单级自养脱氮系统中厌氧氨氧化菌的分子生物学鉴定

Molecular Biology Identification of ANAMMOX Bacteria in Single-stage Autotrophic Nitrogen Removal System

对具有厌氧氨氧化作用的细菌进行更深入的分析有助于了解该菌在生物脱氮过程的应用。对稳定运行、氨氮转化率及总氮去除率分别达到90%及80%左右的单级自养脱氮系统的底部取活性污泥,采用分子生物学方法提取活性污泥细菌总DNA,利用特异引物Pla46rc/Amx820对单级自养脱氮系统中的厌氧氨氧化菌16S rDNA基因进行PCR扩增。扩增产物经克隆、测序及BLAST分析,结果表明该单级自养脱氮系统中存在的厌氧氨氧化菌与Candidatus Kueneniastuttgartiensis和Candidatus Brocadia anammoxidans的16S rDNA序列同源性达99%,进化分析证明与Candidatus Kuenenia stuttgartiensis进化上较为接近。

A deeper analysis of the role of anaerobic ammonium oxidation（ANAMMOX）bacteria will help to understand its application in the biological nitrogen removal.Using the single-stage autotrophic nitrogen removal system with stable operation and 90% of ammonia conversion rate and 80% of total nitrogen removal rate,the total DNA of cultivated sludge taken from the bottom of the system was extracted by molecular biology methods.With the total DNA partial 16S rDNA sequence of ANAMMOX bacteria was amplified by polymerase chain reaction（PCR）with a pair of specific primers Pla46rc/Amx820.Amplified product was cloned,sequenced and BLAST analysis and the result indicated that the sequence has 99% identities with ANAMMOX bacteria Candidatus Kuenenia stuttgartiensis and Candidatus Brocadia anammoxidans.The phylogenetic analysis showed that the anaerobic ammonium-oxidizing bacteria in the single-stage autotrophic nitrogen removal system have closer relationship with Candidatus Kuenenia stuttgartiensis evolutionarily.