基于实时荧光定量PCR技术的油气微生物勘探样品保存研究  

Storage Strategies of Soil Samples for Microbial Prospecting of Oil and Gas Based on Real-time Quantitative PCR

作　　者：王皖蒙[1,2] 许科伟[3] 任春[3] 杨帆[3] 李定龙[1] 贾仲君[2] 

机构地区：[1]常州大学环境与安全工程学院,江苏常州213164 [2]中国科学院南京土壤研究所,南京210008 [3]中国石化石油勘探开发研究院无锡石油地质研究所,江苏无锡214151

出　　处：《土壤》2013年第1期129-134,共6页Soils

基　　金：油气微生物勘探技术及应用研究项目(P11058)资助

摘　　要：针对一个油气藏和两个非油气藏上方的土壤剖面,围绕典型油气指示微生物甲烷氧化细菌,采用分子生态学技术比较了新鲜、自然风干和冷冻干燥3种处理下土壤中甲烷氧化细菌标靶基因pmoA的变化规律,研究了自然风干土壤能否用于油气资源微生物勘探。与新鲜土壤相比,自然风干和冷冻干燥显著降低了不同土壤剖面微生物总DNA含量,甲烷氧化细菌pmoA基因数量最多分别下降了90.7%和77.5%;然而,与非油气藏上方土壤剖面相比,自然风干和冷冻干燥处理后,油气藏上方不同土壤剖面依然检测到了大量的甲烷氧化细菌pmoA基因。本研究中,自然风干或冷冻干燥处理后的土壤适合于油气微生物勘探。Real-time quantitative PCR ofpmoA genes encoding methane monooxygenase of methanotrophs was performed in this stu dy to investigate whether air-dried soil could be used for microbial prospecting for oil and gas resources. Soil samples were collected from the depths of 0-30, 30-60, 60-100 and 100-150 cm from oil and gas fields and from two control fields where no oil and gas reservoirs were observed. The copy number ofpmoA genes was determined by qPCR in soil samples under fresh, air-dried and lyophilized conditions. The amount of DNA extract was apparently lower in air-dried and lyophilized soils than that in fresh soils, and the copy number ofpmoA genes decreased by 90.7% and 77.5%, respectively. Considerable amount ofpmoA genes was detected at soil depth down to 100 cm from oil and gas reservoirs, while pmoA gene was below detection limit in soils samples at depth lower than 30 cm soil samples from control fields. The results of this study suggest that air-dried or lyophilized soils could be used for microbial prospecting for oil and gas reservoirs in the field

关 键 词：油气资源 微生物勘探 自然风干处理 冷冻干燥处理 甲烷氧化细菌 

分 类 号：Q89[生物学] Q7