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首页> 外文期刊>Applied Microbiology >Identification of Nitrogen-Incorporating Bacteria in Petroleum-Contaminated Arctic Soils by Using [15N]DNA-Based Stable Isotope Probing and Pyrosequencing
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Identification of Nitrogen-Incorporating Bacteria in Petroleum-Contaminated Arctic Soils by Using [15N]DNA-Based Stable Isotope Probing and Pyrosequencing

机译：基于[15N] DNA的稳定同位素探测和焦磷酸测序技术鉴定石油污染的北极土壤中的掺氮细菌

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Arctic soils are increasingly susceptible to petroleum hydrocarbon contamination, as exploration and exploitation of the Arctic increase. Bioremediation in these soils is challenging due to logistical constraints and because soil temperatures only rise above 0°C for ～2 months each year. Nitrogen is often added to contaminated soil in situ to stimulate the existing microbial community, but little is known about how the added nutrients are used by these microorganisms. Microbes vary widely in their ability to metabolize petroleum hydrocarbons, so the question becomes: which hydrocarbon-degrading microorganisms most effectively use this added nitrogen for growth? Using [~(15)N]DNA-based stable isotope probing, we determined which taxonomic groups most readily incorporated nitrogen from the monoammonium phosphate added to contaminated and uncontaminated soil in Canadian Forces Station-Alert, Nunavut, Canada. Fractions from each sample were amplified with bacterial 16S rRNA and alkane monooxygenase B ( alkB ) gene-specific primers and then sequenced using lage-scale parallel-pyrosequencing. Sequence data was combined with 16S rRNA and alkB gene C quantitative PCR data to measure the presence of various phylogenetic groups in fractions at different buoyant densities. Several families of Proteobacteria and Actinobacteria that are directly involved in petroleum degradation incorporated the added nitrogen in contaminated soils, but it was the DNA of Sphingomonadaceae that was most enriched in ~(15)N. Bacterial growth in uncontaminated soils was not stimulated by nutrient amendment. Our results suggest that nitrogen uptake efficiency differs between bacterial groups in contaminated soils. A better understanding of how groups of hydrocarbon-degraders contribute to the catabolism of petroleum will facilitate the design of more targeted bioremediation treatments.
机译：随着对北极的勘探和开发的增加，北极土壤越来越容易受到石油烃污染的影响。由于后勤方面的限制，并且由于土壤温度每年仅升高超过0°C约2个月，因此在这些土壤中进行生物修复具有挑战性。通常将氮原位添加到受污染的土壤中以刺激现有的微生物群落，但是对于这些微生物如何利用添加的营养物知之甚少。微生物代谢石油碳氢化合物的能力差异很大，因此问题就变成了：哪种降解碳氢化合物的微生物最有效地利用添加的氮来促进生长？使用基于[〜（15）N] DNA的稳定同位素探测，我们确定了哪类生物分类群最容易从添加到受污染和未受污染的土壤中的磷酸一铵中掺入的氮，添加到加拿大努纳武特的加拿大部队驻地-阿尔特。使用细菌16S rRNA和烷烃单加氧酶B（alkB）基因特异性引物扩增每个样品的馏分，然后使用鼠规模平行焦磷酸测序进行测序。将序列数据与16S rRNA和alkB基因C定量PCR数据结合起来，以测量不同浮力密度的馏分中各种系统发生基团的存在。与石油降解直接相关的变形杆菌属和放线菌属的几个家族在受污染的土壤中掺入了添加的氮，但Sphingomonadaceae的DNA富含〜（15）N。养分改良不会刺激未污染土壤中的细菌生长。我们的结果表明，在受污染的土壤中，不同细菌群体之间的氮吸收效率不同。更好地了解烃类降解物如何促进石油的分解代谢，将有助于设计更有针对性的生物修复方法。

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