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高北极土壤细菌对烃污染及生物修复处理反应的微阵列和实时聚合酶链反应分析

Microarray and real-time PCR analyses of the responses of high-arctic soil bacteria to hydrocarbon pollution and bioremediation treatments.

作者信息

Yergeau Etienne, Arbour Mélanie, Brousseau Roland, Juck David, Lawrence John R, Masson Luke, Whyte Lyle G, Greer Charles W

机构信息

Biotechnology Research Institute, National Research Council of Canada, Montréal, QC, Canada.

出版信息

Appl Environ Microbiol. 2009 Oct;75(19):6258-67. doi: 10.1128/AEM.01029-09. Epub 2009 Aug 14.

Abstract

High-Arctic soils have low nutrient availability, low moisture content, and very low temperatures and, as such, they pose a particular problem in terms of hydrocarbon bioremediation. An in-depth knowledge of the microbiology involved in this process is likely to be crucial to understand and optimize the factors most influencing bioremediation. Here, we compared two distinct large-scale field bioremediation experiments, located at the Canadian high-Arctic stations of Alert (ex situ approach) and Eureka (in situ approach). Bacterial community structure and function were assessed using microarrays targeting the 16S rRNA genes of bacteria found in cold environments and hydrocarbon degradation genes as well as quantitative reverse transcriptase PCR targeting key functional genes. The results indicated a large difference between sampling sites in terms of both soil microbiology and decontamination rates. A rapid reorganization of the bacterial community structure and functional potential as well as rapid increases in the expression of alkane monooxygenases and polyaromatic hydrocarbon-ring-hydroxylating dioxygenases were observed 1 month after the bioremediation treatment commenced in the Alert soils. In contrast, no clear changes in community structure were observed in Eureka soils, while key gene expression increased after a relatively long lag period (1 year). Such discrepancies are likely caused by differences in bioremediation treatments (i.e., ex situ versus in situ), weathering of the hydrocarbons, indigenous microbial communities, and environmental factors such as soil humidity and temperature. In addition, this study demonstrates the value of molecular tools for the monitoring of polar bacteria and their associated functions during bioremediation.

摘要

北极高纬度地区的土壤养分有效性低、水分含量低且温度极低,因此在碳氢化合物生物修复方面存在特殊问题。深入了解这一过程中涉及的微生物学知识,对于理解和优化对生物修复影响最大的因素可能至关重要。在此,我们比较了两个不同的大规模现场生物修复实验,分别位于加拿大北极高纬度地区的阿勒特站(异位方法)和尤里卡站(原位方法)。使用针对在寒冷环境中发现的细菌的16S rRNA基因和碳氢化合物降解基因的微阵列,以及针对关键功能基因的定量逆转录PCR来评估细菌群落结构和功能。结果表明,在土壤微生物学和去污率方面,采样地点之间存在很大差异。在阿勒特土壤中开始生物修复处理1个月后,观察到细菌群落结构和功能潜力迅速重组,以及烷烃单加氧酶和多环芳烃环羟基化双加氧酶的表达迅速增加。相比之下,在尤里卡土壤中未观察到群落结构的明显变化,而关键基因表达在相对较长的滞后期(1年)后增加。这种差异可能是由生物修复处理(即异位与原位)、碳氢化合物的风化、本地微生物群落以及土壤湿度和温度等环境因素的差异造成的。此外,本研究证明了分子工具在生物修复过程中监测极地细菌及其相关功能的价值。

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