基于不同测序技术的生物群落结构及功能菌分析
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摘要
分别采用克隆文库和高通量测序技术,解析生物去除铁锰氨滤池内微生物的群落结构和功能菌多样性,并探讨不同测序手段的差异.高通量测序获得15057条有效序列、共32个分类纲,克隆文库测序涵盖9个具有明确分类地位的纲(75条序列),前者能揭示更为丰富的细菌群落结构多样性.功能菌(铁锰氧化细菌和硝化细菌)分析过程中,一些功能菌属在克隆文库中出现,而在高通量测序中未检测到,反之亦然.与单一的测序手段相比,二者相结合能更好地揭示功能细菌的分布特点.
Effects of the technologies of clone library and high throughout sequencing for revealing microbial communitystructure and functional bacteria in a biofilter which simultaneously removes iron, manganese and ammonia were compared. 15057 sequences with 32 classifications in class level were obtained after the high throughout sequencing process, while there are 9classified groups in clone library. The former revealed more diversity of bacterial community structure. However, in the functional bacteria analysis process, some nitrifiers or iron and manganese bacteria which were detectable in one sequencing analysis process were not appeared in the other process. Comparing with using these two technologies individually in the analysing process, the combined using can compensate for each other and reveal the functional bacteria better.
Effects of the technologies of clone library and high throughout sequencing for revealing microbial communitystructure and functional bacteria in a biofilter which simultaneously removes iron, manganese and ammonia were compared. 15057 sequences with 32 classifications in class level were obtained after the high throughout sequencing process, while there are 9classified groups in clone library. The former revealed more diversity of bacterial community structure. However, in the functional bacteria analysis process, some nitrifiers or iron and manganese bacteria which were detectable in one sequencing analysis process were not appeared in the other process. Comparing with using these two technologies individually in the analysing process, the combined using can compensate for each other and reveal the functional bacteria better.
引文
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[12]Tekerlekopoulou A G,Vayenas D V.Ammonia,iron and manganese removal from potable water using trickling filters[J].Desalination,2007,210(1-3):225-235.
[13]Michalakos G D,Nieva J M,Vayenas D V,et al.Removal of iron from potable water using a trickling filter[J].Water Research,1997,31(5):991-996.
[14]Hope C K,Bott T R.Laboratory modelling of manganese biofiltration using biofilms of Leptothrix discophora[J].Water Research,2004,38(7):1853-1861.
[15]Burger M S,Krentz C A,Mercer S S,et al.Manganese removal and occurrence of manganese oxidizing bacteria in full-scale biofilters[J].Journal of Water Supply Research and Technology,2008,57(5):351-359.
[16]Carmichael M J,Carmichael S K,Santelli C M,et al.Mn(II)-oxidizing bacteria are abundant and environmentally relevant members of ferromanganese deposits in caves of the upper Tennessee River Basin[J].Geomicrobiology Journal,2013,30(9):779-800.
[17]Miller A Z,Hernández-MarinéM,Jurado V,et al.Enigmatic reticulated filaments in subsurface granite[J].Environmental Microbiology Reports,2012,4(6):596-603.
[2]刘涛.基于亚硝化的全程自养脱氮工艺(CANON)效能及微生物特征研究[D].哈尔滨:哈尔滨工业大学,2013.
[3]Regan J M,Harrington G W,Noguera D R.Ammonia-and nitrite-oxidizing bacterial communities in a pilot-scale chloraminated drinking water distribution system[J].Applied and Environmental Microbiology,2002,68(1):73-81.
[4]Siering P,Ghiorse W.Development and application of 16S r RNA-targeted probes for detection of iron-and manganeseoxidizing sheathed bacteria in environmental samples[J].Applied and Environmental Microbiology,1997,63(2):644-651.
[5]Francis C A,Tebo B M.cum A multicopper oxidase genes from diverse Mn(II)-oxidizing and non-Mn(II)-oxidizing Pseudomonas strains[J].Applied and Environmental Microbiology,2001,67(9):4272-4278.
[6]蔡言安,李冬,曾辉平,等.生物滤池净化含铁锰高氨氮地下水试验研究[J].中国环境科学,2014,34(8):1993-1997.
[7]Li X K,Chu Z R,Liu Y J,et al.Molecular characterization of microbial populations in full-scale biofilters treating iron,manganese and ammonia containing groundwater in Harbin,China[J].Bioresource Technology,2013(147):234-239.
[8]Regan J M,Harrington G W,Baribeau H,et al.Diversity of nitrifying bacteria in full-scale chloraminated distribution systems[J].Water Research,2003,37(1):197-205.
[9]Liu T,Li D,Zeng H,et al.Distribution and genetic diversity of functional microorganisms in different CANON reactors[J].Bioresource Technology,2012(123):574-580.
[10]Alawi M,Lipski A,Sanders T,et al.Cultivation of a novel cold-adapted nitrite oxidizing betaproteobacterium from the Siberian Arctic[J].ISME Journal,2007,1(3):256-264.
[11]鲍林林,陈永娟,王晓燕.北运河沉积物中氨氧化微生物的群落特征[J].中国环境科学,2015,35(1):179-189.
[12]Tekerlekopoulou A G,Vayenas D V.Ammonia,iron and manganese removal from potable water using trickling filters[J].Desalination,2007,210(1-3):225-235.
[13]Michalakos G D,Nieva J M,Vayenas D V,et al.Removal of iron from potable water using a trickling filter[J].Water Research,1997,31(5):991-996.
[14]Hope C K,Bott T R.Laboratory modelling of manganese biofiltration using biofilms of Leptothrix discophora[J].Water Research,2004,38(7):1853-1861.
[15]Burger M S,Krentz C A,Mercer S S,et al.Manganese removal and occurrence of manganese oxidizing bacteria in full-scale biofilters[J].Journal of Water Supply Research and Technology,2008,57(5):351-359.
[16]Carmichael M J,Carmichael S K,Santelli C M,et al.Mn(II)-oxidizing bacteria are abundant and environmentally relevant members of ferromanganese deposits in caves of the upper Tennessee River Basin[J].Geomicrobiology Journal,2013,30(9):779-800.
[17]Miller A Z,Hernández-MarinéM,Jurado V,et al.Enigmatic reticulated filaments in subsurface granite[J].Environmental Microbiology Reports,2012,4(6):596-603.