跳到主要內容

簡易檢索 / 詳目顯示

研究生: 林依蓉
Yi-Rong Lin
論文名稱: 多氯聯苯厭氧馴養降解菌群微生物多樣性解析
指導教授: 劉文佐
口試委員:
學位類別: 碩士
Master
系所名稱: 生醫理工學院 - 生命科學系
Department of Life Science
畢業學年度: 89
語文別: 中文
論文頁數: 81
中文關鍵詞: 多氯聯苯厭氧還原性脫氯作用微生物多樣性變性梯度凝膠電泳
外文關鍵詞: PCB, microbial diversity, DGGE, FISH, clone li
相關次數: 點閱:7下載:0
分享至:
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報



  • 第一章前言 1.1 研究緣起…………………………………………………………..1 1.2 研究目的……………………………………………………………4 第二章文獻回顧 2.1 應用微生物降解環境污染物………………………………………6 2.2 厭氧還原脫氯作用…………………………………………………7 2.3 多氯聯苯厭氧生物降解…...………………………………………12 2.4環境微生物分生分析法……………………………………………20 第三章研究方法與原理 3.1實驗方法……………………………………………………………25 3.2方法原理與步驟……………………………………………………31 第四章結果與討論 4.1 不同厭氧狀態和不同基質馴養對多氯聯苯脫氯菌群結構之影響43 4.2 不同多氯聯苯基質長期馴養之菌群結構變化……………………46 4.3 不同多氯聯苯基質馴養之菌群結構組成…………………………52 4.4 以FISH觀察多氯聯苯厭氧馴養菌群的結構組成………………68 第五章結論與建議 5.1 結論…………………………………………………………………71 5.2 建議…………………………………………………………………72 第六章 參考文獻……………………...…………………………………73 圖 目 錄 圖1.1多氯聯苯結構式……………………….……………………..……4 圖1.2環境中多氯聯苯傳播循環途徑……………………………………5 圖2.1厭氧還原性脫氯作用………………………………………………8 圖2.2厭氧脫氯菌群親源樹……………………... …………………….11 圖2.3PCB鄰位脫氯菌群親源樹………………………………………..19 圖 2.4應用分生技術分析環境微生物流程圖……………………………23 圖3.1不同厭氧狀態對基隆河底泥脫氯作用的影響……………………28 圖3.2不同厭樣狀態對脫氯菌群影響之實驗流程………………………29 圖3.3多氯聯苯降解菌群16S rDNA clone library建立流程圖…………30 圖4.1不同厭氧狀態和不同基質馴養之多氯聯苯脫氯菌群結構DGGE指紋譜………………………………………………………………..45 圖4.2多氯聯苯降解降解活性測試………………………………47 圖4.32,2'',5,5''-CB降解菌群DGGE指紋譜……………………………49 圖4.42,2'',5,5''-CB降解菌群DGGE指紋譜菌群結構聚類分析結果…49 圖4.52,2'',4,5,5''-CB脫氯菌群長期馴養之DGGE指紋譜變化………..51 圖4.62,2'',4,5,5''-CB降解菌群DGGE指紋譜菌群結構聚類分析結果..51 圖4.72,4'',5-CB馴養菌群之分子選植DGGE快速大量篩選圖……….54 圖4.82,4'',5-CB馴養菌群之分子選殖分組排序結果……………....…..55 圖4.92,4'',5-CB馴養菌群親源樹………………………………………..56 圖4.10 2,2'',5,5''-CB馴養之菌群親源樹…………………………………...58 圖4.11 以2,2'',4,5,5''-CB馴養之菌群親源….……………………………..60 圖4.12 控制組菌群親源樹...…………………….…………………………62 圖4.13 脫氯菌群親源樹……………………………………………………65 圖4.14 FISH觀察2,2'',4,5,5''-CB馴養菌群結果………………………….70 表 目 錄 表2.1 厭氧還原性脫氯菌株………………………………………………10 表2.2 PCB厭氧降解途徑…………………………………………………14 表2.3para-和meta-PCB降解菌群SSU rRNA序列比對之親源結果…18 表2.4細菌系統分類學之種類與特性……………………………………21 表3.1PCR primers………………………………………………………...33 表3.2DGGE試劑配方……………………………………………………36 表3.3FISH採用的螢光探針……………………………………………..41 表4.1比較四組馴養菌群和控制組菌群之菌群組成分布………………66 表4.2不同含氯有機物厭氧降解菌群分布…………………….…………67 表4.3以定量FISH分析2,2'',4,5,5''-CB馴養菌群的結構組成之結果….67

    Abramowicz, D. A. 1995. Aerobic and anaerobic PCB biodegradation in the environment. Environmental Health Perspective. 103:97-99.
    Amann, R. I., B. J. Binder, R. J. Olson, S.W. Chisholm, R. Devereux, and D.A. Stahl. 1990. Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial population. Applied and Environmental Microbiology. 56:1919-1925.
    Amann, R. I., W. Ludwig, and K.-H. Schleifer. 1995. Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiological Reviews 59:143-169.
    Bedard, D. L., and J. Quensen. 1995. Microbial reductive dechlorination of polychlorinated biphenyls. In “Microbial Transformation and Degradation of Toxic Organic Chemicals”. Young L.Y. and Cernigilia C., Eds., pp. 127-216. Wiley-Liss Division, Joun Wiley and Sons, New York.
    Blackall, L. L., S. Rossetti, C. Christensson, M. Cunningham, P. Hartman, P. Hugenholtz, and V. Tandoi. 1997. The characterization and description of representatives of ‘G’ bacteria from activated sludge plants. Letters of Applied Microbiology. 25:63-69.
    Brosius, J., T. J. Dull, D.D. Sleeter, and H. F. Noller. 1981. Gene organization and primary structure of a ribosomal RNA operon from Escherichia coli. Journal of Molecular Biology. 148: 107-127.
    Brown, J., R.E. Wagner, D. L. Bedard, H. Feng, D. L. Bedard, M. J. Brennan, J. C. Carnahan, and R. J. May. 1987. Environmental dechlorination of PCBs. Environmental Toxicology Chemistry. 6: 579-593.
    Brown, M. P., B. Bush, G. Y. Rhee, and L.Shane. 1988 . PCB dechlorination in Hudson River sediment. Science. 240:1674-1676.
    Brown-Howland, E.B., S.A. Danielsen, and S.A. Nierzwicki-Bauer. 1992. Development of a rapid method for detecting bacterial cells in situ using 16S-targeted probes. BioTechniques. 13:928-933.
    Busse, H.J., E.M.B. Denner, and W. Lubitz. 1996. Classfication and identification of bacteria: current approaches to an old problem. Overview of methods used in bacterial systematics. Journal of Biotechnology. 47:3-38.
    Chang B. V., S. W. Chou, and S. Y. Yuan. 1999. Microbial dechlorination of polychlorinated biphenyls in anaerobic sewage sludge. Chemosphere. 39:45-54.
    Chang, B. V., C. W. Chiang, and S. Y. Yuan. 1998. Dechlorination of pentachlorophenol in anaerobic sewage sludge. Chemosphere. 36:537-545.
    Chang, B. V., K. S. Chen, and S. Y. Yuan. 1995. Dechlorination of 2,4,6-TCP by an anaerobic mixed culture. Chemosphere. 31:3803-3811.
    Chiarenzelli, J. R., R. J. Scrudato, D. E. Rafferty, M. L. Wunderlich, and R. N. Roberts. 1995. Photocatalytic degradation of simulated pesticide rinsates in water and water+soil matrices. Chemosphere. 30:173-185
    Chiavarini, J., R. Scrudato, M. Wunderlich, D. Rafferty, K. Jensen, G. Oenga, R. Roberts, and J. Pagano. Photodecomposition of PCBs absorbed on sediment and industrial waste: Implication for photocatalytic treatment of contaminated solids. Chemosphere. 31:3259-3272.
    Cremisini C., and P. Soldati. 1995Crocetti, G. R., P. Hugenholtz, P. L. Bond, A. Schuler, J. Keller, D. Jenkins, and L. L. Blackall. 2000. Identification of polyphosphate-accumulating organisms and design of 16S rRNA-directed probes for their detection and quantitation. Applied and Environmental Microbiology, 66:1175-1182.
    Deere, D., G. Vesey, N. Ashbold, K.A. Davies, K.L. Williams, and D. Veal. 1998. Evaluation of fluorochromes for flow cytomeric detection of Cryptosporidium parvum oocysts labeled by fluorescence in situ hybridization. Letter of Applied Microbiology. 27:352-356.
    Dojka, M.A., P. Hugenholtz, S. K. Haack, and N. R. Pace .1998. Microbial diversity in a hydrocarbon- and chlorinated-solvent-contaminated aquifer undergoing intrinsic bioremediation. Applied and Environmental Microbiology, 64:3869-3877.
    Fergusion, J. F., and J. M. H. Pietari. 2000. Anaerobic transformation of chlorinated solvents. Environmental pollution. 107: 209-215.
    Ferris, M.J., and G. Muyzer, and D.M. Ward. 1996. Denaturing gradient gel electrophoresis profiles of 16S rRNA-defined populations inhabiting a hot spring microbial mat community. Applied and Environmental Microbiology. 62:340- 346.
    Franks, A. H., H. J. M. Harmsen, G. C. Raangs, G. J. Jansen, F. Schut, and G. W. Welling. 1998. Variations of bacterial populations in human feces measured by fluorescent in situ hybridization with group-specific 16S rRNA-targeted oligonucleotide probes. Applied and Environmental Microbiology. 64:3336-3345.
    Furukawa, K., N. Kimura. 1995. Biochemistry and genetics of PCB metabolism. Environmental Health Perspectives. 103:21-23.
    Furukawa, K.. 1994. Molecular genetics and evolutionary relationship of PCB-degrading baceteria. Biodegradation. 5:289-300.
    Godon, J. J., E. Zumstein, P. Dabert, F. Habouzit, and R. Moletta . 1997. Molecular microbial diversity of an anaerobic digestor as determined by small-subunit rDNA sequence analysis . Applied and Environmental Microbiology. 63: 2802-2813.
    Godon, J.J., E. Zumstein, P. Dabert, F. Habouzit, amd R. Moletta. 1997. Molecular microbial diversity of an anaerobic digestor as determined by small-subunit rDNA sequence analysis. Applied and Environmental Microbiology, 63:2802-2813.
    Hengstmann, U., K. J. Chin, P. H. Janssen, and W. Liesack. 1999. Comparative phylogenetic assignment of environmental sequences of genes encoding 16S rRNA and numerically abundant culturable bacteria from an anoxic rice paddy soil. Applied and Environmental Microbiology. 65:5050-5058.
    Heuer, H., M. Krsek, P. Baker, K. Smalla, and E.M.H. Wellington. 1997. Analysis of actinomycete communities by specific amplification of genes encoding 16S Rrna and gel-electrophoretic separation in denaturing gradients. Applied and Environmental Microbiology. 63:3233-3241.
    Holliger, C., and G. Schraa. 1994. Physiological meaning and potential for application of reductive dechlorination by anaerobic bacteria. FEMS Microbiology Reviews. 15: 297-305.
    Holliger, C., G. Wohlfarth, and G. Diekert. 1999. Reductive dechlorination in the energy metabolism of anaerobic bacteria. FEMS Microbiology Reviews. 22: 383-398.
    Holoman, T. R. P., M. A. Elberson, L. A. Cutter, H. D. May, and K. R. Sowers. 1998. Characterization of a defined 2,3,5,6-tetrachlorbiphenyl-ortho- edchlorinating microbial community by comparative sequence analysis of genes coding for 16S rRNA. Applied and Environmental Microbiology. 64:3359-3367.
    Hou, L. H., and S. K. Dutta. 2000. Phylogenetic characterization of several para- and meta-PCB dechlorinating Clostridium species: 16S rDNA sequence analysis. Letters of Applied Microbiology. 30: 238-243.
    Hugenholtz, P., B. M. Goebel, and N. R. Pace. 1998. Impact of culture-independent studies on the emerging phylogenetic view of bacterial diversity. Journal of Bacteriology. 180: 4765-4774.
    Jurtshuk, R.J., M. Blick, J. Bresser, G.E. Pox, and P. Jurtshuk. 1992. Rapid in situ hybridization technique using 16S rRNA segments for detecting and differentiating the closely related gram-positive organisms Bacillus polymyxa and Bacillus macerans. Applied and Environmental Microbiology. 58:2571- 2578.
    Kumar, S., K. Tamura, and M. Nei. 1993. MEGA:molecular evolutionary genetics analysis, version 1.0. University Park, PA:Pennsylvania State University.
    Kuo, C. E., S. M. Liu, and C. Liu. 1999. Biodegradation of coplanar polychlorinated biphenyls by anaerobic microorganisms from estuarine sediments. Chemosphere. 39:1445-58.
    LaMontagne, M. G., G. J. Daveport, L. H. Hou, and S. K. Dutta. 1998. Identification and analysis of PCB dechlorinating anaerobic enrichments by amplification: accuracy of community structure base on restriction analysis and partial sequencing of 16S rRNA genes. Journal of Applied Microbiology. 84:1156-1162.
    Layton, A. C., M. Muccini, M. M. Chosh, and G. S. Sayler. 1998. Construction of a bioluminescent reporter strain to detect polychlorinated biphenyls. Applied and Environmental Microbiology. 64:5023-5026.
    Lee, N., P.H. Nielsen, K.H. Andreasen, S. Juretschko, J.L. Nielsen, K.-H. Schleifer, and M. Wagner. 1999. Combination of fluorescent in situ hybridization and microautoradiography — a new tool for structure — function analyses in microbial ecology. Applied and Environmental Microbiology. 65:1289-1297.
    Liu, W. T., T. L. Marsh, H. Cheng, and L. J. Forney. 1997. Characterization of microbial diversity by determining terminal restriction fragment length polymorphisms of gene coding 16S rRNA. Applied and Environmental Microbiology. 63: 4516-4522.
    Loffler, F. E., Q. Sun, and J. M. Tiedje. 2000. 16S rRNA gene-based detection of Tetrachloroethene-dechlorinating Desulfuromonas and Dehalococcoides species. Applied and Environmental Microbiology. 66:1369-1374.
    Lonergan, D.J., H. L. Jenter, J. D. Coates, E. J. Phillips, T. M. Schmidt, and D. R. Lovley. 1996. Phylogenetic analysis of dissimilatory Fe(III)-reducing bacteria. Journal of Bacteriology. 178:2402-2408.
    Maltseva, O. V., T. V. Tsoi, J. F. 3rd Quensen, M. Fukuda, and J.M. Tiedje. 1999. Degradation of anaerobic reductive dechlorination products of Aroclor 1242 by four aerobic bacteria. Biodegradation. 10:363-371.
    Manz, W., R. Amann, W. Ludwig, M. Wagner, and K.-H. Schleifer. 1992. Phylogenetic oligonucleotide probes for the major subclasses of Proteobacteria: problems and solutions. Systemic Applied Microbiology. 15:593-600.
    Matthews, J. E. 1994. Handbook of bioremediation. Lewis publishers
    Mohn W. W., and J. M. Tiedje. 1992.Microbial reductive dehalogenation. Microbial Review. 56:482-507.
    Moter, A., C. Hoeing, B.-K. Choi, B. Riep, and U.B. Gobel. 1988. Molecular epidemiology of oral treponemes associated with periodonyal disease. Journal of Clinical Microbiology. 36:1399-1403.
    Muyzer, G., and K. Smalla. 1998. Application of denaturing gradient gel electrophoresis (DGGE) and temperature gradient gel electrophoresis (TGGE) in microbial ecology. Antonie Van Leeuwenhoek. 73:127-141.
    Muyzer, G., and N. B. Ramsing. 1995. Molecular methods to study the organization of microbial communities. Water Science Technology. 32:1-9.
    Muyzer, G., E. C. DeWall, and A. G. Uitterlinden. 1993. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Applied and Environmental Microbiology. 59:695-700.
    Muyzer, G.. 1999. DGGE/TGGE a method for identifying genes from natural ecosystems. Curr Opin Microbiology. 2:317-322.
    Pace, N. R.. 1997. A molecular view of microbial diversity and the biosphere. Science. 276:734-740.
    Poulsen, L.K., G. Ballard, and D.A. Stahl. 1993. Use of rRNA fluorescence in situ hybridization for measuring the activity of single cells in young and established biofilms. Applied and Environmental Microbiology. 59:1354-1360.
    Quensen, J. F. III, J. M. Tiedje, and S. A. Boyd. 1988. Reductive dechlorination of polychlorinated biphenyls by anaerobic microorganism from sediments. Science. 242:752-754.
    Ranjard, L., F. Poly, and S. Nazaret. 2000. Mini-review. Monitoring complex bacterial communities using culture-independent molecular techniques: application to soil environment. Research Microbiology. 151:167-177.
    Rolleke, S., G. Muyzer, C. Wawer, G. Wanner, and W. Lubitz. 1996. Identification of bacteria in a biodegraded wall painting by denaturing gradient gel electrophoresis of PCR-amplified gene fragments coding for 16S rRNA . Applied and Environmental Microbiology.. 62:2059-2065.
    Roller, C., M. Wagner, R. Amann, W. Ludwig, K.-H. Schleifer. 1994. In situ probing of Gram-positive bacteria with high DNA G+C content using 23S rRNA-targeted oligonucleotides. Microbiology. 140:2849-2858.
    Schlotelburg, C., F. von Wintzingerode, R. Hauck, W. Hegemann, and U. B. Gobel. 2000. Bacteria of an anaerobic 1,2-dichloropropane-dechlorinating mixed culture are phylogenetically related to those of other anaerobic dechlorinating consortia. International Journal of Systematic and Evolutionary. 4:1505-1511.
    Stahl, D.A., and R. Amann. 1991. Development and application of nucleic acid probes in bacterial systematics, p. 205-248. In E. Stackebrandt and M. Goodfellow (cd.), Nucleic acid techniques in bacterial systematics. John Wiley & Sons Ltd., Chichester, England.
    Suau, A., R. Bonnet, M. Sutren, J. J. Godon, G. R. Gibson, M. D. Collins, and J. Dore. 1999. Direct analysis of genes encoding 16S rRNA from complex communities reveals many novel molecular species within the human gut. Applied and Environmental Microbiolog. 65:4799-4807.
    Suzuki, M.T., and S. J. Giovannoni. 1996. Bias caused by template annealing in the amplification of mixtures of 16S rRNA genes by PCR. Applied and Environmental Microbiology. 62: 625-630.
    Thompson, J. D., D. G. Higgins, and T. J. Gibson. 1994. CLUSTAL W:improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research. 22:4673-4680.
    Tiedje, J. M., J. F. 3rd Quensen, J. Chee-Sanford, J. P. Schimel, and S.A. Boyd. 1993. Microbial reductive dechlorination of PCBs. Biodegradation. 4:231-240.
    Unterman, R.. 1996. A history of PCB biodegradation. In “Bioremediation Principles and Applications”. Carwford R.L.,and Crawford D.L., Eds., pp. 209-253. Cambridge University Press. New York.
    von Wintzingerode, F., B. Selent, W. Hegemann, and U. B. Gobel. 1999. Phylogenetic analysis of an anaerobic trichlorobenzene-transforming microbial consortium. Applied and Environmental Microbiology. 65:283-286.
    Wackett, L. P., and C. D. Hershberger, 2001. Biocatalysis and Biodegradation. ASM Press. Washington, D. C.
    Wessendorf, M.W., and T.C. Brelje. 1992. Which fluorophore is the brightest? A comparison of the staining obtained using fluorescein, tetramethylrhodamine, lissamine rhodamine, Texas Red, and cyanine 3.18. Histochemistry. 98:81-85.
    Wiegel, J., and Q. Wu. 2000. Microbial reductive dehalogenation of polychlorinated biphenyls. FEMS Microbiology Ecology. 32:1-15
    Wiegel, J., X. Zhang, and Q. Wu. 1999. Anaerobic dehalogenation of hydroxylated polychlorinated biphenyls by Desulfitobacterium dehalogenans. Applied and Environmental Microbiology. 65:2217-2221.
    Woese, C. R. 1987. Bacterial evolution. Microbiology Review. 51:221-271.
    Wu, J.H., W. T. Liu, I. C. Tseng, S.S. Cheng. 2001. Characterization of microbial consortia in a terephthalate-degrading anaerobic granular sludge system. Microbiology. 147:373-382.
    Wu, Q., D. L. Bedard, and J. Wiegel. 1997-a. Temperature determines the pattern of anaerobic microbial dechlorination of Aroclor 1260 primed by 2,3,4,6-tetrachlorobiphenyl in Woods Pond sediment. Applied and Environmental Microbiology. 63:4818-4825.
    Wu, Q., D.L. Bedard, and J. Wiegel. 1997-b. Effect of Incubation Temperature on the Route of Microbial Reductive Dechlorination of 2,3,4,6-Tetrachlorobiphenyl in Polychlorinated Biphenyl (PCB)-Contaminated and PCB-Free Freshwater Sediments. Applied and Environmental Microbiology. 63:2836-2843.
    Yates, J. R., and F. J. Mondello. 1994. Genetic Aspects of PCB Biodegradation. In “Molecular Environmental Biology”. Garte, S. J., Eds., pp:65-81. CRC Press, Inc. U.S.
    Ye, D., J. F. 3rd Quensen, J. M. Tiedje, and S. A. Boyd. 1992. Anaerobic dechlorination of polychlorobiphenyls (Aroclor 1242) by pasteurized and ethanol-treated microorganisms from sediments. Applied and Environmental Microbiolog., 58:1110-1114.
    Yu, Z., and W. W. Mohn. 2001. Bacterial Diversity and Community Structure in an Aerated Lagoon Revealed by Ribosomal Intergenic Spacer Analyses and 16S Ribosomal DNA Sequencing. Applied and Environmental Microbiology. 67:1565-1574.
    Zarda, B., R. Amann, W. Wallner, and K.-H. Schleifer. 1991. Identification of single bacterial cells using digoxigenin-labelled, rRNA-targeted oligonucleotides. Journal of General Microbiology. 137:2823-2830.
    邱子權, 2000. 河川底泥中厭氧微生物對多氯聯苯還原性脫氯之研究。東吳大學微生物學系,碩士論文。
    黃香賓珽, 2001. 單槽連續進流回分式活性污泥系統微生物菌相變化之研究。國立中央大學,碩士論文。
    黃輝榮, 1997. 微生物分解污染物質。環境檢驗通訊雙月刊第17期。
    賴麗瑩, 1999. 談環境荷爾蒙(上)、(下) 。環境檢驗通訊雙月刊第22期

    QR CODE
    :::