基于高通量测序分析的生物修复石油污染土壤菌群结构变化
|
摘要
利用高通量测序技术对微生物修复石油污染土壤过程中的微生物群落结构变化进行研究.结果表明,经修复处理的土壤微生物群落结构及多样性发生明显变化.利用生物强化修复处理(BA)的土壤中,微生物丰富度与均匀度明显降低,土著菌群受到抑制,外加变形菌门(Proteobacteria)成为主要的优势菌门,相对丰度由修复前的37. 44%增加为87. 44%.假单胞菌属(Pseudomonas)成为土壤中的优势菌属,丰度由2. 99%增加为76. 37%;进行生物刺激修复处理的土壤(BS)菌群丰富度和均匀度与原污染土壤相比略有降低.菌群结构组成上,原优势菌门变形菌门(Proteobacteria)丰度由37. 44%降低为10. 90%,厚壁菌门(Firmicutes)丰度由9. 16%增加为35. 32%,属水平上,原优势菌属微小杆菌属(Exiguobacterium)和原小单胞菌属(Promicromonospora)丰度由8. 49%和18. 96%分别降低为2. 19%和14. 97%,诺卡氏菌属(Nocardioides)和芽孢杆菌属(Bacillus)丰度由5. 56%和0. 29%分别增加至28. 95%和22. 70%,成为主要优势菌属.生物强化修复处理引起土壤菌群多样性和结构发生明显变化,生物刺激修复处理可基本保持土壤菌群结构多样性不被破坏,土壤菌群结构的稳定有利于石油烃的生物降解.
The shift in microbial community structure during the bioremediation of oil-polluted soil was analyzed by high-throughput sequencing. The results demonstrated obvious changes in the soil microbial community structure and diversity during bioremediation.The species richness and evenness of the microbial community decreased substantially due to the bioaugmentation treatment.Proteobacteria became the predominant phylum,with a relative increase in abundance from 37. 44% to 87. 44%. Pseudomonas was the most dominant genus,which increased in abundance from 2. 99% to 76. 37%. In the biostimulation treated soil,the relative abundance of Proteobacteria decreased from 37. 44% to 10. 90%,while the phylum Firmicutes increased from 9. 16% to 35. 32%. At the genus level,the relative abundances of Exiguobacterium and Promicromonospora decreased from 8. 49% and 18. 96% to 2. 19%and 14. 97%,respectively. Nocardioides and Bacillus became the dominant genera and increased from 5. 56% and 0. 29% to 28. 95%and 22. 70%,respectively. The results indicated that bioaugmentation substantially influenced the soil microbial diversity and community structure. Additionally,the biostimulation treatment maintained the balance in the soil microbial community structure. The stabilization of bacteria community structure is beneficial to petroleum biodegradation in the soil.
The shift in microbial community structure during the bioremediation of oil-polluted soil was analyzed by high-throughput sequencing. The results demonstrated obvious changes in the soil microbial community structure and diversity during bioremediation.The species richness and evenness of the microbial community decreased substantially due to the bioaugmentation treatment.Proteobacteria became the predominant phylum,with a relative increase in abundance from 37. 44% to 87. 44%. Pseudomonas was the most dominant genus,which increased in abundance from 2. 99% to 76. 37%. In the biostimulation treated soil,the relative abundance of Proteobacteria decreased from 37. 44% to 10. 90%,while the phylum Firmicutes increased from 9. 16% to 35. 32%. At the genus level,the relative abundances of Exiguobacterium and Promicromonospora decreased from 8. 49% and 18. 96% to 2. 19%and 14. 97%,respectively. Nocardioides and Bacillus became the dominant genera and increased from 5. 56% and 0. 29% to 28. 95%and 22. 70%,respectively. The results indicated that bioaugmentation substantially influenced the soil microbial diversity and community structure. Additionally,the biostimulation treatment maintained the balance in the soil microbial community structure. The stabilization of bacteria community structure is beneficial to petroleum biodegradation in the soil.
引文
[1]郝春博,王广才,董健楠,等.石油污染地下水中细菌多样性研究[J].环境科学,2009,30(8):2464-2472.Hao C B,Wang G C,Dong J N,et al. Bacterial biodiversity in the groundwater contaminated by oil[J]. Environmental Science,2009,30(8):2464-2472.
[2] Lim M W,Lau E V,Poh P E. A comprehensive guide of remediation technologies for oil contaminated soil-Present works and future directions[J]. Marine Pollution Bulletin,2016,109(1):14-45.
[3]胡迪,李川,董倩倩,等.油田区土壤石油烃组分残留特性研究[J].环境科学,2014,35(1):227-232.Hu D,Li C,Dong Q Q,et al. Compositions and residual properties of petroleum hydrocarbon in contaminated soil of the oilfields[J]. Environmental Science,2014,35(1):227-232.
[4]王泽华,王峰.低温条件下土壤石油烃微生物修复研究进展[J].环境科学与技术,2017,40(S1):139-144.Wang Z H,Wang F. Review on bioremediation of petroleum hydrocarbon-contaminated soil in cold condition[J].Environmental Science&Technology,2017,40(S1):139-144.
[5]吴蔓莉,陈凯丽,叶茜琼,等.堆肥-生物强化对重度石油污染土壤的修复作用[J].环境科学,2017,38(10):4412-4419.Wu M L,Chen K L,Ye X Q,et al. Remediation of petroleumcontaminated soil using a bioaugmented compost technique[J].Environmental Science,2017,38(10):4412-4419.
[6]杨茜,吴蔓莉,聂麦茜,等.石油污染土壤的生物修复技术及微生物生态效应[J].环境科学,2015,36(5):1856-1863.Yang Q,Wu M L,Nie M Q,et al. Effects and biological response on bioremediation of petroleum contaminated soil[J].Environmental Science,2015,36(5):1856-1863.
[7]吴蔓莉,袁婧,李炜,等.石油污染土壤的微生物修复及土壤微生物活性变化[J].应用与环境生物学报,2016,22(5):878-883.Wu M L,Yuan J,Li W,et al. Bioremediation of petroleum contaminated soil and activity of hydrocarbon degrading bacteria[J]. Chinese Journal of Applied&Environmental Biology,2016,22(5):878-883.
[8] Wu M L, Dick W A,Li W, et al. Bioaugmentation and biostimulation of hydrocarbon degradation and the microbial community in a petroleum-contaminated soil[J]. International Biodeterioration&Biodegradation,2016,107:158-164.
[9] Shahi A,Aydin S,Ince B,et al. Reconstruction of bacterial community structure and variation for enhanced petroleum hydrocarbons degradation through biostimulation of oil contaminated soil[J]. Chemical Engineering Journal,2016,306:60-66.
[10] Liu P W G,Chang T C,Whang L M,et al. Bioremediation of petroleum hydrocarbon contaminated soil:Effects of strategies and microbial community shift[J]. International Biodeterioration&Biodegradation,2011,65(8):1119-1127.
[11]刘艳秋,赵嫣然,刘梦娇,等.外源环烷酸在土壤中的降解过程及对微生物群落结构的影响[J].环境科学,2017,38(11):4756-4762.Liu Y Q,Zhao Y R,Liu M J,et al. Degradation process of exogenous naphthenic acids and their effects on microbial community structure in soil[J]. Environmental Science,2017,38(11):4756-4762.
[12] Pacwa-Pociniczak M, Paza G A, Piotrowska-Seget Z.Monitoring the changes in a bacterial community in petroleumpolluted soil bioaugmented with hydrocarbon-degrading strains[J]. Applied Soil Ecology,2016,105:76-85.
[13]杨萌青,李立明,李川,等.石油污染土壤微生物群落结构与分布特性研究[J].环境科学,2013,34(2):789-794.Yang M Q,Li L M,Li C,et al. Microbial community structure and distribution characteristics in oil contaminated soil[J].Environmental Science,2013,34(2):789-794.
[14] Jiao H H,Bai Z H,Liu Y,et al. Impact of super absorbent polymer and plants on microbial community and petroleum hydrocarbon degradation in contaminated soil[J]. Advanced Materials Research,2013,807-809:353-360.
[15]焦海华,崔丙健,吴尚华,等.紫茉莉对石油污染盐碱土壤微生物群落与石油烃降解的影响[J].环境科学,2015,36(9):3471-3478.Jiao H H,Cui B J,Wu S H,et al. Influence of Mirabilis jalapa Linn. Growth on the microbial community and petroleum hydrocarbon degradation in petroleum contaminated saline-alkali soil[J]. Environmental Science,2015,36(9):3471-3478.
[16] Grant R J,Muckian L M,Clipson N J W,et al. Microbial community changes during the bioremediation of creosotecontaminated soil[J]. Letters in Applied Microbiology,2010,44(3):293-300.
[17] Wang L W,Li F,Zhan Y,et al. Shifts in microbial community structure during in situ surfactant-enhanced bioremediation of polycyclic aromatic hydrocarbon-contaminated soil[J].Environmental Science and Pollution Research,2016,23(14):14451-14461.
[18]叶茜琼.微生物修复对石油烃的去除特性及土壤微生态环境变化研究[D].西安:西安建筑科技大学,2017.
[19]叶茜琼,吴蔓莉,陈凯丽,等.微生物修复油污土壤过程中氮素的变化及菌群生态效应[J].环境科学,2017,38(2):728-734.Ye X Q,Wu M L,Chen K L,et al. Impacts of bioremediation on microbial communities and different forms of nitrogen in petroleum contaminated soil[J]. Environmental Science,2017,38(2):728-734.
[20] Wu M L,Li W,Dick W A,et al. Bioremediation of hydrocarbon degradation in a petroleum-contaminated soil and microbial population and activity determination[J]. Chemosphere,2017,169:124-130.
[21]王鑫,王学江,刘免,等.高效石油降解菌群构建及降解性能[J].海洋环境科学,2014,33(4):576-579,597.Wang X,Wang X J,Liu M,et al. Construction of high-effective microbial consortium and characteristics of oil biodegrading[J].Marine Environmental Science,2014,33(4):576-579,597.
[22]乔俊.油田区石油污染土壤的生物修复及不同修复方法的比较[D].天津:南开大学,2011.
[23] Poi G, Aburto-Medina A, Mok P C, et al. Large scale bioaugmentation of soil contaminated with petroleum hydrocarbons using a mixed microbial consortium[J]. Ecological Engineering,2017,102:64-71.
[24] Li H,Ye D D,Wang X G,et al. Soil bacterial communities of different natural forest types in Northeast China[J]. Plant and Soil,2014,383(1-2):203-216.
[25] Sengupta A,Dick W A. Bacterial community diversity in soil under two tillage practices as determined by pyrosequencing[J].Microbial Ecology,2015,70(3):853-859.
[26] Sun H Y,Deng S P,Raun W R. Bacterial community structure and diversity in a century-old manure-treated agro ecosystem[J].Applied and Environmental Microbiology,2004,70(10):5868-5874.
[27]王光华,刘俊杰,齐晓宁,等. Biolog和PCR-DGGE技术解析施肥对德惠黑土细菌群落结构和功能的影响[J].生态学报,2008,28(1):220-226.Wang G H,Liu J J,Qi X N,et al. Effects of fertilization on bacterial community structure and function in a black soil of Dehui region estimated by Biolog and PCR-DGGE methods[J].Acta Ecologica Sinica,2008,28(1):220-226.
[28]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.
[29] Khan M A I,Biswas B,Smith E,et al. Microbial diversity changes with rhizosphere and hydrocarbons in contrasting soils.[J]. Ecotoxicology and Environmental Safety,2018,156:434-442.
[30]谭银萍,马媛,吕杰.石油污染对土壤细菌结构特征的影响[J].生物技术,2016,26(2):193-198.Tan Y P,Ma Y,Lv J. Effect of oil pollution on soil bacteria structure characteristics[J]. Biotechnology,2016,26(2):193-198.
[2] Lim M W,Lau E V,Poh P E. A comprehensive guide of remediation technologies for oil contaminated soil-Present works and future directions[J]. Marine Pollution Bulletin,2016,109(1):14-45.
[3]胡迪,李川,董倩倩,等.油田区土壤石油烃组分残留特性研究[J].环境科学,2014,35(1):227-232.Hu D,Li C,Dong Q Q,et al. Compositions and residual properties of petroleum hydrocarbon in contaminated soil of the oilfields[J]. Environmental Science,2014,35(1):227-232.
[4]王泽华,王峰.低温条件下土壤石油烃微生物修复研究进展[J].环境科学与技术,2017,40(S1):139-144.Wang Z H,Wang F. Review on bioremediation of petroleum hydrocarbon-contaminated soil in cold condition[J].Environmental Science&Technology,2017,40(S1):139-144.
[5]吴蔓莉,陈凯丽,叶茜琼,等.堆肥-生物强化对重度石油污染土壤的修复作用[J].环境科学,2017,38(10):4412-4419.Wu M L,Chen K L,Ye X Q,et al. Remediation of petroleumcontaminated soil using a bioaugmented compost technique[J].Environmental Science,2017,38(10):4412-4419.
[6]杨茜,吴蔓莉,聂麦茜,等.石油污染土壤的生物修复技术及微生物生态效应[J].环境科学,2015,36(5):1856-1863.Yang Q,Wu M L,Nie M Q,et al. Effects and biological response on bioremediation of petroleum contaminated soil[J].Environmental Science,2015,36(5):1856-1863.
[7]吴蔓莉,袁婧,李炜,等.石油污染土壤的微生物修复及土壤微生物活性变化[J].应用与环境生物学报,2016,22(5):878-883.Wu M L,Yuan J,Li W,et al. Bioremediation of petroleum contaminated soil and activity of hydrocarbon degrading bacteria[J]. Chinese Journal of Applied&Environmental Biology,2016,22(5):878-883.
[8] Wu M L, Dick W A,Li W, et al. Bioaugmentation and biostimulation of hydrocarbon degradation and the microbial community in a petroleum-contaminated soil[J]. International Biodeterioration&Biodegradation,2016,107:158-164.
[9] Shahi A,Aydin S,Ince B,et al. Reconstruction of bacterial community structure and variation for enhanced petroleum hydrocarbons degradation through biostimulation of oil contaminated soil[J]. Chemical Engineering Journal,2016,306:60-66.
[10] Liu P W G,Chang T C,Whang L M,et al. Bioremediation of petroleum hydrocarbon contaminated soil:Effects of strategies and microbial community shift[J]. International Biodeterioration&Biodegradation,2011,65(8):1119-1127.
[11]刘艳秋,赵嫣然,刘梦娇,等.外源环烷酸在土壤中的降解过程及对微生物群落结构的影响[J].环境科学,2017,38(11):4756-4762.Liu Y Q,Zhao Y R,Liu M J,et al. Degradation process of exogenous naphthenic acids and their effects on microbial community structure in soil[J]. Environmental Science,2017,38(11):4756-4762.
[12] Pacwa-Pociniczak M, Paza G A, Piotrowska-Seget Z.Monitoring the changes in a bacterial community in petroleumpolluted soil bioaugmented with hydrocarbon-degrading strains[J]. Applied Soil Ecology,2016,105:76-85.
[13]杨萌青,李立明,李川,等.石油污染土壤微生物群落结构与分布特性研究[J].环境科学,2013,34(2):789-794.Yang M Q,Li L M,Li C,et al. Microbial community structure and distribution characteristics in oil contaminated soil[J].Environmental Science,2013,34(2):789-794.
[14] Jiao H H,Bai Z H,Liu Y,et al. Impact of super absorbent polymer and plants on microbial community and petroleum hydrocarbon degradation in contaminated soil[J]. Advanced Materials Research,2013,807-809:353-360.
[15]焦海华,崔丙健,吴尚华,等.紫茉莉对石油污染盐碱土壤微生物群落与石油烃降解的影响[J].环境科学,2015,36(9):3471-3478.Jiao H H,Cui B J,Wu S H,et al. Influence of Mirabilis jalapa Linn. Growth on the microbial community and petroleum hydrocarbon degradation in petroleum contaminated saline-alkali soil[J]. Environmental Science,2015,36(9):3471-3478.
[16] Grant R J,Muckian L M,Clipson N J W,et al. Microbial community changes during the bioremediation of creosotecontaminated soil[J]. Letters in Applied Microbiology,2010,44(3):293-300.
[17] Wang L W,Li F,Zhan Y,et al. Shifts in microbial community structure during in situ surfactant-enhanced bioremediation of polycyclic aromatic hydrocarbon-contaminated soil[J].Environmental Science and Pollution Research,2016,23(14):14451-14461.
[18]叶茜琼.微生物修复对石油烃的去除特性及土壤微生态环境变化研究[D].西安:西安建筑科技大学,2017.
[19]叶茜琼,吴蔓莉,陈凯丽,等.微生物修复油污土壤过程中氮素的变化及菌群生态效应[J].环境科学,2017,38(2):728-734.Ye X Q,Wu M L,Chen K L,et al. Impacts of bioremediation on microbial communities and different forms of nitrogen in petroleum contaminated soil[J]. Environmental Science,2017,38(2):728-734.
[20] Wu M L,Li W,Dick W A,et al. Bioremediation of hydrocarbon degradation in a petroleum-contaminated soil and microbial population and activity determination[J]. Chemosphere,2017,169:124-130.
[21]王鑫,王学江,刘免,等.高效石油降解菌群构建及降解性能[J].海洋环境科学,2014,33(4):576-579,597.Wang X,Wang X J,Liu M,et al. Construction of high-effective microbial consortium and characteristics of oil biodegrading[J].Marine Environmental Science,2014,33(4):576-579,597.
[22]乔俊.油田区石油污染土壤的生物修复及不同修复方法的比较[D].天津:南开大学,2011.
[23] Poi G, Aburto-Medina A, Mok P C, et al. Large scale bioaugmentation of soil contaminated with petroleum hydrocarbons using a mixed microbial consortium[J]. Ecological Engineering,2017,102:64-71.
[24] Li H,Ye D D,Wang X G,et al. Soil bacterial communities of different natural forest types in Northeast China[J]. Plant and Soil,2014,383(1-2):203-216.
[25] Sengupta A,Dick W A. Bacterial community diversity in soil under two tillage practices as determined by pyrosequencing[J].Microbial Ecology,2015,70(3):853-859.
[26] Sun H Y,Deng S P,Raun W R. Bacterial community structure and diversity in a century-old manure-treated agro ecosystem[J].Applied and Environmental Microbiology,2004,70(10):5868-5874.
[27]王光华,刘俊杰,齐晓宁,等. Biolog和PCR-DGGE技术解析施肥对德惠黑土细菌群落结构和功能的影响[J].生态学报,2008,28(1):220-226.Wang G H,Liu J J,Qi X N,et al. Effects of fertilization on bacterial community structure and function in a black soil of Dehui region estimated by Biolog and PCR-DGGE methods[J].Acta Ecologica Sinica,2008,28(1):220-226.
[28]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.
[29] Khan M A I,Biswas B,Smith E,et al. Microbial diversity changes with rhizosphere and hydrocarbons in contrasting soils.[J]. Ecotoxicology and Environmental Safety,2018,156:434-442.
[30]谭银萍,马媛,吕杰.石油污染对土壤细菌结构特征的影响[J].生物技术,2016,26(2):193-198.Tan Y P,Ma Y,Lv J. Effect of oil pollution on soil bacteria structure characteristics[J]. Biotechnology,2016,26(2):193-198.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |