中文摘要：

目的优化高效甲苯降解菌驯化及筛选方法。方法分别选取啤酒厂、染料厂活性污泥作为驯化对象，采用4种方法分离纯化甲苯降解菌，利用质粒快检鉴定降解性TOL质粒，A600、顶空气相色谱法测定降解菌的生长与降解甲苯能力，筛选出一株高效甲苯降解菌，并利用革兰染色、形态学及16SrDNA分析方法对其进行鉴定。结果染料厂活性污泥较啤酒厂驯化出的细菌种类多；密闭罐培养降解茵生长良好；筛选出一株合有TOL质粒的高效降解菌，经鉴定为铜绿假单胞菌（Pseudomonas aeruginosa）。在28℃、pH7．0，初始甲苯浓度为500mg／L条件下，该菌48h降解率达到99．2％。结论密闭罐培养、质粒快检与顶空气相色谱有机结合在甲苯降解菌的驯化和筛选中具有快速、准确、效果好的特点。

英文摘要：

Objective To establish a highly effective toluene-degrading bacterium acclimation and screening method. Methods Samples were collected from activated sludge in brewery and dye plant. Toluene-degrading bacteria were screened by enrichment and acclimation process, and then were isolated and purified by four methods. TOL plasmid was determined by plasmid rapid detection method. Growths of degrading-bacteria were detected by A6co assay and toluene - degrading abilities were measured by head-space gas chromatography. The highly effective degrading bacteria were isolated and identified by the Gram staining, morphological observation and sequence analysis of 16S rDNA. Results More abundant species of degrading bacteria were isolated from activated sludge in brewery plant than those in the dye plant. Degrading bacteria grew well in a sealed-jar. The highly effective degrading bacterium was identified as Pseudomonas aeruginosa. 99.2%-Toluene were degraded in 48h with initial concentration of 500mg/L at 28℃ , pH 7.0. Conclusion The sealed-jar incubation in combination with plasmid rapid detection and head-space gas chromatography was.applied to identification of degrading bacterium and this method was accurate, time-saving and effective.