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通过变性梯度凝胶电泳(DGGE)分析揭示急性铜暴露对北海沉积物中微生物群落多样性的影响——实验方案的重要性

The effect of an acute copper exposure on the diversity of a microbial community in North Sea sediments as revealed by DGGE analysis--the importance of the protocol.

作者信息

Gillan David C

机构信息

Marine Biology Laboratory, CP160/15, Université Libre de Bruxelles, 50 av. F.D. Roosevelt, Bruxelles B-1050, Belgium.

出版信息

Mar Pollut Bull. 2004 Sep;49(5-6):504-13. doi: 10.1016/j.marpolbul.2004.03.003.

DOI:10.1016/j.marpolbul.2004.03.003
PMID:15325219
Abstract

The aim of the work was to investigate whether the marine bacterial communities in a North Sea sediment with background metal concentrations were affected by an acute copper exposure and if a commonly used molecular technique, denaturing gradient gel electrophoresis (DGGE), was robust enough to investigate the community changes. Sediments (n = 6) were placed in small microcosms and spiked with copper (50 microg/l). Controls were left untreated. After 12 days, bioavailable copper increased up to a factor 2.5 in the sediments. Plate counts and chitinase activity measurements have suggested limited effects of copper on growth rate and cell metabolism. To test the robustness of DGGE three different protocols were used. The three protocols lead to different conclusions. As a whole, it seems that copper had no immediate effect on the genetic diversity of the community. However, copper-sensitive bacterial populations were detected by one of the DGGE protocols. It is concluded that the DGGE approach is a valuable tool to investigate the effect of pollutants on microbial communities only if various DGGE protocols are compared.

摘要

这项工作的目的是调查北海沉积物中背景金属浓度下的海洋细菌群落是否受到急性铜暴露的影响,以及常用的分子技术——变性梯度凝胶电泳(DGGE)是否强大到足以研究群落变化。将沉积物(n = 6)置于小型微观世界中,并添加铜(50微克/升)。对照组不做处理。12天后,沉积物中生物可利用铜增加了2.5倍。平板计数和几丁质酶活性测量表明铜对生长速率和细胞代谢的影响有限。为了测试DGGE的稳健性,使用了三种不同的方案。这三种方案得出了不同的结论。总体而言,铜似乎对群落的遗传多样性没有直接影响。然而,通过其中一种DGGE方案检测到了对铜敏感的细菌种群。得出的结论是,只有比较各种DGGE方案,DGGE方法才是研究污染物对微生物群落影响的有价值工具。

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