荧光定量PCR监测五氯酚对好氧颗粒污泥和活性污泥中氨氧化细菌数量的影响  

Real time PCR quantification of ammonia-oxidizing bacteria in aerobic granular sludge and activated sludge influenced by pentachlorophenol

作　　者：李光伟[1] 刘和[1] 张峰[2] 堵国成[1,2] 陈坚[1,2] 唐德友 李秀芬 

机构地区：[1]江南大学生物工程学院环境生物技术研究室,无锡214036 [2]江南大学工业生物技术教育部重点实验室,无锡214036 [3]不详

出　　处：《微生物学报》2007年第1期136-140,共5页Acta Microbiologica Sinica

基　　金：江苏省自然科学基金创新人才项目资助(BK2005402)~~

摘　　要：通过特异引物扩增环境中氨氧化细菌16S rDNA V2保守区域，将该片段克隆到T-easy载体上，PCR产物经测序和定量PCR扩增体系鉴定，证实PCR扩增产物为氨氧化细菌16S rDNA保守序列，以含该序列的重组质粒作为定量PCR监测氨氧化细菌数量的DNA标准品。用荧光定量PCR技术比较了五氯酚（PCP）对好氧颗粒污泥和活性污泥中氨氧化细菌数量的影响。结果表明，不加PCP的反应器中，好氧颗粒污泥和活性污泥中氨氧化细菌的数量分别为4．28×10^7±5．44×10^6cells／（g干污泥）和2．51×10^9±8．61×10^8cells／（g干污泥）。随着PCP浓度的增加（0—50mg／L），PcP对氨氧化细菌数量的影响不大（P〉0．05），而且，污泥中氨氧化细菌的数量与氨氮的去除率无直接的正相关关系（P〉0．05），PCP主要是抑制氨氧化细菌的代谢活性导致污泥氨氮去除效率降低。The V2 region of the 16S ribosomal DNA of the ammonia-oxidizing bacteria （AOB） was amplified directly from the environmental sample by using the specific PCR primers. The purified PCR product was cloned into T-vector and was identified as 16S rDNA fragment of AOB by sequencing and Real-time PCR method. Then, the recombined plasmid was used as standard molecule sample in Real-time PCR for AOB quantification. The numbers of the AOB were monitored in samples of both aerobic granular sludge and activated sludge influenced by PCP by using Real-time PCR. The results showed that the numbers of AOB in aerobic granular sludge and activated sludge were 4.28 × 10^7 ±5.44 ×10^6cells/g dried sludge and 2.51 × 10^9 ±8.61 × 10^8cells/g dried sludge without PCP in the reactors, respectively. With the increase of PCP concentration （from 0mg/L to 50mg/L）, the numbers of AOB in both types of sludge had no obvious change（ P 〉 0.05）. The numbers of AOB had no obvious correlation with ammonia removal （P 〉 0.05）. The main effect of PCP on AOB in both types of sludge was to inhibit their metabolic activity.

关 键 词：荧光定量PCR 好氧颗粒污泥 活性污泥 氨氧化细菌 五氯酚 

分 类 号：X703[环境科学与工程—环境工程]