Knapik Kamila, Bagi Andrea, Krolicka Adriana, Baussant Thierry
NORCE Environment, NORCE Norwegian Research Centre AS, 4070 Randaberg, Norway.
Microorganisms. 2020 May 15;8(5):744. doi: 10.3390/microorganisms8050744.
The use of natural marine bacteria as "oil sensors" for the detection of pollution events can be suggested as a novel way of monitoring oil occurrence at sea. Nucleic acid-based devices generically called genosensors are emerging as potentially promising tools for in situ detection of specific microbial marker genes suited for that purpose. Functional marker genes are particularly interesting as targets for oil-related genosensing but their identification remains a challenge. Here, seawater samples, collected in tanks with oil addition mimicking a realistic oil spill scenario, were filtered and archived by the Environmental Sample Processor (ESP), a fully robotized genosensor, and the samples were then used for post-retrieval metatranscriptomic analysis. After extraction, RNA from ESP-archived samples at start, Day 4 and Day 7 of the experiment was used for sequencing. Metatranscriptomics revealed that several KEGG pathways were significantly enriched in samples exposed to oil. However, these pathways were highly expressed also in the non-oil-exposed water samples, most likely as a result of the release of natural organic matter from decaying phytoplankton. Temporary peaks of aliphatic alcohol and aldehyde dehydrogenases and monoaromatic ring-degrading enzymes (e.g., , , and clusters) were observed on Day 4 in both control and oil-exposed and non-exposed tanks. Few alkane 1-monooxygenase genes were upregulated on oil, mostly transcribed by families and , together with aromatic ring-hydroxylating dioxygenases, mostly transcribed by . Few transcripts from obligate hydrocarbonoclastic genera of , and were significantly enriched in the oil-treated exposed tank in comparison to control the non-exposed tank, and these were mostly transporters and genes involved in nitrogen and phosphorous acquisition. This study highlights the importance of seasonality, i.e., phytoplankton occurrence and senescence leading to organic compound release which can be used preferentially by bacteria over oil compounds, delaying the latter process. As a result, such seasonal effect can reduce the sensitivity of genosensing tools employing bacterial functional genes to sense oil. A better understanding of the use of natural organic matter by bacteria involved in oil-biodegradation is needed to develop an array of functional markers enabling the rapid and specific in situ detection of anthropogenic pollution.
利用天然海洋细菌作为“油类传感器”来检测污染事件,可被视为一种监测海上油类存在的新方法。一类通常被称为基因传感器的基于核酸的装置,正逐渐成为用于原位检测适合该目的特定微生物标记基因的潜在有前景的工具。功能性标记基因作为与油类相关的基因传感的靶标特别有趣,但其鉴定仍然是一个挑战。在这里,在模拟实际溢油场景的添加油的水箱中采集的海水样本,通过环境样本处理器(ESP,一种全自动化基因传感器)进行过滤和存档,然后将样本用于取回后的宏转录组分析。提取后,实验开始时、第4天和第7天来自ESP存档样本的RNA用于测序。宏转录组学显示,在接触油的样本中,几个KEGG途径显著富集。然而,这些途径在未接触油的水样中也高度表达,很可能是由于腐烂浮游植物释放天然有机物的结果。在第4天,在对照水箱以及接触油和未接触油的水箱中均观察到脂肪族醇和醛脱氢酶以及单芳环降解酶(例如, 、 和 簇)的临时峰值。在油处理的暴露水箱中,与对照未暴露水箱相比,很少有烷烃1-单加氧酶基因上调,主要由 家族和 家族转录,同时芳香环羟基化双加氧酶主要由 转录。与对照未暴露水箱相比,来自专性烃降解属 、 和 的少数转录本在油处理的暴露水箱中显著富集,这些主要是转运蛋白以及参与氮和磷获取的基因。这项研究强调了季节性的重要性,即浮游植物的出现和衰老导致有机化合物的释放,细菌会优先利用这些有机化合物而非油类化合物,从而延迟了后者的过程。因此,这种季节性效应会降低采用细菌功能基因来传感油类的基因传感工具的灵敏度。需要更好地了解参与油类生物降解的细菌对天然有机物的利用情况,以开发一系列功能性标记物,实现对人为污染的快速和特异性原位检测。