生物转鼓过滤器反硝化去除NO过程中微生物群落结构多样性解析  

Microbial Diversity Analysis in Rotating Drum Biofilter for Nitric Oxide Denitrifying Removal

作　　者：陈浚[1] 王智晔[2] 蒋轶锋[1] 钱海丰[1] 沙昊雷[1] 陈建孟[1] 

机构地区：[1]浙江工业大学生物与环境工程学院,杭州310032 [2]浙江大学生命科学学院,杭州310058

出　　处：《环境科学》2008年第4期1092-1098,共7页Environmental Science

基　　金：国家高技术研究发展计划(863)项目(2002AA649310);国家自然科学基金项目(20576124,20776134)

摘　　要：采用PCR-DGGE技术分析了生物转鼓过滤器(RDB)反硝化去除NO过程中的微生物多样性.研究结果表明,RDB中存在16种优势菌,并且沿填料径向不同位置的DGGE图谱差异性不大,细菌群落结构具有较高相似性.通过DGGE图形比较和微生物群落生物多样性指数计算,发现在RDB内加入CuⅡ(EDTA)络合剂后,NO去除率上升,而微生物群落多样性先增加后又下降,但群落结构变化不显著.对DGGE图谱中8条主带进行回收、扩增、克隆和测序,结果显示RDB中微生物群落主要由Cytopahga-Flexibacteria-Bacteroides(CFB)group Bacteroides、β-Proteobacterium、γ-Proteobacterium和Clostridiumsp.组成.反硝化功能与条带G-5(属于γ-Proteobacterium)和G-6、G-8(属于β-Proteobacterium)所代表的菌种相关,相似性≥98%.G-3序列与GenBank数据库中2个最相似序列的相似性只有93%和92%,表明这个条带所代表的微生物可能为新的未培养微生物.PCR-DGGE was applied to analyze the microbial communities in rotating drum biofilter（RDB） for nitric oxide denitrifying removal under anaerobic conditions.The 16S rRNA genes（V3 region） were amplified with two universal primers （338F-GC and 518R）.These amplified DNA fragments were separated by parallel DGGE.The DGGE profiles of biofilm samples from different sections of RDB are similar and about sixteen types of microorganisms are identified in the biofilm samples.These results show that microbial diversity of RDB firstly increases but then decreases with the addition of Cu^Ⅱ（EDTA） and the increase of NO removal efficiency.However,the change of microbial community is not significant during the process.Eight major bands of 16S rRNA genes fragments from DGGE profiles of biofilm samples were further eluted from gel,re-amplified,cloned and sequenced.The sequences of these fragments were compared with the database in GeneBank（NCBI）.The gene analysis of 16S rRNA showed that the major populations are Cytopahga-Flexibacteria-Bacteroides（CFB） group Bacteroides,（β-Proteobacterium,） γ-Proteobacterium and Clostridium sp..In addition,denitrification is related with band G-5,G-6 and G-8.G-5 is affiliated with γ-Proteobacterium. Both G-6 and G-8 are affiliated with β-Proteobacterium.

关 键 词：生物转鼓过滤器 微生物多样性 变性梯度凝胶电泳(DGGE) 16SrRNA 序列测定 

分 类 号：X172[环境科学与工程—环境科学]