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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.
Four mediums of methanotrophic bacteria with different formulations were selected. Simultaneously, samples of oil and gas fields in the Fu’an block and background soil samples from the Sinopec Jiangsu Oilfield were collected and analyzed by plate counting, gene quantification and T-RFLP polymorphism fragment analysis The results showed that under different culture conditions, the amount of methanotrophs in soil samples and the abundance of methanotrophs pmo A gene in the soil samples were all higher than those in the background samples. Can distinguish between oil and gas area and the background area of the number of methanotrophs difference.The abnormal separation index shows that the No. 1 medium in the experimental detection of the abnormal number of microorganisms more highly T-RFLP polymorphism fragment analysis showed that the four medium culture conditions Under the diversity of methanotrophs, the relative abundance of methanogens and methylobacteria indicated slightly higher than that of No. 1 medium, which further proves that Methane 1 The specificity of the culture medium of the oxidative bacteria is strong.This study confirmed the reliability and accuracy of the No. 1 culture medium in the laboratory test and provided the technology for the exploration of oil and gas microbes support.