RAPID: The Microbial Response to the Deepwater Horizon Oil Spill

智力优势：这个快速项目将对路易斯安那州海岸外深水地平线石油泄漏的后果进行一系列微生物和地球化学评估。PI正在他们自2005年以来一直占据的深水地平线附近的微生物观测站(密西西比峡谷118)建立了一个关于漏油前基线微生物和生物地球化学的大型数据库。他们正在对地表水和底层沉积物中的微生物群落结构和功能进行基于分子的基因分析；利用先进的传感器技术(Seaguard系统)对深水羽流进行现场水柱溶解氧和轻烃测量；对漏油事件附近和距离越来越远的沉积物和水进行生物地球化学调查，并在后续航行期间对不同的时间尺度进行调查。对污染和清洁的水和沉积物中的微生物总DNA和RNA进行16S rRNA和功能基因测序将监测受石油影响的微生物群落在组成和活动方面的变化。对聚合酶链式反应扩增的rRNA片段进行高通量焦磷酸测序将使覆盖率增加约10%。三个数量级，并允许检测在传统克隆文库中未被注意到的少数微生物种群。将特别注意石油降解菌在天然样品和实验室进行的时间序列实验中的浓缩，利用特定群体的聚合酶链式反应监测它们的生长，监测随着碳氢化合物降解微生物群落的建立和丰富而发生的地球化学变化，并通过核酸的稳定同位素探测来确定潜在的碳结合途径。综上所述，这一快速项目侧重于通过诊断水柱氧气和轻烃测量确定的深水地平线漏油的空间和时间尺度，对碳氢化合物影响的分子和微生物评估。水柱微生物和溶解气体数据将通过测量沉积厌氧微生物活动的地球化学指标以及对碳氢化合物降解的主要产物DIC、CH4和低分子有机酸的孔隙分析，与对海底沉积物中细菌活动的潜在影响联系起来。PIS正在与乔治亚大学的曼迪·乔伊协调他们的研究。广泛的影响：这个快速项目的结果将确定哪些细菌和古生菌种群，以及以什么顺序对海洋沉积物和水柱中的石油泄漏事件做出反应，以及伴随而来的--通常是有益的--微生物群落及其活动的大规模重组的生物地球化学后果。最重要的是，该项目提供了关于正在展开的石油泄漏的地理和时间尺度的不同海洋生境(深海和浅水海洋沉积物、水柱、水面)的全面微生物和地球化学覆盖范围。这一独立分析将有助于建立一个以观察为基础、以结果为导向的参考数据库，而目前深水地平线事件中感兴趣的各种参与者无法提供这一数据库。PI正在为我们的邮轮和现场工作进行全国宣传，通过邀请国家地理作家(Joel Bourne)乘坐目前的房车Walton Smith邮轮，以及接受科学美国人和国家地理的采访。私人投资机构开发了一个广泛的面向学生的外联部分，通过教学、实验室实习、会议访问和虚拟和真实的邮轮曝光相结合的方式，吸引来自不同背景的本科生进入赠款驱动的研究，并使他们早日开始实质性的、可发表的研究项目。自2005年以来，马滕斯实验室一直在积极地将NSF资助的研究项目与科学教育和外联项目合作，这些项目可以影响和吸引下一代海洋学家。

Intellectual merit: This RAPID project will conduct a time series of microbiological and geochemical assessments of the consequences of the Deepwater Horizon oil spill offshore the Louisiana coast. The PIs are building on a large database of pre-spill baseline microbiology and biogeochemistry at a microbial observatory (Mississippi Canyon 118) near the Deepwater Horizon site they have occupied since 2005. They are applying molecular, gene-based analyses of the microbial community structure and function in surface water and underlying sediments; in situ water column dissolved oxygen and light hydrocarbon measurements using advanced sensor technologies (Seaguard system) for deep water plume tracking; and a biogeochemical survey of the sediments and water in the immediate vicinity of and at increasing distance from the oil spill, and on different time scales during follow-up cruises. 16S rRNA and functional gene sequencing of total microbial DNA and RNA from contaminated and clean water and sediments will monitor how the oil-affected microbial community changes in composition and activity. High-throughput pyrosequencing of PCR-amplified rRNA fragments will increase the coverage by approx. three orders of magnitude, and allow for the detection of minority microbial populations that go unnoticed in conventional clone libraries. Special attention will be paid to the enrichment of oil-degrading bacteria in natural samples and in time-series experiments conducted in the lab, to monitor their growth with group-specific PCR, to monitor geochemical changes concomitant with the establishment and enrichment of a hydrocarbon-degrading microbial community, and to identify potential carbon incorporation pathways with stable isotope probing of nucleic acids. Summarizing, this RAPID project focuses on molecular and microbiological assessments of hydrocarbon impact, across the spatial and time scales of the Deepwater Horizon oil spill as determined by diagnostic water column oxygen and light hydrocarbon measurements. The water column microbiological and dissolved gas data will be linked to potential impacts on the bacterial activity in bottom sediments through measurements of geochemical indicators of sedimentary anaerobic microbial activity, and porewater analyses of DIC, CH4 and low-molecular weight organic acids, the principal products of hydrocarbon degradation. The PIs are coordinating their research with Mandy Joye at the University of Georgia.Broader Impacts: The results of this RAPID project will identify which bacterial and archaeal populations, and in which sequence, respond to oil spill events in marine sediments and the water column, and the attendant - often beneficial - biogeochemical consequences of this massive restructuring of the microbial community and their activities. Most importantly, this project provides comprehensive microbial and geochemical coverage of different marine habitats (deep and shallow marine sediments, water column, surface) on the geographical and time scale of the oil spill as it is unfolding. This independent analysis will contribute to an observation-based and results-oriented reference database that, at present, the various interested players in the Deepwater Horizon saga cannot provide. The PIs are working on national publicity for our cruises and on-site work, by taking a National Geographic writer (Joel Bourne) on the current RV Walton Smith cruise, and interviews with Scientific American and National Geographic. The PIs have developed an extensive student-oriented outreach component that entrains - with a combination of teaching, lab internships, conference visits and virtual as well as real cruise exposure - undergraduates from different backgrounds into grant-driven research and gives them an early start on substantial, publishable research projects. The Martens lab has been active since 2005 in partnering NSF-funded research projects with science education and outreach programs that can influence and attract the next generation of oceanographers.