerals host for considerable quantities of Ag and Bi, are potentially Ag-Bi-enriched in the back-arc hydrothermal field. Moreover, REEs, S and Pb in the OT anglesite are likely to have been leached by fluids from local sub-seafloor volcanic rocks and/or sediments. A knowledge of the anglesite is useful for understanding the influence of volcanic rocks, sediments and altered subducted oceanic plate in hydrothermal systems, showing how trace metals behave during the formation of secondary minerals.

Iron-Copper-Zinc Isotopic Compositions of Andesites from the Kueishantao Hydrothermal Field off Northeastern Taiwan

The studies of iron (Fe), copper (Cu), and zinc (Zn) isotopic compositions in seafloor andesites are helpful in understanding the metal stable isotope fractionation during magma evolution. Here, the Fe, Cu, and Zn isotopic compositions of andesites from the Kueishantao hydrothermal field (KHF) off northeastern Taiwan, west Pacific, have been studied. The majority of δ56Fe values (+0.02‰ to +0.11‰) in the KHF andesites are consistent with those of MORBs (mid-ocean ridge basalts). This suggests that the Fe in the KHF andesites is mainly from a MORB-type mantle. The Fe-Cu-Zn isotopic compositions (δ56Fe +0.22‰, δ65Cu +0.16‰ to +0.64‰, and δ66Zn +0.29‰ to +0.71‰) of the KHF andesites, which are significantly different from those of the MORBs and the continental crust (CC), have a relatively wide range of Cu and Zn isotopic compositions. This is most likely to be a result of the entrainment of the sedimentary carbonate-derived components into an andesitic magma. The recycled altered rocks (higher δ56Fe, lower δ66Zn) could preferentially incorporate isotopically light Fe and heavy Zn into the magma, resulting in relative enrichment of the lighter Fe and heavier Zn isotopes in the andesites. The majority of the δ56Fe values in the KHF andesites are higher than those of the sediments and the local CC and lower than those of the subducted altered rocks, while the reverse is true for δ66Zn, suggesting that the subseafloor sediments and CC materials (lower δ56Fe, higher δ66Zn) contaminating the rising andesitic magma could preferentially incorporate isotopically heavy Fe and light Zn into the magma, resulting in relative enrichment of the heavier Fe and lighter Zn isotopes in the andesites. Thus, the characteristics of the Fe and Zn isotopes in back-arc and island-arc volcanic rocks may also be influenced by the CC and plate subduction components.

Dimethylsulfoniopropionate Biosynthetic Bacteria in the Subseafloor Sediments of the South China Sea.

Dimethylsulfoniopropionate (DMSP) is one of Earth’s most abundant organosulfur molecules, and bacteria in marine sediments have been considered significant producers. However, the vertical profiles of DMSP content and DMSP-producing bacteria in subseafloor sediment have not been described. Here, we used culture-dependent and -independent methods to investigate microbial DMSP production and cycling potential in South China Sea (SCS) sediment. The DMSP content of SCS sediment decreased from 11.25 to 20.90 nmol g–1 in the surface to 0.56–2.08 nmol g–1 in the bottom layers of 8-m-deep subseafloor sediment cores (n = 10). Very few eukaryotic plastid sequences were detected in the sediment, supporting bacteria and not algae as important sediment DMSP producers. Known bacterial DMSP biosynthesis genes (dsyB and mmtN) were only predicted to be in 0.0007–0.0195% of sediment bacteria, but novel DMSP-producing isolates with potentially unknown DMSP synthesis genes and/or pathways were identified in these sediments, including Marinobacter (Gammaproteobacteria) and Erythrobacter (Alphaproteobacteria) sp. The abundance of bacteria with the potential to produce DMSP decreased with sediment depth and was extremely low at 690 cm. Furthermore, distinct DMSP-producing bacterial groups existed in surface and subseafloor sediment samples, and their abundance increased when samples were incubated under conditions known to enrich for DMSP-producing bacteria. Bacterial DMSP catabolic genes were also most abundant in the surface oxic sediments with high DMSP concentrations. This study extends the current knowledge of bacterial DMSP biosynthesis in marine sediments and implies that DMSP biosynthesis is not only confined to the surface oxic sediment zones. It highlights the importance of future work to uncover the DMSP biosynthesis genes/pathways in novel DMSP-producing bacteria.

Insights into the Methanogenic Population and Potential in Subsurface Marine Sediments Based on Coenzyme F430 as a Function-Specific Biomarker.

Coenzyme F430, the prosthetic group of methyl coenzyme M reductase (MCR), is a key compound in methane metabolism. We applied coenzyme F430 as a function-specific biomarker of methanogenesis to subsurface marine sediments collected below the sulfate reduction zone to investigate the distribution and activity of methanogens. In addition, we examined the kinetics of the epimerization of coenzyme F430, which is the first stage of the degradation process after cell death, at various temperatures (4, 15, 34, 60 °C) and pH (5, 7, 9) conditions, which cover in situ conditions of drilled sediments used in this study. The degradation experiments revealed that the kinetics of the epimerization well follow the thermodynamic laws, and the half-life of coenzyme F430 is decreasing from 304 days to 11 h with increasing the in situ temperature. It indicates that the native F430 detected in the sediments is derived from living methanogens, because the abiotic degradation of F430 is much faster than the sedimentation rate and will not be fossilized. Based on coenzyme F430 analysis and degradation experiments, the native form of F430 detected in subseafloor sediments off the Shimokita Peninsula originates from living methanogen cells, which is protected from degradation in cells but disappears soon after cell death. The biomass of methanogens calculated from in situ F430 concentration and F430 contents in cultivable methanogen species decreases by 2 orders of magnitude up to a sediment depth of 2.5 km, with a maximum value at ∼70 m below the seafloor (mbsf), while the proportion of methanogens to the total prokaryotic cell abundance increases with the depth, which is 1 to 2 orders of magnitude higher than expected previously. Our results indicate the presence of undetectable methanogens using conventional techniques.