Collaborative Research: Experimental determination of petroleum biodegradation patterns from a genomically-informed analytical vista

学术价值：这项建议描述了一系列实验室研究，旨在评估在一系列与地球有关的条件下石油生物降解的基因组和分子模式？S，地表和地下。综合的二维气相色谱、傅里叶变换离子回旋共振质谱仪和基于焦磷酸测序的元基因组学的协同应用将提供对石油生物降解和负责微生物的无与伦比的洞察，并使这项工作有别于以往的任何研究。要检验的主要假设如下：1)微生物对碳氢化合物的生物降解同时作用于数千种不同的化合物，而不是严格地以逐步的方式进行。2)微生物群落的碳氢化合物偏好模式和相关基因组势主要受末端接受电子过程的氧化还原电位控制。3)高相对分子质量和极性化合物的生物转化是通过不完全氧化和结构改变实现的，而不是通过完全再矿化。这些假设将通过进行时间序列实验室实验来验证，在这些实验中，石油在不同的电子受体和不同的温度下被生物降解。每个处理内分子的损失和获得模式，以及这些模式和基因组含量在不同处理之间的差异，将为检验这些假说提供数据。预计结果将进一步揭示新的代谢作用和基因组能力，并产生可将环境基因组和石油含量与相关生物过程联系起来的分子降解模式。还将开发和验证新的数据分析工具。广泛影响：这项研究的结果将有助于广泛了解石油生物降解和地球系统中的碳循环，并将在专业艺术家的帮助下，通过流行的渠道广泛传播。从这项研究中获得的知识也将直接转化为联邦机构，包括美国国家海洋和大气局S评估和修复司，以及通过现有的企业关系转化为私营行业。这项研究的直接教育影响包括对高中、本科生和研究生的培训和教育，以及博士后研究人员的高级培训。高中生将通过现有的暑期研究项目纳入其中，该项目针对的是服务不足地区的学生。本科生将通过融入课程作业和REU支持，被纳入拟议研究的所有方面。研究生和博士后研究人员的导师将由私人投资机构提供。

Intellectual merit: This proposal describes a series of laboratory studies designed to assess the genomic and molecular patterns of petroleum biodegradation under a range of conditions relevant to the Earth?s surface and subsurface. The concerted application of comprehensive, two-dimensional gas chromatography, Fourier transform ion cyclotron resonance mass spectrometry, and pyrosequencing-based metagenomics will provide unparalleled insight into petroleum biodegradation and the responsible microbes, and distinguishes this work from any previous studies. The primary hypotheses to be tested are the following: 1) Hydrocarbon biodegradation by microbes acts simultaneously on thousands of distinct compounds, not in a strictly stepwise fashion. 2) Hydrocarbon preference pattern and associated genomic potential for amicrobial community are controlled primarily by the redox potential of the terminal electron-accepting process. 3) High-molecular-weight and polar compounds are bio-transformed through incomplete oxidation and structural alteration, not through complete remineralization. These hypotheses will be tested by conducting time-series laboratory experiments in which petroleum is biodegraded with different electron acceptors and at different temperatures. The loss and gain patterns for molecules within each treatment and differences across treatments in these patterns and in genomic content will provide the data to test these hypotheses. Results are further anticipated to reveal novel metabolic actions and genomic capacity, and yield molecular degradation patterns that can relate environmental genomic and petroleum content to the relevant biological processes. New data analysis tools will also be developed and validated.Broader Impact: Results from this research will contribute broadly to an understanding of petroleum biodegradation and carbon cycling in the Earth system, and will be broadly disseminated through popular outlets with assistance from a professional artist. Knowledge gained from this research will also be translated directly to federal agencies including the NOAA?s Assessment and Restoration Division, as well as to private industry through existing corporate ties. Direct educational impacts of this research include the training and education of high school, undergraduate, and graduate students, as well as the advanced training of postdoctoral researchers. High school students will be incorporated through existing summer research programs targeting students from underserved regions. Undergraduate students will be incorporated into all aspects of the proposed research through integration into coursework and REU support. Mentoring of a graduate student and postdoctoral researchers will be provided by the PIs.