莫莫格湿地石油污染土壤中耐盐碱石油烃降解细菌的降解特性研究
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摘要
原位筛选耐盐碱石油烃降解菌可以用来修复受石油污染的盐碱土壤。本研究从莫莫格湿地的石油污染土壤中,以菲(Phe)作为唯一碳源,分离出一株多环芳烃耐盐碱的Phe高效降解细菌株,通过形态学及16S rRNA基因序列分析,明确其系统发育进化地位;系统研究了该菌株对Phe的降解特性及其降解动力学,为湿地石油污染的治理提供了良好的菌种资源。研究结果表明,耐盐碱的Phe降解细菌S1-8菌株,属于盐单胞菌,命名为Halomonas sp. S1-8; Halomonas sp. S1-8降解Phe的最适盐浓度(Na Cl)为5%,p H为8. 0,温度为30℃。在盐浓度低于5%时,随着盐度的增加,菌株S1-8在盐碱胁迫下对Phe的降解动力学,符合一级降解动力学模型。菌株S1-8对盐浓度和环境酸碱度具有较广的适应范围,分别为Na Cl含量0～30%和p H 6. 0～11. 0。Halomonas sp. S1-8在湿地油田污染的生物修复中具有潜在的应用前景。
In situ screening of saline-alkali-tolerant and petroleum hydrocarbon-degrading bacteria can be used to remediate the saline-alkaline soils contaminated with petroleum. In this study,a polycyclic aromatic hydrocarbon degrading strain was isolated from the oil-contaminated soil of the Momoge wetland with phenanthrene( Phe) as the sole carbon source. The morphology and 16 S rRNA gene sequence analysis were identified. The degradation characteristics and degradation kinetics of the phenanthrene strain were systematically studied,which intended to provide a good solution for the treatment of wetland petroleum pollution. The morphological and phylogenetic tree analysis indicated that the isolated strain belonged to Halomonas,assigned as Halomonas sp. S1-8. The optimum salt concentration of strain S1-8 for Phe degradation was( Na Cl) = 5%,p H = 8. 0,temperature = 30 ℃. The degradation kinetics of strain S1-8 to Phe under saline-alkali stress was in accordance with the first-order degradation kinetics model. Strain S1-8 had a wide range of adaptation to salt concentration and p H,respectively. Therefore,Halomonas sp. S1-8 has a potential application prospect in bioremediation of oil field pollution.
In situ screening of saline-alkali-tolerant and petroleum hydrocarbon-degrading bacteria can be used to remediate the saline-alkaline soils contaminated with petroleum. In this study,a polycyclic aromatic hydrocarbon degrading strain was isolated from the oil-contaminated soil of the Momoge wetland with phenanthrene( Phe) as the sole carbon source. The morphology and 16 S rRNA gene sequence analysis were identified. The degradation characteristics and degradation kinetics of the phenanthrene strain were systematically studied,which intended to provide a good solution for the treatment of wetland petroleum pollution. The morphological and phylogenetic tree analysis indicated that the isolated strain belonged to Halomonas,assigned as Halomonas sp. S1-8. The optimum salt concentration of strain S1-8 for Phe degradation was( Na Cl) = 5%,p H = 8. 0,temperature = 30 ℃. The degradation kinetics of strain S1-8 to Phe under saline-alkali stress was in accordance with the first-order degradation kinetics model. Strain S1-8 had a wide range of adaptation to salt concentration and p H,respectively. Therefore,Halomonas sp. S1-8 has a potential application prospect in bioremediation of oil field pollution.
引文
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[23]张海荣,唐景春,孙克静,等.耐盐碱石油烃降解菌的筛选、鉴定及耐盐碱性研究[J].生物技术通报,2015,31(1):151-159.ZHANG H R,TANG J C,SUN K J,et al.Isolation and identification of saline-alkaline tolerant hydrocarbon-degrading strains and study on their saline-alkaline tolerant characteristics[J].Biotechnology Bulletin,2015,31(1):151-159.
[24]马静.多环芳烃降解菌的筛选、降解机理及降解性能研究[D].大连:大连理工大学,2011.MA J.Isolation of PAHs-degrading bacteria and characterization of the degradation metabolism[D].Dalian:Dalian University of Technology,2011.
[25]李丹丹,徐兴健,翟真浩,等.一株筛选自莫莫格湿地石油污染土壤中的十六烷烃降解细菌的分离鉴定及降解特性研究[J].湿地科学,2017,15(1):85-91.LI D D,XU X J,ZHAI Z H,et al.Isolation and identification an n-Hexadecane bacterial degrader from soil polluted by petroleum oil in Momoge Wetlands and its degradation characteristics[J].Wetland Science,2017,15(1):85-91.
[26]KONG J,WANG H,LIANG L,et al.Phenanthrene degradation by the bacterium Pseudomonas stutzeri JP1 under low oxygen condition[J].International Biodeterioration&Biodegradation,2017,123:121-126.
[27]宋立超.盐土多环芳烃降解菌筛选分离及其污染修复应用基础研究[D].沈阳:沈阳农业大学,2011.SONG L C.Screening of PAHs-degrading microbes in salt soil and basic study of their application on the bioremediation of pollutants[D].Shenyang:Shenyang Agricultural University,2011.
[2]WANG C Y,LIU X,GUO J,et al.Biodegradation of marine oil spill residues using aboriginal bacterial consortium based on Penglai 79-3 oil spill accident,China[J].Ecotoxicology and Environmental Safety,2018,159:20-27.
[3]PRASAD S J,BALAKRISHNAN N,RAHAMAN H,et al.An assessment on oil spill trajectory prediction:Case study on oil spill off Ennore Port[J].Journal of Earth System Science,2018,127(8):111-118.
[4]邱清华,哈尼帕,邓绍云.微生物降解石油烃类污染物的研究进展[J].资源开发与市场,2012,28(2):144-147.QIU Q H,HA,DENG S Y.Research progress on microbial degradation of petroleum hydrocarbon[J].Resource Development and Market,2012,28(2):144-147.
[5]PAN X L,ZHANG D Y,QUAN L.Interactive factors leading to dying off Carex tato in Momoge wetland polluted by crude oil,western Jilin,China[J].Chemosphere,2006,65(10):1772-1777.
[6]OH J Y,CHOI S D,KWON H O,et al.Leaching of polycyclic aromatic hydrocarbons(PAHs)from industrial wastewater sludge by ultrasonic treatment[J].Ultrasonics Sonochemistry,2016,33:61-66.
[7]于一雷,马牧源,徐卫刚,等.碱蓬修复黄河三角洲原油污染土壤试验研究[J].生态环境学报,2018,27(10):1958-1965.YU Y L,MA M Y,XU W G,et al.Study on the remediation of soil contaminated by crude oil using Suaeda salsa in Yellow River Delta[J].Ecology and Environmental Sciences,2018,27(10):1958-1965.
[8]TIAN W J,ZHAO J,ZHOU Y H,et al.Effects of root exudates on gel-beads/reeds combination remediation of high molecular weight polycyclic aromatic hydrocarbons[J].Ecotoxicology and Environmental Safety,2017,135:158-164.
[9]WU M L,CHEN L M,TIAN Y Q,et al.Degradation of polycyclic aromatic hydrocarbons by microbial consortia enriched from three soils using two different culture media[J].Environment Pollution,2013,178:152-158.
[10]SREN T,BRUMMER G W.Fractionated extraction of polycyclic aromatic hydrocarbons(PAHs)from polluted soils:Estimation of the PAHfraction degradable through bioremediation[J].European Journal of Soil Science,2004,55(3):567-578.
[11]ZHANG C,LU J,WU J,et al.Removal of phenanthrene from coastal waters by green tide algae Ulva prolifera[J].Science of the Total Environment,2017,609:1322-1328.
[12]GAO H,ZHANG J H,LAI H X,et al.Degradation of asphaltenes by two Pseudonmoas aeruginosa strains and their effects on physicochemical properties of crude oil[J].International Biodeterioration&Biodegradation,2017,122(6):12-22.
[13]GU Q H,WU Q P,ZHANG J M,et al.Acinetobacter sp.DW-1 immobilized on polyhedron hollow polypropylene balls and analysis of transcriptome and proteome of the bacterium during phenol biodegradation process[J].Scientific Reports,2017,7(1):4863.
[14]CUI Y W,ZHANG H Y,DING J R,et al.The effects of salinity on nitrification using halophilic nitrifiers in a Sequencing Batch Reactor treating hypersaline wastewater[J].Scientific Reports,2016,6:24825.
[15]石伟.极端盐碱土壤细菌的分离筛选及抗盐特性研究[D].哈尔滨:东北林业大学,2011.SHI W.Screening and resistance-analysis of microorganisms in extreme saline soil environments[D].Harbin:Northeast Forestry University,2011.
[16]刘健.胜利油田采油区土壤石油污染状况及其微生物群落结构[D].济南:山东大学,2014.LIU J.Petroleum pollution and the microbial community structure in the soil of Shengli Oilfield[D].Jinan:Shandong University,2014.
[17]梁静南,刘一苇,谢家仪.制备细菌类单细胞生物扫描电镜样品方法的改进[J].电子显微学报,2013,32(3):276-278.LIANG J N,LIU Y W,XIE J Y.The improvement of bacteria sample preparation method for scanning electron microscope observation[J].Journal of Chinese Electron Microscopy Society,2013,32(3):276-278.
[18]LOTFABAD S,GRAY M R.Kinetics of biodegradation of mixtures of polycyclic aromatichydrocarbons[J].Applied Microbiology and Biotechnology,2002,60(3):361-66.
[19]吴盼云.芘降解菌生长及降解数学模型的建立[D].乌鲁木齐:新疆大学,2018.WU P Y.Mathematical models of the growth and degradation of pyrene-degrading bacteria[D].Urumqi:Xinjiang University,2018.
[20]PUGAZHENDI A,QARI H,BASAHI J M A,et al.Role of a halothermophilic bacterial consortium for the biodegradation of PAHs and the treatment of petroleum wastewater at extreme conditions[J].International Biodeterioration and Biodegradation,2017,121:44-54.
[21]周海燕.多环芳烃嗜盐降解菌的分离及芘降解机制的研究[D].北京:清华大学,2015.ZHOU H Y.Isolation of PAHs-degrading halophiles and study on the mechanism of pyrene degradation[D].Beijing:Tsinghua University,2015.
[22]苗朝华.中度嗜盐新种黄河盐单胞菌盐胁迫的适应机制研究[D].北京:中国农业大学,2016.MIAO Z H.Adaptation mechanisms of a novel moderately halophilic species Halomonas huangheensis response to salinity stress[D].Beijing:China Agricultural University,2016.
[23]张海荣,唐景春,孙克静,等.耐盐碱石油烃降解菌的筛选、鉴定及耐盐碱性研究[J].生物技术通报,2015,31(1):151-159.ZHANG H R,TANG J C,SUN K J,et al.Isolation and identification of saline-alkaline tolerant hydrocarbon-degrading strains and study on their saline-alkaline tolerant characteristics[J].Biotechnology Bulletin,2015,31(1):151-159.
[24]马静.多环芳烃降解菌的筛选、降解机理及降解性能研究[D].大连:大连理工大学,2011.MA J.Isolation of PAHs-degrading bacteria and characterization of the degradation metabolism[D].Dalian:Dalian University of Technology,2011.
[25]李丹丹,徐兴健,翟真浩,等.一株筛选自莫莫格湿地石油污染土壤中的十六烷烃降解细菌的分离鉴定及降解特性研究[J].湿地科学,2017,15(1):85-91.LI D D,XU X J,ZHAI Z H,et al.Isolation and identification an n-Hexadecane bacterial degrader from soil polluted by petroleum oil in Momoge Wetlands and its degradation characteristics[J].Wetland Science,2017,15(1):85-91.
[26]KONG J,WANG H,LIANG L,et al.Phenanthrene degradation by the bacterium Pseudomonas stutzeri JP1 under low oxygen condition[J].International Biodeterioration&Biodegradation,2017,123:121-126.
[27]宋立超.盐土多环芳烃降解菌筛选分离及其污染修复应用基础研究[D].沈阳:沈阳农业大学,2011.SONG L C.Screening of PAHs-degrading microbes in salt soil and basic study of their application on the bioremediation of pollutants[D].Shenyang:Shenyang Agricultural University,2011.