Please use this identifier to cite or link to this item:
標題: 重金屬污染農地微生物多樣性及其可能之代謝功能探討
Diversity and Possible Function of Microbial Communities in Heavy Metal Polluted Soil
作者: 張飴璇
Jhang, Yi-Shiuan
關鍵字: PCR-DGGE;重金屬污染農地;amoA gene;nifH gene;alkaline phosphatase gene;功能性基因
出版社: 環境工程學系所
引用: 中文部份 網站資源 中華民國行政院環保署(2008) 行政院環境保護署法規。 中華民國行政院環保署(2008,2009) 土壤及地下水污染整治網。 期刊論文 中華民國行政院環保署 (2004) 土壤及地下水污染整治雙年報。 謝美明,儲三陽. (1986) 大氣中的甲烷. 科學月刊 203. 陳尊賢 (2003) 受重金屬污染農地土壤之整治技術與相關問題分析,台灣土壤及地下水環境保護協會,台北,pp. 2-9. 陳琦玲、廖慶樑、朱戩良、于迺文、葉偉任、蘇雲翰 (2006) 鎘污染農地轉作能源作物可行性評估,彰化縣。 陳慎德 (2003) 淺論我國農地土壤重金屬污染處理之現況與問題,台灣土壤及地下水環境保護協會簡訊,台北,pp. 10-17 劉泰銘 (2006) 土壤不均質性對翻土稀釋整治法之影響.台灣土壤及地下水環境保護協會簡訊,台北,pp. 1-8。 碩士論文 卓昕岑 (2003) 高海拔塔塔加及低海拔福山森林土壤微生物族群研究,國立臺灣大學,碩士論文,台北。 洪鈞煒 (2004) 重金屬鎘污染土壤環境中微生物之分離與鑑定,元智大學,碩士論文,桃園。 翁序伯 (2004) 重金屬污染農地淋洗處理及其土壤性質改變之研究,國立中興大學,碩士論文,台中。 陳雯怡 (2005) 活性污泥銨氧化基因多樣性之研究,中原大學,碩士論文,桃園。 陳賜章 (2002) 台南縣受重金屬污染農地土壤復育成效之追蹤,屏東科技大學,碩士論文,屏東。 葉芷微 (2006) Stenotrophomonas sp. Cd2+ 抗重金屬鎘之機制探討,元智大學,碩士論文,桃園。 蔡書憲 (2006) 福山森林土壤微生物族群、生質量、功能和基因多樣性,臺灣大學,碩士論文,台北。 韓竹婷 (2006) 重金屬污染環境中抗高濃度類金屬碲微生物之分離與探討,元智大學,碩士論文,桃園。 西文部份 Books Alexander, M. (1977) Introduction to Soil Microbiology Krieger Publishing Company. Barber, S.A. (1984) Soil Nutrient Bioavailability. New York: John Wiley & Sons. Madigan, M.T., and Martinko, J.M. (2006) Brock Biology of Microorganisms. New York: Prentice Hall International. Kandeler, E. (2007) Physiological and Biochemical Methods for Studying Soil Biota and Their Function. In Soil Microbiology, Ecology, and Biochemistry. Paul, E.A. (ed). Burlington: Academic Press, pp. 53-80. Killham, K., and Prosser, J.I. (2007) The Prokaryotes. In Soil Microbiology, Ecology, and Biochemistry. Paul, E.A. (ed). Burlington: Academic Press, pp. 119-143. Paul, E.A. (2007) Soil Microbiology, Ecology, and Biochemistry in Perspective. In Soil Microbiology, Ecology, and Biochemistry. Paul, E.A. (ed). Amsterdam: Academic Press, pp. 3-49. Thies, J.E. (2007) Molecular Methods for Studying Soil Ecology. In Soil Microbiology, Ecology, and Biochemistry. Paul, E.A. (ed). Burlington: Academic Press, pp. 85-115. Tisdale, S.L., Nelson, W.L., and Beaton, J.D. (1985) Soil Fertility and Fertilizers. New York: Macmillan Pub. Wuertz, S., and Mergeay, M. (1997) The Impact of Heavy Metals on Soil Microbial Communities and Yheir Activities. In Modern Soil Microbiology Elsas, J.D.v., Trevors, J.T., and Wellington, E.M.H. (eds). New York: MARCEL DEKKER, pp. 607-637. Young, J.P.W. (1992) Biological Nitrogen Fixation New York: Chapman & Hall. Journal Articles Akarsubasi, A.T., Ince, O., Kirdar, B., Oz, N.A., Orhon, D., Curtis, T.P., Head, I.M., and Ince, B.K. (2005) Effect of Wastewater Composition on Archaeal Population Diversity. Water Research 39: 1576-1584. Åkerblom, S., Bååth, E., Bringmark, L., and Bringmark, E. (2007) Experimentally Induced Effects of Heavy Metal on Microbial Activity and Community Structure of Forest Mor Layers. Biology and Fertility of Soils 44: 79-91. Amann, R., Ludwig, W., and Schleifer, K. (1995) Phylogenetic Identification and in Situ Detection of Individual Microbial Cells without Cultivation. microbiology and Molecular Biology Reviews 59: 143-169. Bååth, E. (1989) Effects of Heavy Metals in Soil on Microbial Processes and Populations (a Review). Water, Air, & Soil Pollution 47: 335-379. Bamborough, L., and Cummings, S. (2009) The Impact of Increasing Heavy Metal Stress on the Diversity and Structure of the Bacterial and Actinobacterial Communities of Metallophytic Grassland Soil. Biology and Fertility of Soils 45: 273-280. Bardgett, R.D., and McAlister, E. (1999) The Measurement of Soil Fungal:Bacterial Biomass Ratios as an Indicator of Ecosystem Self-Regulation in Temperate Meadow Grasslands. Biology and Fertility of Soils 29: 282-290. Barkay, T., Tripp, S.C., and Olson, B.H. (1985) Effect of Metal-Rich Sewage Sludge Application on the Bacterial Communities of Grasslands. Applied and Environmental Microbiology 49: 333-337. Beever, R.E., and Burns, D.J.W. (1981) Phosphorus Uptake, Storage and Utilization by Fungi Advances in Botanical Research 8: 127-219. Borneman, J., and Hartin, R.J. (2000) PCR Primers That Amplify Fungal Rrna Genes from Environmental Samples. Applied and Environmental Microbiology 66: 4356-4360. Bossio, D.A., and Scow, K.M. (1998) Impacts of Carbon and Flooding on Soil Microbial Communities: Phospholipid Fatty Acid Profiles and Substrate Utilization Patterns. Microbial Ecology 35: 265-278. Braker, G., Fesefeldt, A., and Witzel, K.-P. (1998) Development of PCR Primer Systems for Amplification of Nitrite Reductase Genes (nirK and nirS) to Detect Denitrifying Bacteria in Environmental Samples. Applied and Environmental Microbiology 64: 3769-3775. Brosius, J., Palmer, M.L., Kennedy, P.J., and Noller, H.F. (1978) Complete Nucleotide Sequence of a 16S Ribosomal RNA Gene from Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America 75: 4801-4805. Caille, O., Rossier, C., and Perron, K. (2007) A Copper-Activated Two-Component System Interacts with Zinc and Imipenem Resistance in Pseudomonas Aeruginosa. Journal of Bacteriology 189: 4561-4568. Calv, L. and Garcia-Gil, L.J. (2004) Use of amoB as a New Molecular Marker for Ammonia-Oxidizing Bacteria. Journal of Microbiological Methods 57: 69-78. Chien, C., Kuo, Y., Chen, C., Hung, C., Yeh, C., and Yeh, W. (2008) Microbial Diversity of Soil Bacteria in Agricultural Field Contaminated with Heavy Metals. Journal of Environmental Sciences 20: 359-363. Chu, H., Fujii, T., Morimoto, S., Lin, X., Yagi, K., Hu, J., and Zhang, J. (2007) Community Structure of Ammonia-Oxidizing Bacteria under Long-Term Application of Mineral Fertilizer and Organic Manure in a Sandy Loam Soil. Applied and Environmental Microbiology 73: 485-491. Dalal, R. (1982) Effect of Plant Growth and Addition of Plant Residues on the Phosphatase Activity in Soil. Plant and Soil 66: 265-269. Deni, J. and Penninckx, M.J. (1999) Nitrification and Autotrophic Nitrifying Bacteria in a Hydrocarbon-Polluted Soil. Applied and Environmental Microbiology 65: 4008-4013. Desai, C., Parikh, R.Y., Vaishnav, T., Shouche, Y.S., and Madamwar, D. (2009) Tracking the Influence of Long-term Chromium Pollution on Soil Bacterial Community Structures by Comparative Analyses of 16S rRNA Gene Phylotypes. Research in Microbiology 160: 1-9. Devergnas, S., Chimienti, F., Naud, N., Pennequin, A., Coquerel, Y., Chantegrel, J., Favier, A., and Seve, M. (2004) Differential Regulation of Zinc Efflux Transporters Znt-1, Znt-5 and Znt-7 Gene Expression by Zinc Levels: A Real-Time Rt-PCR Study. Biochemical Pharmacology 68: 699-709. Dionisi, H.M., Layton, A.C., Harms, G., Gregory, I.R., Robinson, K.G., and Sayler, G.S. (2002) Quantification of Nitrosomonas Oligotropha-Like Ammonia-Oxidizing Bacteria and Nitrospira spp. From Full-Scale Wastewater Treatment Plants by Competitive PCR. Applied and Environmental Microbiology 68: 245-253. Doelman, P. (1985) Resistance of Soil Microbial Communities to Heavy Metals FEMS Symposium: 369-383. Dorigo, U., Volatier, L., and Humbert, J.-F. (2005) Molecular Approaches to the Assessment of Biodiversity in Aquatic Microbial Communities. Water Research 39: 2207-2218. Emtiazi, F., Schwartz, T., Marten, S.M., Krolla-Sidenstein, P., and Obst, U. (2004) Investigation of Natural Biofilms Formed During the Production of Drinking Water from Surface Water Embankment Filtration. Water Research 38: 1197-1206. Fierer, N., Schimel, J.P., and Holden, P.A. (2003) Variations in Microbial Community Composition through Two Soil Depth Profiles. Soil Biology and Biochemistry 35: 167-176. Francis, C.A., Roberts, K.J., Beman, J.M., Santoro, A.E., and Oakley, B.B. (2005) Ubiquity and Diversity of Ammonia-Oxidizing Archaea in Water Columns and Sediments of the Ocean. Proceedings of the National Academy of Sciences of the United States of America 102: 14683-14688. Friedland, A.J., Johnson, A.H., and Siccama, T.G. (1986) Coniferous Litter Decomposition on Camels Hump, Vermont, USA: A Review. . Canadian Journal of Botany 64: 1349-1354. Frostegård, A., Tunlid, A., and Bååth, E. (1993) Phospholipid Fatty Acid Composition, Biomass, and Activity of Microbial Communities from Two Soil Types Experimentally Exposed to Different Heavy Metals. Applied and Environmental Microbiology 59: 3605-3617. Gadd, G.M. (1992) Metals and Microorganisms: A Problem of Definition. FEMS Microbiology Letters 100: 197-203. Garland, J.L. (1996) Patterns of Potential C Source Utilization by Rhizosphere Communities. Soil Biology and Biochemistry 28: 223-230. Garland, J.L., and Mills, A.L. (1991) Classification and Characterization of Heterotrophic Microbial Communities on the Basis of Patterns of Community-Level Sole-Carbon-Source Utilization. Applied and Environmental Microbiology 57: 2351-2359. Gong, P., D, S.S., Sonali, S., W, G.C., and Geoffrey, S. (2002) Assessment of Pollution-Induced Microbial Community Tolerance to Heavy Metals in Soil Using Ammonia-Oxidizing Bacteria and Biolog Assay. Human & Ecological Risk Assessment 8: 1067-1081. Gremion, F., Chatzinotas, A., and Harms, H. (2003a) Comparative 16S rDNA and 16S rRNA Sequence Analysis Indicates That Actinobacteria Might be a Dominant Part of the Metabolically Active Bacteria in Heavy Metal-Contaminated Bulk and Rhizosphere Soil. Environmental Microbiology 5: 896-907. Großkopf, R., Janssen, P.H., and Liesack, W. (1998) Diversity and Structure of the Methanogenic Community in Anoxic Rice Paddy Soil Microcosms as Examined by Cultivation and Direct 16S rRNA Gene Sequence Retrieval. Applied and Environmental Microbiology 64: 960-969. Hackl, E., Zechmeister-Boltenstern, S., Bodrossy, L., and Sessitsch, A. (2004) Comparison of Diversities and Compositions of Bacterial Populations Inhabiting Natural Forest Soils. Applied and Environmental Microbiology 70: 5057-5065. Heuer, H., Krsek, M., Baker, P., Smalla, K., and Wellington, E. (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. Hung, M.-H., Bhagwath, A.A., Shen, F.-T., Devasya, R.P., and Young, C.-C. (2005) Indigenous Rhizobia Associated with Native Shrubby Legumes in Taiwan. Pedobiologia 49: 577-584. Jansen, E., Michels, M., Til, M., and Doelman, P. (1994) Effects of Heavy Metals in Soil on Microbial Diversity and Activity as Shown by the Sensitivity-Resistance Index, an Ecologically Relevant Parameter. Biology and Fertility of Soils 17: 177-184. Janssen, P.H. (2006) Identifying the Dominant Soil Bacterial Taxa in Libraries of 16S rRNA and 16S rRNA Genes. Applied and Environmental Microbiology 72: 1719-1728. Jianping, X. (2006) Microbial Ecology in the Age of Genomics and Metagenomics: Concepts, Tools, and Recent Advances. In Molecular Ecology: Blackwell Publishing Limited, pp. 1713-1731. Kandeler, F., Kampichler, C., and Horak, O. (1996) Influence of Heavy Metals on the Functional Diversity of Soil Microbial Communities. Biology and Fertility of Soils 23: 299-306. Kaur, A., Chaudhary, A., Kaur, A., Choudhary, R., and Kaushik, R. (2005) Phospholipid Fatty Acid-a Bioindicator of Environment Monitoring and Assessment in Soil Ecosystem. Current Science 89: 1103-1112. Kelly, J.J., Häggblom, M.M., and Tate, R.L. (2003) Effects of Heavy Metal Contamination and Remediation on Soil Microbial Communities in the Vicinity of a Zinc Smelter as Indicated by Analysis of Microbial Community Phospholipid Fatty Acid Profiles. Biology and Fertility of Soils 38: 65-71. Khan, M., Zaidi, A., Wani, P., and Oves, M. (2009) Role of Plant Growth Promoting Rhizobacteria in the Remediation of Metal Contaminated Soils. Environmental Chemistry Letters 7: 1-19. Kisand, V., and Wikner, J. (2003) Limited Resolution of 16S rDNA DGGE Caused by Melting Properties and Closely Related DNA Sequences. Journal of Microbiological Methods 54: 183-191. Kowalchuk, G.A., and Stephen, J.R. (2001) Ammonia-Oxizing Bacteria: A Model for Molecular Microbial Ecology. In Annual Review of Microbiology 55:485-529. Lauber, C.L., Strickland, M.S., Bradford, M.A., and Fierer, N. (2008) The Influence of Soil Properties on the Structure of Bacterial and Fungal Communities across Land-Use Types. Soil Biology and Biochemistry 40: 2407-2415. Leckie, S.E. (2005) Methods of Microbial Community Profiling and Their Application to Forest Soils. Forest Ecology and Management 220: 88-106. Leininger, S., Urich, T., Schloter, M., Schwark, L., Qi, J., Nicol, G.W., Prosser, J.I., Schuster, S.C., and Schleper, C. (2006) Archaea Predominate among Ammonia-Oxidizing Prokaryotes in Soils. Nature 442: 806-809. Lorenz, N., Hintemann, T., Kramarewa, T., Katayama, A., Yasuta, T., Marschner, P., and Kandeler, E. (2006) Response of Microbial Activity and Microbial Community Composition in Soils to Long-Term Arsenic and Cadmium Exposure. Soil Biology and Biochemistry 38: 1430-1437. Madejón, E., Burgos, P., López, R., and Cabrera, F. (2001) Soil Enzymatic Response to Addition of Heavy Metals with Organic Residues. Biology and Fertility of Soils 34: 144-150. Marchesi, J.R., Sato, T., Weightman, A.J., Martin, T.A., Fry, J.C., Hiom, S.J., and Wade, W.G. (1998) Design and Evaluation of Useful Bacterium-Specific PCR Primers that Amplify Genes Coding for Bacterial 16S rRNA. Applied and Environmental Microbiology 64: 795-799. McGowan, C., Fulthorpe, R., Wright, A., and Tiedje, J.M. (1998) Evidence for Interspecies Gene Transfer in the Evolution of 2,4-Dichlorophenoxyacetic Acid Degraders. Applied and Environmental Microbiology 64: 4089-4092. Mengoni, A., Barzanti, R., Gonnelli, C., Gabbrielli, R., and Bazzicalupo, M. (2001) Characterization of Nickel-Resistant Bacteria Isolated from Serpentine Soil. Environmental Microbiology 3: 691-698. Mertens, J., Springael, D., De Troyer, I., Cheyns, K., Wattiau, P., and Smolders, E. (2006) Long-Term Exposure to Elevated Zinc Concentrations Induced Structural Changes and Zinc Tolerance of the Nitrifying Community in Soil. In Environmental Microbiology: Blackwell Publishing Limited, pp. 2170-2178. Milling, A., Smalla, K., Maidl, F., Schloter, M., and Munch, J. (2005) Effects of Transgenic Potatoes with an Altered Starch Composition on the Diversity of Soil and Rhizosphere Bacteria and Fungi. Plant and Soil 266: 23-39. Misra, T.K., Brown, N.L., Fritzinger, D.C., Pridmore, R.D., Barnes, W.M., Haberstroh, L., and Silver, S. (1984) Mercuric Ion-Resistance Operons of Plasmid R100 and Transposon Tn501: The Beginning of the Operon Including the Regulatory Region and the First Two Structural Genes. Proceedings of the National Academy of Sciences of the United States of America 81: 5975-5979. Muyzer, G., de Waal, E.C., and Uitterlinden, A.G. (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. Nakas, J., Gould, W., and Klein, D. (1987) Origin and Expression of Phosphatase Activity in a Semi-Arid Grassland Soil. Soil biology & biochemistry 19: 13-18. Ndour, N., Baudoin, E., Guissé, A., Seck, M., Khouma, M., and Brauman, A. (2008) Impact of Irrigation Water Quality on Soil Nitrifying and Total Bacterial Communities. Biology and Fertility of Soils 44: 797-803. Nielsen, A.T., Liu, W.-T., Filipe, C., Grady, L., Jr., Molin, S., and Stahl, D.A. (1999) Identification of a Novel Group of Bacteria in Sludge from a Deteriorated Biological Phosphorus Removal Reactor. Applied and Environmental Microbiology 65: 1251-1258. Nies, A., Nies, D., and Silver, S. (1990) Nucleotide Sequence and Expression of a Plasmid-Encoded Chromate Resistance Determinant from Alcaligenes Eutrophus. Journal of Biological Chemistry 265: 5648-5653. Nies, D.H., Nies, A., Chu, L., and Silver, S. (1989) Expression and Nucleotide Sequence of a Plasmid-Determined Divalent Cation Efflux System from Alcaligenes Eutrophus. Proceedings of the National Academy of Sciences of the United States of America 86: 7351-7355. Oger, C., Berthe, T., Quillet, L., Barray, S., Chiffoleau, J.-F., and Petit, F. (2001) Estimation of the Abundance of the Cadmium Resistance Gene cadA in Microbial Communities in Polluted Estuary Water. Research in Microbiology 152: 671-678. Okano, Y., Hristova, K.R., Leutenegger, C.M., Jackson, L.E., Denison, R.F., Gebreyesus, B., Lebauer, D., and Scow, K.M. (2004) Application of Real-Time PCR to Study Effects of Ammonium on Population Size of Ammonia-Oxidizing Bacteria in Soil. Applied and Environmental Microbiology 70: 1008-1016. Oved, T., Shaviv, A., Goldrath, T., Mandelbaum, R.T., and Minz, D. (2001) Influence of Effluent Irrigation on Community Composition and Function of Ammonia-Oxidizing Bacteria in Soil. Applied and Environmental Microbiology 67: 3426-3433. Panikov, N.S. (1999) Understanding and Prediction of Soil Microbial Community Dynamics under Global Change. Applied Soil Ecology 11: 161-176. Pennanen, T., Frostegård, A., Fritze, H., and Bååth, E. (1996) Phospholipid Fatty Acid Composition and Heavy Metal Tolerance of Soil Microbial Communities Along Two Heavy Metal-Polluted Gradients in Coniferous Forests. Applied and Environmental Microbiology 62: 420-428. Poly, F., Monrozier, L.J., and Bally, R. (2001) Improvement in the RFLP Procedure for Studying the Diversity of nifH Genes in Communities of Nitrogen Fixers in Soil. Research in Microbiology 152: 95-103. Rajapaksha, R.M.C.P., Tobor-Kaplon, M.A., and Baath, E. (2004) Metal Toxicity Affects Fungal and Bacterial Activities in Soil Differently. Applied and Environmental Microbiology 70: 2966-2973. Raskin, L., Stromley, J.M., Rittmann, B.E., and Stahl, D.A. (1994) Group-Specific 16S rRNA Hybridization Probes to Describe Natural Communities of Methanogens. Applied and Environmental Microbiology 60: 1232-1240. Renella, G., Mench, M., Landi, L., and Nannipieri, P. (2005) Microbial Activity and Hydrolase Synthesis in Long-Term Cd-Contaminated Soils. Soil Biology and Biochemistry 37: 133-139. Rodríguez, H., and Fraga, R. (1999) Phosphate Solubilizing Bacteria and Their Role in Plant Growth Promotion. Biotechnology Advances 17: 319-339. Rotthauwe, J., Witzel, K., and Liesack, W. (1997) The Ammonia Monooxygenase Structural Gene amoA as a Functional Marker: Molecular Fine-Scale Analysis of Natural Ammonia-Oxidizing Populations. Applied and Environmental Microbiology 63: 4704-4712. Rusk, J.A., Hamon, R.E., Stevens, D.P., and McLaughlin, M.J. (2004) Adaptation of Soil Biological Nitrification to Heavy Metals. Environmental Science & Technology 38: 3092-3097. Sakurai, M., Wasaki, J., Tomizawa, Y., Shinano, T., and Osaki, M. (2008) Analysis of Bacterial Communities on Alkaline Phosphatase Genes in Soil Supplied with Organic Matter. Soil Science & Plant Nutrition 54: 62-71. Sanadi, D.R. (1982) Mitochondrial Coupling Factor B. Properties and Role in ATP Synthesis. Biochimica et Biophysica Acta 683: 39-56. Sandaa, R.-A., Enger, O., and Torsvik, V. (1999) Abundance and Diversity of Archaea in Heavy-Metal-Contaminated Soils. Applied and Environmental Microbiology 65: 3293-3297. Smit, E., Leeflang, P., Gommans, S., van den Broek, J., van Mil, S., and Wernars, K. (2001) Diversity and Seasonal Fluctuations of the Dominant Members of the Bacterial Soil Community in a Wheat Field as Determined by Cultivation and Molecular Methods. Applied and Environmental Microbiology 67: 2284-2291. Speir, T.W., Kettles, H.A., Parshotam, A., Searle, P.L., and Vlaar, L.N.C. (1999) Simple Kinetic Approach to Determine the Toxicity of As(V) to Soil Biological Properties. Soil Biology and Biochemistry 31: 705-713. Stephen, J., McCaig, A., Smith, Z., Prosser, J., and Embley, T. (1996) Molecular Diversity of Soil and Marine 16S rRNA Gene Sequences Related to Beta-Subgroup Ammonia-Oxidizing Bacteria. Applied and Environmental Microbiology 62: 4147-4154. Stephen, J.R., Chang, Y.-J., Macnaughton, S.J., Kowalchuk, G.A., Leung, K.T., Flemming, C.A., and White, D.C. (1999) Effect of Toxic Metals on Indigenous Soil Beta -Subgroup Proteobacterium Ammonia Oxidizer Community Structure and Protection against Toxicity by Inoculated Metal-Resistant Bacteria. Applied and Environmental Microbiology 65: 95-101. Sun, H.Y., Deng, S.P., and Raun, W.R. (2004) Bacterial Community Structure and Diversity in a Century-Old Manure-Treated Agroecosystem. Applied and Environmental Microbiology 70: 5868-5874. Torsvik, V., and Øvreås, L. (2008) Microbial Diversity, Life Strategies, and Adaptation to Life in Extreme Soils. In Microbiology of Extreme Soils, pp. 15-43. Torsvik, V., Daae, F.L., Sandaa, R.-A., and Øvreås, L. (1998) Novel Techniques for Analysing Microbial Diversity in Natural and Perturbed Environments. Journal of Biotechnology 64: 53-62. Turpeinen, R., Kairesalo, T., and Hägblom, M.M. (2004) Microbial Community Structure and Activity in Arsenic-, Chromium- and Copper-Contaminated Soils. FEMS Microbiology Ecology 47: 39-50. Upchurch, R., Chiu, C.-Y., Everett, K., Dyszynski, G., Coleman, D.C., and Whitman, W.B. (2008) Differences in the Composition and Diversity of Bacterial Communities from Agricultural and Forest Soils. Soil Biology and Biochemistry 40: 1294-1305. Walker, B.H. (1992) Biological Diversity and Ecological Redundancy. Conservation Biology 6: 18-23 . Wang, Y., Shi, J., Wang, H., Lin, Q., Chen, X., and Chen, Y. (2007) The Influence of Soil Heavy Metals Pollution on Soil Microbial Biomass, Enzyme Activity, and Community Composition near a Copper Smelter. Ecotoxicology and Environmental Safety 67: 75-81. Wartiainen, I., Eriksson, T., Zheng, W., and Rasmussen, U. (2008) Variation in the Active Diazotrophic Community in Rice Paddy--nifH PCR-DGGE Analysis of Rhizosphere and Bulk Soil. Applied Soil Ecology 39: 65-75. White, T., Bruns, T., Lee, S., Taylor, J., Innis, M., Gelfand, D., and Shinsky, J. (1990) Amplification and Direct Sequencing of Fungal Ribosomal RNA Genes for Phylogenetics. In PCR Protocols: A Guide to Methods and Applications: Academic Press, pp. 315-322. Yeates, C., Gillings, M., Davison, A., Altavilla, N., and Veal, D. (1998) Methods for Microbial DNA Extraction from Soil for PCR Amplification. Biological Procedures Online 1: 40-47. Zak, J., Willig, M., Moorhead, D., and Wildman, H. (1994) Functional Diversity of Microbial Communities: A Quantitative Approach Soil Biology and Biochemistry 26: 1101-1108.
受重金屬污染農地經整治後,若能將土壤中重金屬濃度降至管制值之下,農地即可解除列管,但污染農地經整治後常因地力下降導致復耕農地收成不佳,往往需要進行土壤改良以恢復地力。土壤微生物族群之代謝功能與土壤肥力息息相關,重金屬會成為微生物長期的生存壓力,微生物族群之代謝也會因重金屬污染而受到抑制。具特殊功能之微生物(如銨氧化菌、溶磷菌等)族群種類如果因整治而下降則可能會造成農地地力無法於短時間恢復之狀況,因此於整治策略制定時須將整治對微生物多樣性之損害狀況納入考量以降低整治成本並提高復耕農地利用效益。在探討整治對重金屬污染農地微生物多樣性之破壞狀況前,需先瞭解重金屬污染農地微生物與具特殊功能之族群多樣性現況,因此本研究以曾受重金屬污染之農地(標號0119及2023)為目標,利用依照演化基因(rRNA gene)與功能性基因(amoA gene、nifH gene及alkaline phosphatase gene)設計之引子結合PCR-DGGE技術對農地土壤中微生物生態進行調查。
實驗結果顯示,以細菌及真菌演化基因設計的引子依PCR-DGGE技術獲得之圖譜有訊號模糊不清的問題,推測部分原因可能為土壤環境核酸多樣性豐富,因此樣本中多樣的核酸序列其解鏈溫度 ( Tm值,melting temperature)範圍過廣,導致Tm值接近的核酸無法在DGGE圖譜上形成清晰亮帶。利用引子344f/522r對土壤中甲烷古細菌多樣性進行分析,結果顯示有9種及6種甲烷古細菌均勻地分布於農地0119及2023地表下0 ~ 30cm土壤中,甲烷古細菌為絕對厭氧菌生長於土壤團粒中央不含氧之區域,其特殊的生長位置可降低重金屬對其族群多樣性之衝擊。利用引子243f/518r對土壤中放線菌族群進行研究後推測,可能因為放線菌k-strategy之生長策略可減緩低濃度之重金屬對其族群多樣性之衝擊,因此土壤中鉻(Cr)、銅(Cu)、鋅(Zn)及鎳(Ni)濃度須超過法規值兩倍以上於DGGE圖譜上放線菌族群多樣性才會產生變化。
以功能性基因分析所獲得之多樣性資料推測因農地2023受重金屬污染嚴重且土壤pH偏酸,因此於農地2023九個採樣點僅有採樣點I (10 ~ 20cm)土壤中發現兩種銨氧化菌(Ammonia-oxidizing bacteria, AOB)存在。農地0119土壤較接近中性外,九採樣點中,採樣點I (0 ~ 10cm)、M (0 ~ 10cm)、O (0 ~ 10cm)及I (10 ~ 20cm)皆有AOB存在,且其多樣性變化推測與土壤中銨濃度變化有關,故建議銨氧化菌族群多樣性變化可以成為評估環境變化之微生物指標。農地2023受重金屬污染嚴重區域之溶磷菌與固氮菌多樣性與其它採樣點明顯不同,且溶磷菌於污染嚴重之採樣點I (0 ~10cm)與I (10 ~ 20cm)中其菌群種類與樣點M及O不同外,溶磷菌多樣性較採樣點M及O豐富,因此農地雖受重金屬污染但具有將磷酸鹽轉型能力之微生物族群仍可維持一定之多樣性,此外隨污染程度加深,具有磷酸鹽轉型能力之微生物種類會產生改變,故推測高濃度的重金屬確實會對溶磷菌群造成影響,但農地土壤仍具一定之磷酸鹽代謝潛力。

In paddy soil, microbial functions and community diversity are closely involved with the in situ soil fertility. Although performing remediation can reduce the concentration of heavy metal, it also leads to poor soil fertility and low harvest. Therefore, the fertility of the remediation soil will still need to be improved if it will still be used for cultivation. Because of heavy metal cannot be degraded by microorganisms, heavy metal become long-term environmental stressed easily and it will inhibit the activity of soil microorganisms in the polluted site. If the specific microbial diversity decline with remediation process, soil fertility will not recover in short-term. For purpose to develop a reliable method to evaluate the damage from the remediation approaches, which may reduce the value of resumed cultiable land, on microbial diversity, molecular tools were applied in this study. In this study, genes diversity, such as rRNA, amoA, nifH and alkaline phosphatase genes, in the heavy metal polluted soil (marked as land 0119 and 2023) were detected using PCR-DDGE methods.
In soil environment existence of high diversity in nucleic acid might leads to the PCR product have wide range of melting temperature (Tm), this has the possibility to cause the fuzzy bands in DGGE fingerprint that cause the poor resolution of PCR-DGGE fingerprints when applying the eubacteria- and fungal- specific primer sets individually. Applying primer pairs 344f/522r to analyze the methanogens archaea diversity, there were nine and six methanogens archaea species distributed in the soil which was 0 to 30 cm under the surface. Using primer pairs 243f/518r to analyze the Actinobacteria community diversity, the results showed the low Acinobacteria diversity in the paddy land 2023, in which heavy metal concentration exceed than the heavy metal criterions published by Taiwan EPA . Furthermore, it was suggested that k- strategy growth mode of Actinobacteria can reduce the impact of heavy metals. Therefore, it was believed the heavy metal would effects the Actinomycetes diversity when polluted soil it's heavy metal concentration higher than the published criterions.
In Ammonia-oxidizing bacteria(AOB) diversity analysis, there were only five samples amplicons were detected, which were 0119 I (0~10cm), 0119 M (0~10cm), 0119 O (0~10cm), 0119 I (10~20cm)and 2023 I (10~20cm). The AOB diversity analysis demonstrated there were two AOB species exited in the land 2023, the more serious heavy metal pollution and lower pH in the paddy land 2023 might be the reason that reduce AOB diversity. Paddy land 0119 variation of AOB diversity may reflect changes ammonium concentration in soil and the variation of ammonium-oxidizing bacteria diversity was suggested to be applied as the microbial indicators for detecting environmental change. In the high heavy metal polluted soil 2023 the phosphate-solubilizing bacteria diversity in sample site I was higher than it in sites M and O and the high concentrations of heavy metals was demonstrated that to be benefit on phosphate-solubilizing bacterial diversity by the results of DGGE analysis in this study, where the phosphate-solubilizing bacteria have high diversity in sample site 2023 I. It was suspected that heavy metal polluted paddy soil still have potential of phosphate metabolism according to the PCR-DGGE analysis based on amoA, nifH and alkaline phosphatase genes. In summary, inherent specific microbial diversity could be used as an indictor to evaluate the damage in the polluted soil.
其他識別: U0005-1008200918191600
Appears in Collections:環境工程學系所

Show full item record

Google ScholarTM


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.