BIOCOMPLEXITY (IDEA): In Situ Measurement of Marine Microbes to Investigate Mechanisms of Community Structure Regulation

生物复杂性（IDEA）：海洋微生物的原位测量以研究群落结构调节机制

• 批准号: 0119915
• 负责人: Robert Olson
• 金额: $ 169.1万
• 依托单位: Woods Hole Oceanographic Institution
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-10-01 至 2007-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0119915&HistoricalAwards=false
• 关键词: BIOCOMPLEXITY; IDEA; Situ; Measurement; Marine

This project is supported by the program Biocomplexity in the Environment, subprogram Instrumentation Development for Environmental Activities (BE-IDEA). The objective is to develop instrumentation and software for automated in situ monitoring of marine microbial communities. Microbes account for almost all the primary productivity and a majority of the biomass in the ocean, and the structure of the microbial community affects higher trophic levels, including species consumed by humans. A fundamental understanding of the complex interaction between physical and biological factors that regulate community structure requires more detailed and sustained observations than are possible with current equipment. This project will develop and deploy in situ instrumentation to monitor individual cells of microscopic plankton. The planned approach will combine in situ flow cytometry and imaging-in-flow techniques to measure the abundance and properties of cells (and other particles) ranging from the smallest phytoplankton (less than 1 um) to large chain diatoms (100 um). An automated image analysis system will be used to determine taxonomic affiliation of each cell for the larger size classes. Smaller cells (which often lack distinctive morphological features) will be evaluated by pulse-shape analysis of individual cell light scattering and fluorescence. For more detailed investigation of particular groups or species, a module capable of applying group-specific rRNA probes (via Peptide Nucleic Acid in situ hybridization) will be developed. The probed samples will be analyzed in real time by the in situ flow cytometer. The same module will also be used to apply fluorescent probes for other properties such as DNA content and cell viability. Software to analyze and integrate the flow cytometric and image data in near-real time will also be developed. The instrument will be based on an in situ flow cytometer developed at WHOI (plus hybridization technology currently under development) and integrated with a commercially available imaging-in-flow system. When completed, the instrument package will be field tested near the Marthas Vineyard Coastal Observatory (MVCO).In combination with physical oceanographic, meteorological, and bulk fluorescence data, individual particle data will facilitate an understanding of processes affecting coastal ecosystems, including the mechanisms regulating phytoplankton blooms, species succession, and trophic interactions. The planned work will form the foundation for an unprecedented long term high resolution time series of natural microbial populations, whose members vary in size by many orders of magnitude. These kinds of observations are critical for evaluating and developing models of long-term ecological change.

该项目由环境中的生物复杂性计划、环境活动仪器开发(BE-IDEA)子计划支持。其目标是开发用于对海洋微生物群落进行自动现场监测的仪器和软件。微生物几乎占据了海洋中所有的初级生产力和大部分生物量，微生物群落的结构影响着更高的营养水平，包括人类消耗的物种。要从根本上理解调节群落结构的物理和生物因素之间的复杂相互作用，需要比目前的设备更详细和持续的观察。该项目将开发和部署现场仪器，以监测微小浮游生物的单个细胞。计划中的方法将结合原位流式细胞术和流动成像技术来测量细胞(和其他颗粒)的丰度和性质，范围从最小的浮游植物(不到1微米)到大的链状硅藻(100微米)。将使用自动图像分析系统来确定较大规模类别的每个细胞的分类从属关系。较小的细胞(通常缺乏明显的形态特征)将通过对单个细胞的光散射和荧光的脉冲形状分析进行评估。为了对特定群体或物种进行更详细的调查，将开发一个能够应用群体特异性rRNA探针(通过肽核酸原位杂交)的模块。被探测的样本将通过原位流式细胞仪进行实时分析。同一模块还将用于应用荧光探针来检测其他特性，如DNA含量和细胞活力。还将开发用于近实时分析和整合流式细胞仪和图像数据的软件。该仪器将基于世界卫生组织开发的原位流式细胞仪(加上目前正在开发的杂交技术)，并与商业上可用的流动成像系统相结合。完成后，该仪器包将在马萨斯葡萄园岛海岸观测站(MVCO)附近进行现场测试。结合物理海洋学、气象学和大宗荧光数据，单个颗粒数据将有助于理解影响沿海生态系统的过程，包括调节浮游植物水华、物种演替和营养相互作用的机制。计划中的工作将为一个史无前例的长期高分辨率自然微生物种群时间序列奠定基础，其成员的大小相差许多数量级。这些类型的观测对于评估和开发长期生态变化模型至关重要。