摘要:Thauera属菌是喹啉降解反应器中比例很高、降解能力强却又难以分离培养的一类细菌。本实验从上海交通大学分子生态学和生态基因组学实验室建立的以喹啉为唯一碳源的厌氧反硝化喹啉降解生物反应器中先后三次利用不同的培养基和培养条件分离微生物。利用Thauera特异性PCR的手段检测分离物中是否含有Thauera属菌。为提高灵敏度，结合采用了nested-PCR的方法进行扩增，对有Thauera阳性信号的样品利用变性梯度凝胶电泳进行纯菌鉴定；还利用ERIC-PCR方法对喹啉反应器的分离菌株进行了分析，并与反应器总DNA的ERIC图谱进行比较，结果本研究未能将反应器中具有优势地位的Thauera细菌分离培养成功，但本研究为进一步的Thauera细菌分离提供了借鉴。本论文还采用ERIC-PCR方法对喹啉降解反应器运行的不同时期样品进行了微生物群落的结构比较，获得了更多关于该反应器的群落结构信息。
关键词：Thauera，喹啉降解，分离培养，变性梯度凝胶电泳，ERIC
Abstract:Thauera genus has been known as one of the functionally important groups in the quinoline-degrading reactor. It occupies a high proportion of the microbial community, and is mostly shown high versatile organic substrate degrading capacity. However, it is very difficult to isolate and culture. In this experiment, different types of media and culture conditions have been designed to isolate microorganisms from the anaerobic denitrifying quinoline-degrading bioreactor in which carbon source is quinoline only for three times in the lab of molecular ecology and ecological genomics of Shanghai Jiao Tong University. Thauera-specific PCR was used to identify Thauera from the isolated samples. Nested-PCR was combined to raise the level of sensitivity. The samples with positive signals were tested with denaturing gradient gel electrophoresis (DGGE) method to confirm whether they were pure colonies or not. ERIC-PCR was used to analyze the samples from the reactor and the profiles were compared with that of the reactor’s total DNA. Although no isolate of Thauera genus, which is one of the dominant genera in the reactor, was successfully isolated, this work provides experience for further isolation. In this study, ERIC-PCR was also used to compare the structures of the microbial community using different reactor samples collected at different times and more information of the structure of the microbial community in the bioreactor was gained.
Key words: Thauera, quinolone degradation, isolation and culture, DGGE, ERIC
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