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揭开海洋中隐匿的参与者——水化学与海洋微生物学指南

Unraveling the unseen players in the ocean - a field guide to water chemistry and marine microbiology.

作者信息

Haas Andreas Florian, Knowles Ben, Lim Yan Wei, McDole Somera Tracey, Kelly Linda Wegley, Hatay Mark, Rohwer Forest

机构信息

Department of Biology, San Diego State University;

Department of Biology, San Diego State University.

出版信息

J Vis Exp. 2014 Nov 5(93):e52131. doi: 10.3791/52131.

Abstract

Here we introduce a series of thoroughly tested and well standardized research protocols adapted for use in remote marine environments. The sampling protocols include the assessment of resources available to the microbial community (dissolved organic carbon, particulate organic matter, inorganic nutrients), and a comprehensive description of the viral and bacterial communities (via direct viral and microbial counts, enumeration of autofluorescent microbes, and construction of viral and microbial metagenomes). We use a combination of methods, which represent a dispersed field of scientific disciplines comprising already established protocols and some of the most recent techniques developed. Especially metagenomic sequencing techniques used for viral and bacterial community characterization, have been established only in recent years, and are thus still subjected to constant improvement. This has led to a variety of sampling and sample processing procedures currently in use. The set of methods presented here provides an up to date approach to collect and process environmental samples. Parameters addressed with these protocols yield the minimum on information essential to characterize and understand the underlying mechanisms of viral and microbial community dynamics. It gives easy to follow guidelines to conduct comprehensive surveys and discusses critical steps and potential caveats pertinent to each technique.

摘要

在此,我们介绍了一系列经过全面测试且标准化的研究方案,适用于偏远海洋环境。采样方案包括评估微生物群落可利用的资源(溶解有机碳、颗粒有机物质、无机养分),以及对病毒和细菌群落的全面描述(通过直接病毒和微生物计数、自发荧光微生物计数以及构建病毒和微生物宏基因组)。我们采用了多种方法,这些方法代表了一个分散的科学学科领域,包括已确立的方案和一些最新开发的技术。特别是用于病毒和细菌群落特征描述的宏基因组测序技术,只是近年来才得以确立,因此仍在不断改进。这导致了目前使用的各种采样和样品处理程序。这里介绍的这套方法提供了一种收集和处理环境样品的最新方法。这些方案涉及的参数产生了表征和理解病毒和微生物群落动态潜在机制所需的最少基本信息。它给出了易于遵循的进行全面调查的指南,并讨论了与每种技术相关的关键步骤和潜在注意事项。

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