College of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510006, People's Republic of China.
Curr Microbiol. 2011 Feb;62(2):374-81. doi: 10.1007/s00284-010-9716-0. Epub 2010 Aug 3.
A 16S rRNA gene-based culture-independent approach was used to study the bacterial and archaeal communities in a sulfide-rich wastewater. Propidium Monoazide (PMA) treatment was applied to limit the analysis to the fraction of viable cells in environment. A total of 104 and 68 clones respective from bacterial clone library and archaeal library were picked and analyzed by restriction fragment length polymorphism (RFLP). 35 RFLP patterns from bacterial clone library and 10 RFLP patterns from archaeal clone library were unique and the respective clones were selected for sequencing. BLAST analysis and RFLP analysis showed that the bacterial clone library mainly consisted of Gammaproteobacteria (73%), Anaerolineae (6%), Bacilli (5%), Deltaproteobacteria (7%), Clostridia (4%), Bacteroidetes (1%), and Chlorobia (1%); Methanomicrobia (99%) and Thermococci (1%) were the only two lineages of the archaeal domains. This study gave a first insight into the overall microbial structure in a cloth printing and dyeing wastewater treatment plant with high concentration of sulfide and increased knowledge on the applicability of the PMA treatment in combination with PCR-based molecular techniques to analyze only viable cells in microbial ecology.
采用基于 16S rRNA 基因的培养非依赖性方法研究了富含硫化物的废水中的细菌和古菌群落。采用吖啶橙（PMA）处理来限制分析仅针对环境中存活细胞的分数。分别从细菌克隆文库和古菌文库中挑取并分析了 104 和 68 个克隆，通过限制性片段长度多态性（RFLP）。从细菌克隆文库中获得了 35 个 RFLP 模式，从古菌克隆文库中获得了 10 个 RFLP 模式，各自的克隆被选择进行测序。BLAST 分析和 RFLP 分析表明，细菌克隆文库主要由γ变形菌（73%）、产甲烷菌（99%）和Thermococci（1%）。本研究首次深入了解了高浓度硫化物的印花和染色废水处理厂中的整体微生物结构，并增加了关于在微生物生态学中仅分析存活细胞时，将 PMA 处理与基于 PCR 的分子技术相结合的适用性的知识。
