中文摘要
本研究以454高通量测序法分析了高效喹啉降解反硝化反应器的微生物群落结构，并分离出1200多株细菌，通过菌落杂交等方法从中筛选出在群落内较优势的菌株，分析了它们的反硝化和喹啉降解能力，发现反应器中不仅具有高度的微生物类群多样性，而且代谢活性也呈多样化。反应器内反硝化菌、好氧非反硝化菌、喹啉降解菌和非降解菌共存。对以Thauera为最优势类群的微生物群落的分析方法进行了创新，建立了Thauera专一性PCR及PCR-DGGE方法。以该法对工业化微生物反应器Thauera类群进行了监测，并在该法指导下，从喹啉降解反应器中成功分离了３株Thauera属细菌。还以PCR-DGGE等方法比较了培养条件对微生物群落结构的影响，为选择更有利于分离获得高效优势细菌的条件奠定了基础。还分析了ERIC-PCR的特定条带作为群落功能菌的分子标记进行群落内重要功能菌的追踪、监测，及功能菌分离。本研究首次系统使用的以群落结构分析获得群落重要功能菌的分子标记，并以之为导向进行功能菌分离的方法（Probing assisted isolation of key player）在环境微生物领域将具有广泛应用。
英文摘要
Microbial community of high efficient denitrifying bioreactor was dissected by 454 high throughput sequencing, and 1200 strains of isolates were isolated. The dominant bacteria were screened by colonies hybridization, and their capacity of denitrification and quinoline degradation was determined. High bacterial phylogenetic and metabolism diversity were found. Due to the high abundance of Thauera existed in the bioreactor, we developed Thauera specific PCR and PCR-DGGE methods. By using these methods we monitored a industrial bioreactor and isolated 3 strains of thauera from quinoline degrading bioreactor. PCR-DGGE was used for the comparison of effects of cultivation condition on the microbial community, and consequently was used as the guild for isolation of key player from community. The specific fragment of ERIC-PCR can also be used as biomarker for the identification, monitor and isolation of key player. This is the first attempt of using the approach of obtaining biomarker of key player from microbial community by comparing the structure of microbial community, and then the biomarker was used for probing assisted isolation of key player.
结题摘要
本研究以454高通量测序法分析了高效喹啉降解反硝化反应器的微生物群落结构，并分离出1200多株细菌，通过菌落杂交等方法从中筛选出在群落内较优势的菌株，分析了它们的反硝化和喹啉降解能力，发现反应器中不仅具有高度的微生物类群多样性，而且代谢活性也呈多样化。反应器内反硝化菌、好氧非反硝化菌、喹啉降解菌和非降解菌共存。对以Thauera为最优势类群的微生物群落的分析方法进行了创新，建立了Thauera专一性PCR及PCR-DGGE方法。以该法对工业化微生物反应器Thauera类群进行了监测，并在该法指导下，从喹啉降解反应器中成功分离了３株Thauera属细菌。还以PCR-DGGE等方法比较了培养条件对微生物群落结构的影响，为选择更有利于分离获得高效优势细菌的条件奠定了基础。还分析了ERIC-PCR的特定条带作为群落功能菌的分子标记进行群落内重要功能菌的追踪、监测，及功能菌分离。本研究首次系统使用的以群落结构分析获得群落重要功能菌的分子标记，并以之为导向进行功能菌分离的方法（Probing assisted isolation of key player）在环境微生物领域将具有广泛应用。