新疆断裂带泉水中细菌群落结构的PCR-DGGE分析
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摘要
乌鲁木齐10号泉起源于天山山脉博格达峰,地下径流于地震断裂带,泉水中硫化氢、甲烷等地球化学元素迁移活跃,为揭示水文地球化学变化对地震断裂带泉水细菌群落结构的影响,本研究对该泉水样品定期采样,以微孔滤膜收集菌体,使用SDS-酶解法提取总DNA,对细菌16S rDNA V3区进行PCR-DGGE分析并对条带测序,条带的光密度信息与监测的地球化学指标进行典型相关分析(CCA)。结果表明B16(Uncultured bacterium)和ε-变性菌纲(epsilon proteobacterium)与氟含量成正相关;黄杆菌B1(Flavobacterium)、绿脓假单胞菌(Pseudomonas aeruginosa)、B10(Uncultured bacterium)和腐生性葡萄球菌(Staphylococcus saprophyticus)与硫化物呈正相关;硫微螺菌(Thiomicrospira arctica)、黄杆菌B3(Flavobacterium)及阿尔莱葡萄球菌(Staphylococ-cus arlettae)与氢气呈正相关。断裂带泉水细菌能够对地层深处渗透出的地球化学元素产生灵敏的应答。
Seismic fault belt is a main passageway of various kinds of gas and geochemistry elements,where are important zone for matters,energy and information transfer between lithosphere and atmosphere.Microbes living in the belt effected intensively by the hydrogeochemical changes caused by earth crust movement.The 10th spring of Urumq originates from Bogeda mountain of Tianshan mountain range,and the groundwater runoff in seismic fault belt.The spring water contains various geochemistry elements,such as sulfide,methane,hydrogen,radon,sulfide,methane,carbon dioxide,helium,fluorine and hydrarg et al,many of which may potentially act as electron donors capable of supporting chemolithotrophy-based primary production.In order to reveal the impact of hydrogeochemical changes to bacterial community in seismic fault belt spring water,thirty-one samples of spring water collected from October 12th to November 11th and their corresponding geochemial parameters were determined.Of them,ten samples were used for bacterial dynamic changing monitoring.The planktonic communities were collected by microporous membrane filtration and then total DNA were extracted by SDS-enzymatic disruption method.The V3 region of the 16S rDNA gene of those samples was analyzed by PCR-DGGE(Denaturing Gradient Gel Electrophoresis) and different bands were excised for sequencing.The DGGE fingerprints combined with hydrogeochamical parameters were analyzed by CCA(Canonical correspondence analysis).The results showed that B16(Uncultured bacterium) and epsilon proteobacterium were positive correlate to Fluorine fluctuation and clustered as group one and two,respectively;B1(Flavobacterium),Pseudomonas aeruginosa,B10(Uncultured bacterium) and Staphylococcus saprophyticus were positive correlate to sulfide change;Thiomicrospira arctica,B3(Flavobacterium) and Staphylococcus arlettae were positive correlated to hydrogen content.The result indicated that bacterial communities in seismic fault belt spring water could response sensitively to hydrogeochemical parameter changes,and this may provide a novel means for crust movement monitoring and earthquake forecast.
Seismic fault belt is a main passageway of various kinds of gas and geochemistry elements,where are important zone for matters,energy and information transfer between lithosphere and atmosphere.Microbes living in the belt effected intensively by the hydrogeochemical changes caused by earth crust movement.The 10th spring of Urumq originates from Bogeda mountain of Tianshan mountain range,and the groundwater runoff in seismic fault belt.The spring water contains various geochemistry elements,such as sulfide,methane,hydrogen,radon,sulfide,methane,carbon dioxide,helium,fluorine and hydrarg et al,many of which may potentially act as electron donors capable of supporting chemolithotrophy-based primary production.In order to reveal the impact of hydrogeochemical changes to bacterial community in seismic fault belt spring water,thirty-one samples of spring water collected from October 12th to November 11th and their corresponding geochemial parameters were determined.Of them,ten samples were used for bacterial dynamic changing monitoring.The planktonic communities were collected by microporous membrane filtration and then total DNA were extracted by SDS-enzymatic disruption method.The V3 region of the 16S rDNA gene of those samples was analyzed by PCR-DGGE(Denaturing Gradient Gel Electrophoresis) and different bands were excised for sequencing.The DGGE fingerprints combined with hydrogeochamical parameters were analyzed by CCA(Canonical correspondence analysis).The results showed that B16(Uncultured bacterium) and epsilon proteobacterium were positive correlate to Fluorine fluctuation and clustered as group one and two,respectively;B1(Flavobacterium),Pseudomonas aeruginosa,B10(Uncultured bacterium) and Staphylococcus saprophyticus were positive correlate to sulfide change;Thiomicrospira arctica,B3(Flavobacterium) and Staphylococcus arlettae were positive correlated to hydrogen content.The result indicated that bacterial communities in seismic fault belt spring water could response sensitively to hydrogeochemical parameter changes,and this may provide a novel means for crust movement monitoring and earthquake forecast.
引文
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[17]Amir M,Gal HR,Bella G,Avihu Y,Erez D,Martin K,Orna D,Yitzhak P.Adaptive prediction of environmental changes by microorganisms.Nature,2009,doi:10.1038/nature08112
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[25]Gao X Q,Wang D,Xu QL,Li YP,Li X Y,Feng X H.Study on the earthquake-reflecting characteristics of sulfide and its earthquake-reflectingmechanism.Earthquake,2001.21(1):91-97.
[26]Gao X Q,Dilixiati,Xu QL,Li Y P,Feng X H,Cui Y.An investigation on the characteristics of earthquake-indicating variations of F-in UrumqNo.9 and No.10 springs.Journal of Seismology,2002,22(1):5-11.
[6]张胜,张翠云,张云,李政红,张明.地质微生物地球化学作用的意义与展望.地质通报,2005,24,10:1027-1031.
[7]高小其,陈华静,杨又陵,李志海,许秋龙.2004年12月26日印尼8.7级强震新疆地区的同震效应.地震地磁观测与研究,2006,27:40-46.
[9]高小其,许秋龙,张学敏,李艳萍,卢静芳,崔勇.乌鲁木齐10号泉流体综合观测及映震灵敏性初析.内陆地震,2000,14(3):243-251.
[22]常桂兰.氡与氡的危害.铀矿地质,2002,18(2):122-128.
[23]李晓辉,申仲华,李建华.水中氡的危害及监管现状.现代预防医学,2005,32(10).1313-1323.
[25]高小其,王道,许秋龙,李艳萍,李新勇,冯晓辉.硫化物的映震特征及其映震机理的研究.地震,2001,21(1):91-97.
[26]高小其,迪里夏提,许秋龙,李艳萍,冯晓晖,崔勇.乌鲁木齐9、10号泉F-映震特征的研究.地震学刊,2002,22(1):5-11.
[2]Kazuya W,Kanako W,Yumiko K.Molecular characterization of bacterial populations in petroleum-contaminated groundwater discharged fromunderground crude oil storage cavities.Applied and Environmental Microbiology,2000,66(11):4803-4809.
[3]BartΙomiej R,Dirk S M.Correlation between microbiological and chemical parameters of some hydrothermal springs in New Mexico.Journal ofHydrology,2003,280:272-284.
[4]Palumbo A V,Schryver J C,Fields MW.Coupling of functional gene diversity and geochemical data from environmental samples.Applied andEnvironmental Microbiology,2004,70(11):6525-6534
[5]Jack C S,Craig C B,Susan MP,Anthony V P,Aaron D P,David C W,James P M,Philip E L.Application of nonlinear analysis methods foridentifying relationships between microbial community structure and groundwater geochemistry.Microbial Ecology,2006,51:177-188.
[6]Zhang S,Zhang C Y,Zhang Y,Li Z H,Zhang M.Geomicrobial geochemical processes:Significance and prospects.Geological Bulletin of China,2005,24,(10):1027-1031.
[7]Gao X Q,Chen H J,Yang Y L,Li Z H,Xu Q L.Coseismal effect of Indonesia M 8.7 earthquake on Dec 26,2004 in Xinjiang region.Seismological and Geomagnetic Observation and Research,2006,27:40-46.
[8]Cinzia F,Alessandro A,Rocco F,Sergio G,Mariano V.Geochemical monitoring of ground waters(1998-2001)at Vesuvius volcano(Italy).Journal of Volcanology and Geothermal Research,2004,133:81-104.
[9]Gao X Q,Xu Q L,Zhang X M,Li Y P,Lu J F,Cui Y.Preliminary analysis on synthetic fluid observation of Urumq spring NO.10 and itsearthquake-reflecting sensitivity.Inland Earthquake,2000,14(3):243-251.
[10]Shaheen B H,Nasreen B,James TH.Depth distribution of microbial diversity in Mono Lake,a meromictic soda Lake in california.Applied andEnvironmental Microbiology,2003,69(2):1030-1042.
[11]Danilo E.PCR-DGGE fingerprinting:novel strategies for detection of microbes in food.Journal of Microbiological Methods,2004,56:297-314.
[12]Keyser M,Witthuhn R C,Lamprecht C,Coetzee MP A,Brizt TJ.PCR-based DGGE fingerprinting and identification of methanogens detected inthree different types of UASB granules.Systematic and Applied Microbiology,2006,29:77-84.
[13]Sambrook J,Fritsch E F,Maniatis T.Molecular Conning a Laboratory Manual.2nd ed∥Hou Y D Translated.Beijing:China SciencePress,1992.
[14]Emilie L,Bndite L,Lo c TH,Rols J L,Frdric G.Analysis of bacterial diversity in river biofilms using16S rDNAPCR-DGGE:methodologicalsettings and fingerprints interpretation.Water Research,2005,39:380-388.
[15]Muyzer G,de Waal E D,Uitterlinden A G.Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis ofpolymerase chain reaction-amplified genes coding for 16S rRNA.Applied and Environmental Microbiology,1993,59:695-700.
[16]Andreas H F,Ines W,Gabriela R,Alexander K T K,Herann S,Martina M B,Romana H,Ulrich S,Gerhard H,Robert L M.Bacterialdynamics in spring water of alpine karst aquifers indicates the presence of stable autochthonous microbial endokarst communities.EnvironmentalMicrobiology,2005,7(8):1248-1259.
[17]Amir M,Gal HR,Bella G,Avihu Y,Erez D,Martin K,Orna D,Yitzhak P.Adaptive prediction of environmental changes by microorganisms.Nature,2009,doi:10.1038/nature08112
[18]Shabir AD,Gijs K,Gerard M.Nested PCR-denaturing gradient gel electrophoresis approach to determine the diversity of sulfate-reducing bacteriain complex microbial communities.Applied and Environmental Microbiology,2005,71(5):2325-2330.
[19]Madigan MT,Martinko J M,Parker J.Brock Biology of Microorganisms.8th ed.Engle-wood Cliffs NJ:Prentice-Hall,1997:871-875.
[20]Longnecker K,Reysenbach A L.Expansion of the geographic distribution of a novel lineage ofε-proteobacteira to a hydrothermal vent site on theSouthern East Pacific Rise.FEMS Microbial Ecology,2001,35:287-293
[21]Lovley D R,Chapelle F H.Deep subsurface microbial processes.Geophysics,1995,33:365-381.
[22]Chang G L.Radon and its hazards.Uranium Geology,2002,18(2):122-128.
[23]Li X H,Shen Z H,Li J H.Radon hazards in water and current supervise.Modem Preventive Medicine,2005,(32):1313-1323.
[24]Nancy N P,Dale TA,Wayne HP,Charles W G,Lyle G W.Characterization of the prokaryotic diversity in cold saline perennial springs of theCanadian high arctic.Applied and Environmental Microbiology,2007:1532-1543.
[25]Gao X Q,Wang D,Xu QL,Li YP,Li X Y,Feng X H.Study on the earthquake-reflecting characteristics of sulfide and its earthquake-reflectingmechanism.Earthquake,2001.21(1):91-97.
[26]Gao X Q,Dilixiati,Xu QL,Li Y P,Feng X H,Cui Y.An investigation on the characteristics of earthquake-indicating variations of F-in UrumqNo.9 and No.10 springs.Journal of Seismology,2002,22(1):5-11.
[6]张胜,张翠云,张云,李政红,张明.地质微生物地球化学作用的意义与展望.地质通报,2005,24,10:1027-1031.
[7]高小其,陈华静,杨又陵,李志海,许秋龙.2004年12月26日印尼8.7级强震新疆地区的同震效应.地震地磁观测与研究,2006,27:40-46.
[9]高小其,许秋龙,张学敏,李艳萍,卢静芳,崔勇.乌鲁木齐10号泉流体综合观测及映震灵敏性初析.内陆地震,2000,14(3):243-251.
[22]常桂兰.氡与氡的危害.铀矿地质,2002,18(2):122-128.
[23]李晓辉,申仲华,李建华.水中氡的危害及监管现状.现代预防医学,2005,32(10).1313-1323.
[25]高小其,王道,许秋龙,李艳萍,李新勇,冯晓辉.硫化物的映震特征及其映震机理的研究.地震,2001,21(1):91-97.
[26]高小其,迪里夏提,许秋龙,李艳萍,冯晓晖,崔勇.乌鲁木齐9、10号泉F-映震特征的研究.地震学刊,2002,22(1):5-11.
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