PROJECT DESCRIPTION. TO INCREASE THE IMPACT OF THE ASTROBIOLOGY WORK CONDUCTED IN MY LAB AND TO HELP TO DRIVE THE FIELD OF ASTROBIOLOGY FORWARD WITH NEW CONCEPTS TECHNOLOGIES AND METHODS I REQUEST SUPPORT TO EXPAND OUR CURRENT LABORATORY CAPABILITIES BY ACQUIRING ALL NECESSARY INSTRUMENTATION TO PERFORM STABLE ISOTOPE PROBING (SIP) OF RNA AND DNA POOLS. SIP WHEN COMBINED WITH METAGENOMIC AND METATRANSCRIPTOMIC APPROACHES WILL ENABLE RESEARCH AIMED AT ESTABLISHING DIRECT LINKS BETWEEN CELLULAR METABOLISM ANDTHE ENZYMES AND TAXA INVOLVED IN THESE PROCESSES IN NATURAL SYSTEMS. IN PARTICULAR BY FOCUSING ON HYDROGEN (H2) BASED PRIMARY PRODUCTION IN EARLY EARTH ANALOG ENVIRONMENTS USING SIP BASED TECHNIQUES WE WILL PROVIDE NEW INSIGHTS INTO ENERGY FLOW IN MODERN DAY ANCESTORS OF PRIMITIVE FORMS OF LIFE THAT LIKELY INHABITED EARLY EARTH AND WHICH MAY INHABIT OTHER PLANETARY BODIES. MOREOVER THISTECHNOLOGY WILL FACILITATE THE ADVANCEMENT OF AN UNDEREXPLORED AREA OF ASTROBIOLOGICAL INTEREST ENERGY FLOW BETWEEN POPULATIONS AND THE ESTABLISHMENT OF COMPLEX MINERAL-SUPPORTED ECOSYSTEMS.NUMEROUS STUDIES ARE CURRENTLY PLANNED OR IN THE PROCESS OF BEING CONDUCTED BY MEMBERS OF MY RESEARCH GROUP THAT AIM TO TAKE ADVANTAGE OF SIP TECHNOLOGY UNDERSCORING MY LAB GROUPS COLLECTIVE ENTHUSIASM AND INTEREST IN INTEGRATING THIS TECHNOLOGY INTO THEIR THESIS PROJECTS. MOREOVER THESE STUDIES WILL PROVIDE A MECHANISM TO FORGE NEW COLLABORATIONS AND TO EXPAND UPON ONGOING AND FUTURENASA ASTROBIOLOGY PROJECTS INCLUDING THOSE THAT FOCUS ON CHEMOSYNTHETIC LIFE IN HOT SPRINGS (11-EXO11-0035) SUBSURFACE PERIDOTITES (NNA15BB02A) AND SUBGLACIAL ENVIRONMENTS (PROPOSAL PENDING). TOGETHER THESE PROJECTS WILL MAKE SIGNIFICANT CONTRIBUTIONS TOWARDLINKING MICROBIAL TAXA WITH FUNCTION IMPROVING UNDERSTANDING OF HOW RESOURCE AVAILABILITY SHAPES INTERSPECIES INTERACTIONS HOW THESE INTERACTIONS AFFECT EMERGENT ECOSYSTEM LEVEL FUNCTIONS AND HOW THESE INTERACTIONS UNDERPIN COEVOLUTIONARY DYNAMICS. DEVELOPINGTHIS TECHNOLOGY IN MY LAB AND APPLYING TO OUR RESEARCH QUESTIONS WILL POSITION ME TO MORE EFFECTIVELY COMPETE FOR ASTROBIOLOGY FOCUSED FEDERAL FUNDING.RELEVANCE. THE EXTENT TO WHICH WE DEFINE PLANETARY HABITABILITY IS CONTINGENT ON LIFES ABILITY TO DIVERSIFY INTO NEW ECOLOGICAL NICHES OVER PLANETARY TIME SCALES. BIOLOGICAL DIVERSIFICATION ON EARTH IS THE RESULT OF MYRIAD COMPLEX INTERACTIONS BETWEEN LIFE AND ITS SURROUNDINGS THAT HAVE PLAYED OUT OVER BILLIONS OF YEARS OF EARTH HISTORY. TO THIS END THE INTERACTIONS BETWEEN LIFE AND ITS ENVIRONMENT DEFINE PLANETARY HABITABILITY AND THUS FORM THE BASIS FOR OUR SEARCH FOR LIFE IN THE UNIVERSE. THE STUDIES AND TOOLS PROPOSED HERE PROVIDE A ROBUST MEANS TO EXAMINE THE INTERACTIONS BETWEEN LIFE AND ITS SURROUNDINGS AND TO DETERMINE HOW THESE INTERACTIONS CHANGE WITH NUTRIENT AMENDMENT SUCH AS H2. SUCH STUDIES WHEN APPLIED TO COMMUNITIES INHABITING EARLY EARTH ANALOGS AS PROPOSED HERE ADDRESS THE 2015 NASA STRATEGY DOCUMENT FOCAL AREA ON EARLY LIFE AND INCREASING COMPLEXITY SINCE THEY AIM TO STUDY HOW H2 DEPENDENT PRIMARY PRODUCERS AND PIONEER ORGANISMS COLONIZE ENVIRONMENTS ALLOWING FOR THE ASSEMBLY OF MORE COMPLEX COMMUNITIES. MOREOVER PLANNED STUDIES ARE ALIGNED WITH THE FOCAL AREA THE CO-EVOLUTION OF LIFE AND THE PHYSICAL ENVIRONMENT SINCE THEY FOCUS ON HOW GRADIENTS IN LITHOGENIC H2 INFLUENCE THE EXTENT AND DIVERSITY OF ECOSYSTEMS DEPENDENT ON THIS ENERGY. FINALLY OUR WORK IS CLOSELY ALIGNED WITH RESEARCH AIMED AT ADDRESSING SYNTHESIS AND FUNCTION OF MACROMOLECULES IN THE ORIGIN OF LIFE SINCE THE PROJECT FOCUSES ON METABOLIC PROCESSES (H2 METABOLISM CO2 FIXATION) AND MACROMOLECULES (E.G. [NIFE]-HYDROGENASES) THAT LIKELY PLAYED A KEY ROLE INLIFES ORIGINS.