基于16S rDNA不同靶序列对厌氧ABR反应器微生物多样性分析的影响

在反硝化抑制硫酸盐还原菌的研究中,探讨了不同引物扩增16S rDNA靶序列对厌氧ABR反应器的活性污泥DGGE图谱多样性的影响,从反应器中提取污泥总DNA,以4对通用引物341F/534R、968F/1401R、63F/534R、341F/926R扩增16S rDNA序列,对指纹图谱的分辨率和种群多样性进行分析.研究表明,采用PBS洗涤污泥和超声振荡有利于硫酸盐还原污泥总DNA提取;不同引物DGGE图谱分析,群落多样性存在显著的差异,341F/534R和968F/1401R的靶序列分离效果较好,341F/926R分离效果一般,63F/534R分离的效果最差.341F/534R的DGGE图谱中条带丰富,多样性最好,968F/1401R的DGGE图谱次之,341F/926R的DGGE图谱条带一般,63F/534R图谱条带最少,多样性也较差.建议DGGE分析厌氧活性污泥样品时,同时采用引物341F/534R和968F/1401R是比较适宜的.

Influence of Different 16S rDNA Target Sequence on Analysis of Microbial Diversity in Anaerobic ABR Reactor

In the study on denitrification inhibiting sulfate reducing bacterium, it was discussed for the influence of 16S rDNA target sequence from different primers on DGGE fingerprinting from the anaerobic active sludge diversity in ABR reactor. The sludge DNA in the reactor was isolated, four sets primers 341F/534R, 968F/1401R, 63F/534R, and 341F/926R was used to amplify 16S rDNA, and the resolution of DGGE fingerprinting, community diversity were analyzed. The result indicated that it was beneficial to the DNA extraction from sulfate reducing sludge by PBS washing sludge and sonic oscillation; by analysis of DGGE from different primers, there were obvious differences in community diversity. The target sequences from primers 341F/534R and 968F/1401R were isolated relatively well, that from primers 341F/926R was in common, and that from primers 63F/534R was the worst. The DGGE fingerprinting bands from primers 341F/534R were abundant with the best diversity, while that from 968F/1401R was not as well as the former, that from 341F/926R was in common, and that from 63F/534R was the least with worse diversity than the others. Primers 341F/534R and 968F/1401R are recommended to be used simultaneously to analyze anaerobic active sludge by DGGE.