PCR-DGGE法用于活性污泥系统中微生物群落结构变化的解析

Application of polymerase chain reaction-denaturing gradient gel electro-(phoresis) (PCR-DGGE) to the analysis of changes of microbial ecological communities in activated sludge systems

应用PCR- DGGE方法,对在相同的操作条件下分别用低温菌和常温菌接种的两套活性污泥系统中的微生物群落结构的动态变化进行了追踪。研究结果表明:由于工艺和操作条件相同,两系统的微生物群落结构的相似性随着运行时间的增加而增加。PCR-

Population dynamics in two activated sludge systems inoculated with seed sludge cultivated respectively at a low temperature ((5±1)℃) and normal temperature ((20±1)℃) were followed under the same operation conditions using the PCR-DGGE method. The genomic DNA was extracted from activated sludge using phenol-chloroform. PCR amplification of the extracted microbial 16S rRNA gene fragments was performed using primers GC341F and 518R. The result of agarose gel (1.8%) electrophoresis shows that the PCR products were about 230bp in length. Using PCR products, DGGE was performed on a DGGE analyzer with a 10% (w/v) loading acrylamide (39:1 acrylamide-bisacrylamide) gel under a 30% to 50% linear gradient of denaturant. Some bands appeared in all samples, while the others were specific only in specific samples. Despite the difference of inoculums, however, the bacterial community structures in the two systems were quite similar. The similarity of community structures in the two systems increased gradually from 41.6% (the first day), 50.2% (the fourth day) up to 59.3% (the twentieth day) with the prolongation of operational period. The PCR-DGGE method was a useful tool for the analysis of population dynamics in biological systems.