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실규모 하수처리 생물반응기에서 발견되는 암모니아산화균 군집조성 및 특징
박희등,Park, Hee-Deung 한국미생물학회 2009 미생물학회지 Vol.45 No.2
질소제거 하수고도처리공정에서 암모니아산화균은 질소제거에 핵심 역할을 하는 독립영양세균이다. 하수처리 생물반응기에는 다양한 암모니아산화균이 서식하며 군집조성도 시간에 따라 변화한다. 본 연구에서는 생물반응기의 운전인자 및 환경조건이 암모니아산화균 군집구조의 조성과 다양성에 영향을 미친다는 가설을 설정하였다. 이 가설을 검증하기 위해 질산화 반응이 활발한 포항, Palo Alto, Nine Springs, Marshall 하수처리장 활성슬러지 생물반응기로부터 암모니아산화균의 ammonia monooxygenase subunit A 유전자 clone library를 제작하였다. 하수처리 생물반응기에는 Nitrosomonas europaea, N. oligotropha, N.-like, Nitrosospira lineage에 속하는 암모니아산화균이 주로 발견되었으며, N. communis, N. marina, N. cryotolerans lineage에 속하는 암모니아산화균은 주종을 이루지 못했다. 암모니아산화균 군집조성은 하수처리장별로 차이를 보였는데, 포항, Palo Alto, Marshall 하수처리장에서는 N. oligotropha lineage에 속하는 암모니아산화균이 가장 빈번히 발견되었고, Nine Springs 하수처리장에서는 N. europaea lineage에 속하는 암모니아산화균이 주종을 이루었다. 한편, 암모니아산화균 군집조성과 생물반응기 운전인자(HRT, SRT, MLSS) 및 환경조건(온도, pH, COD, $NH_3$, $NO_3{^-}$)의 연관성은 다변수 통계분석법인 Redundancy Analysis 방법을 이용하여 분석하였다. 그 결과, 생물반응조의 COD와 $NO_3{^-}$ 농도가 하수처리 생물반응기에서 암모니아산화균 군집구조를 결정하는 통계학적으로 유의한 변수로 나타났다. Ammonia-oxidizing bacteria (AOB) are chemolithoautotrophs that play a key role in nitrogen removal from advanced wastewater treatment processes. Various AOB species inhabit and their community compositions vary over time in the wastewater treatment bioreactors. In this study, a hypothesis that operational and environmental conditions affect both the community compositions and the diversity of AOB in the bioreactors was proposed. To verify the hypothesis, the clone libraries based on ammonia monooxygenase subunit A were constructed using activated sludge samples from aerobic bioreactors at the Pohang, the Palo Alto, the Nine Springs, and the Marshall wastewater treatment plants (WWTPs). In those bioreactors, AOB within the Nitrosomonas europaea, N. oligotropha, N.-like, and Nitrosospira lineages were commonly found, while AOB within the N. communis, N. marina, and N. cryotolerans lineages were rarely detected in the samples. The AOB community structures were different in the bioreactors: AOB within the N. oligotropha lineage were the major microorganisms in the Pohang, the Palo Alto, and the Marshall WWTPs, while AOB within the N. europaea lineage were dominant in the Nine Springs WWTP. The correlations between the AOB community compositions of the wastewater treatment bioreactors and their operational (HRT, SRT, and MLSS) and environmental conditions (temperature, pH, COD, $NH_3$, and $NO_3{^-}$) were evaluated using a multivariate statistical analysis called the Redundancy Analysis (RDA). As a result, COD and $NO_3{^-}$ concentrations in the bioreactors were the statistically significant variables influencing the AOB community structures in the wastewater treatment bioreactors.
박희등(Hee Deung Park),박정구(Jeong Ku Park),이재선(Jae Seon Lee),심웅섭(Woong Seop Sim) 한국식물학회 1995 Journal of Plant Biology Vol.38 No.4
Binary vectors, pBI-ActR1, pBI-ActF1 and pBSH-ActR1, were constructed using pGA642, the replication origin of pTil2 and the rice actin promoter. The sizes of pBI-ActR1, pBI-ActF1 and pBSH-ActR1 were 12.9 kb, 13.2kb and 11.95 kb, respectively. These vectors containing a rice actin promoter followed by a GUS structural gene could induce strongly the expression of GUS gene in transformed rice cells. Rice explants form 3-4 day old seedlings after germination were cocultured with A. tumefaceins harboring pBI-ActR1, pBI-ActF1 or pBSH-ActR1, and then GUS expression in the explants was assayed. Transformation of rice explants by these binary vectors was tissue-specific, such that the meristematic regions of shoot apex, root and hypocotyl were transformed by these binary vectors.
16S rRNA 유전자 기반의 Pyrosequencing을 이용한 하수처리시설 생물반응기의 세균군집구조 분석
김택승,김한신,권순동,박희등,Kim, Taek-Seung,Kim, Han-Shin,Kwon, Soon-Dong,Park, Hee-Deung 한국미생물학회 2010 미생물학회지 Vol.46 No.4
서로 다른 처리공정으로 운영되는 4개의 하수시설을 대상으로 16S rRNA 유전자 기반의 pyrosequencing을 이용해 활성슬러지 하수처리 생물반응기의 세균군집구조를 분석하였다. 활성 슬러지에는 Rhodocyclales, Burkholderiales, Sphingobacteriales, Myxococcales, Xanthomonadales, Acidobacteria group 4, Anaerolineales, Methylococcales, Nitrospirales, Planctomycetales 목에 속하는 염기서열이 전체의 54-68%를 차지해, 소수의 세균 분류군이 활성슬러지 세균군집의 대부분을 차지하고 있었다. 이들 소수 세균 분류군의 조성은 처리장별로 차이가 있었으며, 하수처리장의 운전조건 및 환경조건에 영향을 받는 것으로 추측되었다. 또한, 활성슬러지는 매우 다양한 세균 종을 가지는 것으로 관찰되었는데(Chao1 richness estimate: 1,374-2,902 operational taxonomic units), 대부분의 다양성은 희귀 종에 기인한 것으로 나타났다. 특히, 분리막으로 운영되는 하수처리시설에서 높은 다양성을 나타내었는데, 처리공정이 매우 긴 고형물체류시간으로 운영되어 느리게 성장하는 다양한 세균이 서식하는데 용이하기 때문인 것으로 판단되었다. High-throughput pyrosequencing 기술을 이용하여 활성슬러지 세균군집을 처리장별로 비교 분석한 본 연구는 향후 활성슬러지 미생물의 생태학적 특성을 보다 잘 이해하고 하수처리공정을 개선하는데 도움이 될 것이다. Bacterial community composition in activated sludge wastewater treatment bioreactors were analyzed using 16S rRNA gene-based pyrosequencing for the four different wastewater treatment processes. Sequences within the orders Rhodocyclales, Burkholderiales, Sphingobacteriales, Myxococcales, Xanthomonadales, Acidobacteria group 4, Anaerolineales, Methylococcales, Nitrospirales, and Planctomycetales constituted 54-68% of total sequences retrieved in the activated sludge samples, which demonstrated that a few taxa constituted majority of the activated sludge bacterial community. The relative ratio of the order members was different for each treatment process, which was assumed to be affected by different operational and environmental conditions of each treatment process. In addition, activated sludge had very diverse bacterial species (Chao1 richness estimate: 1,374-2,902 operational taxonomic units), and the diversity was mainly originated from rare species. Particularly, the bacterial diversity was higher in membrane bioreactor than conventional treatment processes, and the long solids retention time of the operational strategy of the membrane bioreactor appeared to be appropriate for sustaining diverse slow growing bacteria. This study investigating bacterial communities in different activated sludge processes using a high-throughput pyrosequencing technology would be helpful for understanding microbial ecology in activated sludge and for improving wastewater treatment in the future.