PCR-DGGE分析啤酒废水生物处理工艺的微生物区系

应用基于16s rDNA的PER-DGGE(变性梯度凝胶电泳)技术对啤酒废水"水解酸化 SBR"工艺的微生物多样性进行研究.分别取水解酸化池与SBR池中不同深度以及不同处理时段的活性污泥,提取样品总DNA,通过PER扩增、变性梯度凝胶电泳,将16S rDNA(V3区)的PER扩增片段割胶克隆测序确定样品中的微生物群落,与筛选出的高效菌株进行对比分析.结果表明,水解酸化池中的微生物群落随深度的改变,在结构组成和种群数量上均有较大差异,2 m深处微生物群落相对丰富,优势条带突出;SBR池不同深度微生物种群结构一致,沉淀期、进水期和曝气期不同处理时段的微生物种类一致,但优势菌群不同;所测序列v2、23、25、31、h5和15号分别与菌株uncultured Thermotogales sp.Comamonas sp.WT OTU1、Agrobaaerium tumefaciens、Bacillus subtilis、Bdellovibrio bacteriovorus、Comamonas testosteroni有高度同源性,活性污泥样品中的优势条带与高效菌株的序列不同,表明筛选出的高效菌并非为实际处理过程中的优势菌.

Microbial Community Structure of Beer Wastewater Treatment Plants Analyzed by PCR-DGGE Technique

The microbial community structure of beer wastewater treatment plants with hydrolyze-acidification and SBR process was studied by PCR amplification and denaturing gradient gel electrophoresis (DGGE) based 16S rDNA. Activated sludge samples were collected from both hydrolyze-acidification tank and SBR tank at different depth and disposal period. The total DNA was extracted and the 16S rDNA was amplified by universal primer. The microbial community structure was analyzed by denaturing gradient gel electrophoresis, compared with the DGGE band pattern of the screened high efficient bacteria. The PCR products were cloned and sequenced to analyze the microbial community in phylogenic evolution. The DGGE pattern showed that the microbial community in hydrolyze-acidification tank changed with the variation of depth both in microbial structure and population, and at the depth of two-meter the microbial community was relatively rich, the dominant bands were notable. The microbial community in SBR tank at different depths and different disposal periods (precipitation period, inflow period, aeration period) were consistent respectively, but the dominant bands were different. Sequences of y2, 23, 25, 31, h5, 15 yielded high homogeneity with the strains uncultured Thermotogales sp., Comamonas sp. WT OTU1, Agrobacterium tumefaciens, Bacillus subtilis, Bdellovibrio bacteriovorus, Comamonas testosteroni from GenBank. The 16S rDNA sequences of the dominant bands were not completely same with that of the high efficient bacteria, which indicated that the strains screened were not the dominant strains in actual process.