典型油田多环芳烃污染对土壤反硝化微生物群落结构的影响

油田区土壤具有潜在的PAHs(polycyclic aromatic hydrocarbons)污染风险,而以硝酸根为电子受体的反硝化作用可能在PAHs的厌氧代谢中起到重要作用.以具有50多年历史的江汉油田区域为对象,从该油田的油井口附近采集了9个土壤样品,编号为JH-1~JH-9,以反硝化相关的nir K(Cu-亚硝酸还原酶基因)和nirS(细胞色素cd1-亚硝酸还原酶基因)为分子标识,通过定量PCR及克隆文库结合T-RFLP(terminal-restriction fragment length polymorphism)的方法,研究典型油田区土壤反硝化微生物的群落结构,并探讨其与土壤环境因子之间的关系.结果表明,该油田区土壤中nirK基因的丰度高于nirS基因,PAHs含量最高的土壤样品(JH-4)中反硝化功能基因nir K和nir S的丰度均最低,相关性分析表明,土壤nir K及nirS基因的丰度均与土壤PAHs含量呈显著负相关(nirK:R2=0.54,P0.05;nirS:R~2=0.58,P0.05).克隆文库及T-RFLP的结果则表明,该油田土壤中nirK基因的群落组成在不同样品间的变异较大,且PAHs含量最高的JH-4中该基因的群落组成与其它各样品有明显的不同,RDA(redundancy analysis)的分析结果进一步表明除有效氮、有效磷外,土壤PAHs含量也是影响nirK型反硝化微生物群落组成的重要因子.相较于nirK,该油田区土壤中nirS基因的群落组成在不同样品间的差异较小,但发现nirS型假单胞菌的丰度与土壤PAHs含量呈正相关,表明具备较强有机污染物降解能力的假单胞菌属可能在该区域土壤PAHs的反硝化代谢中起到重要作用.

Effects of PAHs Pollution on the Community Structure of Denitrifiers in a Typical Oilfield

Agricultural soils in the oilfields have the potential risk of PAHs (polycyclic aromatic hydrocarbons) pollution, and the denitrification process with nitrate as the terminal electron acceptor might be important for soil PAHs elimination under anaerobic condition. In this study, 9 soil samples listed as JH-1 to JH-9 were collected from the JiangHan oilfield with a history of more than 50 years. Using the functional genes (nirK: Cu-nitrite reductase gene; nirS: cd₁-nitrite reductase gene) involved in denitrification as biomarkers, the community structure of soil denitrifiers was investigated by quantitative-PCR and T-RFLP (terminal-restriction fragment length polymorphism) combined with clone library, and the relationship between soil properties and community structure of soil denitrifers was discussed. The result indicated that the copy numbers of nirK were higher than those of nirS in all soil samples, and the lowest copy numbers of nirK and nirS were both detected in the JH-4 with the highest PAHs content. Meanwhile, the correlation analysis also showed a negative correlation between the copy numbers of those functional genes and soil PAHs content (nirK: R²=0.54, P<0.05; nirS: R²=0.58, P<0.05). Furthermore, the result of T-RFLP indicated that the nirK community structures in different soil samples varied significantly, which was obviously unique in the sample (JH-4) with the highest PAHs content. The subsequent RDA (redundancy analysis) also demonstrated that soil PAHs content as well as the available nitrogen and phosphorus belonged to the most important factors affecting the nirK community structure in this oilfield soil. Compared with nirK, little variation was shown about the nirS community structure among the soil samples. However, the abundance of nirS-harboring pseudomonas had a remarkably positive relation with the soil PAHs content, which indicated that pseudomonas, a well known bacterial genus with strong ability to degrade organic pollutants, might be an essential driver for PAHs degradation via denitrification process in this oilfield soil.