基于高通量测序技术的老化油泥对大庆油田土壤 细菌多样性的影响

全文下载：   20240307.pdf

文章编号： 1672-6987（2024）03-0060-09； DOI： 10.16351/j.1672-6987.2024.03.007

王卉卉， 匡少平， 王炳辰（青岛科技大学 环境与安全工程学院， 山东 青岛 266042）

摘要： 采集大庆油田不同泥龄老化油泥污染土壤样本，采用高通量测序技术对样本细菌多样性特征开展研究，结合土壤理化性质分析，揭示老化油泥对大庆油田土壤细菌多样性的影响效应，并筛选确定样本中的优势特色耐油菌，为老化油泥污染土壤的原位微生物修复提供理论依据和数据支持。理化性质分析结果表明，污染土壤pH值下降，由碱性（844）变为中性（747~785），总有机碳（TOC，2012~2835 g·kg-1）和总石油烃（TPHs，2 475~2 955 mg·kg-1）含量均显著升高，有效磷（AP）和有效氮（AN）急剧降低。Illumina Miseq高通量测序结果显示，老化油泥对土壤细菌多样性和群落结构具有显著影响。土壤受老化油泥污染后，厚壁菌减少了约85%，变形菌（Proteobacteria）和芽单胞菌（Gemmatimonadetes）的相对丰度显著增加至4663%±465%和1411%±153%，且污染时间越长，对变形菌的促进作用越大。细菌科和属水平凸显了Woeseiaceae（γ-变形菌）和JTB255-MBG（变形菌门-Woeseiaceae菌科）的丰度增长，且其相对丰度与油泥污染时间呈显著正相关。高通量测序结果揭示，老化油泥污染一方面抑制了土壤中许多重要细菌的生长，另一方面也衍生和促进了具有耐油特性的其他细菌的生长发育。其中，变形菌门是石油污染土壤中最主要的优势菌门，它与所包含的耐油细菌科、属形成了“变形菌门-Woeseiaceae菌科-JTB255-MBG菌属”的优势耐油细菌群落结构。本研究成果可为老化油泥污染土壤的原位微生物修复提供理论依据与支持。

关键词： 高通量测序； 细菌多样性； 老化油泥； 土壤

中图分类号： X 122文献标志码： A

引用格式： 王卉卉， 匡少平， 王炳辰. 基于高通量测序技术的老化油泥对大庆油田土壤细菌多样性的影响［J］. 青岛科技大学学报(自然科学版), 2024, 45（3）： 60-68.

WANG Huihui, KUANG Shaoping, WANG Bingchen. Bacterial community structure of aged oil sludge contaminated soil in daqing oil field revealed by illumina high-throughput sequencing ［J］. Journal of Qingdao University of Science and Technology（Natural Science Edition), 2024， 45（3）： 60-68.

Bacterial Community Structure of Aged Oil Sludge Contaminated Soil in

Daqing Oil Field Revealed by Illumina High-throughput Sequencing

WANG Huihui, KUANG Shaoping, WANG Bingchen

(College of Environment and Safety Engineering,Qingdao University of Science and Technology, Qingdao 266042, China)

Abstract: In this study, aged oil sludge contaminated soil samples were collected from Daqing oil filed and characteristics of bacterial diversity in the samples were studied by High-Throughput Sequencing method. Combined with the analysis of soil physical and chemical properties, bacterial community structures and the dominant or core oil resistant bacteria in the oil contaminated soils were obtained. The results showed that the pH value of the contaminated soil decreased from alkaline (844) to neutral (747—785).The contents of total organic carbon (TOC, 2012—2835 g·kg-1) and total petroleum hydrocarbons (TPHs, 2 475—2 955 mg·kg-1) significantly increased, while available phosphorus (AP) and available nitrogen (AN) decreased sharply. The internal transcribed spacer (ITS) sequencing by Illumine Miseq platform at each taxonomic level revealed notable toxicological effect of aged oil sludge on soil bacterial diversity and community structure. Sequence analysis showed 85% reduction of Firmicutes and the relative abundance of Proteobacteria and Gemmatimonadetes increased to 4663%±465% and 1411%±153%, respectively. The relative abundance of Proteobacteria showed strong positive correlation with pollution time. Woeseiaceae (γ-Proteobacteria) and JTB255-MBG (Proteobacteria-Woeseiaceae) are core bacterial at family and genus level. Results of high-throughput sequencing revealed that the aged oil contaminaiton not only inhibited the growth of many important bacteria in soil, but also promoted or developed the growth of other bacteria with oil resistivity. Proteobacteria is the most dominant phyla in oil-contaminated soils. And “Proteobacteria-Woeseiaceae-JTB255-MBG” is the oil-resistant bacteria system formed in aged oil contaminated soils. Results of this study can provide theoretical basis and support for in situ bioremediation of oil contaminations in soil.

Key words: high throughput sequencing; bacterial diversity; aged oil sludge; soil

收稿日期： 2023-05-04

基金项目： 山东省重大科技创新工程项目（2021CXGC011206）.

作者简介： 王卉卉（1984—），女，讲师.