克隆文库方法分析厌氧氨氧化反应器中细菌群落结构

Analysis on Bacterial Diversity of an Anaerobic Ammonium-oxidizing Reactor by Use of 16S rDNA Clone Library

为了认识低基质浓度污水厌氧氨氧化(ANAMMOX)脱氮过程的生物学机制,为ANAMMOX脱氮工艺的优化提供理论依据,通过构建细菌16S rDNA(约1 500 bp)克隆文库和浮霉菌特有16S rDNA(约830 bp)克隆文库对ANAMMOX脱氮反应器中活性污泥的细菌群落结构进行分析。从细菌16S rDNA克隆文库中随机挑选到160个克隆子,共31个分类单元(OTU),与GenBank数据库比对结果表明,厌氧颗粒污泥中的细菌群落具有丰富的多样性,包括:变形菌门(Proteobacteria)、拟杆菌门(Bacteroidete)、硝化螺旋菌门(Nitrospira)、酸杆菌门(Acidobacteria)、绿弯菌门(Chloroflexi)、Candidate division OP10、浮霉菌门(Planctomycetes)和未知菌,其中,变形菌门和拟杆菌门为优势菌群,分别占41.9%和34.2%。在浮霉菌特有16S rDNA克隆文库的40个克隆子中,34个克隆子属于CandidatusKuenenia属的厌氧氨氧化细菌,它们是ANAMMOX脱氮过程的主要功能菌。

Anaerobic ammonium-oxidizing （ANAMMOX）de-nitrification reactor, featuring an activated sludge process o~ low substrate concentration, has been known as a cost/effective approach to de-nitrification of wastewater. To investigate its biological mechanism, a bacterial 16S rDNA clone（ 14 500 bp）library and a Planctomycetes-specific bacterial 16S rDNA （830 bp）library were constructed in laboratory focusing on the total and the functional bacterial diversity of activated sludge. The results indicated that bacterial diversity of the anaerobic granular sludge was very high with the communities including Proteobacteria, Bacteroidete, Nitrospira, Acidobacteria, Chloroflexi, Candidate division OP10 and Planctomycetes, inter alia, Proteobacteria and Bacteroidete were the dominant ones. Furthermore,34 clones of the 40 clones from the Planctomycetes-specific 16S rDNA library were proved to be anaerobic ammonium-oxidizing bacteria that belong to Candidatus Kuenenia, the main functional bacteria for ANAMMOX de-nitrification.