CAREER:Fractured Shales as Portals into Cryptic Carbon and Nitrogen Cycling in the Deep Biosphere

职业：破裂页岩作为深层生物圈隐秘碳和氮循环的门户

• 批准号: 1847684
• 负责人: Michael Wilkins
• 金额: $ 55.72万
• 依托单位: Colorado State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-15 至 2025-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1847684&HistoricalAwards=false
• 关键词: CAREER; Fractured; Shales; Portals; into

Microbial communities in terrestrial environments catalyze myriad biogeochemical cycles with global implications. However, in many ecosystems the combination of biological and chemical complexity can preclude a true mechanistic understanding of how microorganisms interact, are affected by viruses, and exchange metabolites. These kinds of insights are critical for our ability to manipulate microbiomes for beneficial outcomes, and better predict how biogeochemical cycles might change under varying environmental conditions. Deep fractured shales host more constrained microbial consortia that are more amenable to interrogation, and offer opportunities for new scientific insights and development of tools that can be applied to increasingly complex systems. Indeed, the investigator has previously revealed that the cycling of methylamines--simple carbon and nitrogen compounds--is a conserved metabolic strategy that fuels methylotrophic methanogenesis in deep shale formations across the Appalachian Basin. Work outlined in this proposal will determine the extent of functional conservation across geographically distinct shales, and use an extensive library of deep biosphere microbial isolates to investigate microbial metabolic interdependencies that enable persistence of life in these extreme environments. The role of viruses in driving microbial community dynamics and metabolite cycling will also be investigated. Finally, novel tools and insights developed in the shale ecosystem will be applied to a more complex wetland system to further illuminate the extent of methylamine metabolism in terrestrial environments. This project will train a graduate student and offer opportunities to undergraduate researchers. The involvement of industrial partners will enable internship placements for graduate students, and contribute to curriculum materials aimed at informing students of diverse career opportunities in subsurface science. Given the level of public interest in hydraulic fracturing and deep biosphere topics, a series of outreach events at local organizations are also planned.Genome-resolved metagenomic and metabolomic analyses, coupled with laboratory experimentation, has revealed that methylamine cycling in deep terrestrial shales is a conserved functional process across the Appalachian Basin that enables microbial communities to persist for many hundreds of days. Using multiomic approaches, this work will expand investigations into western US shale plays, and determine how functional traits are altered under varying salinity and temperature conditions. Genomic insights into metabolite exchange and methane isotope fractionation in these systems will be tested in the laboratory using environmental isolates and high-pressure incubation apparatus that enables in situ conditions (e.g., pressure, temperature, salinity) to be recreated. Novel analytical tools including high-pressure real-time NMR will be used to track the cycling of substrates at high temporal resolution. The ability of novel and abundant viral populations to mediate carbon and nitrogen cycling in the deep biosphere will also be investigated using a combination of bioinformatic tools and laboratory experimentation. Data resulting from these studies will offer unique insights into the functioning of microbial and viral populations, and the impact of microbial metabolism on poorly-resolved carbon and nitrogen cycles in the deep terrestrial subsurface.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

陆地环境中的微生物群落催化无数的生物地球化学周期具有全球影响。但是，在许多生态系统中，生物学和化学复杂性的组合可以排除对微生物如何相互作用，受病毒和交换代谢产物影响的真正机械理解。这类见解对于我们操纵微生物的有益结果的能力至关重要，并且可以更好地预测生物地球化学周期在不同的环境条件下如何变化。深层断裂的页岩容纳更受限制的微生物联盟，这些财团更适合审讯，并为新的科学见解和开发工具提供了机会，这些工具可用于日益复杂的系统。实际上，研究者先前已经揭示了甲胺的循环 - 简单的碳和氮化合物 - 是一种保守的代谢策略，可以在阿巴拉契亚盆地深层页岩中燃料甲基营养的甲烷生成。该提案中概述的工作将确定在地理上不同的页岩中的功能保护程度，并使用深广的生物圈微生物分离株的广泛文库来研究微生物代谢相互依存关系，从而在这些极端环境中能够持久生活。还将研究病毒在驱动微生物群落动力学和代谢产物循环中的作用。最后，在页岩生态系统中开发的新工具和见解将应用于更复杂的湿地系统，以进一步阐明陆地环境中甲胺代谢的程度。该项目将培训研究生，并为本科研究人员提供机会。工业合作伙伴的参与将为研究生提供实习安排，并为旨在告知学生地下科学中不同职业机会的课程材料。鉴于公众对液压破裂和深层生物圈主题的兴趣水平，还计划了一系列在当地组织进行的外展活动。基因组分辨的宏基因组和代谢组分析，再加上实验室实验，与实验室实验相结合，已经揭示了跨陆地式的甲基甲基甲基甲基胺在跨层次上的甲基甲基甲基苯丙胺构成，这是跨层次的造成式供应，该过程是在跨层次的基础上，该过程是在跨越的官能化过程。几天。使用多媒体方法，这项工作将扩大对美国西部页岩戏剧的研究，并确定在不同的盐度和温度条件下的功能性状如何改变。这些系统中对代谢产物交换和甲烷同位素分馏的基因组洞察力将在实验室中使用环境分离株和高压孵化装置进行测试，这些孵化设备可以重现原位条件（例如压力，温度，盐分）。包括高压实时NMR在内的新型分析工具将用于跟踪高时间分辨率的底物的循环。还将使用生物信息学工具和实验室实验的组合研究新的和丰富的病毒群体介导碳和氮循环的能力。这些研究产生的数据将为微生物和病毒群体的功能提供独特的见解，以及微生物代谢对深陆地地下中碳和氮循环差的碳和氮循环的影响。该奖项反映了NSF的法定任务，并通过评估了基金会的智力效果，并通过评估了cr依基金会和广泛的影响。

项目成果

Comparative geochemistry of flowback chemistry from the Utica/Point Pleasant and Marcellus formations

• DOI: 10.1016/j.chemgeo.2020.120041
• 发表时间: 2021-03
• 期刊: Chemical Geology
• 影响因子: 3.9
• 作者: S. Welch;J. Sheets;R. Daly;Andrea J. Hanson;Shikha Sharma;T. Darrah;J. Olesik;A. Lutton;P. Mouser;K. Wrighton;M. Wilkins;T. Carr;D. Cole

Identification of Persistent Sulfidogenic Bacteria in Shale Gas Produced Waters

• DOI: 10.3389/fmicb.2020.00286
• 发表时间: 2020-02-21
• 期刊: FRONTIERS IN MICROBIOLOGY
• 影响因子: 5.2
• 作者: Cliffe, Lisa;Nixon, Sophie L.;Lloyd, Jonathan R.
• 通讯作者: Lloyd, Jonathan R.

Ecological Assembly Processes Are Coordinated between Bacterial and Viral Communities in Fractured Shale Ecosystems

• DOI: 10.1128/msystems.00098-20
• 发表时间: 2020-03-01
• 期刊: MSYSTEMS
• 影响因子: 6.4
• 作者: Danczak, R. E.;Daly, R. A.;Wilkins, M. J.
• 通讯作者: Wilkins, M. J.

Genome-Resolved Metagenomics Extends the Environmental Distribution of the Verrucomicrobia Phylum to the Deep Terrestrial Subsurface

• DOI: 10.1128/msphere.00613-19
• 发表时间: 2019-11-01
• 期刊: MSPHERE
• 影响因子: 4.8
• 作者: Nixon, Sophie L.;Daly, Rebecca A.;Wrighton, Kelly C.
• 通讯作者: Wrighton, Kelly C.

Microbial colonization and persistence in deep fractured shales is guided by metabolic exchanges and viral predation.

• DOI: 10.1186/s40168-021-01194-8
• 发表时间: 2022-01-16
• 期刊: Microbiome
• 影响因子: 15.5
• 作者: Amundson KK;Borton MA;Daly RA;Hoyt DW;Wong A;Eder E;Moore J;Wunch K;Wrighton KC;Wilkins MJ
• 通讯作者: Wilkins MJ