RAPID: The Microbial Response to the Deepwater Horizon Oil Spill

知识价值：这个RAPID项目将对路易斯安那州近海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。自2005年以来，石油公司一直在深水地平线附近的一个微生物观测站（密西西比峡谷118号）建立一个大型的泄漏前微生物基线和生物地球化学数据库。他们正在对地表水和下层沉积物中的微生物群落结构和功能进行分子、基因分析；利用先进的传感器技术（Seaguard系统）进行深水羽流跟踪的现场水柱溶解氧和轻烃测量；以及在后续巡航中，在不同的时间尺度上，对漏油点附近和离漏油点越来越远的沉积物和水进行生物地球化学调查。16S rRNA和功能基因测序将监测石油影响的微生物群落组成和活性的变化。pcr扩增的rRNA片段的高通量焦磷酸测序将增加大约的覆盖范围。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。将特别关注天然样品和实验室时间序列实验中石油降解细菌的富集，利用群体特异性PCR监测其生长，监测与烃类降解微生物群落建立和富集相关的地球化学变化，并利用核酸稳定同位素探测识别潜在的碳结合途径。总的来说，RAPID项目侧重于碳氢化合物影响的分子和微生物评估，通过诊断水柱氧和轻烃测量来确定深水地平线漏油的空间和时间尺度。通过测量沉积厌氧微生物活性的地球化学指标，以及对DIC、CH4和低分子量有机酸（烃降解的主要产物）的孔隙水分析，水柱微生物和溶解气体数据将与对底部沉积物细菌活性的潜在影响联系起来。pi正在与乔治亚大学的曼迪·乔伊协调他们的研究。更广泛的影响：这个RAPID项目的结果将确定哪些细菌和古细菌种群，以哪种顺序，对海洋沉积物和水柱中的溢油事件做出反应，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，该项目提供了全面的微生物和地球化学覆盖不同的海洋栖息地（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，因为它正在展开。这种独立的分析将有助于建立一个基于观察和以结果为导向的参考数据库，这是目前深水地平线事件中各种感兴趣的参与者无法提供的。私人助理正在为我们的游轮和现场工作进行全国宣传，通过邀请国家地理杂志的作家（乔尔·伯恩）参加目前的RV沃尔顿·史密斯游轮，并接受科学美国人和国家地理杂志的采访。ppi已经开发了一个广泛的以学生为导向的扩展组件，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，吸引来自不同背景的本科生参与资助驱动的研究，并使他们尽早开始实质性的，可发表的研究项目。自2005年以来，马丁实验室一直积极参与美国国家科学基金会资助的科学教育和推广项目，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.