胜利油田油泥沙细菌和古菌多样性研究
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摘要
胜利油田某油泥沙是含有石油烃、水和泥沙的混合物,也含有丰富的微生物资源。采用454焦磷酸,并结合生物信息学分析了油泥沙中的细菌和古菌群落结构,发现油泥沙中的细菌群落主要由Psychrobacter(16.6%),Pseudomonas(14.7%),Rhodobacteraceae(6.5%),Thiobacillus(6.0%)和unclassified Actinomycetales(4.4%)组成,这些细菌大部分属于好氧和兼性好氧菌。古菌主要属于产甲烷古菌,主要由Methanohalobium(77.7%),Methanosaeta(5.4%),Methermicoccus(3.5%),Methanobrevibacter(2.8%),Methanobacterium(4.0%),Methanomethylovorans(1.8%)和Methanosphaera(1.7%),表明产甲烷古菌以甲基营养型为主,同时也存在少量的乙酸营养型和氢营养型产甲烷古菌。这些研究为今后认识和分离新型石油微生物资源提供了指导和帮助。
Oily sludge in oilfield is a complex mixture containing crude oil,water and sediments,in which diversity of microbial communities could be found. In this study,454-pyrosequencing technology was adopted to analyze the structures of bacterial community and archaea community in oily sludge of Shengli oilfield. Combined with bioinformatics analysis,it was revealed that the bacterial community was mainly composed of Psychrobacter( 16. 6%), Pseudomonas( 14. 7%),Rhodobacteraceae( 6. 5%),Thiobacillus( 6. 0%) and unclassified Actinomycetales( 4. 4%),most of them belong to aerobic and facultative aerobic bacteria. And the archaea were mainly methanogenic,including Methanohalobium( 77. 7%),Methanosaeta( 5. 4%),Methermicoccus( 3. 5%),Methanobrevibacter( 2. 8%),Methanobacterium( 4. 0%),Methanomethylovorans( 1. 8%) and Methanosphaera( 1. 7%),suggesting that methylotrophs dominated the methanogenic archaea,followed by acetotrophic and hydrogenotrophic methanogens.
Oily sludge in oilfield is a complex mixture containing crude oil,water and sediments,in which diversity of microbial communities could be found. In this study,454-pyrosequencing technology was adopted to analyze the structures of bacterial community and archaea community in oily sludge of Shengli oilfield. Combined with bioinformatics analysis,it was revealed that the bacterial community was mainly composed of Psychrobacter( 16. 6%), Pseudomonas( 14. 7%),Rhodobacteraceae( 6. 5%),Thiobacillus( 6. 0%) and unclassified Actinomycetales( 4. 4%),most of them belong to aerobic and facultative aerobic bacteria. And the archaea were mainly methanogenic,including Methanohalobium( 77. 7%),Methanosaeta( 5. 4%),Methermicoccus( 3. 5%),Methanobrevibacter( 2. 8%),Methanobacterium( 4. 0%),Methanomethylovorans( 1. 8%) and Methanosphaera( 1. 7%),suggesting that methylotrophs dominated the methanogenic archaea,followed by acetotrophic and hydrogenotrophic methanogens.
引文
[1]吴丽华,王志强.胜利油田的油泥沙现状及处理工艺探讨[J].油气田环境保护,2005,8(4):23-25.
[2]中华人民共和国环境保护部令和中华人民共和国国家发展和改革委员会.国家危险废物名录[S].北京,2008.
[3]刘五星,骆永明,滕应,李振高,吴龙华.我国部分油田土壤及油泥的石油污染初步研究[J].土壤,2007,39(2):247-251.
[4]李冰,谢卫红,朱景义.中国石油油田含油污泥处理现状[J].石油规划设计,2009,20(4):18-20.
[5]陆秀君,郭书海,孙清,梁成华.石油污染土壤的修复技术研究现状及展望[J].沈阳农业大学学报,2003,34(1):63-67.
[6]Head I M,Jones D M,Larter S R.Biological activity in the deep subsurface and the origin of heavy oil[J].Nature,2003,426(6964):344-352.
[7]承磊,仇天雷,邓宇,张辉.油藏厌氧微生物研究进展[J].应用与环境生物学报,2006,12(5):740-744.
[8]Orphan V J,Taylor L T,Hafenbradl D,Delong E F.Culture-dependent and culture-independent characterization of microbial assemblages associated with hightemperature petroleum reservoirs[J].Appl Environ Microbiol,2000,66(2):700-711.
[9]Pham V D,Hnatow L L,Zhang S,Fallon R D,Jackson S C,Tomb J F,De Long E F,Keeler S J.Characterizing microbial diversity in production water from an Alaskan mesothermic petroleum reservoir with two independent molecular methods[J].Environ Microbiol,2009,11(1):176-187.
[10]Whitman W B,Coleman D C,Wiebe W J.Prokaryotes:The unseen majority[J].PNAS,1998,95(12):6578-6583.
[11]Claesson M J,Wang Q,O'Sullivan O,Greene-Diniz R,Cole J R,Ross R P,O'Toole P W.Comparison of two next-generation sequencing technologies for resolving highly complex microbiota composition using tandem variable 16S rRNA gene regions[J].Nucleic Acids Res,2010.
[12]Ye L,Zhang T.Bacterial communities in different sections of a municipal wastewater treatment plant revealed by 16S r DNA 454 pyrosequencing[J].Appl Microbiol Biotechnol,2013,97(6):2681-2690.
[13]Lu Y,Lueders T,Friedrich M W,Conrad R.Detecting active methanogenic populations on rice roots using stable isotope probing[J].Environ Microbiol,2005,7(3):326-336.
[14]Lueders T,Manefield M,Friedrich MW.Enhanced sensitivity of DNA-and rRNA-based stable isotope probing by fractionation and quantitative analysis of isopycnic centrifugation gradients[J].Environmental Microbiology,2004,6(1):73-78.
[15]Schloss P D,Westcott S L,Ryabin T,Hall J R,Hartmann M,Hollister E B,Lesniewski R A,Oakley B B,Parks D H,Robinson C J,Sahl JW,Stres B,Thallinger G G,Van Horn D J,Weber C F.Introducing mothur:open-source,platform-independent,community-supported software for describing and comparing microbial communities[J].Appl Environ Microbiol,2009,75(23):7537-7541.
[16]Schloss P D,Gevers D,Westcott S L.Reducing the effects of PCR amplification and sequencing artifacts on16S rRNA-based studies[J].PLo S ONE,2011,6(12):e27310.
[17]Wang Q,Garrity G M,Tiedje J M,Cole J R.Nave bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy[J].Appl Environ Microbiol,2007,73(16):5261-5267.
[18]Juni E,Heym G.Psychrobacter immobilis gen.nov.,sp.nov.:Genospecies Composed of Gram-Negative,Aerobic,Oxidase-Positive Coccobacilli[J].Int J Syst Evol Microbiol,1986,36(3):388-391.
[19]Baik K S,Park S C,Lim C H,Lee K H,Jeon D Y,Kim C M,Seong C N.Psychrobacter aestuarii sp.nov.,isolated from a tidal flat sediment[J].Int J Syst Evol Microbiol,2010,60(7):1631-1636.
[20]Bakermans C,AyaladelRío H L,Ponder M A,Vishnivetskaya T,Gilichinsky D,Thomashow MF,Tiedje JM.Psychrobacter cryohalolentis sp.nov.and Psychrobacter arcticus sp.nov.,isolated from Siberian permafrost[J].Int J Syst Evol Microbiol,2006,56(6):1285-1291.
[21]Romanenko L A,Tanaka N,Frolova G M,Mikhailov V V.Psychrobacter fulvigenes sp.nov.,isolated from a marine crustacean from the Sea of Japan[J].Int J Syst Evol Microbiol,2009,59(6):1480-1486.
[22]Yassin A F,Busse H J.Psychrobacter lutiphocae sp.nov.,isolated from the faeces of a seal[J].Int J Syst Evol Microbiol,2009,59(8):2049-2053.
[23]Yoon J H,Lee C H,Yeo S H,Oh T K.Psychrobacter aquimaris sp.nov.and Psychrobacter namhaensis sp.nov.,isolated from sea water of the South Sea in Korea[J].Int J Syst Evol Microbiol,2005,55(3):1007-1013.
[24]孙玉萍,王红英,刘素辉,倪萍,马海梅.新疆油污土壤中石油烃降解菌筛选及鉴定[J].中国公共卫生,2011,27(10):1340-1342.
[25]王福强,李凤超,李彦芹,何炜,康现江.原油降解菌的筛选鉴定及降解特性的研究[J].环境科学与技术,2010,12:39-42.
[26]任红燕,宋志勇,李霏霁,汪卫东,张晓君,赵立平.胜利油藏不同时间细菌群落结构的比较[J].微生物学通报,2011,38(4):561-568.
[27]Microbiology of Hydrocarbon-Degrading Pseudomonas//Palleroni N J,Pieper D H,Moore ERB Handbook of Hydrocarbon and Lipid Microbiology[J].Springer Berlin Heidelberg,2010:1787-1798.
[28]Hirota K,Yamahira K,Nakajima K,Nodasaka Y,Okuyama H,Yumoto I.Pseudomonas toyotomiensis sp.nov.,a psychrotolerant facultative alkaliphile that utilizes hydrocarbons[J].Int J Syst Evol Microbiol,2011,61(8):1842-1848.
[29]Ma Y,Wang L,Shao Z.Pseudomonas,the dominant polycyclic aromatic hydrocarbon-degrading bacteria isolated from Antarctic soils and the role of large plasmids in horizontal gene transfer[J].Environ Microbiol,2006,8(3):455-465.
[30]Yoon J H,Kang S S,Cho Y G,Lee S T,Kho Y H,Kim C J,Park Y H.Rhodococcus pyridinivorans sp.nov.,a pyridine-degrading bacterium.Int.J.Syst[J].Evol Microbiol,2000,50(6):2173-2180.
[31]Bicca F C,Fleck L C,Ayub M A Z.Production of biosurfactant by hydrocarbon degrading Rhodococcus ruber and Rhodococcus erythropolis[J].Revista de Microbiologia,1999,30:231-236.
[32]Kelly D P,Wood A P.Reclassification of some species of Thiobacillus to the newly designated genera Acidithiobacillus gen.nov.,Halothiobacillus gen.nov.and Thermithiobacillus gen.nov[J].Int J Syst Evol Microbiol,2000,50(2):511-516.
[33]Robertson L,Kuenen JG The Prokaryotes.The Genus Thiobacillus[J].Springer New York,2006:812-827.
[34]Findley J,Appleman M D,Yen T F.Degradation of Oil Shale by Sulfur-Oxidizing Bacteria[J].Applied Microbiology,1974,28(3):460-464.
[35]Dubinsky E A,Conrad M E,Chakraborty R,Bill M,Borglin S E,Hollibaugh J T,Mason O U,M,Piceno Y,Reid F C,Stringfellow W T,Tom L M,Hazen T C,Andersen G L.Succession of Hydrocarbon-Degrading Bacteria in the Aftermath of the Deepwater Horizon Oil Spill in the Gulf of Mexico[J].Environ Sci Technol,2013,47(19):10860-10867.
[36]Gerdes B,Brinkmeyer R,Dieckmann G,Helmke E.Influence of crude oil on changes of bacterial communities in Arctic sea-ice[J].FEMS Microbiol Ecol,2005,53(1):129-139.
[37]Kostka J E,Prakash O,Overholt W A,Green S J,Freyer G,Canion A,Delgardio J,Norton N,Hazen TC,Huettel M.Hydrocarbon-Degrading Bacteria and the Bacterial Community Response in Gulf of Mexico Beach Sands Impacted by the Deepwater Horizon Oil Spill[J].Appl Environ Microbiol,2011,77(22):7962-7974.
[38]Mortazavi B,Horel A,Beazley M J,Sobecky P A.Intrinsic rates of petroleum hydrocarbon biodegradation in Gulf of Mexico intertidal sandy sediments and its enhancement by organic substrates[J].J Hazard Mater,2013,244-245(0):537-544.
[39]Zhilina T N,Zavarzin G A.Methanohalobium evestigatus,n.gen.,n.sp.The extremely halophilic methanogenic Archaebacterium[J].Dokl Akad Nauk SSSR(In Russian),1987,293:464-468.
[40]Charakhchyan I,Mityushina L,Kuznetsova V,Belyaev S.The morphology and ultrastructure of Methanococcoides euhalobius and the role of calcium in its morphogenesis.Microbiology,1989,58(5):667-672.
[41]Cheng L,Qiu T L,Yin X B,Wu X L,Hu G Q,Deng Y,Zhang H.Methermicoccus shengliensis gen.nov.,sp.nov.,a thermophilic,methylotrophic methanogen isolated from oil-production water,and proposal of Methermicoccaceae fam.nov[J].Int J Syst Evol Microbiol,2007,57(12):2964-2969.
[42]Davidova I A,Harmsen H J M,Stams A J M,Belyaev S S,Zehnder A J B.Taxonomic description of Methanococcoides euhalobius and its transfer to the Methanohalophilus genus[J].Antonie Leeuwenhoek,1997,71(4):313-318.
[43]Mori K,Iino T,Suzuki K i,Yamaguchi K,Kamagata Y.Aceticlastic and Na Cl-requiring Methanogen“Methanosaeta pelagica”sp.nov.,Isolated from Marine Tidal Flat Sediment[J].Appl Environ Microbiol,2012,78(9):3416-3423.
[44]Patel G B,Sprott G D.Methanosaeta concilii gen.nov.,sp.nov.(“Methanothrix concilii”)and Methanosaeta thermoacetophila nom.rev.,comb.nov[J].Int J Syst Bacteriol,1990,40(1):79-82.
[45]Cheng L,Ding C,Li Q,He Q,Dai L-r,Zhang H.DNASIP reveals that Syntrophaceae play an important role in methanogenic hexadecane degradation[J].PLo S ONE,2013,8(7):e66784.
[46]Cheng L,He Q,ding C,Dai L-r,Li Q,Zhang H.Novel bacterial groups dominate in a thermophilic methanogenic hexadecane-degrading consortium FEMS Microbiol[J].Ecol,2013,85(3):568-577.
[2]中华人民共和国环境保护部令和中华人民共和国国家发展和改革委员会.国家危险废物名录[S].北京,2008.
[3]刘五星,骆永明,滕应,李振高,吴龙华.我国部分油田土壤及油泥的石油污染初步研究[J].土壤,2007,39(2):247-251.
[4]李冰,谢卫红,朱景义.中国石油油田含油污泥处理现状[J].石油规划设计,2009,20(4):18-20.
[5]陆秀君,郭书海,孙清,梁成华.石油污染土壤的修复技术研究现状及展望[J].沈阳农业大学学报,2003,34(1):63-67.
[6]Head I M,Jones D M,Larter S R.Biological activity in the deep subsurface and the origin of heavy oil[J].Nature,2003,426(6964):344-352.
[7]承磊,仇天雷,邓宇,张辉.油藏厌氧微生物研究进展[J].应用与环境生物学报,2006,12(5):740-744.
[8]Orphan V J,Taylor L T,Hafenbradl D,Delong E F.Culture-dependent and culture-independent characterization of microbial assemblages associated with hightemperature petroleum reservoirs[J].Appl Environ Microbiol,2000,66(2):700-711.
[9]Pham V D,Hnatow L L,Zhang S,Fallon R D,Jackson S C,Tomb J F,De Long E F,Keeler S J.Characterizing microbial diversity in production water from an Alaskan mesothermic petroleum reservoir with two independent molecular methods[J].Environ Microbiol,2009,11(1):176-187.
[10]Whitman W B,Coleman D C,Wiebe W J.Prokaryotes:The unseen majority[J].PNAS,1998,95(12):6578-6583.
[11]Claesson M J,Wang Q,O'Sullivan O,Greene-Diniz R,Cole J R,Ross R P,O'Toole P W.Comparison of two next-generation sequencing technologies for resolving highly complex microbiota composition using tandem variable 16S rRNA gene regions[J].Nucleic Acids Res,2010.
[12]Ye L,Zhang T.Bacterial communities in different sections of a municipal wastewater treatment plant revealed by 16S r DNA 454 pyrosequencing[J].Appl Microbiol Biotechnol,2013,97(6):2681-2690.
[13]Lu Y,Lueders T,Friedrich M W,Conrad R.Detecting active methanogenic populations on rice roots using stable isotope probing[J].Environ Microbiol,2005,7(3):326-336.
[14]Lueders T,Manefield M,Friedrich MW.Enhanced sensitivity of DNA-and rRNA-based stable isotope probing by fractionation and quantitative analysis of isopycnic centrifugation gradients[J].Environmental Microbiology,2004,6(1):73-78.
[15]Schloss P D,Westcott S L,Ryabin T,Hall J R,Hartmann M,Hollister E B,Lesniewski R A,Oakley B B,Parks D H,Robinson C J,Sahl JW,Stres B,Thallinger G G,Van Horn D J,Weber C F.Introducing mothur:open-source,platform-independent,community-supported software for describing and comparing microbial communities[J].Appl Environ Microbiol,2009,75(23):7537-7541.
[16]Schloss P D,Gevers D,Westcott S L.Reducing the effects of PCR amplification and sequencing artifacts on16S rRNA-based studies[J].PLo S ONE,2011,6(12):e27310.
[17]Wang Q,Garrity G M,Tiedje J M,Cole J R.Nave bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy[J].Appl Environ Microbiol,2007,73(16):5261-5267.
[18]Juni E,Heym G.Psychrobacter immobilis gen.nov.,sp.nov.:Genospecies Composed of Gram-Negative,Aerobic,Oxidase-Positive Coccobacilli[J].Int J Syst Evol Microbiol,1986,36(3):388-391.
[19]Baik K S,Park S C,Lim C H,Lee K H,Jeon D Y,Kim C M,Seong C N.Psychrobacter aestuarii sp.nov.,isolated from a tidal flat sediment[J].Int J Syst Evol Microbiol,2010,60(7):1631-1636.
[20]Bakermans C,AyaladelRío H L,Ponder M A,Vishnivetskaya T,Gilichinsky D,Thomashow MF,Tiedje JM.Psychrobacter cryohalolentis sp.nov.and Psychrobacter arcticus sp.nov.,isolated from Siberian permafrost[J].Int J Syst Evol Microbiol,2006,56(6):1285-1291.
[21]Romanenko L A,Tanaka N,Frolova G M,Mikhailov V V.Psychrobacter fulvigenes sp.nov.,isolated from a marine crustacean from the Sea of Japan[J].Int J Syst Evol Microbiol,2009,59(6):1480-1486.
[22]Yassin A F,Busse H J.Psychrobacter lutiphocae sp.nov.,isolated from the faeces of a seal[J].Int J Syst Evol Microbiol,2009,59(8):2049-2053.
[23]Yoon J H,Lee C H,Yeo S H,Oh T K.Psychrobacter aquimaris sp.nov.and Psychrobacter namhaensis sp.nov.,isolated from sea water of the South Sea in Korea[J].Int J Syst Evol Microbiol,2005,55(3):1007-1013.
[24]孙玉萍,王红英,刘素辉,倪萍,马海梅.新疆油污土壤中石油烃降解菌筛选及鉴定[J].中国公共卫生,2011,27(10):1340-1342.
[25]王福强,李凤超,李彦芹,何炜,康现江.原油降解菌的筛选鉴定及降解特性的研究[J].环境科学与技术,2010,12:39-42.
[26]任红燕,宋志勇,李霏霁,汪卫东,张晓君,赵立平.胜利油藏不同时间细菌群落结构的比较[J].微生物学通报,2011,38(4):561-568.
[27]Microbiology of Hydrocarbon-Degrading Pseudomonas//Palleroni N J,Pieper D H,Moore ERB Handbook of Hydrocarbon and Lipid Microbiology[J].Springer Berlin Heidelberg,2010:1787-1798.
[28]Hirota K,Yamahira K,Nakajima K,Nodasaka Y,Okuyama H,Yumoto I.Pseudomonas toyotomiensis sp.nov.,a psychrotolerant facultative alkaliphile that utilizes hydrocarbons[J].Int J Syst Evol Microbiol,2011,61(8):1842-1848.
[29]Ma Y,Wang L,Shao Z.Pseudomonas,the dominant polycyclic aromatic hydrocarbon-degrading bacteria isolated from Antarctic soils and the role of large plasmids in horizontal gene transfer[J].Environ Microbiol,2006,8(3):455-465.
[30]Yoon J H,Kang S S,Cho Y G,Lee S T,Kho Y H,Kim C J,Park Y H.Rhodococcus pyridinivorans sp.nov.,a pyridine-degrading bacterium.Int.J.Syst[J].Evol Microbiol,2000,50(6):2173-2180.
[31]Bicca F C,Fleck L C,Ayub M A Z.Production of biosurfactant by hydrocarbon degrading Rhodococcus ruber and Rhodococcus erythropolis[J].Revista de Microbiologia,1999,30:231-236.
[32]Kelly D P,Wood A P.Reclassification of some species of Thiobacillus to the newly designated genera Acidithiobacillus gen.nov.,Halothiobacillus gen.nov.and Thermithiobacillus gen.nov[J].Int J Syst Evol Microbiol,2000,50(2):511-516.
[33]Robertson L,Kuenen JG The Prokaryotes.The Genus Thiobacillus[J].Springer New York,2006:812-827.
[34]Findley J,Appleman M D,Yen T F.Degradation of Oil Shale by Sulfur-Oxidizing Bacteria[J].Applied Microbiology,1974,28(3):460-464.
[35]Dubinsky E A,Conrad M E,Chakraborty R,Bill M,Borglin S E,Hollibaugh J T,Mason O U,M,Piceno Y,Reid F C,Stringfellow W T,Tom L M,Hazen T C,Andersen G L.Succession of Hydrocarbon-Degrading Bacteria in the Aftermath of the Deepwater Horizon Oil Spill in the Gulf of Mexico[J].Environ Sci Technol,2013,47(19):10860-10867.
[36]Gerdes B,Brinkmeyer R,Dieckmann G,Helmke E.Influence of crude oil on changes of bacterial communities in Arctic sea-ice[J].FEMS Microbiol Ecol,2005,53(1):129-139.
[37]Kostka J E,Prakash O,Overholt W A,Green S J,Freyer G,Canion A,Delgardio J,Norton N,Hazen TC,Huettel M.Hydrocarbon-Degrading Bacteria and the Bacterial Community Response in Gulf of Mexico Beach Sands Impacted by the Deepwater Horizon Oil Spill[J].Appl Environ Microbiol,2011,77(22):7962-7974.
[38]Mortazavi B,Horel A,Beazley M J,Sobecky P A.Intrinsic rates of petroleum hydrocarbon biodegradation in Gulf of Mexico intertidal sandy sediments and its enhancement by organic substrates[J].J Hazard Mater,2013,244-245(0):537-544.
[39]Zhilina T N,Zavarzin G A.Methanohalobium evestigatus,n.gen.,n.sp.The extremely halophilic methanogenic Archaebacterium[J].Dokl Akad Nauk SSSR(In Russian),1987,293:464-468.
[40]Charakhchyan I,Mityushina L,Kuznetsova V,Belyaev S.The morphology and ultrastructure of Methanococcoides euhalobius and the role of calcium in its morphogenesis.Microbiology,1989,58(5):667-672.
[41]Cheng L,Qiu T L,Yin X B,Wu X L,Hu G Q,Deng Y,Zhang H.Methermicoccus shengliensis gen.nov.,sp.nov.,a thermophilic,methylotrophic methanogen isolated from oil-production water,and proposal of Methermicoccaceae fam.nov[J].Int J Syst Evol Microbiol,2007,57(12):2964-2969.
[42]Davidova I A,Harmsen H J M,Stams A J M,Belyaev S S,Zehnder A J B.Taxonomic description of Methanococcoides euhalobius and its transfer to the Methanohalophilus genus[J].Antonie Leeuwenhoek,1997,71(4):313-318.
[43]Mori K,Iino T,Suzuki K i,Yamaguchi K,Kamagata Y.Aceticlastic and Na Cl-requiring Methanogen“Methanosaeta pelagica”sp.nov.,Isolated from Marine Tidal Flat Sediment[J].Appl Environ Microbiol,2012,78(9):3416-3423.
[44]Patel G B,Sprott G D.Methanosaeta concilii gen.nov.,sp.nov.(“Methanothrix concilii”)and Methanosaeta thermoacetophila nom.rev.,comb.nov[J].Int J Syst Bacteriol,1990,40(1):79-82.
[45]Cheng L,Ding C,Li Q,He Q,Dai L-r,Zhang H.DNASIP reveals that Syntrophaceae play an important role in methanogenic hexadecane degradation[J].PLo S ONE,2013,8(7):e66784.
[46]Cheng L,He Q,ding C,Dai L-r,Li Q,Zhang H.Novel bacterial groups dominate in a thermophilic methanogenic hexadecane-degrading consortium FEMS Microbiol[J].Ecol,2013,85(3):568-577.