Pseudomonas putida GM6的gfp基因标记及在活性污泥中的定殖  

gFP GENE-LABELING AND COLONIZATION OF PSEUDOMONAS PUTIDA GM6 IN ACTIVATED SLUDGE

作　　者：蔡天明[1] 管莉菠[1] 李顺鹏[1] 

机构地区：[1]南京农业大学生命科学学院微生物系

出　　处：《土壤学报》2007年第3期516-521,共6页Acta Pedologica Sinica

基　　金：国家自然科学基金项目(30500010;50308011)资助

摘　　要：强化生物除磷（EBPR）工艺在废水生物除磷中广为应用,但由于对EBPR工艺的微生物学和分子机理尚不了解,对大规模生活污水处理厂EBPR工艺处理效果突然恶化和运行不稳定等状况难以预测和控制。本研究利用本实验室分离鉴定的高效聚磷菌Pseudomonas putida GM6,来进行投菌试验以快速恢复和强化活性污泥的除磷能力。为了解GM6菌株在装置中的定殖情况,首先,通过三亲结合对Pseudomonas putida GM6成功进行了gfp基因标记;其次,将标记菌株GMTR投至实验室规模除磷效果不好的序批式反应器（SBR）装置中,研究GMTR在活性污泥中的定殖情况,投菌初期,GMTR占污泥细菌总数的1.5%～3%;至21d时,GMTR所占比例升至9.2%,GMTR在活性污泥中得以定殖。与此同时,研究了投菌后R2的聚磷能力变化,投菌5d后,磷的去除率逐渐提高,运行21d时磷的去除率升至96%,28d后出水浓度稳定在0.2mgL^-1左右。结果表明,Pseu-domonas putida GM6能快速启动强化废水生物除磷功能。这为下一步的大规模的工程化应用提供了科学依据。Although the enhanced biological phosphorus removal （EBPR） process is widely applied in wastewater treatment to remove phosphorus biologically, it is difficult to predict and control its effect, which is unstable and would often decline suddenly in large-scale municipal sewage treatment plants, on account of lack of knowledge about microbiological and molecular mechanisms of the EBPR process. An experiment was carried out using Pseudomoncts putida GM6, a strain of high-efficiency phosphorus-accumulating bacteria isolated and identified in the lab, to quickly reactivate the phosphorus removing capacity of sludge. In order to learn how GM6 colonize in the reactor, Pseudomonas putida GM6 was first labeled with gfp gene through triparent conjugation, and then introduced into a sequencing batch reactor （SBR） installation, low in phosphorus removing efficiency in the lab to investigate how GMTR colonize in activated sludge. The initial count of GMTR introduced into the sludge accounted for 1.5% ～ 3% of the total count of bacteria in the sludge, and rose up to 9.2% after 21 days, indicating GMTR colonized firmly in the sludge. At the same time, variation of phosphorus-accumulating capacity of R2 was studied after inoculation. It was found that phosphorus removal rate gradually increased 5 days after inoculation, and up to 96% at D 21. And phosphate concentration in the effluent lingered around 0.2 mg L^-1 after 28 days. The findings demonstrate that the strain of Pseudomonas putida GM6 is quick to initiate its biological phopshorus removing function in wastewater and may serve as scientific basis for large-scale engineering application.

关 键 词：GFP 定殖 序批式反应器(SBR) 强化生物除磷(EBPR) 

分 类 号：S154.39[农业科学—土壤学]