利用PCR-DGGE分析未开发油气田地表微生物群落结构  

Microbial Community Structure Analysis of Unexploited Oil and Gas Fields by PCR-DGGE

作　　者：满鹏[1] 齐鸿雁[1] 呼庆[1] 马安周[1] 白志辉[1] 庄国强[1] 

机构地区：[1]中国科学院生态环境研究中心环境生物技术研究室,北京100085

出　　处：《环境科学》2012年第1期305-313,共9页Environmental Science

基　　金：国家自然科学基金项目(20977106);中国科学院知识创新工程项目(kzcx1-yw-06-03)

摘　　要：利用PCR-DGGE和克隆测序技术,分别对未开发油田、气田和非油气田对照区域,地表30、60、100、150、200 cm深度的土壤微生物分布进行研究,目的在于了解未开发油气田区域土壤微生物的分布特征,寻找潜在的油气资源指示菌.结果表明,不同深度土壤样品间的菌群相似度很低(26～69.9).在150 cm和200 cm处,DGGE图谱具有更好的丰富度(≥19)、多样性(≥2.69)和均匀度(≥0.90).油田区域富含γ-Proteobacteria(75%),此外还包括α-Proteobacteria、Actinobacteria、Acidobacteria,其相似菌株主要为石油相关菌和烃降解菌,如食烷菌、噬甲基菌.气田含有α、β、γ、δ-Proteobacteria和Bacteroidetes,γ-Proteobacteria含量较小(24%),所获16S rDNA条带相似菌株大多不具有烃降解能力,部分具有甲烷相关性,如甲基孢囊菌.在此类研究中,150 cm、200 cm更适于作为统一取样深度进行大范围取样调查;甲基孢囊菌可用来作为潜在的气田指示菌,食烷菌和噬甲基菌可作为潜在的油田指示菌,但仍需进一步地大范围取样验证.Microbial communities of different depths （30, 60, 100, 150, 200cm） from the unexploited oilfield, gas field and control area were studied by PCR-DGGE and sequencing methods. The objectives of this study were to understand the microbial distribution in the regions of unexploited oil and gas fields, and to investigate the potential microbial indicators of oil and gas resources. The results showed that the Dice coefficients between different depths were very low （26-69.9）. The microbial communities in the soil of 150 cm and 200 cm depth had greater richness （S≥19）, diversity （H≥2.69） and evenness （E≥0.90）. The results of sequencing demonstrated that the bands from oilfield were mainly grouped into α-Proteobacteria, γ-Proteobacteria, Actinobacteria, Acidobacteria with the predominance of γ-Proteobacteria（75%）. Most of the bands were related to oil-associated and hydrocarbon degrading bacteria, such as Methylophaga and Alcanivorax. While the gas field had α, β, γ, δ-Proteobacteria and Bacteroidetes, and γ-Proteobacteria accounted for only 24%. More strains showed relativity to methanotrophs, such as Methylocystaceae. Thus, 150 cm and 200 cm were more suitable as the oil-gas exploration sampling depth. Methylocystaceae may act as potential indicators for gas resources, Methylophaga and Alcanivorax for oil.

关 键 词：未开发油气田 PCR—DGGE 16S RDNA 微生物多样性 指示性微生物 

分 类 号：X172[环境科学与工程—环境科学]