应用PCR-DGGE技术解析MBR中微生物群落多样性

Application of PCR-DGGE to Analysis of Microbial Community Diversity in MBRs

为了揭示膜生物反应器中活性污泥的微生物多样性，取样处理不同水质的膜生物反应器中的活性污泥，通过细胞裂解直接提取其中的基因组DNA，以细菌通用引物进行16SrRNA基因V3高变异区域PCR扩增，再将PCR产物（约240bp）进行变性梯度凝胶电泳分离（变性剂梯度为30％～60％），获得表征污泥中微生物群落特征的DNA指纹图谱．研究表明，不同的膜生物反应器中既存在着共同的微生物种属也有各自特异的种属、相同的微生物种群在不同反应器中的优势地位不同，各自特有的微生物群落则是由于水质的不同经过长期演替而来．

In order to reveal microbial diversity of activated sludge in membrane bioreactor （ MBR ） , sludges in MBRs treating different wastewater were sampled. The genomic DNA of microbial communities was extracted directly by cell lysis .After the 16S rRNA genes （ V3 region ） were amplified by using the universal primers, the polymerase chain reaction （ PCR ） products separated by denaturing gradient gel electrophoresis （ DGGE ） were about 240 bp. The denaturant gradient was from 30% to 60%. The profile of DGGE showed that different sludges had different DNA finger printings. There exist some common and specific bands in both MBRs. The amount of the common microorganisms were different in the two MBRs. The specific microorganisms were gradually become the predominant communities after a long period of cultivation and acclimatization because of the different water quality.