PCR-DGGE解析好氧-沉淀-厌氧工艺微生物群落多样性

Study on microbial diversity of oxic-settling-anaerobic process using PCR-DGGE of 16S rDNA

为研究好氧-沉淀-厌氧(OSA)污泥减量工艺微生物群落结构和多样性,以及运行条件对微生物群落的影响,使用16S rDNA的PCR-DG-DGGE图谱分析方法结合条带割胶回收DNA序列比对,对OSA工艺微生物群落特征进行分析。DG-DGGE图谱聚类分析和多样性指数分析表明,OSA工艺微生物群落多样性比传统活性污泥工艺更加丰富。增加Ns和提高废水水质的复杂性,都能使Shannon指数略有提高,微生物多样性增加,但优势微生物种群基本不受影响。DG-DGGE图谱中优势条带序列分析表明,OSA工艺中的回收到的11个优势菌与α-proteobacte-ria,β-proteobacteria,CFB-group bacteria种属的微生物有很近的亲缘关系,β-proteobacteria约占63.6%。其中6个优势菌与强化生物除磷系统、反硝化系统中出现的微生物同源性非常高,证实了在OSA系统中存在的慢速生长微生物-反硝化菌和聚磷菌。

16S rDNA based PCR amplification and double gradient denaturing gradient gel electrophoresis（PCR-DG-DGGE） were used to monitor the microbial diversity in the oxic-settling-anaerobic（OSA） process for exploring the effect of operation conditions on microbial community structure.The DG-DGGE profiles and Shannon indices（SI） showed that there were more abundant communities in the OSA system than that in the conventional activated sludge（CAS） process.Shannon indices were increased slightly by increasing Ns or more complex wastewater quality.However,the predominant bacteria were not affected.11 sequences of excised dominant bands are affiliated with α-proteobacteria、β-proteobacteria、CFB-group bacteria,respectively.β-proteobacteria is prominent subclass,accounting for 63.6%.Six clones show relatively high similarity to uncultured bacteria isolated from the EBPR sludge and denitrifying sludge.The results showed there were slow growers-denitrifying bacteria and phosphorus accumulating organisms in the OSA process.