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

智力优点：这项建议描述了一系列的实验室研究，旨在评估基因组和分子模式的石油生物降解的一系列条件下有关的地球？的表面和地下。全面的二维气相色谱、傅立叶变换离子回旋共振质谱和基于焦测序的宏基因组学的协同应用将为石油生物降解和负责微生物提供无与伦比的洞察力，并将这项工作与以往的任何研究区分开来。待检验的主要假设如下：1）微生物对烃的生物降解同时作用于数千种不同的化合物，而不是以严格的逐步方式。2)微生物群落的碳氢化合物偏好模式和相关的基因组潜力主要由末端电子接受过程的氧化还原电位控制。3)高分子量和极性化合物通过不完全氧化和结构改变而不是通过完全生物化进行生物转化。这些假设将通过进行时间序列的实验室实验进行检验，在实验中，石油在不同的电子受体和不同的温度下发生生物降解。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的处理间差异将提供数据来测试这些假设。结果进一步预期揭示新的代谢作用和基因组能力，并产生分子降解模式，可以将环境基因组和石油含量与相关的生物过程。更广泛的影响：这项研究的结果将广泛有助于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协助下通过大众媒体广泛传播。从这项研究中获得的知识也将直接翻译给包括NOAA在内的联邦机构。的评估和恢复司，以及私营企业通过现有的公司关系。这项研究的直接教育影响包括高中，本科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夏季研究计划，针对来自服务不足地区的学生纳入。本科生将通过融入课程和REU支持纳入拟议研究的各个方面。PI将为研究生和博士后研究人员提供指导。

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.