密云水库入库河流水体总细菌和反硝化菌群落组成与结构
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摘要
研究密云水库入库河流水体中总细菌和反硝化菌的群落组成与差异性,为水库的安全管理与水质保障提供基础数据和科学依据。选择对密云水库水质影响最大的5条入库河流(潮河、白河、清水河、对家河和白马关河),对水体中的总细菌和氮循环功能微生物反硝化菌进行了聚合酶链式反应(PCR)扩增与高通量测序分析。5条入库河流中全细菌主要隶属于6个门,而以Proteobacteria(变形菌门)为优势种群;反硝化菌在门水平的分布则相对简单,主要以Proteobacteria和少量Deferribacteres(脱铁杆菌门)构成。不同入库河流全细菌的群落结构具有空间差异,其中水文特征较为接近的白河和潮河全细菌群落结构具有较高相似性,与其他3条入库河流群落差别较大;水体中全细菌的Shannon多样性和Chao1丰富度均表现出较高水平。氮磷污染较重的对家河和潮河反硝化菌群落结构比较类似,其他3条入库河流之间则相对分散;对家河全细菌和反硝化菌的多样性和丰富度表现出较低水平。清水河水体中的全细菌和反硝化菌群落结构都较少受到已分析的环境因子影响,可能与该河道的人工改造有关。河流水体中全细菌以及关键功能微生物群落特征受到河流的水文、形态和理化指标的综合影响,可以作为水体环境与生态状况的重要指示项目,在今后的水库水环境保护与管理中应予以重视。
Miyun Reservoir is the largest surface drinking water source for Beijing City.Precipitation recharge and water diversion from south to north contribute,but inflow from tributaries is the primary source of water to Miyun Reservoir and the primary determinant of reservoir water quality.The microbial community of rivers is an important indicator of water quality and ecological succession.To provide basic data and a scientific basis for reservoir management and water quality protection,we analyzed the community structure of the total bacteria and denitrifying bacteria in Miyun Reservoir tributaries by polymerase chain reaction(PCR)amplification and high throughput sequencing technology.Correlation of the microbial community with environmental parameters was examined using principal component analysis(PCA).On April 19,2016,water samples for microbial analysis were collected from five tributaries(Chaohe River,Baihe River,Qingshui River,Duijia River and Baimaguan River)and physicochemical parameters for each sampling site were determined.The bacteria collected belonged primarily to six phyla(Proteobacteria,Bacteroidetes,Cyanobacteria,Actinobacteria,Firmicutes,Verrucomicrobia),with absolute dominance by Proteobacteria in all five tributaries.Denitrifying bacteria sequences were in two phyla,Proteobacteria and a small fraction from Deferribacteres.The bacterial and denitrifying bacterial community structure presented obvious spatial variation.Baihe River and Chaohe River,with similar hydrological properties,displayed high similarity in bacterial composition and were very different from the other three rivers.The Shannon diversity and Chao1 richness of the bacterial communities in Baihe River and Chaohe River were high,but low in Duijia River.Duijia River and Chaohe River,with high pollutant loading of N and P,exhibited similar compositions of denitrifying bacteria.PCA results indicate that pH,TOC,NH_3-N and Cl-were the primary environmental parameters affecting the bacterial community,while DO,Chla,TP and NH_3-N were the primary environmental parameters affecting the denitrifying bacteria community.However,neither bacterial community correlated well with environmental variables in Qingshui River.The community structure of bacteria in the five tributaries is affected by a combination of hydrological and physicochemical properties and is a good indicator of water quality and trophic status.We recommend that bacterial monitoring be given more attention in reservoir management.
Miyun Reservoir is the largest surface drinking water source for Beijing City.Precipitation recharge and water diversion from south to north contribute,but inflow from tributaries is the primary source of water to Miyun Reservoir and the primary determinant of reservoir water quality.The microbial community of rivers is an important indicator of water quality and ecological succession.To provide basic data and a scientific basis for reservoir management and water quality protection,we analyzed the community structure of the total bacteria and denitrifying bacteria in Miyun Reservoir tributaries by polymerase chain reaction(PCR)amplification and high throughput sequencing technology.Correlation of the microbial community with environmental parameters was examined using principal component analysis(PCA).On April 19,2016,water samples for microbial analysis were collected from five tributaries(Chaohe River,Baihe River,Qingshui River,Duijia River and Baimaguan River)and physicochemical parameters for each sampling site were determined.The bacteria collected belonged primarily to six phyla(Proteobacteria,Bacteroidetes,Cyanobacteria,Actinobacteria,Firmicutes,Verrucomicrobia),with absolute dominance by Proteobacteria in all five tributaries.Denitrifying bacteria sequences were in two phyla,Proteobacteria and a small fraction from Deferribacteres.The bacterial and denitrifying bacterial community structure presented obvious spatial variation.Baihe River and Chaohe River,with similar hydrological properties,displayed high similarity in bacterial composition and were very different from the other three rivers.The Shannon diversity and Chao1 richness of the bacterial communities in Baihe River and Chaohe River were high,but low in Duijia River.Duijia River and Chaohe River,with high pollutant loading of N and P,exhibited similar compositions of denitrifying bacteria.PCA results indicate that pH,TOC,NH_3-N and Cl-were the primary environmental parameters affecting the bacterial community,while DO,Chla,TP and NH_3-N were the primary environmental parameters affecting the denitrifying bacteria community.However,neither bacterial community correlated well with environmental variables in Qingshui River.The community structure of bacteria in the five tributaries is affected by a combination of hydrological and physicochemical properties and is a good indicator of water quality and trophic status.We recommend that bacterial monitoring be given more attention in reservoir management.
引文
北京市水务局,2017.北京市水务统计年鉴(2016)[M].北京:中国水利水电出版社.
康丽娟,曹勇,付融冰,等,2016.泵站放江对景观河道微生物指标的影响[J].水生态学杂志,37(1):47-52.
寇文伯,黄正云,张杰,等,2015.鄱阳湖湖泊细菌群落组成及结构---以松门山为例[J].生态学报,35(23):7608-7614.
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王敏,李玉仙,常思博,等,2016.密云水库水源水微生物群落解析[J].中国给水排水,32(21):63-65.
原媛,汪善全,竺建荣,等,2007.密云水库微生物相变化对水质及嗅味的影响[J].环境科学研究,20(6):8-13.
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Burgin A J,S K Hamilton,2007.Have we overemphasized the role of denitrification in aquatic ecosystems-A review of nitrate removal pathways[J].Frontiers in Ecology and the Environment,5(2):89-96.
Edgar R C,2010.Search and clustering orders of magnitude faster than BLAST[J].Bioinformatics,26(19):2460.
Fuhrman J A,2009.Microbial community structure and its functional implications[J].Nature,459(7244):193-199.
James E,M Joyce,2004.Assessment and Management of Watershed Microbial Contaminants[J].Critical Reviews in Environmental Science and Technology,34(2):109-139.
Jetten M S M,2008.The microbial nitrogen cycle[J].Environmental Microbiology,10(11):2903-2909.
Lindstr9m E S,M Forslund,G Algesten,et al,2006.External control of bacterial community structure in lakes[J].Limnology and Oceanography,51(1):339-342.
Magoc T,S L Salzberg,2011.FLASH:fast length adjustment of short reads to improve genome assemblies[J].Bioinformatics,27(21):2957-2963.
Mori H,F Maruyama,H Kato,et al,2013.Design and Experimental Application of a Novel Non-Degenerate Universal Primer Set that Amplifies Prokaryotic 16SrRNAGenes with a Low Possibility to Amplify Eukaryotic rRNA Genes[J].DNA Research,21(2):217-227.
Pujalte M J,T Lucena,M A Ruvira,et al,2014.The Family Rhodobacteraceae[M].Berlin Heidelberg:Springer.
Reddy K,W Patrick J,C Lindau,1989.Nitrification-denitrification at the plant root-sediment interface in wetlands[J].Limnology and Oceanography,34(6):1004-1013.
Rosch C,A Mergel,H Bothe,2002.Biodiversity of Denitrifying and Dinitrogen-Fixing Bacteria in an Acid Forest Soil[J].Applied and Environmental Microbiology,68(8):3818-3829.
Tago Y,A Yokota,2004.A floe-forming bacterium isolated from activated sludge[J].Journal of General&Applied Microbiology,50(5):243-248.
Tiedje J M,1988.Ecology of denitrification and dissimilatory nitrate reduction to ammonium[J].Biology of anaerobic microorganisms,717:179-244.
康丽娟,曹勇,付融冰,等,2016.泵站放江对景观河道微生物指标的影响[J].水生态学杂志,37(1):47-52.
寇文伯,黄正云,张杰,等,2015.鄱阳湖湖泊细菌群落组成及结构---以松门山为例[J].生态学报,35(23):7608-7614.
任丽娟,何聃,邢鹏,等,2013.湖泊水体细菌多样性及其生态功能研究进展[J].生物多样性,21(4):421-432.
王敏,李玉仙,常思博,等,2016.密云水库水源水微生物群落解析[J].中国给水排水,32(21):63-65.
原媛,汪善全,竺建荣,等,2007.密云水库微生物相变化对水质及嗅味的影响[J].环境科学研究,20(6):8-13.
朱亮,孙凌宇,储如花,等,2013.城市纳污河流沉积物微生物群落结构特征[J].水科学进展,24(1):132-137.
赵荫薇,蔡妙英,钱兵,等,1986.城市河流的微生物生态研究[J].微生物学报,26(3):12-17.
Abell G C J,D J Ross,J P Keane,et al,2013.Nitrifying and denitrifying microbial communities and their relationship to nutrient fluxes and sediment geochemistry in the Derwent Estuary,Tasmania[J].Aquatic Microbial Ecology,70(1):63-75.
Burgin A J,S K Hamilton,2007.Have we overemphasized the role of denitrification in aquatic ecosystems-A review of nitrate removal pathways[J].Frontiers in Ecology and the Environment,5(2):89-96.
Edgar R C,2010.Search and clustering orders of magnitude faster than BLAST[J].Bioinformatics,26(19):2460.
Fuhrman J A,2009.Microbial community structure and its functional implications[J].Nature,459(7244):193-199.
James E,M Joyce,2004.Assessment and Management of Watershed Microbial Contaminants[J].Critical Reviews in Environmental Science and Technology,34(2):109-139.
Jetten M S M,2008.The microbial nitrogen cycle[J].Environmental Microbiology,10(11):2903-2909.
Lindstr9m E S,M Forslund,G Algesten,et al,2006.External control of bacterial community structure in lakes[J].Limnology and Oceanography,51(1):339-342.
Magoc T,S L Salzberg,2011.FLASH:fast length adjustment of short reads to improve genome assemblies[J].Bioinformatics,27(21):2957-2963.
Mori H,F Maruyama,H Kato,et al,2013.Design and Experimental Application of a Novel Non-Degenerate Universal Primer Set that Amplifies Prokaryotic 16SrRNAGenes with a Low Possibility to Amplify Eukaryotic rRNA Genes[J].DNA Research,21(2):217-227.
Pujalte M J,T Lucena,M A Ruvira,et al,2014.The Family Rhodobacteraceae[M].Berlin Heidelberg:Springer.
Reddy K,W Patrick J,C Lindau,1989.Nitrification-denitrification at the plant root-sediment interface in wetlands[J].Limnology and Oceanography,34(6):1004-1013.
Rosch C,A Mergel,H Bothe,2002.Biodiversity of Denitrifying and Dinitrogen-Fixing Bacteria in an Acid Forest Soil[J].Applied and Environmental Microbiology,68(8):3818-3829.
Tago Y,A Yokota,2004.A floe-forming bacterium isolated from activated sludge[J].Journal of General&Applied Microbiology,50(5):243-248.
Tiedje J M,1988.Ecology of denitrification and dissimilatory nitrate reduction to ammonium[J].Biology of anaerobic microorganisms,717:179-244.