Comparison and validation of sampling strategies for the molecular microbial analysis of surface microlayers
Comparison and validation of sampling strategies for the molecular microbial analysis of surface microlayers
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DOI：
10.3354/ame01330
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发表时间：
2009-01-01
影响因子：
1.4
通讯作者：
Murrell, J. Colin
中科院分区：
文献类型：
--
作者：
Cunliffe, Michael;Harrison, Emma;Murrell, J. Colin
有一系列的采样方法可用于采样海面微层，包括网屏（MS），玻璃板（GP），疏水（聚四氟乙烯，聚四氟乙烯）和亲水（PC，聚碳酸酯）膜。我们比较了采用上述采样方法采集的表层微层样品和地下水（0.4 m）中的细菌群落结构。从泰恩河口采集样本，采用细菌16S rRNA基因DGGE进行分析。用GP和MS采集的样品与地下水样品相似。聚四氟乙烯和PC采集的样品与GP和MS采集的样品和地下样品相比存在明显差异。膜样品在实际海面微层的建议深度范围内，而GP和MS收集更深的样品，因此也含有地下水。因此，有明确的证据表明GP和MS采样器不足以代表表面微层。对太平洋3个地点的表层微层和亚表层样品中细菌和古细菌群落进行了比较。采用细菌和古细菌16S rRNA基因DGGE对样品进行分析。在瓦胡岛两侧的两个地点采集的海洋表面微层样品中的细菌群落彼此之间的相似性比其同源地下样品更相似。古细菌群落未表现出明显的表面微层效应。这里提供的数据清楚地表明，细菌对这种海面微层环境有显著的反应，并进一步支持PC在海面微层取样中的使用。
A range of sampling methods are available to sample the sea surface microlayer, including mesh screens (MS), glass plates (GP), hydrophobic (PTFE, polytetrafluoroethylene) and hydrophilic (PC, polycarbonate) membranes. We compared bacterial community structures in surface microlayer samples and in subsurface water (0.4 m) collected using the above sampling methods. Samples were collected from the Tyne Estuary and analysed using DGGE of bacterial 16S rRNA genes. Samples collected using GP and MS were similar to subsurface water samples. Samples collected using PTFE and PC showed distinct differences compared to samples collected using GP and MS and subsurface samples. Membranes sample within the proposed depth range of the actual sea surface microlayer while GP and MS collect deeper samples which therefore also contain subsurface water. There is therefore clear evidence that GP and MS samplers under-represent the surface microlayer. Bacterial and archaeal communities were compared in sea surface microlayer and subsurface samples collected with PC from 3 sites in the Pacific Ocean. Samples were analysed using DGGE of bacterial and archaeal 16S rRNA genes. Bacterial communities in samples collected from the sea surface microlayer at 2 sites on either side of Oahu Island were more similar to each other than to their cognate subsurface samples. Archaeal communities did not show a distinct surface microlayer effect. Data presented here clearly indicate that Bacteria respond significantly to this sea surface microlayer environment and further support the use of PC in sampling surface microlayers.