甲烷氧化菌培养基特异性分析  

Analysis on the specificity of methane-oxidizing bacteria media

作　　者：杨帆[1,2] 沈忠民[1] 梅泽 

机构地区：[1]成都理工大学,油气藏地质及开发工程国家重点实验室,成都610059 [2]中国石化石油勘探开发研究院无锡石油地质研究所,无锡214151 [3]江南大学环境与土木工程学院,无锡214112

出　　处：《应用与环境生物学报》2016年第6期1127-1133,共7页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金项目(41202241)资助~~

摘　　要：选取4种不同配方的甲烷氧化菌培养基,同时采集中石化江苏油田富安区块油气区和背景区地表土壤样品,利用平板计数法、基因定量和T-RFLP多态性片段分析技术分析和评估其培养效果.实验结果表明,在不同配方培养基培养条件下,油气区土壤样品中甲烷氧化菌数量及扩培后的甲烷氧化菌pmo A基因丰度均高于背景区样品,4种培养基均能区分油气区和背景区甲烷氧化菌数量差异.异常分离指数表明1号培养基在实验检测中对微生物数量异常分离度更高.T-RFLP多态性片段分析表明,4种培养基培养条件下的甲烷氧化菌多样性程度均较高,1号培养基对应的优势油气指示微生物甲基球菌和甲基暖菌相对丰度略高,从甲烷氧化菌群落多样性角度进一步证明了1号甲烷氧化菌培养基的特异性较强.本研究确认了1号培养基在实验检测中的可靠性和准确性,为油气微生物勘探提供了技术支撑.This research tried to analyze and evaluate four methane oxidizing bacteria culture medium with different formula using plate count method, quantitative gene technology and T-RFLP analysis. We collected soil samples of oil-gas area and background area in Fuan Block. The experimental result showed that both the number of methane oxidizing bacteria and its associated pmoA gene abundance in bacterial liquid were higher in oil-gas field soil sample than in background soil sample under different media culture conditions. The difference in number of methane oxidizing bacteria between oil-gas field soil and background field soil was independent of the culture medium. ASI demonstrated that microbial anomalies could be easily identified with No.1 culture medium. T-RFLP analysis showed high biodiversity of methane oxidizing bacteria with all four culture media. Under the culture condition of No.1 culture medium, the abundance of metbylococcus and methylocaldum, the two dominant oil-gas indicator microorganisms, was slightly higher than other communities, indicating the effectiveness of No.1 culture medium. This study confirms the reliability and accuracy of No.1 culture medium, and provides technical support for microbial prospecting of oil and gas.

关 键 词：培养基 平板计数法 甲烷氧化菌 pmoA基因 多样性分析 

分 类 号：P618.13[天文地球—矿床学] P632.4[天文地球—地质学]