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1. A PCR-Based Survey of Methane-Cycling Archaea in Methane-Soaked Subsurface Sediments of Guaymas Basin, Gulf of California

   Hinkle, J.E.. ; Mara, P. ; Beaudoin, D. ; Edgcomb, V. ; Teske, A. ( December 2023 , Microorganisms)

   The Guaymas Basin in the Gulf of California is characterized by active seafloor spreading, the rapid deposition of organic-rich sediments, steep geothermal gradients, and abundant methane of mixed thermogenic and microbial origin. Subsurface sediment samples from eight drilling sites with distinct geochemical and thermal profiles were selected for DNA extraction and PCR amplification to explore the diversity of methane-cycling archaea in the Guaymas Basin subsurface. We performed PCR amplifications with general (mcrIRD), and ANME-1 specific primers that target the alpha subunit of methyl coenzyme M reductase (mcrA). Diverse ANME-1 lineages associated with anaerobic methane oxidation were detected in seven out of the eight drilling sites, preferentially around the methane-sulfate interface, and in several cases, showed preferences for specific sampling sites. Phylogenetically, most ANME-1 sequences from the Guaymas Basin subsurface were related to marine mud volcanoes, seep sites, and the shallow marine subsurface. The most frequently recovered methanogenic phylotypes were closely affiliated with the hyperthermophilic Methanocaldococcaceae, and found at the hydrothermally influenced Ringvent site. The coolest drilling site, in the northern axial trough of Guaymas Basin, yielded the greatest diversity in methanogen lineages. Our survey indicates the potential for extensive microbial methane cycling within subsurface sediments of Guaymas Basin. 

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2. Guaymas Basin Tectonics and Biosphere

   https://doi.org/10.14379/iodp.proc.385.2021  

   Teske, A. ; Lizarralde, D. ; Höfig, T.W. ( September 2021 , Proceedings of the International Ocean Discovery Program)

   null (Ed.)

   International Ocean Discovery Program Expedition 385 drilled organic-rich sediments and intruded sills in the off-axis region and axial graben of the northern spreading segment of Guaymas Basin, a young marginal seafloor spreading system in the Gulf of California. Guaymas Basin is characterized by high heat flow and magmatism in the form of sill intrusions into sediments, which extends tens of kilometers off axis, in contrast with the localized volcanism found at most mid-ocean ridge spreading centers. Sill intrusions provide transient heat sources that mobilize buried sedimentary carbon, in part as methane and other hydrocarbons, and drive hydrothermal circulation. The resulting thermal and geochemical gradients shape abundance, composition, and activity of the deep subsurface biosphere of the basin. Drill sites extend over a broad region of Guaymas Basin. Adjacent Sites U1545 and U1546, located ~52 km northwest of the northern Guaymas Basin axial graben, recovered sediment successions to ~540 meters below seafloor (mbsf) (equivalent to the core depth below seafloor, Method A [CSF-A] scale), including a thin sill (a few meters thick) drilled near the bottom of Site U1545 and a massive sill (~355–430 mbsf) at Site U1546 that chemically and physically affects the surrounding sediments. Sites U1547 and U1548, located ~27 km northwest of the axial graben, were drilled to investigate an active sill-driven hydrothermal system evident at the seafloor as an 800 m wide, circular bathymetric high called Ringvent because of its outline of a ring of active vent sites. Ringvent is underlain by a thick sill at shallow depth (Site U1547). Geothermal gradients steepen toward the Ringvent periphery (Holes U1548A–U1548C), and the zones of authigenic carbonate precipitation and of highest microbial cell abundance correspondingly shallow toward the periphery. The underlying sill was drilled several times and yielded diverse igneous rock textures, sediment/sill interfaces, and alteration minerals in veins and vesicles. The Ringvent sill became the target of an integrated, interdisciplinary sampling and research effort that included geological, geochemical, and microbiological components. The thermal, lithologic, geochemical, and microbiological contrasts between the northwestern sites (U1545 and U1546) and the Ringvent sites (U1547 and U1548) form the core scientific observations informing the direct influence of sill-sediment interaction. These observations are supplemented by results from sites that exhibit persistent influence of thermally equilibrated sill intrusions, including supporting long-lived methane cold seeps, as observed at off-axis Sites U1549 and U1552, and the persistent geochemical record of hydrocarbon formation near the sill/sediment contact, as observed at the northern axial trough Site U1550, which confirms observations from Deep Sea Drilling Project (DSDP) Leg 64. Drilling at Site U1551 ~29 km southeast of the axial graben was not successful due to unstable shallow sands, but it confirmed the dominant influence of gravity-flow sedimentation processes southeast of the axial graben. The scientific outcomes of Expedition 385 will (1) revise long-held assumptions about the role of sill emplacement in subsurface carbon mobilization versus carbon retention, (2) comprehensively examine the subsurface biosphere of Guaymas Basin and its responses and adaptations to hydrothermal conditions, (3) redefine hydrothermal controls on authigenic mineral formation in sediments, and (4) yield new insights into the long term influence of sill-sediment interaction on sediments deposited at the earliest stages of seafloor spreading, that is, when spreading centers are proximal to a continental margin. The generally high quality and high degree of completeness of the shipboard data sets present opportunities for inter- and multidisciplinary collaborations during shore-based studies. In comparison to DSDP Leg 64 to Guaymas Basin in 1979, continuous availability of sophisticated drilling strategies (e.g., the advanced piston corer [APC] and half-length APC systems) and numerous analytical innovations greatly improved sample recovery and scientific yield, particularly in the areas of organic geochemistry and microbiology. For example, microbial metagenomics did not exist 40 y ago. However, these technical refinements do not change the fact that Expedition 385 in many respects builds on the foundations of understanding laid by Leg 64 drilling in Guaymas Basin. 

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3. Expedition 385 Preliminary Report: Guaymas Basin Tectonics and Biosphere

   https://doi.org/10.14379/iodp.pr.385.2020  

   Teske, A. ; Lizarralde, D. ; Höfig, T.W. ( December 2020 , Preliminary report)

   null (Ed.)

   International Ocean Discovery Program (IODP) Expedition 385 drilled organic-rich sediments with sill intrusions on the flanking regions and in the northern axial graben in Guaymas Basin, a young marginal rift basin in the Gulf of California. Guaymas Basin is characterized by a widely distributed, intense heat flow and widespread off-axis magmatism expressed by a dense network of sill intrusions across the flanking regions, which is in contrast to classical mid-ocean ridge spreading centers. The numerous off-axis sills provide multiple transient heat sources that mobilize buried sedimentary carbon, in part as methane and other hydrocarbons, and drive hydrothermal circulation. The resulting thermal and geochemical gradients shape abundance, composition, and activity of the deep subsurface biosphere of the basin. Drill sites extend over the flanking regions of Guaymas Basin, covering a distance of ~81 km from the from the northwest to the southeast. Adjacent Sites U1545 and U1546 recovered the oldest and thickest sediment successions (to ~540 meters below seafloor [mbsf]; equivalent to the core depth below seafloor, Method A [CSF-A] scale), one with a thin sill (a few meters in thickness) near the drilled bottom (Site U1545), and one with a massive, deeply buried sill (~356–430 mbsf) that chemically and physically affects the surrounding sediments (Site U1546). Sites U1547 and U1548, located in the central part of the northern Guaymas Basin segment, were drilled to investigate a 600 m wide circular mound (bathymetric high) and its periphery. The dome-like structure is outlined by a ring of active vent sites called Ringvent. It is underlain by a remarkably thick sill at shallow depth (Site U1547). Hydrothermal gradients steepen at the Ringvent periphery (Holes U1548A–U1548C), which in turn shifts the zones of authigenic carbonate precipitation and of highest microbial cell abundance toward shallower depths. The Ringvent sill was drilled several times and yielded remarkably diverse igneous rock textures, sediment–sill interfaces, and hydrothermal alteration, reflected by various secondary minerals in veins and vesicles. Thus, the Ringvent sill became the target of an integrated sampling and interdisciplinary research effort that included geological, geochemical, and microbiological specialties. The thermal, lithologic, geochemical, and microbiological contrasts between the two deep northwestern sites (U1545 and U1546) and the Ringvent sites (U1547 and U1548) form the scientific centerpiece of the expedition. These observations are supplemented by results from sites that represent attenuated cold seepage conditions in the central basin (Site U1549), complex and disturbed sediments overlying sills in the northern axial trough (Site U1550), terrigenous sedimentation events on the southeastern flanking regions (Site U1551), and hydrate occurrence in shallow sediments proximal to the Sonora margin (Site U1552). The scientific outcomes of Expedition 385 will (1) revise long-held assumptions about the role of sill emplacement in subsurface carbon mobilization versus carbon retention, (2) comprehensively examine the subsurface biosphere of Guaymas Basin and its responses and adaptations to hydrothermal conditions, (3) redefine hydrothermal controls of authigenic mineral formation in sediments, and (4) yield new insights into many geochemical and geophysical aspects of both architecture and sill–sediment interaction in a nascent spreading center. The generally high quality and high degree of completeness of the shipboard datasets present opportunities for interdisciplinary and multidisciplinary collaborations during shore-based studies. In comparison to Deep Sea Drilling Project Leg 64 to Guaymas Basin in 1979, sophisticated drilling strategies (for example, the advanced piston corer [APC] and half-length APC systems) and numerous analytical innovations have greatly improved sample recovery and scientific yield, particularly in the areas of organic geochemistry and microbiology. For example, microbial genomics did not exist 40 y ago. However, these technical refinements do not change the fact that Expedition 385 will in many respects build on the foundations laid by Leg 64 for understanding Guaymas Basin, regardless of whether adjustments are required in the near future. 

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4. Well-hidden methanogenesis in deep, organic-rich sediments of Guaymas Basin

   https://doi.org/10.1038/s41396-023-01485-y  

   Bojanova, Diana P. ; De Anda, Valerie Y. ; Haghnegahdar, Mojhgan A. ; Teske, Andreas P. ; Ash, Jeanine L. ; Young, Edward D. ; Baker, Brett J. ; LaRowe, Douglas E. ; Amend, Jan P. ( August 2023 , The ISME Journal)

   Deep marine sediments (>1mbsf) harbor ~26% of microbial biomass and are the largest reservoir of methane on Earth. Yet, the deep subsurface biosphere and controls on its contribution to methane production remain underexplored. Here, we use a multidisciplinary approach to examine methanogenesis in sediments (down to 295 mbsf) from sites with varying degrees of thermal alteration (none, past, current) at Guaymas Basin (Gulf of California) for the first time. Traditional (13C/12C and D/H) and multiply substituted (13CH3D and 12CH2D2) methane isotope measurements reveal significant proportions of microbial methane at all sites, with the largest signal at the site with past alteration. With depth, relative microbial methane decreases at differing rates between sites. Gibbs energy calculations confirm methanogenesis is exergonic in Guaymas sediments, with methylotrophic pathways consistently yielding more energy than the canonical hydrogenotrophic and acetoclastic pathways. Yet, metagenomic sequencing and cultivation attempts indicate that methanogens are present in low abundance. We find only one methyl-coenzyme M (mcrA) sequence within the entire sequencing dataset. Also, we identify a wide diversity of methyltransferases (mtaB, mttB), but only a few sequences phylogenetically cluster with methylotrophic methanogens. Our results suggest that the microbial methane in the Guaymas subsurface was produced over geologic time by relatively small methanogen populations, which have been variably influenced by thermal sediment alteration. Higher resolution metagenomic sampling may clarify the modern methanogen community. This study highlights the importance of using a multidisciplinary approach to capture microbial influences in dynamic, deep subsurface settings like Guaymas Basin.

    

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5. Heterogeneity of methane seep biomes in the Northeast Pacific

   https://doi.org/10.1016/j.dsr2.2017.10.016  

   Seabrook, S. ( January 2018 , Deep sea research. Part II, Topical studies in oceanography)

   Methane seeps provide biogeochemical and microbial heterogeneity in deep-sea habitats. In the Northeast (NE) Pacific Ocean recent studies have found an abundance of seeps at varying spatial separations and within distinct biogeochemical environments ranging in oxygen, depth, and temperature. Here, we examine eight newly discovered seeps and two known seeps covering 800 km and varying across 2000 m water depth to identify: (1) novel megafaunal communities in this geographical region; (2) variations in the microbiome of seep habitats across the margin; (3) spatial and biogeochemical drivers of microbial diversity at seeps. In addition to authigenic carbonates, clam beds, microbial mats, and exposed hydrates - we also observed Siboglinidae tube worm bushes and an anomalous deep-sea barnacle adding to the overall habitats known from the NE Pacific. The microbial communities showed high variability in their spatial distribution and community structure. The seep communities formed distinct groups that included multiple groups of anaerobic methane oxidizing Archaea (ANME; 1, 2ab, 2c, and 3), often co-occurring within one site – however, there were also other sites with clearly dominant members (e.g. ANME-1s at Nehalem Bank). Sulfide oxidizers were dominated by the non-mat forming Campylobacterales and even though vertical gradients in redox potential typify seep sediments, in two cases there was not a significant change in community structure across the top five cm of sediment. We posit that these patterns were driven by ‘bubble-turbation,’ and bioirrigation by megafauna. A surprising latitudinal trend was observed in species diversity and richness with increasing richness significantly correlated to increasing latitude. Overall, our results demonstrate that heterogeneity is ubiquitous in the seep biome, spanning all faunal classes, and that the understanding of seeps and the drivers of the community structure can be improved by studying seeps at a range of spatial scales. 

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