RAPID: The Microbial Response to the Deepwater Horizon Oil Spill

智力优点：该RAPID项目将对路易斯安那州海岸近海深水地平线石油泄漏的后果进行时间序列的微生物和地球化学评估。自2005年以来，PI一直在深水地平线现场附近的微生物观测站（密西西比峡谷118）建立泄漏前基线微生物学和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和基因的分析;使用先进的传感器技术对水柱溶解氧和轻烃进行现场测量（Seaguard系统）用于深水羽流跟踪;并对石油泄漏附近和距离石油泄漏越来越远的沉积物和水进行生物地球化学调查，并在后续巡航期间以不同的时间尺度进行。对污染和清洁水及沉积物中微生物总DNA和RNA的16 S 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。PCR扩增的rRNA片段的高通量焦磷酸测序将使覆盖率增加约10%。三个数量级，并且允许检测在常规克隆文库中未被注意到的少数微生物群体。将特别注意石油降解细菌在自然样品中的富集和在实验室进行的时间序列实验，以监测其生长与组特异性聚合酶链反应，监测地球化学变化伴随着烃降解微生物群落的建立和富集，并确定潜在的碳掺入途径与稳定同位素探针的核酸。总而言之，该RAPID项目侧重于通过诊断水柱氧和轻烃测量确定的深水地平线石油泄漏的空间和时间尺度上的烃影响的分子和微生物评估。将通过测量沉积物厌氧微生物活动的地球化学指标，以及对DIC、CH 4和低分子量有机酸（碳氢化合物降解的主要产物）进行孔隙水分析，将水柱微生物和溶解气体数据与对底部沉积物细菌活动的潜在影响联系起来。PI正在与格鲁吉亚大学的Mandy Joye协调他们的研究。更广泛的影响：这个快速项目的结果将确定哪些细菌和古细菌种群以及它们的顺序对海洋沉积物和水柱中的石油泄漏事件作出反应，以及伴随着微生物群落及其活动的这种大规模重组的生物地球化学后果--通常是有益的。最重要的是，该项目提供了全面的微生物和地球化学覆盖不同的海洋生境（深海和浅海沉积物，水柱，表面）的地理和时间尺度的石油泄漏，因为它是发展。这一独立分析将有助于建立一个以观测为基础、注重成果的参考数据库，目前，深水地平线佐贺的各有关方面无法提供这一数据库。PI正在为我们的游轮和现场工作进行全国宣传，通过在目前的RV Walton Smith游轮上邀请国家地理作家（Joel Bourne），并接受科学美国人和国家地理的采访。PI已经开发了一个广泛的以学生为导向的外展部分，通过教学，实验室实习，会议访问和虚拟以及真实的巡航曝光的组合，吸引来自不同背景的本科生进入赠款驱动的研究，并让他们早日开始实质性的，可持续的研究项目。自2005年以来，Martens实验室一直积极参与NSF资助的研究项目，并与科学教育和推广计划合作，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.