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Deep Life Community -‐ The Deep Carbon Observatory – 2014-‐2015 ALFRED P. SLOAN FOUNDATION www.sloan.org | proposal guidelines PROPOSAL COVER SHEET Project Information Principal Investigator Mitchell L. Sogin, Ph.D. 7 MBL Street, Woods Hole MA 02543 [email protected] 5085661468 Grantee Organization: Woods Hole Amount Requested: st Requested Start Date: January 1 , 2014 st Requested End Date: December 31 , 2015 Deep Life Community-‐ The Deep Carbon Observatory—2014-‐2015 Project Goal The Deep Life Community (DLC) seeks to map the abundance and diversity of subsurface marine and continental microorganisms in time and space as a function of their phylogenomic and biogeochemical properties, and to explore their roles and interactions with the deep carbon cycle. Objectives This proposal will strengthen our nascent research and coordination organization dedicated to achieving DLC’s decadal goals. It will extend molecular studies to a greater number of samples from high-‐value marine and continental sites in order to describe diversity, distribution and functional adaptations of deep life. It will explore life’s interplay with geological processes in the deep subsurface including studies of microbial activities and distributions in hydrogen-‐rich habitats, which favor abiogenic synthesis of methane and higher hydrocarbons. We will: a) Explore the limits of deep life using improved life detection capabilities, b) Develop and apply tracer approaches to track the flow of carbon into biomolecules and cells, and c) Measure the interrelationship between composition of carbonaceous materials and deep life. To achieve these objectives, the DLC must seek additional resources and engage the best deep-‐life researchers from around the globe. Proposed Activities The Census of Deep Life (CoDL) will support marker gene analyses for 1600 new samples or metagenomic analyses of as up to 100 samples. The DLC will cohost a workshop with the Deep Energy Community (DEC) on abiotic H2 generation and will collaborate with DEC on methane isotopologues in biologically produced methane spanning temperatures from 20°C to 120°C. The DLC will use stable-‐ and radioisotope probing to trace the utilization and assimilation of carbon compounds in subsurface environments. The DLC will support laboratory activities and expenses associated with the preparation of compelling research proposals, workshops with the International Ocean Discovery Program (IODP) and the International Continental Scientific Drilling Program (ICDP) for developing biological drilling initiatives, and co-‐funded meetings with the NASA Astrobiology Institute (NAI) or the Center for Dark Energy Biosphere Investigations (C-‐DEBI) to develop new collaborations. This proposal will support DCL-‐wide meetings in 2014 and 2015, and will support DCL-‐wide participation at the 2015 Deep Carbon Observatory (DCO) “all hands” meeting. Expected Products Expected products of these proposed activities include publications about deep subsurface microbes and microbial mediated processes on the cycling of carbon; online molecular data describing deep life diversity and processes; new research proposals to leverage Sloan funding; and DLC-‐dedicated field missions with participation from other DCO communities. Expected Outcomes This proposed activities will generate new insights about diversity, evolution, and processes that govern deep life and its role in the cycling of deep carbon. Finally, it will expand the constituency of the DLC and will enlighten the public about microbes deep underground. PROPOSAL: DEEP LIFE COMMUNITY - THE DEEP CARBON OBSERVATORY Project Advocates Co-Principle Investigators: Mitchell L. Sogin ([email protected]) Kai-Uwe Hinrichs ([email protected]) Investigators: Douglas H. Bartlett ([email protected]) Antje Boetius ([email protected]) Frederick S. Colwell ([email protected]) Isabelle Daniel ([email protected]) Steven D’Hondt ([email protected]) Thomas L. Kieft ([email protected]) Matthew O. Schrenk ([email protected]) Advocates: Fumio Inagaki ([email protected]) Roland Winter ([email protected]) Deep Life Community - The Deep Carbon Observatory – 2014-2015 TABLE OF CONTENT 1 - INTRODUCTION - STATE OF DEEP LIFE RESEARCH AND KEY QUESTIONS ……………….……..4 2 - DEEP LIFE COMMUNITY ORGANIZATION – PARTICIPANT QUALIFICATIONS…………………….6 3 - DLC RESEARCH ACCOMPLISHMENTS – 2012-2013…………………... ……………………….7 3.1 - Progress by the Census of Deep Life.(CoDL) …………..…………………………..7 3.2 - Progress by Deep Life I: Microbial Carbon Transformations in Rock-Hosted Deep Subsurface Habitats Project (RHC) ...………………………………………………8 4 - THE PROPOSED PROJECT: DEEP LIFE RESEARCH AND COORDINATION NETWORK……………...9 4.1 - CENSUS OF DEEP LIFE – PHASE II-III………………………………………………..10 4.2 - DEEP LIFE II: ROCK-HOSTED COMMUNITIES………………………………………..12 4.2.1 - Metagenomics and Deep Carbon Cycling ……………...………………….12 4.2.2 - Abiotic H2, abiotic organic C, and origins of life. ……...………………….13 4.2.3 - Integration of data sets between studies. ……...…………………………...14 4.3 - LINKING DEEP LIFE TO DEEP ENERGY COMMUNITY INVESTIGATIONS. …………......14 4.4 - NEW FRONTIERS IN DEEP LIFE ………………………………...15 4.5 - PARTNERSHIPS THAT BUILD THE DEEP LIFE COMMUNITY………………………...…17 4.6- SUMMER SCHOOL DEEP MARINE BIOSPHERE ……….………………………..…....….21 4.7 - MBL INFORMATICS COURSE ……………...………………………………..……….21 4.8 - DATA SCIENCE …………………..………………………………………………....22 4.9 – ENGAGEMENT……………………………………………………………………....22 5 - OUTPUT OF RESEARCH PROJECT.………………………………………………………….......22 6 - BUDGET JUSTIFICATION AND LEVERAGING……………………………………..……….…….23 7 - PRIOR SLOAN SUPPORT…………………...………………………..…………….…….......23(7) 8 - BUDGET………………...………………...………………...………………...…………….…24 9 - CURRICULUM VITAE…………………...……………...………………...……….………........25 A - APPENDICES………………...………………...………………...………………...………..…38 A.1 - REFERENCES CITED………………...………………...………………...…………...38 A.2 - DECADAL GOALS………………...………………...………………...……………..39 A.3 - FUNDING THAT SUPPORTS ONE OR MORE DEEP LIFE’S DECADAL GOALS…………. 40 A.3.1 - Funded Projects………………...………………...…..……………...……40 A.3.2 - Pending Leveraged Proposals………………...………..………................42 A.4 - DEEP LIFE COMMUNITY PUBLICATIONS……………………………………...…….43 A.5 - POST DOCTORAL SUPPORT………………...………………...………………......…47 A.6 - MANAGEMENT PLAN ………………...………………...………………................. 48 A.6.1 - MEMBERSHIP OF DEEP LIFE SCIENTIFIC STEERING COMMITTEE……...…...48 A.6.2 – SCIENTIFIC STEERING COMMITTEE ACTIVITIES………………...……....... 48 3 Deep Life Community - The Deep Carbon Observatory – 2014-2015 PROPOSAL: DEEP LIFE COMMUNITY - THE DEEP CARBON OBSERVATORY 1 - INTRODUCTION - STATE OF DEEP LIFE RESEARCH AND KEY QUESTIONS. Studies of terrestrial and marine sediments reveal subsurface microbial ecosystems that harbor ≥1029 organisms with total carbon content possibly equaling all surface life [1, 2]. These deep and dark biological reservoirs may extend to six km or more beneath the seafloor and continental surface. Instead of tapping into solar power, members of deep subsurface microbial communities harvest energy from geofuels or buried, refractory detrital matter, exploiting small disequilibria between chemical redox states in order to drive the synthesis of macromolecules and biological reproduction. These processes lead to large-scale transformations of inorganic and organic compounds with an attendant impact on the deep carbon cycle. Within the deep biosphere, some organisms survive and grow near the interface of the abiotic and biotic realms, where high pressures, elevated temperatures and energy limitations require the ability to tolerate and adapt to extreme conditions. How deep subsurface microbes differ from surface and near surface organisms remains unknown. Adaptations of deep life to high pressures and temperatures, limited energy resources, diffusion limited transport of nutrients and other environmental factors may require different rule sets that govern primary production, competition, succession, dispersal, and their mode of evolution. First order questions about organisms that inhabit this largely unexplored environment include: Who is there? How is deep life distributed and what is its biomass? How do they survive? How quickly do they reproduce and what provides a source of energy? What factors govern their dispersal patterns and how do they rapidly adapt to a changing environment? What is their impact on the deep carbon cycle? The Deep Life Community (DLC), which explores the evolutionary and functional diversity of Earth’s deep biosphere and its interactions with the carbon cycle, embraces three primary Decadal Goals: 1) Determine the processes that define the diversity and distribution of deep life 4 Deep Life Community - The Deep Carbon Observatory – 2014-2015 as it relates to the carbon cycle; 2) Determine the environmental limits of deep life; and 3) Determine the interactions between deep life and carbon cycling. Each of these goals addresses aspects of Quantities, Movements, Origins, and Forms (QMOF): Quantities (concentrations of cells that constitute deep life and its carbon content), Movements (Dispersal and Distribution of microbial life and its carbon substrates), Origins (Abiotic vs Biotic sources of substrates fueling deep life) and Forms (Different genotypes and phenotypes, as well as compositional and redox states of carbon) of deep life. Our research strategy will leverage DLC’s investment in conducting a global census over time and space for all three domains of life and viruses in both the marine and continental subsurface. Through omics we seek to explore mechanisms that shape microbial evolution and dispersal in the deep biosphere and to identify the ecological rules that shape community structures. Studies of the environmental limits of life employ a combination of laboratory and field experiments to determine physical and chemical extremes that are compatible with life. These measurements will inform modeling studies and potentially will provide clues about differences between the biotic/abiotic interface that may have played a role in the origins of life. Finally, Deep Life will explore patterns and mechanisms of bioticallymediated carbon transformations in the subsurface and the interaction of these processes with the surface world. As described below, the DLC has progressed towards exploring the diversity and distribution of life in marine and continental deep subsurface environments. Laboratory experiments have investigated pressure limits on microbial survival, and omic studies have provided insights about the role of microbes in the deep carbon cycle. Despite these achievements, the DLC faces daunting challenges that require funding beyond what the Sloan Foundation can provide. This proposal will continue building the DLC, provide resources for research coordination activities, and support activities that can lead to newly funded initiatives. 5 Deep Life Community - The Deep Carbon Observatory – 2014-2015 2 - DEEP LIFE COMMUNITY ORGANIZATION – PARTICIPANT QUALIFICATIONS. The 2010 DCO Deep Life Workshop on Catalina Island California convened by Mitchell Sogin, Katrina Edwards and Steve D’Hondt set DLC’s initial agenda. Microbiologists, biogeochemists, and geochemists outlined research goals in the white paper Deep Subsurface Microbiology and the Deep Carbon Observatory [3]. Major recommendations of the report included surveying the deep biosphere through a Census of Deep Life (CoDL), investigating deep life’s evolutionary history, exploring the global distribution of microbes in the deep biosphere, inventorying their metabolic character, identifying how they have adapted to high pressures and high temperatures, and determining the influence on microbes on the deep carbon cycle in both continental and marine subsurface habitats. Fredrick Colwell and Mitchell Sogin developed a CoDL proposal that the Alfred P. Sloan Foundation funded in late 2010 and the DCO recruited Isabelle Daniel and Mitchell Sogin to serve as Co-Chairs of the DLC. In June 2011, the steering committee for Deep Life met in Bremen to review DLC research proposals solicited by the DCO secretariat. The DLC identified the most important common themes in the proposals and charged Matt Schrenk and Isabelle Daniel with organizing the Deep Life I: Microbial Carbon Transformations in Rock-Hosted Deep Subsurface Habitats Project (RHC). This multi-investigator project (with 11 laboratories spanning 7 countries) seeks to describe and quantify the metabolic activities of microorganisms in the rock-hosted subsurface biosphere. The project includes observational studies of key understudied deep subsurface environments coupled with experimental investigations aimed at elucidating the physiological underpinnings of microbial adaptations to these environments. This research focused on the hypothesis that rock-hosted deep biosphere communities use abiotically-sourced carbon compounds and that unique high-pressure, hydrogen-enriched environments shape the deep biosphere communities. In early 2013, Kai-Uwe Hinrichs joined Mitchell Sogin as Co-Chair of the DLC. Isabelle Daniel remains on the DLC 6 Deep Life Community - The Deep Carbon Observatory – 2014-2015 steering committee but has also assumed the chair of the Deep Energy Community. The section CURRICULUM VITAE provides c.v’s of the advocates for this proposal including DLC Steering Committee members who have led major efforts in Deep Life research projects. 3 - DLC RESEARCH ACCOMPLISHMENTS – 2012-2013. The CoDL and RHC have synergies that have contributed to remarkable progress over 18 months. Seventeen research groups received important data from the initial phase of CoDL’s marker gene (rRNA) survey. A second phase enabled by reduced sequencing costs added sites to the census and offered opportunities for shotgun metagenome surveys. Eleven laboratories participated in the RHC project. APPENDIX A.4 provides a list of publications, manuscripts in review, and manuscripts in preparation. APPENDIX A.3 cites leveraging activities that have supported DLC science. 3.1 - PROGRESS BY THE CENSUS OF DEEP LIFE (CODL) (F. Colwell, M. Sogin) • Desulforudis audaxviator represents a “keystone species” in subsurface samples from South African gold mines, seafloor crustal materials, and Great Basin wells and springs. • A subglacial Icelandic microbiome lacks Archaea and consists of only five microaerophilic and anaerobic chemolithoautotrophs [4]. This unique subsurface microbial community does not receive input of energy from the surface (i.e., sunlight or photosynthetically-derived organic matter) and could serve as an analogue for life in the subsurface of other planets. • Firmicutes dominate deep methane hydrate zones in the sediments of the Indian Ocean including zones above, within and below strata that contained hydrates [5]. The absence of Archaea in these samples - and by inference, methanogens – agrees with prior reports that concentrations of methane producing microbes must be very low in hydrate-bearing sediments • Pilot scale geological sequestration of CO2 [6] will utilize deep basalt aquifers that host diverse bacterial and archaeal communities. Microbes at the depth of planned CO2 injection 7 Deep Life Community - The Deep Carbon Observatory – 2014-2015 are related to taxa that use hydrogen and single-carbon compounds. The metabolic activity of these communities could impact long-term stability of carbon sequestered underground. • High-resolution mineral colonization experiments in the sub-seafloor igneous crust reveal that mineralogy and mineral chemistry dictate the types and diversity of attached microbes (Smith et al. in review). Geological formations rich in olivine (e.g., slow-spreading ridges) serve as hotspots for biological activity in oceanic crust, where biogeochemical processes on crustal minerals rather than transported products of photosynthesis fuel microbial growth. 3.2 - PROGRESS BY DEEP LIFE I: MICROBIAL CARBON TRANSFORMATIONS IN ROCK-HOSTED DEEP SUBSURFACE HABITATS PROJECT (RHC)– (Matt Schrenk, Isabelle Daniel, T Kieft) • Molecular diversity studies, supported by measurements of H2, CH4 - rich and pH, of timeseries samples from active serpentinization environments of uplifted ultramafic oceanic crust at the Coast Range Ophiolite Microbial Observatory (CROMO) in northern California identified Betaproteobacteria and Clostridales as key taxa. Collaborations with the CoDL and DOE community sequencing program have identified pathways such as assimilation of small organic acids and carbon monoxide that putatively control the exchange of carbon and energy between the deep Earth and the surface environment. • Coupled molecular and 13 C tracer studies of microbial communities in diffuse hydrothermal fluids collected from the world’s deepest known hydrothermal vents at the Mid Cayman Rise (MCR) in the Caribbean Sea documented microbial and functional diversity for abundant taxa related to Methanococcales, Archaeoglobales, and Epsilonproteobacteria. Diffuse fluid samples from vents located on basaltic and ultramafic substrates between 2000 and 5000 m water depth hosted taxa typical of more well-known high temperature systems at the deeper site (Piccard) and taxa that may be involved in methane production at the shallower ultramafic rock site (Von Damm). These data will provide insight into links between the microbial 8 Deep Life Community - The Deep Carbon Observatory – 2014-2015 communities and their geochemical environment. Gene sequencing and stable isotope tracer experiments indicated that methanogenesis from formate occurs at the ultramafic hosted sites, and is absent from the deeper, higher temperature basalt hosted sites such as Piccard. • Fluorescently activated cell-sorting and taxonomic-screening of 10 samples from deep fractures in Precambrian Shield environments of South Africa, Finland, and Canada identified close relatives of Candidatus Desulforudis audaxviator, which dominated communities in earlier studies of South African gold mines. To understand the ecology, population genetics, and biogeography of these microbes in the deep subsurface, the JGI Community Sequencing Program, will generate draft genome sequences for 200 single-cell genomic preparations. • The RHC project has isolated and characterized 5-10 new strains of novel, thermophilic, piezophilic organisms from the MCR and the Precambrian Shield environments. One of the strains isolated from the MCR grows well at 120 MPa (corresponding to 12 km water depth!). Physiologic and genomic characterization of these isolates will provide insight into the adaptations of microorganisms to deep subsurface, high-pressure environments. • Biophysical and biochemical studies of Shewanella oneidensis adapted in the lab to 2.5 GPa (comparable to depths of ~75 km), identified two pressure domains. Beyond 500 MPa, the physiology of the P-adapted S. oneidensis appears to be very different. Quasi Elastic Neutron Scattering experiments to 200 MPa revealed that the mobility of water across cell membranes decreases slightly with pressure but is many orders of magnitude less than the decrease in metabolic activity measured over the same pressure range using X-ray spectroscopy. 4 - THE PROPOSED PROJECT: DEEP LIFE RESEARCH AND COORDINATION NETWORK. The Deep Life Community steering committee has developed a research agenda / networking plan that leverages advances from the CoDL and the RHC project, and outlines opportunities to engage initiatives beyond the current umbrella of the Deep Carbon Observatory. This plan 9 Deep Life Community - The Deep Carbon Observatory – 2014-2015 incorporates new elements that address key objectives in each of the three primary Decadal Goals (APPENDIX A.2). Beyond scheduled meetings and modest continued funding for the Census of Deep Life, the DLC must seek sources of funding from other foundations and agencies. Over the next two years, the DLC will strategically deploy limited resources to support community building (networking), workshops, logistical expenses of writing new proposals e.g. travel to other laboratories, small-scale laboratory and field experiments (including the collection of samples of opportunity and collaborative initiatives with other DCO communities), and the purchase of affordable equipment or accessories that would further the research agenda of the DLC. The collective goal is to advance deep life science through discovery based research and enablement through new successful research proposals. The appendix A.6.2 describes the mechanisms and guidelines for submitting ideas to the DLC steering committee and the criteria that we will use to render decisions about resource deployment. Unlike more traditional pilot project opportunities, decisions about support will largely hinge upon the potential impact on success of new funding proposals to other agencies or foundations. 4.1 - CENSUS OF DEEP LIFE – PHASE II-III – (A. Boetius, R. Colwell, S. D’Hondt, M. Sogin). The Census of Deep Life (CoDL), seeks to identify the diversity and distribution of microbial life in continental and marine deep subsurface environments. This project directly addresses deep life’s Decadal Goal I to develop a global 3-D census of diversity in continental and marine deep subsurface environments and to explore mechanisms that govern microbial evolution and dispersal in the deep biosphere. It contributes to Decadal Goal II by informing us about the limits of life and metabolic properties necessary to adapt to deep subsurface environments. CoDL activities integrate closely with the RHC project by providing a taxonomic framework for describing and comparing deep subsurface communities. With the introduction of shotgun metagenomics, the CoDL will address key functional questions about deep life. 10 Deep Life Community - The Deep Carbon Observatory – 2014-2015 During Phase I of the CoDL, 17 laboratories submitted samples for analysis. Phase II of the project accepted a second round of samples from 12 research groups (in progress) and Phase III (supported through this proposal) will support a larger number of samples but at significantly reduced costs. Samples for analysis will continue to include drill cores of rock and sediment, water pumped from newly drilled wells, seeps in mines, crustal fluids, in situ colonized media from CORKS as well as samples of opportunity from other DCO communities. CoDL’s general experimental strategy relies upon massively parallel DNA sequencing for 1) “marker gene” studies that inform us about the relative abundance and kinds of microbes in a sample, and 2) shotgun metagenomics investigations that capture information about the metabolic potential of microbial communities. Both extract information from microbial DNA from deep subsurface samples. Our original strategy employed pyrosequencing technology but the CoDL transitioned to the Illumina MiSeq platform, which can reduce sequencing costs/sample by ~100 fold. The Phase 3 CoDL project will accept many more samples with an investment of less than $100,000. The CoDL portal on the web site VAMPS (http://vamps.mbl.edu) will provide access to the data and a wide range of analytical capabilities including the ability to process data through the QIIME paradigm [7]. Cross project synthesis activities through formation of a collaborating working group will provide descriptions of distribution and dispersal patterns for microbes in the deep subsurface. For metagenomic analyses, the CoDL will take advantage of an Illumina HiSeq platform, which provides ~6 fold coverage for any 2-4 Mbp genome that accounts for >0.2% of the microbial genomes in a sample. This information has the potential to provide detailed genotypic descriptions of the metabolic and functional capabilities of typical communities from deep subsurface environments. By coupling estimates of community diversity from marker gene analyses with shotgun metagenomics, we will be able to titrate the amount of shot-gun 11 Deep Life Community - The Deep Carbon Observatory – 2014-2015 metagenomic sequencing required to infer the functional properties of microbial populations in deep subsurface communities. With appropriate sample preparation to isolate viral particles, this technology will support shotgun metagenomic analysis of viral populations. 4.2 - DEEP LIFE II: ROCK-HOSTED COMMUNITIES. Projects listed below are being generously supported during the current funding cycle. In the 2014-2015 funding period, we anticipate that the activities by the RHC will be increasingly supported by leveraged funds. The DLC will deploy additional resources as requested to complete tasks necessary for data synthesis and for securing funding from outside sources. 4.2.1 - METAGENOMICS AND DEEP CARBON CYCLING (M. Schrenk, T. Kieft). In just a few short years, the CoDL and RHC studies have dramatically improved our understanding of microbial diversity in deep subsurface environments, spanning both marine and terrestrial realms and advanced our understanding of the interplay between abiotic, geological processes operating at depth and deep life, capitalizing upon coincident advances made in the technology of gene sequencing. These studies have fed into complementary “–omics” approaches, including metagenomics, transcriptomics, and single cell genomes which have enabled a broad view of microbial metabolic potential, activity, and population genetics in subsurface environments. Focused efforts have made headway in the collection and preparation of samples from low biomass environments and assembly from short sequence reads. Coupled with leveraged resources, such as the DOE Joint Genome Institute Community Sequencing Program, these efforts have begun to pay dividends in terms of the accumulation of massive amounts of genomic data. Automated sequence annotation, using programs such as the MG-RAST server have permitted the initial high-throughput analysis of these datasets. However, the accurate annotation of these genes requires verification with reference organisms harboring sequences coding for bonafide functions. Further, a sequences-specific phylogenomic approach allows for analysis of 12 Deep Life Community - The Deep Carbon Observatory – 2014-2015 the evolutionary relatedness of these microbial communities. Alignments have been constructed for several genes expected to be critical for carbon cycling in the deep subsurface (e.g. methanogenesis, carbon fixation, formate assimilation) and will be expanded to include other genes related to carbon processing. These alignments will facilitate the rapid analysis of new metagenomics datasets, expected to be a part of the next phase of the CoDL, and for phylogenetic analyses relating deep life across habitats. Genomic information gathered by the RHC will address central goals in the DLC’s agenda but will require additional efforts in annotation and phylogenetic reconstruction. We anticipate several requests for modest support to resolve these questions. 4.2.2 - ABIOTIC H2, ABIOTIC ORGANIC C, AND ORIGINS OF LIFE. (M. Schrenk, T. Kieft, I. Daniel, D. Bartlett). In certain subsurface settings, deep life is independent of photosynthetically generated energy and instead relies on abiotic H2 derived from either rock-water reactions [8] or radiolysis of water [9], or on simple organic C compounds generated abiotically via FischerTropsch-type reactions [10]. Subsurface habitats harboring these chemosynthetic ecosystems fueled by abiogenic H2 have even been suggested as sites for the origins and early evolution of life [11]. The RHC project has facilitated a range of research projects that investigate the physiology and ecology of hydrogen-utilizing subsurface microorganisms, including those influenced by serpentinization (CROMO and Cayman Ridge) and radiolysis (Witwatersrand Basin, South Africa), and the DEC focuses on the abiogenic production of organic compounds. We will strengthen this framework through investigations that link the process of carbon assimilation in abiotic H2-generating environments with analysis of microbial genomes. The work will build upon the RHC’s and DLC’s expertise in stable isotope probing, isotope analysis of various carbon pools from bulk to cellular and molecular levels, and single-cell genomics and will link studies of carbon reservoirs and fluxes conducted by DE, e.g., at the CROMO site in 13 Deep Life Community - The Deep Carbon Observatory – 2014-2015 California, USA, and the Bay of Prony site in New Caledonia. We seek to identify microbes that utilize abiogenically produced H2 and carbon substrates, and to obtain genetic and biochemical information to reveal evolutionary relationships and metabolic pathways. Beyond explaining how microorganisms in the deep subsurface assimilate carbon, these analyses will provide a framework for inferring relationships between the biochemistry of extant systems and origins of life scenarios in the deep subsurface. 4.2.3 - INTEGRATION OF DATA SETS BETWEEN STUDIES (M. Schrenk, M. Sogin, I. Daniel). The RHC sub-projects will continue to integrate with each other and the CoML. The Census of Deep Life will anchor taxonomic descriptions and comparison of deep subsurface ecosystems. Measurements using similar protocols for stable isotope tracer measurements across each of the field sites, will allow comparison of the rates and types of operative metabolism. The sharing of samples and microbial cultures among research teams will facilitate comparative analyses into a larger synthesis that describes carbon cycling by the rock-hosted microbial biosphere. We seek to compare microbiomes across all different subsurface realms, including continental vs marine, rock hosted vs sedimentary, organic rich vs organic poor, hot vs cold. 4.3 - LINKING DEEP LIFE TO DEEP ENERGY COMMUNITY INVESTIGATIONS. During the past few years, an important synergy has developed between projects pursued in the Rock-Hosted Communities (RHC) project and the Deep Energy Community (DEC) of the DCO. The RHC project has made headway in understanding microbial activities and distributions in hydrogenrich subsurface habitats, which favor the abiogenic synthesis of methane and higher hydrocarbons. In parallel, the DEC has initiated a range of investigations, including fieldwork in both marine and terrestrial settings and laboratory experiments, to identify sources and signatures of methane and other hydrocarbons. The process of serpentinization, the aqueous alteration of ultramafic rocks characteristics of Earth’s upper mantle has become a centerpiece in both types 14 Deep Life Community - The Deep Carbon Observatory – 2014-2015 of investigations. Likewise, the joint interest in carbon transformations and microbial communities in subsurface zones associated with the biotic-abiotic interface unifies both DCO communities and will provide a breeding ground for future collaboration. Several areas of focus have emerged. Studies in both Deep Life and Deep Energy have strived to inventory microbial processes and their impact upon carbon biogeochemistry in subseafloor rocks from locations such as the Mid-Cayman Rise and the Mid Atlantic Ridge. Studies initiated through the CoDL and the DEC are examining microbes in shallow sea hydrothermal chimneys in New Caledonia venting volatile-rich, high pH fluids. Scientists in both communities took part in the Oman Drilling Workshop, co-sponsored by ICDP and DCO, in Palisades, NY in September 2012. Laboratory experiments in both communities are serving to constrain the geochemical processes and signatures associated with abiogenic hydrocarbon production, and how they are intertwined with the deep biosphere. These same processes may ultimately be operative at the depth limits to the biosphere in the uppermost mantle and lower crust, and in forearc basins associated with subduction zones. Serpentinization-associated processes may play important roles in influencing the exchange of carbon and energy between the deep Earth and the biosphere. Furthermore, these mineral catalytic systems are believed to have played a role in the origins and early evolution of life. Serpentinizing environments inhabited by microorganisms may contain biochemical relicts of ancient prebiotic processes. We propose to build upon these efforts with a series of strategic investments, e.g., in the form of collaborations between DLC and DEC, including: • Co-hosting a workshop with the DEC on the topic of abiotic H2 generation and the associated potential to support life. This workshop will highlight both geochemical and biochemical aspects of the process and its impact on the carbon cycle through time and be hosted at a site near a serpentinizing ophiolite (e.g. Portugal, Northern Italy). (I. Daniel, M. Schrenk) 15 Deep Life Community - The Deep Carbon Observatory – 2014-2015 • Collaboration on methane isotopologues (K.-U. Hinrichs; with B. Sherwood Lollar, DE) – DLC will collaborate with DEC on the examination of clumped isotopes in biologically produced methane covering a temperature range from 20°C to 120°C. A good fraction of this temperature range overlaps with temperatures at which methane may be formed abiotically in subsurface systems. Examination of the associated signals will be crucial for the development of methane’s clumped isotopes as an indicator of its formation pathway. • Cataloguing microbial abundance and diversity (M. Schrenk, R. Colwell, T. Kieft, D. Bartlett, I. Daniel) in sites that are currently being investigated by the DEC, but which lack a microbiological component (e.g. gas fluxes studies of serpentinizing ophiolites, carbonated seafloor rocks, active drilling projects, etc.). Cultivation efforts will allow physiological studies at high pressures in the laboratory. 4.4 - NEW FRONTIERS IN DEEP LIFE (K.-U. Hinrichs). The DLC has identified a number of key tasks of direct relevance to our ability to reach our decadal goals. For example, we must improve life detection capabilities, develop new tracer approaches to track the flow of carbon into biomolecules and cells, and advance our ability to measure the interrelationship between composition of carbonaceous materials and deep life. The DLC Steering community will encourage submission of ideas for modest short-term support that will address these and other relevant / meritorious efforts with high potential to attract new funding from other agencies and foundations. (see section A. 6.2.). • Investments in stable- and radioisotope probing experiments, for example in the context of the RHC project, to trace the utilization and assimilation of carbon compounds commonly found in subsurface environments (e.g. CO2, formate, acetate, methane) linked with single-cellgenomic approaches to query their genetic potential and diversity. 16 Deep Life Community - The Deep Carbon Observatory – 2014-2015 • Generation of biochemical profiles from novel microbial isolates obtained from subsurface ecosystems, including lipid biomarkers, sporulation-related molecules, and Raman spectroscopic signatures that can inform and expand upon geochemical studies [12]. • Improving life detection capabilities in order to differentiate between live and dead cells, to develop and implement molecular markers diagnostic of active cells, and to reduce detection limits of cellular and molecular life markers. Future initiatives targeting the biotic-abiotic interface (see below) will inherently depend on these improvements. • Advancing our quantitative and qualitative understanding of the relationship between deep life and cycling of carbon. For example, the impact of deep life on the fluxes and redox state of carbon entering subduction zones [13] as well as the influence of deep subseafloor life on oceanic carbon budgets are poorly constrained. Likewise, the biochemical mechanisms associated with the microbial utilization of kerogen, Earth’s largest organic carbon pool, are essentially unknown. We hypothesize that structural properties of kerogen strongly influence the release kinetics of metabolizable smaller carbon compounds and are thus intimately linked to the activity of deep heterotrophic life. The DLC will support field and experimental studies targeting the detection, biochemical mechanisms and kinetics of microbially-induced structural modification of organic matter in the subsurface using modern mass spectrometry, spectroscopy, NMR, rate assays, metabolomics and genomics approaches. As described above under 4 - THE PROPOSED PROJECT: DEEP LIFE RESEARCH AND COORDINATION NETWORK, initiating and developing these frontiers may require seed funding to capture preliminary data for new applications to foundations and agencies. Appendix A.6.2 STEERING COMMITTEE ACTIVITIES. Expanding Deep Life Support; for selection mechanism and criteria. 4.5 – STRATEGIC PARTNERSHIPS THAT BUILD THE DEEP LIFE COMMUNITY. The Integrated Ocean Drilling Program (IODP) and the International Continental Scientific Drilling Program 17 Deep Life Community - The Deep Carbon Observatory – 2014-2015 (ICDP) support expensive scientific field projects that drill into the subsurface. IODP and ICDP have sponsored drilling for biologically useful samples that led to important findings that appeared in high profile research papers. The path to funding drilling from these organizations is arduous and highly competitive. We envision multiple avenues for DCO to work with IODP and ICDP to foster further deep carbon-relevant studies that involve deep life. These range from small studies with short lead times to long-term development of a transformational combined marine/continental drilling projects that address DL Decadal Goals. Deep Life proposes to leverage IODP and ICDP funding in support of biological studies in three ways: • DLC support (up to $10K inclusive of travel, supplies e.g. drilling fluid tracers and lab expenses) for PIs and/or their students to join near term deep drilling projects (<1 year in the future) that have a high potential to generate biologically useful samples, but that currently have no participating biologists on board. This modest level of support will supplement existing dedicated funding opportunities on the national level for post-expedition research available in several countries (e.g., small science-directed grants from U.S. Science Support Program with the Consortium of Ocean Leadership) and could also allow support of DLC investigators to participate in non-IODP/non-ICDP deep drilling programs that access the subsurface (e.g., US Department of Energy related projects). • Workshops to develop new proposals. Deep Life will fund workshops either directly at ~$30k or with ~$15k to match IODP or ICDP funds, e.g. for so-called Magellan Workshops to develop deep biosphere-inspired drilling projects that often result in full drilling proposals to IODP or ICDP, or to initiate / strengthen the microbiological component of existing proposals previously designed from a geological perspective. These will be multidisciplinary but will have deep life and deep carbon as central themes. Smaller workshops will support participation of ~five DLC members to develop projects of opportunity in the framework of 18 Deep Life Community - The Deep Carbon Observatory – 2014-2015 IODP via so-called ancillary project letters (APL) that seek to take advantage of scheduled deep drilling expeditions with no obvious link to DL but which could be extended by a DLdedicated hole. We will also support joint workshops with the NASA Astrobiology Institute (NAI) and the Center for Dark Energy Biosphere Investigations (C-DEBI) to develop collaborative projects that span marine and continental subsurface systems. • We will also explore the concept of merging IODP and ICDP support for drilling an onshoreoffshore transect to address scientific questions that straddle continental marine boundaries. IODP and ICDP achieved this while coring the New Jersey Margin, [14] and scientific questions involving processing of carbon in the deep biosphere across marine/continental gradients could warrant such an ambitious drilling effort. As just one example, high-latitude sites where permafrost and methane hydrates sensitive to temperature change link the deep biosphere to the planet’s surface along a continental/marine transect [15]. The DLC will start this long-term (ca. 6-year) team-building, fund-raising effort by co-organizing a workshop and soliciting funding and participation from other DCO Communities, as well as IODP, ICDP, NAI, and C-DEBI to develop collaborative projects that span marine and continental subsurface systems. Products from the workshops include accelerated involvement of DLC researchers in ongoing expedition planning, alignment of expedition research questions with the DL Decadal goals, identification of essential scientific disciplines and key teams for addressing those questions, and a workable plan for funding a PI or small group of PIs (e.g., from NSF, the Moore Foundation, the Keck Foundation) to develop full proposals to IODP and ICDP and to advocate with these organizations and funding agencies for support. DLC will assign liaisons with IODP and ICDP who will maintain an active list of current projects from these two programs that could serve the DLC and vice versa. 19 Deep Life Community - The Deep Carbon Observatory – 2014-2015 • With participation of representatives from the DE and RF communities, the DLC will prepare and lead a deep ocean drilling proposal (submission of a pre-proposal is schedule for Oct 1, 2013) to utilize DV Chikyu for probing the presumed biotic/abiotic temperature interface in deeply buried, geothermally heated subseafloor sediments in the Shikoku Basin off Japan. At two sites that have been previously drilled during ODP Leg 190 (Sites 1173 and 1174), the critical temperature zone between 80 and 140°C could be probed over a several hundred meters at a total subseafloor depth of less than 1 km. Previous initial studies of deep life at these sites by John Parkes and colleagues [16, 17] have demonstrated overlapping zones of deep life and abiotic, thermal processes that supply substrates and thereby potentially stimulate microbial communities. Based on existing cellular count data [18] however, the subsurface strata in which sterilization is likely to occur has not been possible due to high detection limits on the order of around 105 cells/mL. This location provides an excellent opportunity to determine the temperature limit in a sedimentary subseafloor microbial ecosystem and characterize the zones where life goes extinct. A dedicated expedition with a science team dedicated to DL and DE objectives would benefit from the advances in deep life investigative strategies made during the last decade. Fundamental research goals could be accomplished at this location; these include “constraining the T-limit of life in the sedimenthosted subseafloor; studying the relationship of abiotic chemical reactions at high T and biology; the physical and chemical characterization of horizons where life becomes extinct. Constraining the deepest limits of life is one of the most fundamental questions for the DLC community (Decadal Goal 2). The discoveries resulting from this would be of great interest to geoscientists, life scientists and astrobiologists and the public. • Partnerships with private industry. Both the mining and oil and gas industries offer opportunities for exploring deep microbial life by providing access to sample cores, drilling 20 Deep Life Community - The Deep Carbon Observatory – 2014-2015 fluids, or resources for analysis. For example, S. D’Hondt and M. Sogin are working with ExxonMobil Upstream Research to develop an exploratory project to study microbial community structure in a wide range of subsurface and surface environments. The project will utilize sequencing technologies described for the CoDL to analyze samples from globally distributed reference sites collected by different IODP expeditions and oceanographic expeditions. ExxonMobil Upstream Research will provide $250,000 for a pilot project and if successful will significantly expand the project over the next decade. 4.6. - SUMMER SCHOOL DEEP MARINE BIOSPHERE. DL co-chair Kai-Uwe Hinrichs will organize the 2014 ECORD (European Consortium of Ocean Research Drilling” summer school “Subseafloor Biosphere: recent advances and future challenges” at MARUM, University of Bremen. This 8th annual ECORD summer school will provide a unique program of training for 30 international graduate students and postdocs working in the broader DLC. The program consists of one week of lectures by international experts, many of them directly associated with DLC, and a second week of “virtual ship” experience with hands-on laboratory work that includes methods such a microbial cell counts and sediment core descriptions, supervised by scientist and staff from the Bremen IODP core repository (leveraging: $32,000 in funds by ECORD and MARUM to support travel of international lecturers and the virtual ship experience. 4.7 - MBL INFORMATICS COURSE. M.L. Sogin will direct the course “Strategies and Techniques for Analyzing Microbial Population Structures”. This 10-day immersion course will focus on design and analysis of marker gene and shotgun metagenomic projects for advanced graduate students, postdoctoral fellows and established investigators who seek to leverage the power of massively-parallel DNA sequencing technologies for investigations of microbial communities from a range of environments that span the deep subsurface to the human microbiome. 21 Deep Life Community - The Deep Carbon Observatory – 2014-2015 4.8 - DATA SCIENCE. Mitch Sogin serves as the liaison to the Data Science Advisory Committee led by Peter Fox. The DLC stands ready to contribute meta data and information about samples and projects to the Data Science Group. The CoDL and associated genomic investigations require data capabilities that extend beyond the scope of the Data Science Group. Instead, the WEB site VAMPS (http://vamps.mbl.edu) provides an analytical platform for analysis and visualization of marker gene surveys. It offers a portal that links all metadata including detailed project descriptions to the large molecular datasets. It also provides functional ties to the QIIME analysis pipeline and soon will link into the Earth Microbiome Project through VAMPS and QIIME. Shotgun metagenomic data will be submitted by users for analysis on the MGRAST platform [19]. The DLC will follow a data release policy that requires release of data within 9 months of generation of immediate release of data that appears in published manuscripts. 4.9 – ENGAGEMENT. Steve D’Hondt serves as the Liaison to the Engagement team led by Sara Hickox, both at URI-GSO. He will convey information about scientific developments/successes, new collaborations, workshops, and DLC meetings to the Engagement team. Steve D’Hondt has led IODP expeditions and is co-PI on the C-DEBI Science and Technology Center. 5 - OUTPUT OF RESEARCH PROJECT • Expanded opportunities for students and postdocs to study deep life • Two intensive training courses (informatics and deep marine biosphere) • Expanding the international DL community to include the best deep life scientists • Development of novel methods for identifying biosignatures in the deep subsurface • Quantification of live vs. dead, active vs. inactive, and vegetative cells vs. spores • Document and characterize abiotically derived carbon assimilation in serpentinization environments in collaboration with the Deep Energy Directorate including methane isotopologue studies to discriminate biotic from abiotic methane in the deep subsurface • Document how deep subsurface life adapts and evolves through evolutionary genomics 22 Deep Life Community - The Deep Carbon Observatory – 2014-2015 • Inventory of organic C forms in select deep marine and continental sites • Identification of microbes metabolizing those organic C compounds • Extend the CoDL to include other sites and those under investigation by DCO communities. • Addition of a microbiological component to non-biologically oriented subsurface projects • New biology-driven IODP and ICDP deep scientific drilling projects • Will perform a cross project synthesis of life’s distribution in the deep subsurface • Publication of key scientific findings in open access journals 6 - BUDGET JUSTIFICATION AND LEVERAGING. In contrast to the DLC’s research activities enabled by generous funding from the Alfred P. Sloan Foundation, this proposal describes a strategy for securing resources necessary to realize Deep Life’s primary Decadal Goals within the next eight to ten years. It builds upon research success over the past two years and the growing interest in Deep Life. We describe a science plan that will require significantly greater resources – some of which are already in hand. The Deep Life has community has identified ~$26,000,000 to fund science that addresses some of our primary decadal goals (See Appendix A.3). The budget in this proposal will sustain CoDL activities, build the Deep Life community through a modest size annual meeting, fund Steering committee activities, support workshops and data synthesis groups, and participation in the 2015 DCO meeting. We have also reserved significant resources for to support travel, workshops, and small-scale science projects for Post Doctoral Students, Graduate Students and Investigators who seek to advance the decadal goals of the DLC. The BUDGET provides full details and Appendix A.6.2 STEERING COMMITTEE ACTIVITIES describes the rationale for allocating resources, the application mechanism, review criteria and the review process for the funding of DLC activities. 7 - PRIOR SLOAN SUPPORT: See - 3 – DLC RESEARCH ACCOMPLISHMENTS – 2012-2013. page 7 23 Deep Life Community - The Deep Carbon Observatory – 2014-2015 8 - BUDGET 9 - CURRICULUM VITAE Kai-Uwe Hinrichs Ph.D. – Co-PI. Dean and Professor, Department of Geosciences; Head, Organic Geochemistry Group (Hinrichs Lab), MARUM Center for Marine Environmental Sciences, University of Bremen, Germany Education and Training 1994 Diploma (equiv. to M.Sc.), Chemistry, Institute for Chemistry and Biology of the Marine Environment (ICBM), U Oldenburg, Germany. 1997 Ph.D., ICBM, U Oldenburg, Germany, thesis in Organic Geochemistry. Appointments 1994-1997 Research Assistant, ICBM, University of Oldenburg 1997-2000 Postdoctoral Investigator/Fellow, Dept. of Geology & Geophysics, WHOI 2000-2002 Assistant Scientist, tenure-track, Dept. of Geology & Geophysics, WHOI 2002-2004 Professor (C3, tenured), Dept. of Geosciences, U Bremen 2004-2010 Adjunct Scientist, Dept. of Geology & Geophysics, WHOI 2004-present Full Professor (W3 with tenure), Dept. of Geosciences, U Bremen Honors 2000 2009 2011 Fellow, Hanse Institute of Advanced Studies, Delmenhorst, Germany Advanced Grant by the European Research Council, 2.9 M€ Gottfried Wilhelm Leibniz Price (Germany’s most prestigious res. prize), 2.5M€ Representative Publications (senior/first author contributions only, out of currently 99 peer-reviewed) Hinrichs, KU, Hayes, JM, Sylva, SP, Brewer, PG, and DeLong, EF Methane-consuming archaebacteria in marine sediments. Nature, 398, 802-805. (1999) Hinrichs KU, Hmelo LR, and Sylva, SP Molecular fossil record of elevated methane levels in late Pleistocene coastal waters, Science, 299, 1214-1217. (2003) Sturt, HF, Summons, RE, Smith, KJ, Elvert, M, and Hinrichs, KU Intact polar membrane lipids in prokaryotes and sediments deciphered by ESI-HPLC-MSn – new biomarkers for biogeochemistry and microbial ecology. Rap Comm Mass Spectr, 18, 617-628. (2004) Hinrichs, KU, Hayes, JM, Bach, W, Spivack, A, Hmelo, LR, Holm, N, Johnson, CG, Sylva, SP Biological formation of ethane and propane in the deep marine subsurface. PNAS, 103, 1468414689. (2006) Biddle, JF, Lipp, JS, Lever, M, Lloyd, K, Sørensen, K, Anderson, R, Fredricks, HF, Elvert, M, Kelly, TJ, Schrag, DP, Sogin, ML, Brenchley, JE, Teske, A, House, CH, Hinrichs, KU Heterotrophic Archaea dominate sedimentary subsurface ecosystems off Peru. PNAS, 103, 38463851. (2006) Lipp, JS, Morono, Y, Inagaki, F, Hinrichs, KU Significant contribution of Archaea to extant biomass in marine subsurface sediments. Nature, 454, 991-994. (2008). 24 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Heuer, VB, Pohlman, JW, Torres, ME, Elvert, M, Hinrichs, KU The stable carbon isotope biogeochemistry of acetate and other dissolved carbon species in deep sub-seafloor sediments at the northern Cascadia Margin. Geochim Cosmochim Acta, 73, 3323-3336. (2009) Schubotz, F, Wakeham, SG, Lipp, JS, Fredricks, HF, Hinrichs, KU Detection of microbial biomass by intact membrane lipid analysis in the water column and surface sediments of the Black Sea, Env Microbiol, 11, 2720-2734. (2009) Sepúlveda, JC, Wendler, J, Summons, RE, Hinrichs, KU Rapid resurgence of marine productivity at the Cretaceous-Paleogene mass extinction event, Science, 326, 129-132. (2009) Schubotz, F, Lipp, JS, Elvert, M, Hinrichs, KU, Stable carbon isotopic compositions of intact polar lipids reveal complex carbon flow patterns among hydrocarbon degrading microbial communities at the Chapopote asphalt volcano, Geochim Cosmochim Acta, 75, 4399-4415. (2011) Schmidt, F, Koch, B, Elvert, M, Schmidt, G, Witt, M, Hinrichs, KU Diagenetic transformation of dissolved organic nitrogen compounds under contrasting sedimentary redox conditions in the Black Sea, Env Sci Tech, 45, 5223-5229. (2011) Rossel, PE, Elvert, M, Ramette, A, Boetius, A, Hinrichs, KU Factors controlling the distribution of anaerobic methanotrophic communities in marine environments: evidence from intact polar membrane lipids, Geochim Cosmochim Acta, 75, 164-184. (2011) Kellermann, MY, Wegener, G, Elvert, M, Yoshinaga, MY, Lin, YS, Holler, T, Mollar, XP, Knittel, K, Hinrichs, KU Autotrophy as predominant mode of carbon fixation in thermophilic anaerobic methane-oxidizing microbial communities, PNAS, 109, 19321-19326. (2012) Liu, XL, Lipp, JS, Simpson, JS, Lin, YS, Summons, RE, Hinrichs, KU Mono- and dihydroxyl glycerol gibiphytanyl glycerol tetraethers in marine sediments: identification of both core and intact polar lipid forms, Geochim Cosmochim Acta, 89, 102-115. (2012) Lin, YS, Lipp, JS, Elvert, M, Holler, T, Hinrichs, KU Assessing production of the ubiquitous archaeal diglycosyl tetraether lipids in marine subsurface sediment using intramolecular stable isotope probing, Environ Microbiol, 15, 1634-1646. (2013) Lever, MA, Rouxel, O, Alt, JC, Shimizu, N, Ono, S, Coggon, RM, Shanks, WC, Lapham, L, Elvert, M, Prieto-Mollar, X, Hinrichs, KU, Inagaki, F, Teske, A Evidence for microbial carbon and sulfur cycling in deeply buried ridge flank basalt, Science, 339, 1305-1308. (2013) Xie, S, Lipp, JS, Wegener, G, Ferdelman, TG, Hinrichs, KU, Turnover of microbial lipids in the deep biosphere and growth of benthic archaeal populations, PNAS, 110, 6010-6014. (2013) Mitchell L. Sogin, Ph.D Co-PI. Director, Bay Paul Center, Marine Biological Laboratory, Woods Hole, MA Education and Training 1967 University of Illinois, Urbana B.S. Chemistry and Microbiology 1969 University of Illinois, Urbana M.S. Industrial Microbiology 1972 University of Illinois, Urbana Ph.D., Microbiology & Molecular Biology 25 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Appointments 1972-1976 National Jewish Center, Denver, CO. Postdoctoral NIH Fellowship 1976-1989 Senior Staff Scientist, National Jewish Center, Denver, CO. 1980-1986 Assist. Professor, University of Colorado Health Sciences Center 1987-1989 Assoc. Professor, University of Colorado Health Sciences Center 1986-1999 Associate Fellow, Canadian Institute for Advanced Research 1997-1998 Visiting Miller Research Professor, UC Berkeley 1989-present Senior Scientist, Marine Biological Laboratory, Woods Hole, MA 1996-present Director of Josephine Bay Paul Center for Comparative Molecular Biology and Evolution at the MBL. 2004-present Professor (MBL), Department of Molecular Biology, Cell Biology and Biochemistry, Brown University, Providence, RI Honors 1992 1993 1995 9/96-present 1998-present 1998-present 2007 Division Lecturer - American Society for Microbiology Stoll Stunkard Award - American Society of Parasitologists Elected Chairman - Division R, American Society of Microbiologists Fellow of the American Academy of Microbiology Fellow of the American Academy of Arts and Sciences Fellow of the American Association for the Advancement of Science American Society for Microbiology – Roger Porter Award. Representative Publications (230 TOTAL) Biddle, J.F., J.S. Lipp, M.A. Lever, K.G. Lloyd, K.B. Sørensen, R. Anderson, H.F. Fredricks, M. Elvert, T.J. Kelly, D.P. Schrag, M.L. Sogin, J.E. Brenchley, A. Teske, C.H. House and K.-U. Hinrichs. Heterotrophic Archaea dominate sedimentary subsurface ecosystems off Peru. Proc. Natl. Acad. Sci. USA 103(10): 3846-3851 (2006). Sogin, M.L., H.G. Morrison, J.A. Huber, D. Mark Welch, S.M. Huse, P.R. Neal, J.M. Arrieta, and G.J. Herndl. Microbial diversity in the deep sea and the under-explore "rare biosphere". Proc. Natl. Acad. Sci. USA 103(32): 12115-12120 (2006) Morrison, H.G., A.G. McArthur, F.D. Gillin, S.B. Aley, R.D. Adam, G.J. Olsen, A.A. Best, Z. Cande, F. Chen, M.J. Cipriano, B.J. Davids, S.C. Dawson, H.G. Elmendorf, A.B. Hehl, M.E. Holder, S.M. Huse, U.U. Kim, E. Lasek-Nesselquist, G. Manning, A. Nigam, J.E. Nixon, D. Palm, N.E. Passamaneck, A. Prabhu, C.I. Reich, D.S. Reiner, J. Samuelson, S. G.Svard, M.L.Sogin. Genomic Minimalism in the Early Diverging Intestinal Parasite Giardia lamblia. SCIENCE 317:1921-1926. (2007) Huber, J.A., D.M. Welch, H.G. Morrison, S.M. Huse, P.R. Neal, D.A. Butterfield and M.L. Sogin. Microbial Population Structures in the Deep Marine Biosphere. SCIENCE 318:97-100 (2007) Santelli, C.A., B.N. Orcutt, E. Banning, W. Bach, C.L. Moyer, M.L. Sogin, K. J. Edwards. Abundance and diversity of microbial life in the ocean crust, Nature 453: 653-656 (2008). Huber, J.A., H.G. Morrison, S.M. Huse, P.R. Neal, M.L. Sogin, and D.B. Mark Welch. Effect of PCR amplicon size on assessments of clone library microbial diversity and community structure. Environmental Microbiology 11(5): 1292-1302 (2009). Bodaker, I, I. Sharon, M.T. Suzuki, R. Feingersch, M. Shmoish, E. Andreishcheva, M.L. Sogin, 26 Deep Life Community - The Deep Carbon Observatory – 2014-2015 M. Rosenberg, M.E. Maguire, S. Belkin, A. Oren, O. Be´ja. Comparative community genomics in the Dead Sea: an increasingly extreme environment. ISME J. 4: 399-407 (2009) Brazelton, W.J., M.L. Sogin, and J.A. Baross, Multiple scales of diversification within a single population of archaea in a hydrothermal vent biofilm. Environmental Microbiology, 2(2): 236242 (2010). Hasegawa, Y., J.L. Mark Welch, A.Valm, C. Rieken, M.L. Sogin, and G. Borisy. Imaging marine bacteria with unique 16S rRNA V6 sequences by fluorescence in situ hybridization and spectral analysis. 27(3): 251-260 (2010) Brazelton, W.J., K.A. Ludwig, M.L. Sogin, E.N. Andreishcheva, D.S. Kelley, C. Shen, R.L. Edwards, J.A. Baross. Archaea and bacteria with surprising microdiversity show shifts in dominance over 1000-year time scales in hydrothermal chimneys. Proc. Natl. Acad. Sci. USA, 107: 1612-1617 (2010). Huse, S.M., D. Mark Welch, H. G. Morrison, M.L. Sogin, Ironing Out the Wrinkles in the Rare Biosphere. Environmental Microbiology 12(7): 1889-1898 (2010). Huber, J., Cantin, H., Huse, S., Mark Welch, D., Sogin, M.L., Butterfield, D. Isolated communities of Epsilon-proteobacteria in hydrothermal vent fluids of the Mariana Arc seamounts. FEMS Microbiology Ecology, 73:538-549 (2010). Zinger L, L.A. Amaral-Zettler, J.A. Fuhrman, M.C. Horner-Devine, S.M. Huse, D.B. Mark Welch, J.B.H. Martiny, M.L. Sogin, A. Boetius, A. Ramette. Global Patterns of Bacterial BetaDiversity in Seafloor and Seawater Ecosystems PLoS ONE 6(9): e24570. doi:10.1371/journal.pone.0024570 (2011). Gobet, A., S.I. Bo ̈er, S.M. Huse, J.E.E. van Beusekom, C. Quince, M.L. Sogin, A. Boetius A., Ramette, A. Diversity and dynamics of rare and of resident bacterial populations in coastal sands. ISME Journal, 6:542-553 (2012). Siam, R., G.A. Mustafa, H. Sharaf, A. Moustafa, A. Ramadan, A. Antunes, V.B. Bajic, U. Stingl, N.G.R. Marsis, M.J.L. Coolen, M.L. Sogin, A.J. Ferreira, H. El-Dorry. Unique Prokaryotic Consortia in Geochemically Distinct Sediments from Red Sea Atlantis II and Discovery Deep Brine Pools. PLOS One. 7(8), e42872 (2012). Amend, A.S., T.A. Oliver, L.A. Amaral-Zettler, A. Boetius, J.A. Fuhrman, M. Claire HornerDevine, S.M. Huse, D.B. Mark Welch, A.C. Martiny, A. Ramette, L. Zinger, M.L. Sogin, J.B.H. Martiny.. Macroecological patterns of marine bacteria on a global scale. Journal of Biogeography. DOI: 10.1111/jbi.12034 (2012). Sul, W.J., T.A. Oliver, H.W. Ducklow, L.A. Amaral-Zettler, M.L. Sogin. Marine bacteria exhibit a bipolar distribution. PNAS. 110(6) 2342-2347 (2013). Eren, A. M. H.G. Morrison, S.M. Huse and M.L. Sogin, DRISEE Overestimates Errors in Metagenomic Sequencing Data, Briefings in Bioinformatics. In press (2013). Eren, A.M., J.H. Vineis, H.G. Morrison and M.L. Sogin. A Filtering Method to Generate High Quality Short Reads Using Illumina Paired-End Technology. PLOS ONE. In press (2013) Douglas H. Bartlett Ph.D. Co-I. 27 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Professor, Scripps Institution of Oceanography, La Jolla, CA Education and Training 1979 Valparaiso University, Biology B.S. 1985 University of Illinois, Molecular Biology Ph.D. Appointments 1985-1987 The Agouron Institute, Postdoctoral Scholar 1987-1989 The Agouron Institute, Research Scientist 1989-1995 Scripps Institution of Oceanography, Assistant Professor 1995-2001 Scripps Institution of Oceanography, Associate Professor 2001-Present Scripps Institution of Oceanography, Professor Representative Publications Vezzi, A., Campanaro, S., D’Angelo, M., Simonato, F., Vitulo, N., Lauro, F. M., Cestaro, A., Malacrida, G., Simionati, B., Cannata, N., Romualdi, C., Bartlett D. H., and Valle, G. Life at depth: Photobacterium profundum genome sequence and expression analysis. Science 307:14591463. 2005. Lauro, F. M., Tran, K. , Vezzi, A., Vitulo, N., Valle, G. , Bartlett, D. H. Large-scale transposon mutagenesis of Photobacterium profundum SS9 reveals new genetic loci important for growth at low temperature and high pressure. J. Bacteriol. 190:1699-1709. 2008. Nagata, T., Tamburini, C., Arístegui, J., Baltar, F., Bochdansky, A., Fonda-Umani, S., Fukuda, H., Gogou, A., Hansell, D. A., Hansman, R. J., Herndl, G. J., Panagiotopoulos, C., Reinthaler, T., Sohrin, R., Verdugo, P., Yamada, N., Yamashita, Y., Yokokawa, T., Bartlett, D. H. Emerging concepts on microbial processes in the bathypelagic ocean–ecology, biogeochemistry, and genomics. Deep-Sea Research II 57: 1519-1536. 2010. Yoshioka, H., Maruyama, A., Nakamura, T., Higashi, Y., Fuse, H., Sakata, S., Bartlett, D. H. Activities and distribution of methanogenic and methane-oxidizing microbes in marine sediments from the Cascadia Margin. Geobiology 8:223-233. 2010. Eloe, E. A., Malfatti, F., Gutierrez, J., Hardy, K., Schmidt, W. E., Pogliano, K., Pogliano, J., Azam, F. and D. H. Bartlett.. Isolation and characterization of the first psychropiezophilic Alphaproteobacterium. Appl. Environ. Microbiol. 77:8145- 8153. 2011 Oger, P., Sokolova, T. G., Kozhevnikova, D. A., Chernyh, N. A., Bartlett, D. H., BonchOsmolovskaya, E. A., Lebedinsky, A. V. Complete genome sequence of the hyperthermophilic archaeon Thermococcus sp. AM4 capable of organotrophic growth and growth at the expense of hydrogenogenic or sulfidogenic oxidation of carbon monoxide. J. Bacteriol. 193:7019-7020. 2011. Eloe, E. A., Fadrish, D. W., Novotny, M., Zeigler Allen, L., Kim, M., Lombardo, M.- J., YeeGreenbaum, J., Yooseph, S., Allen, E. A., Lasken, R., Williamson, S. J., Bartlett, D. H. Going deeper: metagenome of a hadopelagic microbial community. PLoS ONE. 6: e20388. 2011. Eloe, E., Shulse, C. N., Fadrosh,D. W., Williamson, S. J., Allen, E. E. and Bartlett, D. H. Compositional differences in particle-associated and free-living microbial assemblages from an extreme deep-ocean environment. Environ. Microbiol. Reports. 3: 449–458. 2011. Campanaro, S, DePascale, F., Telatin, A., Schiavon, R., Bartlett, D. H. and Valle, G. The transcriptional landscape of the deep-sea bacterium Photobacterium profundum in both a toxR 28 Deep Life Community - The Deep Carbon Observatory – 2014-2015 mutant and its parental strain. BMC Genomics 13: article 5672012. Filip Meersman, Isabelle Daniel, Douglas H. Bartlett, Roland Winter, Rachael Hazael and Paul F. McMillan. High-pressure biochemistry and biophysics. In Deep Carbon (R. M. Hazen, ed.). Mineralogical Society of America and The Geochemical Society (Chantilly VA), pp. 607-648. 2013. Antje Boetius Ph.D. Co-I. Head of the Helmholtz - Max Planck Research Group on Deep Sea Ecology and Technology, Bremerhaven/Bremen, Germany Education and Training 1992 Diploma in Biology, Hamburg University, Germany 1996 Ph.D., Bremen University, Germany 2001 Professor for Microbiology, Jacobs Univ. Bremen, Germany Appointments 2008-present Leader HGF MPG Research Group, Alfred Wegener Institute Helmholtz Centre for Polar and Marine Research 2008-present Full Professor of Geomicrobiology at Bremen University 2003-present Group leader, Microbial Habitat Group, Max Planck Institute for Marine Microbiology 2008 Full Professor of Microbiology at Jacobs University Bremen 2003-2007 Associate Professor of Microbiology at Jacobs University Bremen 2001-2003 Assistant Professor of Microbiology at Intenational University Bremen, 1999-2000 Postdoc at Max Planck Institute for Marine Microbiology 1996-1999 Postdoc at Institute for Baltic Sea Research, Warnemünde 1993-1996 Research Assistant (PhD) at AWI, Bremerhaven 1989-1990 Laboratory Assistant at Scripps Institution of Oceanography, La Jolla, USA Honors 2012 2011 2010 2009 2009 2006 2004 Heinrich-Hertz Professor of the KIT Member of the Academy of Sciences and Literature Mainz External Scientific Member of the Max Planck Society Member of the National Academy of Sciences (Leopoldina) Gottfried-Wilhelm-Leibniz Price of the DFG Medaille de la Societe d’Oceanographie de France Guest Professor of the University Paris 6 (UPMC) Representative Publications (163 total) Treude T, Knittel K, Blumenberg M, Seifert R, Boetius A., Subsurface microbial methanotrophic mats in the Black Sea. AEM 71: 6375-6378. (2005) Inagaki F, Kuypers MM, Tsunogai U, Ishibashi J, Nakamura K, Treude T, Ohkubo S, Nakaseama M, Gena K, Chiba H, Hirayama H, Nunoura T, Takai K, Jørgensen BB, Horikoshi K, Boetius A Microbial community in a sediment-hosted CO2 lake of the southern Okinawa Trough hydrothermal system. PNAS 103 (38), 13899-13900 (2006) 29 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Niemann H, Lösekann T, de Beer D, Elvert M, Nadalig T, Knittel K, Amann R, Sauter EJ, Schlüter M, Klages M, Foucher JP, Boetius A., Novel microbial communities of the Haakon Mosby mud volcano and their role as methane sink. Nature, (443) 854-858. (2006) Nauhaus K., Albrecht M., Elvert M., Boetius A., Widdel F., In Vitro cell growth of marine archaeal-bacterial consortia by anaerobic oxidation of methane with sulphate. Environmental Microbiology, 9(1), 187–196. (2007) Jørgensen BB, Boetius A., Feast and famine – microbial life in the deep-sea bed. Nature Microbiology Reviews, 5, 770-781. (2007) Omoregie EO, Mastalerz V, de Lange G, Straub KL, Kappler A, Røy H, Stadnitskaia A, Foucher JP, Boetius AB., Biogeochemistry and Community Composition of Iron- and SulfurPrecipitating Microbial Mats at the Chefren Mud Volcano (Nile Deep Sea Fan, Eastern Mediterranean). Appl Environ Microbiol 74 (10): 3198–3215. (2008). Wegener, G; Boetius, A., An experimental study on short-term changes in the anaerobic oxidation of methane in response to varying methane and sulfate fluxes. BIOGEOSCIENCES 6 (5): 867-876 (2009). Omoregie E. O., Niemann, H., Mastalerz, V., de Lange, G., Stadnitskaia, A., Mascle, J., Foucher , J.P. Boetius, A., Microbial methane oxidation and sulfate reduction at cold seeps of the deep Eastern Mediterranean Sea. Marine Geology. 261:114-127. (2009). Felden J, Wenzhöfer F, Feseker T, Boetius A., Transport and consumption of oxygen and methane in different habitats of the Håkon Mosby Mud Volcano. Limnology and Oceanography 55(6), 2010, 2366–2380. (2010). Wei C-L, Rowe GT, Escobar-Briones E, Boetius A, Soltwedel T, et al. Global Patterns and Predictions of Seafloor Biomass Using Random Forests. PLoS ONE 5(12): e15323. doi:10.1371/journal.pone.0015323. (2010). Holler T., Widdel F., Knittel K., Amann R., Kellermann M., Hinrichs K.U., Teske A., Boetius A., Wegener G., Thermophilic anaerobic oxidation of methane by marine microbial consortia. ISME doi:10.1038/ismej.2011.77. (2011). Zinger L, Amaral-Zettler LA, Fuhrman JA, Horner-Devine MC, Huse SM, Mark Welch DB, Martiny JBH, Sogin M, Boetius A, Ramette A., Global patterns of bacterial beta-diversity in seafloor and seawater ecosystems. PLoS ONE 6(9): e24570. (2011). Holler, T; Wegener, G; Niemann, H; Deusner, C; Ferdelman, TG; Boetius, A; Brunner, B; Widdel, F., Carbon and sulfur back flux during anaerobic microbial oxidation of methane and coupled sulfate reduction. PNAS 108:52 E1484-E1490 DOI: 10.1073/pnas.1106032108. (2011). Wegener G, Bausch M, Holler T, Thang NM, Prieto Mollar X, Kellermann MY, Hinrichs KU, Boetius A., Assessing sub-seafloor microbial activity by combined stable isotope probing with deuterated water and 13C-bicarbonate. Environmental Microbiology, DOI: 10.1111/j.14622920.2012.02739.x. (2012). Felden, J., Lichtschlag, A., Wenzhöfer, A., de Beer, F., Feseker, D., Pop Ristova, T., P., de Lange, G., Boetius, A. Limitations of microbial hydrocarbon degradation at the Amon Mud Volcano (Nile Deep Sea Fan). Biogeosciences 10, 3269–3283. (2013). 30 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Frederick S. Colwell, Ph.D. Co-I. Professor, Oregon State University Education and Training 1977 B.A., Biology, Whitman College, Walla Walla, WA 1982 M.S., Microbiology, Northern Arizona University, Flagstaff, AZ 1986 Ph.D., Microbiology, Virginia Tech, Blacksburg, VA Appointments 1986-1988 Postdoctoral Fellow, Biotechnol. Dept., INL 1988-1990 Scientist, Biotechnol. Dept., INL 1990-1992 Senior Scientist, Biotechnol. Dept., INL 1992-1994 Scientific Specialist, Biotechnol. Dept., INL 1994-1998 Advisory Scientist, Biotechnol. Dept., INL 1998-2006 Consulting Scientist, Biotechnol. Dept., Idaho National Laboratory (INL) 2006-present Professor, College of Oceanic & Atmos. Sci., Oregon State Univ. Representative Publications Reed, D.W., Y. Fujita, M. Delwiche, D.B. Blackwelder, P.P. Sheridan, T. Uchida, and F. Colwell. Microbial communities from methane hydrate-bearing deep marine sediments in a forearc basin. Appl. Environ. Microbiol. 68: 3759-3770. 2002. Mikucki, J.A., Y. Liu, M. Delwiche, F.S. Colwell, and D.R. Boone. Isolation of a methanogen from deep marine sediments that contain methane hydrates, and description of Methanoculleus submarinus sp. nov. Appl. Environ. Microbiol. 69: 3311-3316. 2003. Colwell, F., R. Matsumoto, and D.W. Reed. A review of the gas hydrates, geology, and biology of the Nankai Trough. Chem. Geol. 205: 391-404. 2004. Inagaki, F. T. Nunoura, S. Nakagawa, A Teske, M. Lever, A. Lauer, M. Suzuki, K. Takai, M. Delwiche, F.S. Colwell, K.H. Nealson, K. Horikoshi, S. D’Hondt, and B.B. Jørgensen. Biogeographical distribution and diversity of microbes in methane hydrate-bearing deep marine sediments on the Pacific Ocean Margin. Proc. Nat. Acad. Sci. USA. 103: 2815-2820. 2006. Colwell, F.S., S. Boyd, M.E. Delwiche, D.W. Reed, T.J. Phelps, and D.T. Newby. Estimates of biogenic methane production rates in deep marine sediments at Hydrate Ridge, Cascadia Margin. Appl. Environ. Microbiol. 74: 3444-3452. 2008. Caldwell, S.L., J.R. Laidler, E.A. Brewer, J.O. Eberly, S.C. Sandborgh, and F.S. Colwell. Anaerobic oxidation of methane: Mechanisms, bioenergetics, and the ecology of associated microorganisms. Environ. Sci. Technol. 42: 6791-6799. 2008. Briggs, B.R., J. Pohlman, M. Torres, M. Riedel, E. Brodie, and F.S. Colwell. Macroscopic biofilms of the anaerobic oxidation of methane consortia in subseafloor sediment fractures. Appl. Environ. Microbiol. 77: 6780-6787. 2011. Gu, G., G.R. Dickens, G. Bhatnagar, F. Colwell, G. Hirasaki, and W.G. Chapman. Abundant early Palaeogene marine gas hydrates despite warm deep ocean temperatures. Nature – Geoscience. DOI: 10.1038/NGEO1301. 2011. Briggs, B.R., F. Inagaki, Y. Morono, T. Futagami, C. Huguet, A. Rosell-Mele, T. Lorenson, and 31 Deep Life Community - The Deep Carbon Observatory – 2014-2015 F.S. Colwell. Bacterial dominance in subseafloor sediments characterized by gas hydrates. FEMS Microbiol. Ecol. 81: 88-98. 2012. Edwards, K.J., K. Becker, and F. Colwell. The deep, dark energy biosphere: Intraterrestrial life on Earth. Ann. Rev. Earth Planet. Sci. 40: 551-568. 2012. Isabelle Daniel, Ph.D Co-PI. Professor University Claude Bernard Lyon 1 Education and Training 1991 Teaching degree (Agrégation) in Earth and life Sciences 1992 Master in Earth Sciences (ENS Lyon, University of Rennes 1) 1995 Ph.D. Geology, University Lyon 1 2002 Habilitation, University Lyon 1 Appointments 1996-2004 Assistant professor, University Lyon 1, Dept of Earth Sciences 2004-2010 Professor of Mineralogy, University Lyon 1, Dpt of Earth Science 2010–2013 Chair of the Department of Earth Sciences, University Lyon 1 Honors 2008-2010 Junior fellow of the Institut Universitaire de France Fellow of the Mineralogical Society of America Representative Publications (57 TOTAL) Auzende, A.L., I. Daniel, C. Lemaire, B. Reynard, F. Guyot High-pressure behavior of serpentine minerals: a Raman spectroscopic study, Physics Chemistry of Minerals, 31, 269-277. (2004) Perrillat, J.P., I. Daniel, K.T. Koga, B. Reynard, H. Cardon, W.A. Crichton (2005) Kinetics of antigorite dehydration: a real-time X-ray diffraction study, Earth and Planetary Science Letters 236:899-913. (2005) Daniel, I., Oger, P.M. and Winter, R. Origins of life and biochemistry under high-pressure conditions. Chemical Society Reviews, 35, 858-875. (2006) Hilairet, N., B. Reynard, Y. Wang, I. Daniel, S. Merkel, N. Nishiyama, S. Petitgirard Highpressure creep of serpentine, interseismic deformation and initiation of subduction, Science, 318, 1910-1913. (2007) Picard, A., I. Daniel, G. Montagnac, P.M. Oger In situ monitoring by quantitative Raman spectroscopy of alcoholic fermentation by Saccharomyces cerevisiae under high pressure, Extremophiles, 11, 445-452. (2007) Chollet, M., Daniel, I., Koga, K.T., Petitgirard, S., and Morard, G. Kinetics and mechanism of antigorite dehydration: implications for subduction zone seismicity. Journal of Geophysical Research, 116, B04203. (2011) Picard, A., I. Daniel et al., Monitoring microbial redox transformations of metal and metalloid elements under high pressure using in situ X-ray absorption spectroscopy. Geobiology, 9, doi: 10.1111/j.1472-4669.2010.00270.x. (2011) 32 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Feuillie, C., Merheb, M., Gillet, B., Montagnac, G., Hänni, C., and Daniel, I. Enzyme-free detection and quantification of double-stranded nucleic acids. Analytical and Bioanalytical Chemistry, 404(2), 415-422. (2012) Picard, A., Testemale, D., Hazemann, J.-L., and Daniel, I. The influence of high hydrostatic pressure on bacterial dissimilatory iron reduction. Geochimica Cosmochimica Acta, 88, 120– 129. (2012) Feuillie, C., Daniel, I., Michot, L., and Pedreira Segade, U. Adsorption of ribonucleotides to FeMg-Al rich swelling clays. Geochimica Cosmochimica Acta, (2013) in press. Steven D’Hondt, Ph.D. Co-I. Professor, Graduate School of Oceanography (GSO), University of Rhode Island (URI), Narragansett, RI Education and Training 1984 Stanford University, Geology, B.S. 1990 Princeton University, Geological and Geophysical Sciences, Ph.D. Appointments 1989-1995 Assistant Professor, URI, GSO 1995-2000 Associate Professor, URI, GSO 2011-2012 Interim Dean, URI, GSO 2000-present Professor, University of Rhode Island, Graduate School of Oceanography Representative Publications (10 examples of ~80). D'Hondt, S., P. Donaghay, J.C. Zachos, D. Luttenberg, and M. Lindinger, Organic carbon fluxes and ecological recovery from the Cretaceous/Tertiary mass extinction, Science 282, 276-279. 1998. Rutherford, S.D., S. D'Hondt, and W. Prell, Environmental controls on the geographic distribution of zooplankton diversity, Nature 400, 749-753. 1999. D'Hondt, S., Rutherford, S., Spivack, A.J. Metabolic activity of the subsurface biosphere in deep-sea sediments, Science 295: 2067-2070. 2002. D’Hondt, S, Jørgensen, B.B., Miller, D.J., and 32 others. Distributions of microbial activities in deep subseafloor sediments, Science 306: 2216-2221. 2004. Jørgensen, B.B., and S. D’Hondt, A starving majority deep beneath the seafloor, Science 314, 932-934. 2006. D’Hondt, S., and 11 others. Subseafloor sedimentary life in the South Pacific gyre, PNAS 106(28): 11651-11656. 2009. D’Hondt, S., F. Inagaki, C.A. Alvarez Zarikian, and the Expedition 329 Scientists, South Pacific Gyre Subseafloor Life, Proceedings IODP, 329: Tokyo (Integrated Ocean Drilling Program Management International, Inc.). doi:10.2204/ iodp.proc.329.2011. 2011. Kallmeyer, J., R. Pockalny, R. Adhikari, D.C. Smith and S. D’Hondt, Global distribution of subseafloor sedimentary biomass, Proceedings of the National Academy of Science (PNAS) 109(40), 16213-16216. 2012. 33 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Røy, H., J. Kallmeyer, R.R. Adhikari, R. Pockalny, B.B Jørgensen and S. D’Hondt, Aerobic microbial respiration in 86-million-year-old deep-sea red clay, Science 336 (6083), 922-925, DOI: 10.1126/science.1219424. 2012. Lomstein, B.A., A.T. Langerhuus, S. D’Hondt, B.B. Jørgensen and A.J. Spivack, Spore abundance, microbial growth and necromass turnover in deep subseafloor sediment, Nature 484, 101–104, doi:10.1038/nature10905. 2012. Thomas L. Kieft Ph.D. Co-I. Department of Biology, New Mexico Institute of Mining and Technology (New Mexico Tech) / Socorro, New Mexico Education and Training 1973 Carleton College, B.A. Biology 1978 New Mexico Highlands University M.S. Biology 1983 University of New Mexico, Ph.D. Biology Appointments 09/83 - 08/85 01/97 - 12/97 01/05 - 07/05 08/05 - present 08/85 - present Post-doc, Plant and Soil Biology, Univ. of California, Berkeley, CA Sabbatical leave, Northwest National Laboratory, Richland, WA Sabbatical leave, Los Alamos Nat’l Lab, Los Alamos, NM Adjunct Professor, Hydrology Program, New Mexico Tech Faculty member (Professor, 1993 - present), Biol. Dept., NM Tech Representative Publications Kieft, T.L., S.M. McCuddy, T.C. Onstott, M. Davidson, L.-H. Lin, B. Mislowac, L. Pratt, E. Boice, B. Sherwood Lollar, J. Lippmann-Pipke, S.M. Pfiffner, T.J. Phelps, T. Gihring, D. Moser, A. van Heerden. Geochemically generated, energy-rich substrates and indigenous microorganisms in deep, ancient groundwater. Geomicrobiol. J. 22:325-335. 2005. Gihring, T., D.P. Moser, L.-H. Lin, M. Davidson. T.C. Onstott, L. Morgan, M. Milleson, T. L. Kieft, E. Trimarco, D.L. Balkwill, M.E. Dollhopf. The distribution of microbial taxa in the subsurface water of the Kalahari Shield, South Africa. Geomicrobiol. J. 23:415-430. 2006. Kieft, T. L., Phelps, T. J., J. K. Fredrickson. Drilling, coring, and sampling subsurface environments. pp. 799-817, In: Manual of Environmental Microbiology, Third Edition. Hurst, C.J. (Ed.), ASM Press, Washington, DC. 2007. Onstott, T.C., L.-H. Lin, M. Davidson, B. Mislowac, M. Borcsik, J. Hall, G. Slater, J. Ward, B. Sherwood Lollar, J. Lippmann-Pipke, E. Boice, L. Pratt, B. S. Pfiffner, D. Moser, T. Gihring, T. L. Kieft, T. J. Phelps, E. van Heerden, D. Litthauer, M. DeFlaun, and R. Rothmel. The origin and age of biogeochemical trends in deep fracture water of the Witwatersrand basin, South Africa. Geomicrobiol. J. 23:369-414. 2006. Sahl, J.W., R. Schmidt, E.D. Swanner, K.W. Mandernack, A.S. Templeton, T.L. Kieft, R.L. Smith, W.E. Sanford, R.L. Callaghan, J.B. Mitton, and J.R. Spear. Subsurface microbial diversity in deep-granitic fracture water in Colorado. Appl. Environ. Microbiol. 74:143-152. 2008. Kminek, G., J.D. Rummel, C.S. Cockell , R. Atlas, N. Barlow, D. Beaty, W. Boynton, M. Carr, S. Clifford, C.A. Conley, A.F. Davila, A. Debus, P. Doran, M. Hecht, J. Heldmann, J. Helbert, 34 Deep Life Community - The Deep Carbon Observatory – 2014-2015 V. Hipkin, G. Horneck, T.L. Kieft, G. Klingelhoefer, M. Meyer, H. Newsom, G.G. Ori, J. Parnell, D. Prieur, F. Raulin, D. Schulze-Makuch, J.A. Spry, P.E. Stabekis, E. Stackebrandt, J. Vago, M. Viso, M. Voytek, L. Wells, F. Westall. Report of the COSPAR mars special regions colloquium. Adv. Space Res. 46:811-829. 2010. Tang, H., P. Zhang, T.L. Kieft, S.J. Ryan, S.M. Baker, W.P. Wiesmann, S. Rogelj Antibacterial action of a novel functionalized chitosan-arginine against gram-negative bacteria. Acta Biomaterialia 6:2562-2571. 2010. Silver, B.J., T.C. Onstott, G. Rose, L.-H., Lin, C. Ralston, B. Sherwood-Lollar, S. M. Pfiffner, T. L. Kieft, S. McCuddy. 2010. In situ cultivation of subsurface microorganisms in a deep mafic sill: implications for SLiMEs. Geomicrobiol. J. 27:329-348. 2010. Kieft, T.L. Sampling the Deep Sub-surface using drilling and coring techniques. pp. 3427-3441, In: Microbiology of Hydrocarbons and Lipids. K.N. Timmis (Ed.), Springer Verlag, Berlin. 2010. Davidson, M.M., B.J. Silver, T.C. Onstott, D.P. Moser, T.M. Gihring, L.M. Pratt, E.A. Boice, B. Sherwood Lollar, J. Lippmann-Pipke, S.M. Pfiffner, T.L. Kieft, W. Seymore, C. Ralston. Planktonic microbial diversity reflects geochemistry of subsurface fluid-filled fractures, Evander Basin, South Africa. Geomicrobiol. J. 18:275-300. 2011. Matthew O. Schrenk Ph.D. Co-I. Assistant Professor, East Carolina University, Department of Biology, Howell Science Complex, S303 Education and Training 1998 B.Sc. in Geology & Geophysics and S. Asian Studies, University of WisconsinMadison 2001 M.Sc.in Oceanography, University of Washington 2005 Ph.D. in Oceanography, Certificate in Astrobiology, University of Washington 2005-2008 Postdoctoral appointment in Astrobiology, Carnegie Institution for Science Appointments 1996-1998 Undergraduate Research Assistant, University of Wisconsin-Madison, Department of Geology and Geophysics 2000, 2013 Consultant, American Museum of Natural History, New York, NY 1998-2005 Research/Teaching Assistant, University of Washington, School of Oceanography 2005-2007 Postdoctoral Fellow, NASA Astrobiology Institute 2007-2008 Postdoctoral Associate, Carnegie Institution of Washington, Geophysical Laboratory and Department of Terrestrial Magnetism 2008-2013 Assistant Professor, Department of Biology, Adjunct Professor, Department of Geological Sciences East Carolina University, Greenville, NC 2013-present Assistant Professor, Department of Geological Sciences & Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI Representative Publications Schrenk, M.O., K.J. Edwards, R.M. Goodman, R.J. Hamers, and J.F. Banfield. Distribution of 35 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Thiobacillus ferrooxidans and Leptospirillum ferrrooxidans: implications for generation of acidic mine drainage. Science. 279:1519-1522. 1998. Schrenk, M.O., D.S. Kelley, J.R. Delaney, and J.A. Baross. Incidence and diversity of microorganisms within the walls of an active black smoker hydrothermal chimney. Appl. Environ. Microbiol. 69(6): 3580-3592. 2003. Schrenk, M.O., S.A. Bolton, D.S. Kelley, and J.A. Baross. Low archaeal diversity linked to subseafloor geochemical processes at the Lost City Field, Mid Atlantic Ridge. Environ. Microbiol. 6(10):1086-1095. 2004. Kelley, D.S., J. Karson, G. Früh-Green, D. Yoerger, T. Shank, D. Butterfield, J. Hayes, M.O. Schrenk, E. Olson, G. Proskurowski, M. Jakuba, A. Bradley, B. Larson, K. Ludwig, D. Glickson, K. Buckman, A.S., Bradley, W. Brazelton, K. Roe, M. Elend, A. Delacour, S. Bernasconi, m. Lilley, J. Baross, R. Summons, S. Sylva. A Serpentinite-hosted ecosystem: The Lost City Hydrothermal Field. Science. 307: 1428-1434. 2005. Brazelton, W.J. , M.O. Schrenk, D.S. Kelley, J.A. Baross. Methane and sulfur metabolizing microbial communities dominate in the Lost City Hydrothermal Field ecosystem. Appl. Environ. Microbiol. 72(9):6257-6270. 2006. Schrenk, M.O., J.A. Huber, K.J. Edwards. Microbial Provinces in the Subseafloor. Annual Review of Marine Science. 2:279-304. 2010. Jiao, Y., G.D. Cody, A.K. Harding, P. Wilmes, M. Schrenk, K.E. Wheeler, J.F. Banfield, M.P. Thelen. Characterization of Extracellular Polymeric Substances from Acidophilic Microbial Biofilms. Appl. Environ. Microbiol. 76: 2916-2922. 2010. Brazelton, W.J., B. Nelson, M.O. Schrenk. Metagenomic evidence of H2 oxidation and H2 production by serpentinite-hosted subsurface microbial communities. Frontiers in Microbiology. 2:doi:10.3389/fmicb.2011.00268. 2012. Schrenk, M.O., W.J. Brazelton, S.Q. Lang. Serpentinization, carbon, and deep life. Reviews in Mineralogy and Geochemistry. 75:575-606. 2013. Brazelton, W.J., P.L. Morrill, N. Szponar, M.O. Schrenk. Microbial communities associated with subsurface geochemical processes in continental serpentinite springs. Appl. Environ. Microbiol. 7: 2013. In press. A - APPENDICES A.1 - REFERENCES CITED A.1 - REFERENCES CITED 1. Lomstein, B.A., et al., Endospore abundance, microbial growth and necromass turnover in deep sub-seafloor sediment. Nature, 2012 . 484(7392): p. 101-4. 2. Whitman, W.B., D.C. Coleman, and W.J. Wiebe, Prokaryotes: the unseen majority. Proc Natl Acad Sci U S A, 1998. 95(12): p. 6578-83. 3. Sogin, M.L., K.T. Edwards, and S. D'Hondt, 2010. Deep Subsurface Microbiology and the Deep Carbon Observatory, http://codl.coas.oregonstate.edu/documents/Deep_Life_White_Paper.pdf2010. 36 Deep Life Community - The Deep Carbon Observatory – 2014-2015 4. Marteinsson, V.T., et al., Microbial communities in the subglacial waters of the Vatnajokull ice cap, Iceland. . ISME, 2012. 5. Briggs, B.R., et al., Bacterial dominance in subseafloor sediments characterized by methane hydrates. FEMS Microbiol. Ecol. , 2012. 81: p. 88-98. 6. Lavalleur, H.J. and F.S. Colwell, Microbial characterization of basalt formation waters targeted for geological carbon sequestration. FEMS Microbiol Ecol, 2013. 7. Kuczynski, J., et al., Using QIIME to analyze 16S rRNA gene sequences from microbial communities. Curr Protoc Microbiol, 2012. Chapter 1: p. Unit 1E 5. 8. Kelley, D.S., et al., A serpentinite-hosted ecosystem: the Lost City hydrothermal field. Science, 2005. 307(5714): p. 1428-34. 9. Lin, L.H., et al., Long-term sustainability of a high-energy, low-diversity crustal biome. Science, 2006. 314(5798): p. 479-82. 10. Sherwood Lollar, B., et al., Isotopic signatures of CH4 and higher hydrocarbon gases from Precambrian Shield sites: A model for abiogenic polymerization of hydrocarbons. Geochmica et Cosmochimica Acta 2008. 72. 11. Horsfield, B., et al., Living microbial ecosystems within the active zone of catagenesis: Implications for feeding the deep biosphere. Earth and Planetary Science Letters, 2006. 246(1): p. 55-69. 12. Menez, B., V. Pasini, and D. Brunelli, Life in the hydrated suboceanic mantle. Nature Geoscience, 2012. 5(2): p. 133-137. 13. Hayes, J.M. and J.R. Waldbauer, The carbon cycle and associated redox processes through time. Philos Trans R Soc Lond B Biol Sci, 2006. 361(1470): p. 931-50. 14. Preliminary Report: Integrated Ocean Drilling Program expedition 313. 2010; Available from: http://publications.iodp.org/preliminary_report/313/313pr_4.htm. 15. Ruppel, C. Catching Climate Change in Progress: Drilling on Circum-Arctic Shelves and Upper Continental Slopes; San Francisco, California, 10–11 December 2011. in Scientific Drilling for Climate Related Objectives on Arctic Ocean Margins. 2011. San Francisco, CA: EOS. 16. Parkes, R.J., et al., Temperature activation of organic matter and minerals during burial has the potential to sustain the deep biosphere over geological time scales. Organic Geochemistry, 2007. 38: p. 845-852. 17. Horsfield, B., et al., eds. The geobiosphere. Continental Scientific Drilling: A Decade of Progress and Challenges for the Future, ed. U. Harms, C. Koeberl, and M.D. Zoback. 2007, Springer: Berlin-Heidelberg. 163-212. 18. Moore, G.F., et al., Structural Setting of the Leg 190 Muroto Transect. Proc. ODP, Init. Repts College Station, TX (Ocean Drilling Program), 2001. 190. 19. Glass, E.M., et al., Using the metagenomics RAST server (MG-RAST) for analyzing shotgun metagenomes. Cold Spring Harb Protoc, 2010. 2010(1): p. pdb prot5368. A.2 - DECADAL GOALS 37 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Overall Summary of Deep Life Deep Life exerts a vital influence on Earth's subsurface carbon fluxes and reservoirs. It exploits Earth's deep energy at the intersection between abiotic and biotic realms. The Deep Life Community will map the abundance and diversity of subsurface marine and continental microorganisms in time and space as a function of their phylogenomic and biogeochemical properties, and their interactions with deep carbon. By integrating in situ and in vitro assessments of biomolecules, cells, communities, process rates and subsurface habitats using advanced measurement, imaging, and cultivation technologies, we will describe the environmental limits to deep life, its survival, metabolism and reproduction. The resulting data will inform experiments and models that seek to measure Deep Life’s impact on the carbon cycle, to constrain biologically-mediated structural alteration of deep reservoirs of carbon and other elements, and to define the deep biosphere’s relation to the surface world. Decadal Goals: DEEP LIFE: The Deep Life Community will explore the evolutionary and functional diversity of Earth’s deep biosphere and its interaction with the carbon cycle. I. Determine the processes that define the diversity and distribution of deep life as it relates to the carbon cycle. Examples of research focus include: • • • Conduct a global 3-D census over time of biological diversity (Bacteria, Archaea, Eukarya, viruses) in continental and marine deep subsurface environments. Investigate whether specific mechanisms govern microbial evolution and dispersal in the deep biosphere. Determine what ecological rules explain deep microbial community structure, e.g. spatial and temporal scales of community turnover, the role of the rare biosphere, and the effects of limited dispersal. II. Determine the environmental limits of deep life. For example we will: • • • Probe and test life’s response to physical and chemical extremes using observation, experimentation in the laboratory and in the field, and modeling. Explore what genomes can tell us about the limits and possible origins of life. Establish a bio-energetic framework for understanding the limits and adaptation of life in the deep subsurface. III. Determine the interactions between deep life and carbon cycling on Earth. For example we will: • • • Determine the principal pathways of carbon transformations in the subsurface and quantify the rates of these reactions. Characterize transitions between abiotic and biotic realms. Quantify how these processes interact with the surface world. A.3 - FUNDING THAT SUPPORTS ONE OR MORE OF DEEP LIFE’S DECADAL GOALS A.3.1 - Funded Projects: Bartlett, D. Deep Trench single-cell genomics. National Science Foundation, 9/1/08-8/31/13, $658,000 38 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Bartlett, D. Active microbial populations at depth. NASA, 8/10/10-8/9/14, 686,212 Bartlett, D. Deep-sea Trench Landers. The Prince Albert II of Monaco foundation, 1/1/1212/31/14, $600,000 Boetius, A. Gottfried-Wilhelm Leibniz Award 2009, 2009-2016, Deutsche Forschungsgemeinschaft, 2.5 M€ (approx. 30% will be invested towards DL research) Boetius, A. ERC-Adv. Investigator Grant ABYSS; 3.5 Mio€, appr. 30% spent on questions addressing carbon cycling and microbial biodiversity at the boundary between surface and subsurface life in polar realms. Boetius, A. DeBeer D. EU ESONET/EMSO seafloor observatories network; 500k€; 2008-2012 LOOME mudvolcano observatory Colwell, F.S., Microbiological Studies of Geological Systems Exposed to Supercritical CO2, DOE-NETL, 4/1/12-11/30/13. $97,000 Colwell, F.S., Experimental Characterization of Microbial Communities in Geological Materials Exposed to Hydrofracturing Fluids, DOE-NETL, 1/1/13-11/30/13. $80,000 Daniel, I. (PI), Hazael, R. (Co-I), Picard, A. (Co-I), Foglia, F. (Co-I), 6 days ESRF beamtime 06/2013, $63,000 Daniel, I. (PI), Reynard, B. (Co-I), Andreani, M. (Co-I). Lyon Institute of the Origins equipment grant 2013-2015, $368,000 Daniel, I. (PI), Feuillie, C. (Co-I), Pedreira Segade, U. (Co-I), Michot, L. (Co-I), Serpentines and the origin of the genetic material, CNRS-CNES interdisciplinary program, 2009 - 2016, $252,000 Daniel, I. Institut Universitaire de France. 2009-2013, $158,000 D’Hondt, S. (PI), Radiolytic hydrogen and microbial life in subseafloor sediment and basalt, NASA Astrobiology: Exobiology and Evolutionary Biology, 2011-present, $173,246. D’Hondt, S. (PI), Quantification of contamination potential in South Pacific Gyre sediment, Consortium for Ocean Leadership/USSSP, 2011-present, $15,000. D’Hondt, S. (PI) and A.J. Spivack (Co-PI), Collaborative Research: IODP Expedition 329 Objective Research on Supply of H2 by Water Radiolysis in Subseafloor Sediment of the South Pacific Gyre, NSF Ocean Drilling, 2011-present, Collaboration with R. Murray (Boston University), URI portion is $185,507. D’Hondt, S. (PI). U.S. Science Support Program Salary for IODP Expedition 329, USSSP/IODP, 2010-present, $196,127. D’Hondt, S. (PI), and J. Sauvage (Co-PI), IODP Expedition 337 Shimokita Coalbed Biosphere USSSP Support for Justine Sauvage, Consortium for Ocean Leadership, 2012-2013, $12,000. D’Hondt, S. (PI), and E.A. Walsh (Co-PI). Microbial Community Composition of the Bering Sea Site U1344, Consortium for Ocean Leadership/USSSP, 2010-present, $15,000. D’Hondt, S. (Co-I), Sogin, M. (Co-I). Microbial Community Structure of Subsurface Marine Environments. 8/1/13-12/30/13. $250,000 39 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Edwards, K.J. (PI), J. Cowen (Co-PI), S. D’Hondt (Co-PI), A. Fisher (Co-PI) and G. Wheat (CoPI), Center for Dark Energy Biosphere Investigations, Science and Technology Centers: Integrative Partnerships program, 2010-2015, $25M (URI portion is $1.25M). Foglia, F. (Co-I), McMillan, P.F. (Co-I), Hazael, R. (Co-I), Forsyth, T. (Co-I), Simeoni, G. (CoI), Appavou, M-S. (Co-I), Meersman, F. (Co-I), Intra-cellular and trans-membrane H2O diffusion and chemical exchange processes at high pressure, 9 days FRMII and ISIS neutron days 20122013, $94,600 Hinrichs, K.-U., Gottfried-Wilhelm Leibniz Award 2011, 2011-2018, Deutsche Forschungsgemeinschaft, 2.5 M€ (approx.. 50% will be invested towards DL research) Hinrichs, K.-U., DARCLIFE, European Research Council, 2010-2015. 2.908 M€ Hinrichs, K.-U., Timothy Ferdelman, Michael Friedrich, Sabine Kasten, MARUM-GB2: Biogeochemical processes fueling sub-seafloor life: Transformation of C, S, Fe; 2012-2017, Deutsche Forschungsgemeinschaft. 537 k€ (multiple Co-Is, only Hinrichs Lab portion indicated) Hinrichs, K.-U., Molecular-isotopic studies of microbial processes and organic matter in the subseafloor coalbed biosphere of Shimokita (IODP Exp. 337), 2012-2015, Deutsche Forschungsgemeinschaft, 231.75 k€ Hornbach, M. (PI), F.S. Colwell (Co-PI), Gas Hydrate Dynamics on the Alaskan Beaufort Continental Slope: Modeling and Field Characterization. DOE - Office of Fossil Energy, 10/1/12-9/30/15. $1,105,000 Itavaara, M. Deep Metapathway 2012-2015. Finnish Academy grant. 560.000 euros. Kieft, T. Determination of Regional Fluid Flow and gas flux rates for a continental plateau using cosmogenic and radiogenic noble gas isotopes., 03/01/12 – 02/28/14, NSF Hydrology Program $24,820 Kieft, T. (Co-I), Onstott, T. (Co-I), ETBC: Collaborative Research: Deep Crustal Biosphere Microbial Cycling of Carbon. National Science Foundation. 10/01/10 – 09/30/13, $638,590 Morgan-Smith, D. Pressure Resuscitation of the Deep Subsurface Biosphere. C-DEBI postdoctoral fellowship with M. Schrenk (ECU) and D. Bartlett (SIO). $58,000. Schrenk, M. (Co-I), Brazelton W., (Co-I). Metagenome- and Metatranscriptome- enabled Investigations of Carbon and Hydrogen Flux through the Serpentinite-hosted Subsurface Biosphere. DOE Joint Genome Institute, Community Sequencing Program. Stepanauskas, R. (PI), Emerson, D. (Co-I), Itavaara, M. (Co-I), Kieft, T. (Co-I), Lau, M. (Co-I), Moser, D. (Co-I), Moyer, C. (Co-I), Onstott, T.C. (Co-I), Orcutt, B. (Co-I), Wommack, E. (CoI), Bomberg, M. (Co-I). Enigmatic life underground: Large-scale single cell genomics of deep subsurface microorganisms. DOE Joint Genome Institute, Community Sequencing Program. Stepanauskas, R., Illumina MiSeq Grant. http://www.illumina.com/landing/miseqgrant/index.ilmn. $150,000 A.3.2 - Pending Leveraged proposals Cardace, D. How Deep Does Life Go? NSF-CAREER. $399,997.. 40 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Colwell, F.S., Biogeochemistry of Carbon Sequestration in Columbia River Basalts, Subcontract from Pacific Northwest National Laboratory; $24,000 Daniel, I. X-ray Proposal for 5-6 days beamtime at the ESRF D’Hondt, S. (PI), and J. Sauvage (Co-PI), Microbial Energetics in Subseafloor Hydrocarbon Reservoirs, Shimokita Peninsula, Consortium of Ocean Leadership, pending, $12,000. D’Hondt, S., J.B. Kirkpatrick, A. Abrajevitch, F. Colwell, H. Cypionka, B. Engelen, S. Gallagher, C. Hubert, F. Inagaki, J. Kallmeyer, Y. Morono, R.W. Murray, B. Opdyke and R. Pockalny. Nature and origin of subseafloor life in Mesozoic sediment of the Scott Plateau, Integrated Ocean Drilling Program (IODP) Ancillary Program Letter 830 (submitted April 1, 2013). Huber, J.A (PI), Stepanauskas, R. (Co-I). Deciphering metabolic and evolutionary processes at the upper temperature limits of life in subseafloor archaea using single cell genomics" NASA Exobiology and Evolutionary Biology Program. $649,581. Kelley, K., D. Cardace, and S. Carey.. Acquisition of a Fourier-Transform Infrared Spectrometer for igneous petrology, volcanology, and geobiology research. NSF-EAR IF, $155,522. Kieft, T. (PI), Mike Pullin (Co-PI) Characterization of dissolved organic carbon in deep crustal fracture water using solid state nuclear magnetic resonance analysis, Environmental Molecular Sciences Laboratory (EMSL) at DOE’s Pacific Northwest National Lab, proposal for facility access. Malinverno, A., (PI), Colwell, F.S. (Co-PI) IODP – Constraining Methane Cycling in Continental Margins: A Combined Microbiological, Geochemical, and Modeling Approach Proposal for ship time. Pockalny, R., and S. D’Hondt, EarthCube RCN: EarthCube Building Blocks: Crowdsourcing Geoscientific Data Curation, Collaboration with USC (prime sponsor: NSF), Submitted May 22 2013), URI portion = $133,745. Ruppel, C. (PI), Colwell, F.S. (Co-PI). Alaskan Beaufort Margin: Investigating the Impact of Warming Since the Last Glacial Maximum on Climate-Sensitive Sediments in the Arctic, IODP – Proposal for ship time. Schrenk, M. Microbial Activities in Serpentinized Rocks-Impacts upon Global and Regional Carbon Exchange. DOE Early Career Research Program. $750,000. Spivack, A.J. (PI), S. D’Hondt (Co-PI) and R. Pockalny (Co-PI), North Atlantic Meridional Circulation during the Last Glacial Maximum: Density Structure and Pre-formed Nitrate, submitted Feb 1 2013, $606,432. Wildenshild, D. (PI), Colwell, F.S. (Co-PI) Development of a State-of-the-Art High-Resolution Microtomography Facility Customized for Dynamic (4D) Imaging, NSF Major Research Instrumentation, $1,332,000 Winter, R. Exploring the Dynamical Landscape of Biomolecular Systems by Pressure Perturbation (FOR 1979 )", German Science Foundation (DFG) final decision pending involving 9 research groups from Germany 2.4 M Euro funding for three years. A.4 - DEEP LIFE COMMUNITY PUBLICATIONS 41 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Anderson, R.E., Brazelton, W.J., Baross, J.A. The deep virosphere: assessing the viral impact on microbial community dynamics in the deep subsurface. MSA volume "Deep Carbon", (R. M. Hazen, ed.). Mineralogical Society of America and The Geochemical Society (Chantilly VA), pp 649-675. 2013. Andreani, M., I. Daniel, and M. Pollet-Villard, Aluminum speeds up the hydrothermal alteration of olivine. American Mineralogist, 2013. in press. Bennett, S.A., Coleman, M., Huber, J.A., Reddington, E., Kinsley, J.C., McIntyre, C., Seewald, J.S., and C.R. German. Trophic regions of a hydrothermal plume dispersing away from an ultramafic-hosted vent-system: Von Damm vent-site, Mid-Cayman Rise. Geochemistry Geophysics Geosystems. 14:317-327. 2013. Brazelton, W.J.. P.L. Morrill, N. Szponar, M.O. Schrenk. Microbial communities associated with subsurface geochemical processes in continental serpentinite springs. Appl. Environ. Microbiol. 79(13):3906-3916. (2013). Breier, J.A., Gomez-Ibanez, D., Reddington, E., Huber, J.A., and D. Emerson. A precision multisampler for deep-sea hydrothermal microbial mat studies. Deep-Sea Research Part I: Oceanographic Research Papers.70:83-90. 2012 Briggs, B.R., F. Inagaki, Y. Morono, T. Futagami, C. Huguet, A. Rosell-Mele, T.D. Lorenson, F.S. Colwell. Bacterial dominance in subseafloor sediments characterized by methane hydrates. FEMS Microbiol. Ecol. 81: 88-98. 2012. Campanaro, S, DePascale, F., Telatin, A., Schiavon, R., Bartlett, D. H. and Valle, G. The transcriptional landscape of the deep-sea bacterium Photobacterium profundum in both a toxR mutant and its parental strain. BMC Genomics 13:567. 2012. Colwell, F.S. , Dhondt, S. Nature and extent of the deep biosphere. MSA volume "Deep Carbon", (R. M. Hazen, ed.). Mineralogical Society of America and The Geochemical Society (Chantilly VA), pp. 547-574. 2013. Daniel, I. and H.G.M. Edwards, Raman spectroscopy in Biogeology and Astrobiology, in Raman Spectroscopy Applied to Earth Sciences and Cultural Heritage J. Dubessy, M.C. Caumont, and F. Rull, Editors. The Mineralogical Society of Great Britain & Ireland. 2012. Davydov, D. R., Sineva, E. V., Davydov, N. Y., Bartlett, D. H., and Halpert, J. R. 2013. CYP261 enzymes from deep sea bacteria: A clue to conformational heterogeneity in cytochromes P450. Biotechnology and Applied Biochemistry 60:30-40. 2013. D’Hondt, S., Subsurface sustenance, News and Views, Nature Geoscience 6, 426–427, doi:10.1038/ngeo1843. 2013. D’Hondt, S., F. Inagaki, C Alvarez Zarikian and the IODP Expedition 329 Scientists, IODP Expedition 329: Life and habitability beneath the seafloor of the South Pacific Gyre, Scientific Drilling 15, 4-10. 2013. Dunlea, A.G., R.W. Murray, R.N. Harris, M.A. Vasiliev, H. Evans, A.J. Spivack, and S. D’Hondt, Assessment and Use of NGR Instrumentation on the JOIDES Resolution to Quantify U, Th, and K Concentrations in Marine Sediment, Scientific Drilling 15, 57-63. 2013. Edwards K.T., K. Becker, and F. Colwell. The Deep, Dark Energy Biosphere: Intraterrestrial Life on Earth. Ann. Rev. Earth Planet Sci. 40: 551-568. 2012. 42 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Eloe, E.A., Shulse, C. N., Fadrosh,D. W., Williamson, S. J., Allen, E. E. and Bartlett, D. H. Compositional differences in particle-associated and free-living microbial assemblages from an extreme deep-ocean environment. Environ. Microbiol. Reports. 3: 449–458. 2011. Eloe, E.A., Fadrish, D. W., Novotny, M., Zeigler Allen, L., Kim, M., Lombardo, M.-J., YeeGreenbaum, J., Yooseph, S., Allen, E. A., Lasken, R., Williamson, S. J., Bartlett, D. H. Going deeper: metagenome of a hadopelagic microbial community. PLoS ONE. 6: e20388. 2011. Eloe, E.A., Malfatti, F., Gutierrez, J., Hardy, K., Schmidt, W. E., Pogliano, K., Pogliano, J., Azam, F. and D. H. Bartlett. Isolation and characterization of the first psychropiezophilic Alphaproteobacterium. Appl. Environ. Microbiol. 77:8145-8153. 2011. Feuillie, C., et al., A novel SERRS sandwich-hybridization assay to detect specific DNA target. 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Global distribution of subseafloor sedimentary biomass, Proceedings of the National Academy of Science 109(40), 16213-16216. 2012. Kapoor, S. Triola, G., Vetter, I., Erlkamp, M., Waldmann, H., Winter, R., Revealing Conformational Substates of Lipidated N-Ras Protein by Pressure Modulation, Proc. Natl. Acad. Sci. U.S.A. 109: 460-465. (2012). Kapoor, S., Werkmüller, A., Goody, R. S., Waldmann, H., and Winter, R., Pressure Modulation of Ras-Membrane Interactions and Intervesicle Transfer, J. Am. Chem. Soc. 135 6149-6156 (2013). Kellermann, M.Y., et al., Autotrophy as a predominant mode of carbon fixation in thermophillic anaerobic methane-oxidizing microbial communities. Proceedings of the National Academy of Sciences of the United States of America, 109(47): p. 19321-26. 2012. Lavalleur, H.J., F. Colwell. Microbial characterization of basalt formation waters targeted for geological carbon sequestration. FEMS Microbiol. Ecol. DOI: 10.1111/1574-6941.12098. 2013. Lever, M.A., et al., Evidence for microbial carbon and sulfur cycling in deeply buried ridge flank basalt. Science, 339(6125): p. 1305-08. 2013. 43 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Lin, Y.S., et al., Towards constraining H2 concentration in subseafloor sediment: A proposal for combined analysis by two distinct approaches. Geochimica et Cosmochimica Acta, 77(0): p. 186201. 2012. Lin, Y.S., et al., Assessing production of the ubiquitous archaeal diglycosyl tetraether lipids in marine subsurface sediment using intramolecular stable isotope probing. Environmental Microbiology, 15(5): p. 1634-46. 2013. Lomstein, B.A., A.T. Langerhuus, S. D’Hondt, B.B. Jørgensen and A.J. Spivack, Spore abundance, microbial growth and necromass turnover in deep subseafloor sediment, Nature 484, 101–104, doi:10.1038/nature10905. 2012. Lloyd, K.G., Schreiber L., Petersen D.G., Kjeldsen K., Lever M.A., Stepanauskas R., Richter M., Kleindienst S, Lenk S, Schramm A, Jorgensen BB. Predominant archaea in marine sediments degrade detrital proteins. Nature 496:215-218. (2013). Lucas, S., Han, J., Lapidus, A., Cheng, J.- F., Goodwin, L.A., Pitluck, S., Peters, L., Mikhailova, N., Teshima, H., Detter, J. C., Han, C., Tapia, R., Land, M., Hauser, L., Kyrpides, N. C., Ivanova, N., Pagani, I., Vannier, P., Oger, P., Bartlett, D. H., Noll, K. M., Woyke T., and Jebbar, M. Complete genome sequence of the thermophilic piezophilic heterotrophic bacterium Marinitoga piezophila KA3. J. Bacteriol. 194:5974-5975. 2012. Marteinsson, V.T., Runarsson, A., Stefansson, A., Thorsteinsson, T., Johannesson, T., Magnusson, S.H., Reynisson, E., Einarsson, B., Wade, N., Morrison, H.G., Gaidos, E. Microbial communities in the subglacial waters of the Vatnajokull ice cap, Iceland. ISME J. DOI: 10.1038/ismej.2012.97. 2012. Meersman, F., Daniel I., Bartlett D., Winter R., Hazael R., and P.F. McMillan. High pressure biochemistry and biophysics. MSA volume "Deep Carbon", (R. M. Hazen, ed.). 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Wegener, G., et al., Assessing sub-seafloor microbial activity by combined stable isotope probing with deuterated water and 13C-bicarbonate. Environmental Microbiology, 14(6): p. 1517-27. 2012. Xie, S., et al., Turnover of microbial lipids in the deep biosphere and growth of benthic archaeal populations. Proceedings of the National Academy of Sciences of the United States of America, 110(15): p. 6010-14. 2013. Zhai, Y., and Winter, R., Effect of Molecular Crowding on the Temperature-Pressure Stability Diagram of Ribonuclease A, ChemPhysChem 14: 386-393. (2013). 45 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Lavalleur, H., C. Verba, C.R. Disenhof, W.K. O’Connor, and F.S. Colwell. Changes in native microbial communities exposed to geological carbon sequestration conditions in basalts. Internat. J. Greenhouse Gas Control. In review. Onstott, T.C., Aubrey, A.D., Kieft, T.L., Silver, B.J., Phelps, T.J., van Heerden, E., Opperman, D.J. and Bada, J.L. Is the Depth Limit of the Subsurface Biosphere Constrained by Aspartic Acid Racemization? Proc. Nat. Acad. Sci. In review. Smith, A., G. Flores, M. Fisk, F. Colwell, A. Thurber, O. Mason, and R. Popa. Deep crustal communities of the Juan de Fuca Ridge are governed by mineralogy. Science. In review. Méhay, S., G. L. Früh-Green, S. Q. Lang, S.M. Bernasconi, W.J. Brazelton, M. O. Schrenk, P. Schaeffer, P. Adam. Record of archaeal activity at the serpentinite-hosted Lost City Hydrothermal Field. Geobiology. In review. A.5 - POST DOCTORAL SUPPORT William Brazelton ECU Melitza CrespoMedina ECU Priya Narasingarao SIO Rachael Hazael UCL Fabrizia Foglia UCL Dani Morgan-Smith ECU Maggie Lau Princeton Borja Linage UFS Malin Bomberg VTT Julie Revillaud MBL Woo Jun Sul MBL Schrenk Schrenk Bartlett McMillan McMillan Schrenk/Bartlett Onstott van Heerden Itavaara Huber Sogin A.6 - MANAGEMENT PLAN A.6.1 - MEMBERSHIP OF DEEP LIFE SCIENTIFIC STEERING COMMITTEE Kai-Uwe Hinrichs – Co-Chair Dean and Professor, Department of Geosciences and Head, Organic Geochemistry Group MARUM Center for Marine Environmental Sciences, University of Bremen, Germany Mitchell L. Sogin – Co-Chair Director of the Josephine Bay Paul Center for Comparative Molecular Biology and Evolution. Marine Biological Laboratory, Woods Hole MA Professor, Molecular and Cellular Biology, Brown University, Providence RI Douglas H. Bartlett Professor, Scripps Institution of Oceanography, La Jolla, CA Antje Boetius Professor, HGF-MPG Group for Deep Sea Ecology and Technology 46 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Alfred Wegener Institute for Polar and Marine Research in the Helmholtz Association, Max Planck Institute for Marine Microbiology, Bremen Germany, Professor of Geomicrobiology, University Bremen Frederick S. Colwell Professor, Oregon State University, Corvallis, OR Isabelle Daniel Professor, University Claude Bernard Lyon 1, Lyon, France Steven D’Hondt Professor, Graduate School of Oceanography, University of Rhode Island, Narragansett, RI Fumio Inagaki Group Leader/Senior Scientist Geomicrobiology Group, Kochi Institute for Core Sample Research and Submarine Resources Research Project. Japan Agency for Marine-Earth Science and Technology (JAMSTEC), Japan Thomas L. Kieft Professor, New Mexico Institute of Mining and Technology, Socorro, New Mexico Matthew O. Schrenk Assistant Professor, East Carolina University, Greenville, NC Roland Winter Professor, Technical University Dortmund, Germany A.6.2 - STEERING COMMITTEE ACTIVITIES Meeting schedule. The Co-Chairs Kai-Uwe Hinrichs and Mitchell L. Sogin have formed a partnership to co-lead the Deep Life Scientific Steering Committee (SSC). As part of our efforts to expand internationally, we have invited Fumio Inagaki of JAMSTEC to serve on the DLC SSC. Drs. Hinrichs and Sogin will schedule Scientific Steering Committee meetings (a minimum of one dedicated in-person DLC SSC meeting each year, quarterly video conferences, and inperson meetings linked to the 2014 and 2015 Deep Life Community Meeting and the 2015 DCO “all-hands” meetings). Either Hinrichs or Sogin or (when possible) both will represent the DLC at in person and teleconferencing DCO Executive Committee meetings. The entire Steering Committee will participate in the planning of DLC annual meetings, workshops, and organization of special sessions at national and international meetings and conferences. 47 Deep Life Community - The Deep Carbon Observatory – 2014-2015 Liaison activities and resource management. Sogin will serve as the liaison to the DCO Data Science Team and D’Hondt will serve as liaison to the DCO Engagement Team (See 4.7 DATA SCIENCE, p. 21, and 4.8 ENGAGEMENT, p. 22. The Co-Chairs will assume fiduciary responsibility for managing DLC resources from the Sloan Foundation in consultation with SSC members who will provide leadership in developing proposals that seek new resources for scientific exploration that addresses the DLC Decadal Goals. Expanding Deep Life Support. The DLC has articulated an ambitious science agenda that will require resources that extend well beyond funding provided by the Sloan Foundation. The collection of samples and their interrogation through cultivation, chemical characterization and genomic analysis requires major investments that cannot be satisfied through the funding of salaries for a few post doctoral and graduate students over the next two years. Instead the DLC must extract information from its existing complex datasets and develop techniques and strategies for taking deep life research to the next level. At the DLC May, 2013 meeting in Portland ORE, with extensive community input the DL steering committee settled upon a strategy that will enable generation of new information from which to leverage new research proposals. As described under 4 - THE PROPOSED PROJECT: DEEP LIFE RESEARCH AND COORDINATION NETWORK, the DLC has set aside resources that will enable activities – both scientific investigations and synthesis activities, that will serve a key role in developing new proposals. The budget has set aside >45% of our direct costs to support DLC activities that lay the groundwork for new proposals. Unlike grants from a funding agency or foundation, these resources will have a short spending period of <= 6 months during which they should address the following criteria: a) Requests for support must address unresolved questions related to deep life and its decadal goals; b) Requests must identify the potential for attracting additional funding 48 Deep Life Community - The Deep Carbon Observatory – 2014-2015 resources and the description must outline a plan for seeking new research support; c) the amount of support can range from $1000 to as large as ~$25,000 and can be submitted at any time between January 1, 2014 and July 1, 2015; d) The supported activity must conclude within a six-month time-frame. Other criteria that will contribute to an application’s success include: e) the potential for leveraging other funding; f) fostering collaborative science; g) supporting synthesis of knowledge across the different themes; h) support for projects too risky for funding agencies; j) funding projects of opportunity that require immediate resources; i) projects that will enhance international programs and collaborations. By way of example, two days before the final draft for this this proposal was completed, Tom Kieft and Mike Pullin of New Mexico Tech were approved for a Rapid Access (30-day) project at the Environmental Molecular Sciences Laboratory (EMSL) at DOE’s Pacific Northwest National Lab titled Characterization of dissolved organic carbon in deep crustal fracture water using solid state nuclear magnetic resonance analysis. The project will use solid state NMR (~200 hours instrument time) to structurally examine organic matter extracted from deep (0.7-3.5 km) fracture water in South Africa. The deep water samples were collected with partial support from the DCO DL RHC project; the collaboration with EMSL is a direct result of the May 2013 DCO DL meeting in Portland. Kieft will have the opportunity to submit an application for DL support that could lead to securing major funding (NSF, DOE, etc.) for expansion to other analytical approaches (GC-MS, GC-Fourier transform-ion cyclotron resonance MS, etc.) and other sites (continental and marine). This provides an example of how DLC resources can both leverage recent awards from agencies and foundations as well as provide resources to support the submission of new applications for expanded research capabilities. 49 Deep Life Community - The Deep Carbon Observatory – 2014-2015 The DLC will accept 3-4 page applications for such support without specific deadlines throughout 2013-2015. The Deep Life section on the DCO website (http://www.deepcarbon.net/content/deep-life) will disseminate this information and we will use e-mail lists to notify members of the DLC of this opportunity. In addition we will share announcements of these opportunities with allied research initiatives such as C-DEBI. The Steering Committee (exclusive of members who participate in authorship of the proposed activity under evaluation or who have an inherent conflict of interest) will review the merits of proposals and render recommendations for funding to the Co-Chairs during in person meetings or quarterly video-conferences. Decisions by the Steering committee for supporting an activity will consider its impact on one or more DCO DL Decadal Goals; building the DL – DCO community, the potential for leveraging other funding; their influence on fostering collaborative science and synthesis of knowledge across the different themes; proposed activities too risky for funding agencies; novel opportunities with immediate funding requirements; and activities that will enhance international programs and collaborations. 50