克拉玛依石油污染土壤微生物群落结构及其代谢特征
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摘要
为了分析克拉玛依油区内土壤中正构烷烃含量间的差异,微生物群落生理多样性、微生物代谢活性在不同石油污染梯度土壤中的变化规律。本研究采用GC、平板稀释法、Biolog微平板技术探讨了土壤微生物群落特征在3种不同污染程度下的变化情况。研究表明,石油污染土壤烷烃含量与微生物代谢活性呈显著负相关(r=-0.783, p<0.05)。随着石油污染程度增加微生物数量呈下降趋势,不同石油污染土壤中细菌数量占决定优势,细菌>真菌>放线菌。不同石油污染土壤微生物群落对6大碳源的利用体现出差异。主成分分析(PCA)表明,清洁土壤与石油污染土壤对底物利用有明显差异。石油污染严重土样碳源利用率为"酯类>酸类>胺类>氨基酸类>单糖/糖苷/聚合糖类>醇类"。本研究成果为后期修复污染土壤时调整投入的碳源底物等提供科学帮助。
In order to analyze the difference of n-alkanes content in the oil-contaminated soil, the physiological diversity of microbial community and the metabolic activity of microbial metabolism in oil-contaminated soil with gradient of oil pollution in Karamay oil region. The changes of soil microbial community characteristics were analyzed by GC, plate dilution method and Biolog microplate technique in three different levels of pollution changes. The results showed that the alkane content in the oil contaminated soil was negatively correlated with the microbial metabolic activity(r=-0.783, p<0.05). The microbes decreased with the increasing of petroleum pollution,and the number of bacteria in different oil-contaminated soils was the dominant factor, bacteria >fungi >actinomycetes. Utilization of six carbon sources was different in the distinct oil contaminated soil. Principal component analysis(PCA) showed that there was a significant difference in substrate utilization between clean soil and petroleum contaminated soil. In the heavy contaminated areas, utilization rate of carbon source was Ester >Acid>Amine>Amino acid>Monosaccharide/Glycoside/Polysaccharide>Alcohol. The results of this study provided scientific help for the adjustment of the carbon source substrates, which will be putting into the remediation of contaminated soils in the later period.
In order to analyze the difference of n-alkanes content in the oil-contaminated soil, the physiological diversity of microbial community and the metabolic activity of microbial metabolism in oil-contaminated soil with gradient of oil pollution in Karamay oil region. The changes of soil microbial community characteristics were analyzed by GC, plate dilution method and Biolog microplate technique in three different levels of pollution changes. The results showed that the alkane content in the oil contaminated soil was negatively correlated with the microbial metabolic activity(r=-0.783, p<0.05). The microbes decreased with the increasing of petroleum pollution,and the number of bacteria in different oil-contaminated soils was the dominant factor, bacteria >fungi >actinomycetes. Utilization of six carbon sources was different in the distinct oil contaminated soil. Principal component analysis(PCA) showed that there was a significant difference in substrate utilization between clean soil and petroleum contaminated soil. In the heavy contaminated areas, utilization rate of carbon source was Ester >Acid>Amine>Amino acid>Monosaccharide/Glycoside/Polysaccharide>Alcohol. The results of this study provided scientific help for the adjustment of the carbon source substrates, which will be putting into the remediation of contaminated soils in the later period.
引文
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Chen L.H.,Sun W.H.,Li H.L.,Yang Q.,and Zhang X.,2016,Biological degradation and transformation characteristics of different components in petroleum hydrocarbon by oil-degradation bacteria,Huanjing Kexue Xuebao(Acta Scientiae Circumstantiae),36(1):124-133(陈丽华,孙万虹,李海玲,杨琴,张璇,2016,石油降解菌对石油烃中不同组分的降解及演化特征研究,环境科学学报,36(1):124-133)
Chodak M.,Klimek B.,Azarbad H.,and Jazwa M.,2015,Functional diversity of soil microbial communities under Scots pine,Norway spruce,silver birch and mixed boreal forests,Pedobiologia J.Soil Eco.,58(2/3):81-88
Feigl V.,Ujaczki E.,Vaszita E.,and Molnár M.,2017,Influence of red mud on soil microbial communities:Application and comprehensive evaluation of the Biolog EcoPlate approach as a tool in soil microbiological studies,Sci.Total Environ.,595:903-911
Gryta A.,Frac M.,and Oszust K.,2014,The application of the Biolog EcoPlate approach in ecotoxicological evaluation of dairy sewage sludge,Appl.Biochem.Biotechnol.,174(4):1434-1443
Kenarova A.,Radeva G.,Traykov I.,and Boteva S.,2014,Community level physiological profiles of bacterial communities inhabiting uranium mining impacted sites,Ecotoxicol.Environ.Saf.,100:226-232
Li X.L.,2017,Influence of oil pollution on soil microbial community diversity,Shengwu Gongcheng Xuebao(Chinese Journal of Biotechnology),33(6):968-975(李晓楼,2017,石油污染对土壤微生物群落多样性的影响,生物工程学报,33(6):968-975)
Liang J.F.,Yang J.K.,Yang Y.,Chao Q.F.,Yin Y.L.,and Zhao Y.G.,2016,Effect of environmental factors on bacterial community structure in petroleum contaminated soil of Karamay oil field,Weishengwu Xuebao(Acta microbiologica Sinica),56(8):1301-1313(梁建芳,杨江科,杨杨,晁群芳,殷亚兰,赵亚光,2016,克拉玛依油田石油污染土壤细菌群落结构与环境因子的关系,微生物学报,56(8):1301-1313)
Manli W.,Wei L.,Dick W.A.,Xiqiong Y.,Kaili C.,Kost D.,and Liming C.,2017,Bioremediation of hydrocarbon degradation in a petroleum-contaminated soil and microbial population and activity determination,Chemosphere,169:124-130
Mummey D.L.,Stahl P.D.,and Buyer J.S.,2002,Microbia biomarkers as an indicator of ecosystem recovery following surface mine reclamation,Applied Soil Ecology,21(3):251-259
Paolis D.M.R.,and Lippi D.,2008,Use of metabolic and molecular methods for the identification of a strain isolated from paper affected by foxing,Microbiol.Res.,163(2):121-131
Selmants P.C.,Hart S.C.,Boyle S.I.,and Stark J.M.,2005,Red alder(Alnus rubra)alters community-level soil microbial function in conifer forests of the Pacific Northwest,USASoil Biology and Biochemistry,37(10):1860-1868
Shen W.H.,Zhu N.W.,Yin F.H.,Wang H.J.,and Dang Z.,2015Study on combined bio-indicators in ecotoxicity monitoring of oil-contaminated soil during bioremediation.Shengta Huanjing Xuebao(Ecology and Environmental Sciences)24(9):1560-1569(沈伟航,朱能武,尹富华,王华金,党志2015,微生物修复石油污染土壤的生态毒性指示,生态环境学报,24(9):1560-1569)
Singh M.P.,2009,Application of Biolog FF MicroPlate for substrate utilization and metabolite profiling of closely related fungi,Journal of Microbiological Methods,77(1):102-108
Song J.Y.,Chen M.M.,Zhang H.T.,Yang J.P.,Wu B.C.,and Liu S.M.,2013,Metabolic characteristics of oil contaminated soil microbial communities,Huanjing Kexue Yu Jishu(Environmental Science and Technology),36(10):63-66(宋佳宇,陈梅梅,张海涛,杨玖坡,吴百春,刘思敏,2013,石油污染土壤微生物群落代谢特征分析,环境科学与技术,36(10):63-66)
Song S.,Chen X.M.,Xiao W.,Wu D.,Hao X.C.,Zhang X.H.Zhang Q.,and Luo X.G.,2016,Response of soil cultivatable microorganisms to uranium contamination based on BI-OLOG fingerprint analysis method,Henong Xuebao(Journal of Nuclear Agricultural Sciences),30(6):1169-1177(宋收,陈晓明,肖伟,伍迪,郝希超,张祥辉,张倩,罗学刚2016,基于BIOLOG指纹解析土壤可培微生物对铀污染的响应,核农学报,30(6):1169-1177)
Stroud J.L.,Paton G.I.,and Semple K.T.,2007,Microbe-aliphatic hydrocarbon interactions in soil:implications for biodegradation and bioremediation,J.Appl.Microbiol.,102(3)1239-1253
Sutton N.B.,Maphosa F.,Morillo J.A.,Al-Soud W.A.,Langenhoff A.A.M.,Grotenhuis T.,Rijnaarts H.H.M.,and Smidt H.2013,Impact of long-term diesel contamination on soil microbial community structure,Appl.Environ.Microbiol.,79(2):619-630
Tan Y.P.,Ma Y.,and Lv J.,2016,Effect of oil pollution on soi bacteria structure characteri stics,Shengwu Jishu(Biotechnology),26(2):193-198(谭银萍,马媛,吕杰,2016,石油污染对土壤细菌结构特征的影响,生物技术,26(2):193-198)
Wan C.L.,Yang X.,Du M.A.,Yu C.G.,and Li G.Y.,2010,Characteristics of bacterial community in long-term oil polluted soil,Shiyou Xuebao(Acta Petrolei Sinica),26(6):928-933(万春黎,杨雪,杜茂安,于春光,李观元,2010,石油污染土壤中细菌群落结构特征,石油学报,26(6):928-933)
Wang R.G.,Wang M.,Niu X.W.,and Tang J.C.,2010,Determination of total petroleum hydrocarbons content in soil by ultrasonic-soxhlet extraction-gravimetric analysis,Fenxi Huaxue(Chinese Journal of Analytical Chemistry),38(3):417-420(王如刚,王敏,牛晓伟,唐景春,2010,超声-索氏萃取-重量法测定土壤中总石油烃含量,分析化学,38(3):417-420)
Yang M.Q.,Li L.M.,Li C.,and Li G.H.,2013,Microbial community structure and distribution characteristics in oil contaminated soil,Huanjing Kexue(Environmental Science),34(2):789-794(杨萌青,李立明,李川,李广贺,2013,石油污染土壤微生物群落结构与分布特性研究,环境科学,34(2):789-794)
Yang Q.,Wu M.L.,Nie M.Q.,Wang T.T.,and Zhang M.H.,2015,Effects and biological response on bioremediation of petroleum contaminated soil,Huanjing Kexue(Envioronmental Science),36(5):1856-1863(杨茜,吴蔓莉,聂麦茜,王婷婷,张明辉,2015,石油污染土壤的生物修复技术及微生物生态效应,环境科学,36(5):1856-1863)
Yue B.B.,Li X.,Ren F.F.,Meng F.J.,Yin P.D.,Zhang H.H.,and Sun G.Y.,2011,Carbon source utilization characteristics of microbial communities in a petroleum-contaminated soil in Daqing oil field,Northeast China,Yingyong Shengtai Xuebao(The Journal of Applied Ecology),22(12):3259-3264(岳冰冰,李鑫,任芳菲,孟凡娟,尹鹏达,张会慧,孙广玉,2011,石油污染地土壤微生物群落的碳源利用特性,应用生态学报,22(12):3259-3264)
Zhang J.,Hu W.,Liu Y.Z.,Ge G.,and Wu L.,2015,Response of soil microbial functional diversity to different land-use types in wetland of Poyang Lake,China,Shengtai Xuebao(Acta Ecologica Sinica),35(4):965-971(张杰,胡维,刘以珍,葛刚,吴兰,2015,鄱阳湖湿地不同土地利用方式下土壤微生物群落功能多样性,生态学报,35(4):965-971)
Zhang Q.,Song J.M.,Peng Q.C.,Li X.G.,Yuan H.M.,Li N.,Duan L.Q.,Qu B.X.,and Wang Q.D.,2017,Distribution and sources of n-alkanes in surface seawater of Jiaozhou bay,Huanjing Kexue(Environmental Science),38(7):2763-2772(张倩,宋金明,彭全材,李学刚,袁华茂,李宁,段丽琴,曲宝晓,王启栋,2017,胶州湾表层海水中的正构烷烃及其来源解析,环境科学,38(7):2763-2772)
Zhao Y.G.,Duan W.W.,Chao Q.F.,Liang J.F.,Xu M.,and Xiao L.M.,20 17,Microbial groups and metabolic activation of oil-contaminated soil in Karamay,Huanjing Kexue Yu Jishu(Environmental Science&Technology),40(4):7-12(赵亚光段魏魏,晁群芳,梁建芳,徐苗,肖璐梅,2017,克拉玛依石油污染土壤微生物类群及其代谢活性研究,环境科学与技术,40(4):7-12)
Zhen L.S.,Gu J.,Hu T.,Lv R.,Jia F.A.,Liu C.,and Li Y.,2015Microbial community structure and metabolic characteristics of oil-contaminated soil in the Loess Plateau,Shengtai Xuebao(Acta Ecologica Sinica),35(17):5703-5710(甄丽莎谷洁,胡婷,吕睿,贾凤安,刘晨,李燕,2015,黄土高原石油污染土壤微生物群落结构及其代谢特征研究,生态学报,35(17):5703-5710)
Zuo L.M.,Ma X.Y.,Li Z.M.,Yu R.H.,Wang Y.,Feng L.,Liu H.,and Peng Z.H.,2015,Research on Interaction between Petroleum hydrocarbon and degrading bacteria consortium in soil,Anquan Yu Huanjing Gongcheng(Safety and Environmental Engineering),22(4):63-68(左丽敏,马晓阳,李智民,余荣华,王燕,冯亮,刘慧,彭正华,2015,中国土壤中石油烃与降解菌群的相互作用研究,安全与环境工程,22(4):63-68)
Chen L.H.,Sun W.H.,Li H.L.,Yang Q.,and Zhang X.,2016,Biological degradation and transformation characteristics of different components in petroleum hydrocarbon by oil-degradation bacteria,Huanjing Kexue Xuebao(Acta Scientiae Circumstantiae),36(1):124-133(陈丽华,孙万虹,李海玲,杨琴,张璇,2016,石油降解菌对石油烃中不同组分的降解及演化特征研究,环境科学学报,36(1):124-133)
Chodak M.,Klimek B.,Azarbad H.,and Jazwa M.,2015,Functional diversity of soil microbial communities under Scots pine,Norway spruce,silver birch and mixed boreal forests,Pedobiologia J.Soil Eco.,58(2/3):81-88
Feigl V.,Ujaczki E.,Vaszita E.,and Molnár M.,2017,Influence of red mud on soil microbial communities:Application and comprehensive evaluation of the Biolog EcoPlate approach as a tool in soil microbiological studies,Sci.Total Environ.,595:903-911
Gryta A.,Frac M.,and Oszust K.,2014,The application of the Biolog EcoPlate approach in ecotoxicological evaluation of dairy sewage sludge,Appl.Biochem.Biotechnol.,174(4):1434-1443
Kenarova A.,Radeva G.,Traykov I.,and Boteva S.,2014,Community level physiological profiles of bacterial communities inhabiting uranium mining impacted sites,Ecotoxicol.Environ.Saf.,100:226-232
Li X.L.,2017,Influence of oil pollution on soil microbial community diversity,Shengwu Gongcheng Xuebao(Chinese Journal of Biotechnology),33(6):968-975(李晓楼,2017,石油污染对土壤微生物群落多样性的影响,生物工程学报,33(6):968-975)
Liang J.F.,Yang J.K.,Yang Y.,Chao Q.F.,Yin Y.L.,and Zhao Y.G.,2016,Effect of environmental factors on bacterial community structure in petroleum contaminated soil of Karamay oil field,Weishengwu Xuebao(Acta microbiologica Sinica),56(8):1301-1313(梁建芳,杨江科,杨杨,晁群芳,殷亚兰,赵亚光,2016,克拉玛依油田石油污染土壤细菌群落结构与环境因子的关系,微生物学报,56(8):1301-1313)
Manli W.,Wei L.,Dick W.A.,Xiqiong Y.,Kaili C.,Kost D.,and Liming C.,2017,Bioremediation of hydrocarbon degradation in a petroleum-contaminated soil and microbial population and activity determination,Chemosphere,169:124-130
Mummey D.L.,Stahl P.D.,and Buyer J.S.,2002,Microbia biomarkers as an indicator of ecosystem recovery following surface mine reclamation,Applied Soil Ecology,21(3):251-259
Paolis D.M.R.,and Lippi D.,2008,Use of metabolic and molecular methods for the identification of a strain isolated from paper affected by foxing,Microbiol.Res.,163(2):121-131
Selmants P.C.,Hart S.C.,Boyle S.I.,and Stark J.M.,2005,Red alder(Alnus rubra)alters community-level soil microbial function in conifer forests of the Pacific Northwest,USASoil Biology and Biochemistry,37(10):1860-1868
Shen W.H.,Zhu N.W.,Yin F.H.,Wang H.J.,and Dang Z.,2015Study on combined bio-indicators in ecotoxicity monitoring of oil-contaminated soil during bioremediation.Shengta Huanjing Xuebao(Ecology and Environmental Sciences)24(9):1560-1569(沈伟航,朱能武,尹富华,王华金,党志2015,微生物修复石油污染土壤的生态毒性指示,生态环境学报,24(9):1560-1569)
Singh M.P.,2009,Application of Biolog FF MicroPlate for substrate utilization and metabolite profiling of closely related fungi,Journal of Microbiological Methods,77(1):102-108
Song J.Y.,Chen M.M.,Zhang H.T.,Yang J.P.,Wu B.C.,and Liu S.M.,2013,Metabolic characteristics of oil contaminated soil microbial communities,Huanjing Kexue Yu Jishu(Environmental Science and Technology),36(10):63-66(宋佳宇,陈梅梅,张海涛,杨玖坡,吴百春,刘思敏,2013,石油污染土壤微生物群落代谢特征分析,环境科学与技术,36(10):63-66)
Song S.,Chen X.M.,Xiao W.,Wu D.,Hao X.C.,Zhang X.H.Zhang Q.,and Luo X.G.,2016,Response of soil cultivatable microorganisms to uranium contamination based on BI-OLOG fingerprint analysis method,Henong Xuebao(Journal of Nuclear Agricultural Sciences),30(6):1169-1177(宋收,陈晓明,肖伟,伍迪,郝希超,张祥辉,张倩,罗学刚2016,基于BIOLOG指纹解析土壤可培微生物对铀污染的响应,核农学报,30(6):1169-1177)
Stroud J.L.,Paton G.I.,and Semple K.T.,2007,Microbe-aliphatic hydrocarbon interactions in soil:implications for biodegradation and bioremediation,J.Appl.Microbiol.,102(3)1239-1253
Sutton N.B.,Maphosa F.,Morillo J.A.,Al-Soud W.A.,Langenhoff A.A.M.,Grotenhuis T.,Rijnaarts H.H.M.,and Smidt H.2013,Impact of long-term diesel contamination on soil microbial community structure,Appl.Environ.Microbiol.,79(2):619-630
Tan Y.P.,Ma Y.,and Lv J.,2016,Effect of oil pollution on soi bacteria structure characteri stics,Shengwu Jishu(Biotechnology),26(2):193-198(谭银萍,马媛,吕杰,2016,石油污染对土壤细菌结构特征的影响,生物技术,26(2):193-198)
Wan C.L.,Yang X.,Du M.A.,Yu C.G.,and Li G.Y.,2010,Characteristics of bacterial community in long-term oil polluted soil,Shiyou Xuebao(Acta Petrolei Sinica),26(6):928-933(万春黎,杨雪,杜茂安,于春光,李观元,2010,石油污染土壤中细菌群落结构特征,石油学报,26(6):928-933)
Wang R.G.,Wang M.,Niu X.W.,and Tang J.C.,2010,Determination of total petroleum hydrocarbons content in soil by ultrasonic-soxhlet extraction-gravimetric analysis,Fenxi Huaxue(Chinese Journal of Analytical Chemistry),38(3):417-420(王如刚,王敏,牛晓伟,唐景春,2010,超声-索氏萃取-重量法测定土壤中总石油烃含量,分析化学,38(3):417-420)
Yang M.Q.,Li L.M.,Li C.,and Li G.H.,2013,Microbial community structure and distribution characteristics in oil contaminated soil,Huanjing Kexue(Environmental Science),34(2):789-794(杨萌青,李立明,李川,李广贺,2013,石油污染土壤微生物群落结构与分布特性研究,环境科学,34(2):789-794)
Yang Q.,Wu M.L.,Nie M.Q.,Wang T.T.,and Zhang M.H.,2015,Effects and biological response on bioremediation of petroleum contaminated soil,Huanjing Kexue(Envioronmental Science),36(5):1856-1863(杨茜,吴蔓莉,聂麦茜,王婷婷,张明辉,2015,石油污染土壤的生物修复技术及微生物生态效应,环境科学,36(5):1856-1863)
Yue B.B.,Li X.,Ren F.F.,Meng F.J.,Yin P.D.,Zhang H.H.,and Sun G.Y.,2011,Carbon source utilization characteristics of microbial communities in a petroleum-contaminated soil in Daqing oil field,Northeast China,Yingyong Shengtai Xuebao(The Journal of Applied Ecology),22(12):3259-3264(岳冰冰,李鑫,任芳菲,孟凡娟,尹鹏达,张会慧,孙广玉,2011,石油污染地土壤微生物群落的碳源利用特性,应用生态学报,22(12):3259-3264)
Zhang J.,Hu W.,Liu Y.Z.,Ge G.,and Wu L.,2015,Response of soil microbial functional diversity to different land-use types in wetland of Poyang Lake,China,Shengtai Xuebao(Acta Ecologica Sinica),35(4):965-971(张杰,胡维,刘以珍,葛刚,吴兰,2015,鄱阳湖湿地不同土地利用方式下土壤微生物群落功能多样性,生态学报,35(4):965-971)
Zhang Q.,Song J.M.,Peng Q.C.,Li X.G.,Yuan H.M.,Li N.,Duan L.Q.,Qu B.X.,and Wang Q.D.,2017,Distribution and sources of n-alkanes in surface seawater of Jiaozhou bay,Huanjing Kexue(Environmental Science),38(7):2763-2772(张倩,宋金明,彭全材,李学刚,袁华茂,李宁,段丽琴,曲宝晓,王启栋,2017,胶州湾表层海水中的正构烷烃及其来源解析,环境科学,38(7):2763-2772)
Zhao Y.G.,Duan W.W.,Chao Q.F.,Liang J.F.,Xu M.,and Xiao L.M.,20 17,Microbial groups and metabolic activation of oil-contaminated soil in Karamay,Huanjing Kexue Yu Jishu(Environmental Science&Technology),40(4):7-12(赵亚光段魏魏,晁群芳,梁建芳,徐苗,肖璐梅,2017,克拉玛依石油污染土壤微生物类群及其代谢活性研究,环境科学与技术,40(4):7-12)
Zhen L.S.,Gu J.,Hu T.,Lv R.,Jia F.A.,Liu C.,and Li Y.,2015Microbial community structure and metabolic characteristics of oil-contaminated soil in the Loess Plateau,Shengtai Xuebao(Acta Ecologica Sinica),35(17):5703-5710(甄丽莎谷洁,胡婷,吕睿,贾凤安,刘晨,李燕,2015,黄土高原石油污染土壤微生物群落结构及其代谢特征研究,生态学报,35(17):5703-5710)
Zuo L.M.,Ma X.Y.,Li Z.M.,Yu R.H.,Wang Y.,Feng L.,Liu H.,and Peng Z.H.,2015,Research on Interaction between Petroleum hydrocarbon and degrading bacteria consortium in soil,Anquan Yu Huanjing Gongcheng(Safety and Environmental Engineering),22(4):63-68(左丽敏,马晓阳,李智民,余荣华,王燕,冯亮,刘慧,彭正华,2015,中国土壤中石油烃与降解菌群的相互作用研究,安全与环境工程,22(4):63-68)