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定量分析石油污染的海洋港口沉积物中异化(双)亚硫酸盐还原酶基因的表达

Quantifying expression of a dissimilatory (bi)sulfite reductase gene in petroleum-contaminated marine harbor sediments.

作者信息

Chin Kuk-Jeong, Sharma Manju L, Russell Lyndsey A, O'Neill Kathleen R, Lovley Derek R

机构信息

Department of Microbiology, University of Massachusetts, Amherst, MA 01003, USA.

出版信息

Microb Ecol. 2008 Apr;55(3):489-99. doi: 10.1007/s00248-007-9294-2. Epub 2007 Sep 5.

Abstract

The possibility of quantifying in situ levels of transcripts for dissimilatory (bi)sulfite reductase (dsr) genes to track the activity of sulfate-reducing microorganisms in petroleum-contaminated marine harbor sediments was evaluated. Phylogenetic analysis of the cDNA generated from mRNA for a ca. 1.4 kbp portion of the contiguous dsrA and dsrB genes suggested that Desulfosarcina species, closely related to cultures known to anaerobically oxidize aromatic hydrocarbons, were active sulfate reducers in the sediments. The levels of dsrA transcripts (per mug total mRNA) were quantified in sediments incubated anaerobically at the in situ temperature as well as in sediments incubated at higher temperatures and/or with added acetate to increase the rate of sulfate reduction. Levels of dsrA transcripts were low when there was no sulfate reduction because the sediments were depleted of sulfate or if sulfate reduction was inhibited with added molybdate. There was a direct correlation between dsrA transcript levels and rates of sulfate reduction when sulfate was at ca. 10 mM in the various sediment treatments, but it was also apparent that within a given sediment, dsrA levels increased over time as long as sulfate was available, even when sulfate reduction rates did not increase. These results suggest that phylogenetic analysis of dsr transcript sequences may provide insight into the active sulfate reducers in marine sediments and that quantifying levels of dsrA transcripts can indicate whether sulfate reducers are active in particular sediment. Furthermore, it may only be possible to use dsrA transcript levels to compare the relative rates of sulfate reduction in sediments when sulfate concentrations, and possibly other environmental conditions, are comparable.

摘要

对石油污染的海洋港口沉积物中异化型(双)亚硫酸盐还原酶(dsr)基因转录本的原位水平进行定量,以追踪硫酸盐还原微生物活性的可能性进行了评估。对连续的dsrA和dsrB基因约1.4 kbp部分的mRNA产生的cDNA进行系统发育分析表明,与已知厌氧氧化芳烃的培养物密切相关的脱硫弧菌属物种是沉积物中活跃的硫酸盐还原菌。在原位温度下厌氧培养的沉积物中,以及在较高温度下和/或添加乙酸盐以提高硫酸盐还原速率的沉积物中,对dsrA转录本水平(每微克总mRNA)进行了定量。当没有硫酸盐还原时,dsrA转录本水平较低,这是因为沉积物中的硫酸盐已耗尽,或者如果添加钼酸盐抑制了硫酸盐还原。在各种沉积物处理中,当硫酸盐浓度约为10 mM时,dsrA转录本水平与硫酸盐还原速率之间存在直接相关性,但同样明显的是在给定的沉积物中,只要有硫酸盐,dsrA水平就会随时间增加,即使硫酸盐还原速率没有增加。这些结果表明,对dsr转录本序列进行系统发育分析可能有助于深入了解海洋沉积物中活跃的硫酸盐还原菌,并且对dsrA转录本水平进行定量可以表明硫酸盐还原菌在特定沉积物中是否活跃。此外,只有当硫酸盐浓度以及可能的其他环境条件相当时,才有可能使用dsrA转录本水平来比较沉积物中硫酸盐还原的相对速率。

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