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論文名稱 Title |
以植物修復技術處理遭受重金屬鎘污染土壤之研究 Treatment of Cadmium Contaminated Soil by Phytoremediation |
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系所名稱 Department |
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畢業學年期 Year, semester |
語文別 Language |
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學位類別 Degree |
頁數 Number of pages |
106 |
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研究生 Author |
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指導教授 Advisor |
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召集委員 Convenor |
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口試委員 Advisory Committee |
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口試日期 Date of Exam |
2005-12-26 |
繳交日期 Date of Submission |
2006-01-10 |
關鍵字 Keywords |
菌根菌、根圈微生物、重金屬鎘、植物修復 PCR-DGGE, dehydrogenase, cadmium, rhizosphere microorganisms, phytoremediation |
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統計 Statistics |
本論文已被瀏覽 5651 次,被下載 3621 次 The thesis/dissertation has been browsed 5651 times, has been downloaded 3621 times. |
中文摘要 |
本研究嘗試以植物修復技術來處理遭受鎘污染土壤。研究分為兩階段實驗,第一階段實驗中,先從文獻中選出兩種耐重金屬植物:培地茅(Vetiveria zizanioides)、銀葉鳳尾蕨(Pteris ensiformis cv. 'Victoriae')及重金屬污染現地之長梗滿天星(Alternanthera philoxeroides (Mart.) Griseb)三種陸生植物試驗耐受鎘污染土壤(10、20及30 mg/kg)測試,實驗進行九週,觀察其生長狀況及植物體內鎘累積的量。實驗結果顯示培地茅的生長狀況良好,且每盆平均鎘累積量為高於其他兩種植物種。因此篩選出培地茅做為第二階段實驗植物種。 在第二階段實驗中,將進一步針對培地茅,種植於較高鎘污染濃度土壤(30及50 mg/kg)中,進行植物修復。經過210天盆栽實驗,結果顯示,培地茅有助於鎘污染土壤中之微生物相(總生菌、真菌及放線菌)生長、去氫酶酵素活性的增加及降低有效性鎘的量,此外,菌根菌感染率的增加,有幫助培地茅抵抗重金屬鎘逆境之效果。 實驗最後,利用分子生物技術PCR/DGGE方法,觀察土壤中微生物之多樣性,由DGGE剖面圖顯示,在遭受鎘污染濃度30 m/kg,且以沒種植物最為豐富,微生物在此一濃度下並無抑制作用,同樣鎘濃度30 mg/kg下,有栽種植物菌相較少,猜測有植物根系的存在,可能利於某些特定微生物之生長,使得豐富度相對減少。 在第210天時,培地茅種植於土壤中鎘濃度30 mg/kg及50 mg/kg分別從26.9 mg/kg降為19.0 mg/kg;44.8 mg/kg降為31.2 mg/kg去除率均在30%左右,效果沒預期好,此外,植物從土壤移除重金屬鎘效果,在30與50 mg/kg鎘污染土壤中分別為7.8及8.9%,所以在未來應用於現場時,可搭配超累積重金屬鎘植物種,增加土壤中鎘之移除能力。 |
Abstract |
In this study we attempt to use phytoremediation techniques to treat the contaminated soil of cadmium. The experiment is divided into two stages. In the first stage, we selected three different species of plants which could tolerate heavy-metals: vetiver (Vetiveria zizanioides), Pteris ensiformis cv. 'Victoriae' according to the past records, and Alternanthera philoxeroides (Mart.) Griseb, which were sampled from the metal contaminated site in Hunei, Kaohsiung county. These three species were planted in three pots with 10, 20 and 30 mg Cd kg-1 in soil respectively. After 9 weeks of the growth, the vetiver was found accumulating the highest Cd and grew better than the other two species. Therefore, we selected the species of vetiver in the second stage of experiment. First, the species of vetiver was planted in the pots with concentrations of 30 and 50 mg Cd kg-1 in soil respectively. Then the pots were put in the greenhouse for incubation. After the test was run for 210 days, we found that the species of vetiver was helpful in the increasing the number of species and amounts of each species of microbe ( total bacteria, fungi and actinomycete ), as well as dehydrogenase activity. Meanwhile, it was effective to decrease the bioavailability of cadmium. In addition, the infection rate of mycorrhizal fungi was increased , which showed that the species of vetiver could resist the cadmium stress in soils and stimulate the soil fertility. Finally, we use molecular biotechniques of PCR-DGGE to observe the microbial diversity in the contaminated soil. We found that the pots with 30 mg Cd kg-1 in soil had more number of bands than the pots with different Cd concentrations in soil, while the pots without vegetation was found more fruitful than vegetated pots. These experimental results indicated that the pots planted with the species of vetiver under this situation would help some special microorganisms to grow, and thus that the microbial diversity was reduced. The results also showed that the pots planted with vetiver with initial cadmium concentrations of 30 and 50 mg Cd kg-1 respectively, in soil exhibited the degradation rate of about 30 percent for both. It was not satisfied to this result in this study. However, the phytoextraction rates of cadmium were measured equal to 7.8 and 8.9 percent, respectively. According to these results, we suggested that the plant, which could hyperaccumulate heavy metals, might be used to increase the removable ability of cadmium in the future. |
目次 Table of Contents |
目 錄 誌 謝……………………………………………………………………I 中文要……………………………………………………………….II 英文摘要………………………………………………………………III 表目錄…………………………………………………………………….IX 圖目錄………………………………………………………………………X 第一章 前言 1 1.1 研究動機 1 1.2 研究目的 2 第二章 文獻回顧 3 2.1 重金屬污染對環境的影響 3 2.1.1 重金屬污染來源及危害 3 2.1.2 鎘對環境造成影響 3 2.2 重金屬鎘特性 6 2.2.1 基本物理化學性質介紹及用途 6 2.2.2 鎘在土壤的特性 6 2.3 台灣地區重金屬污染等級分類及管制標準 8 2.4 試驗植物介紹 10 2.5 重金屬污染整治復育技術介紹 12 2.6 植物修復技術介紹 13 2.6.1 植物選擇 16 2.6.2 根圈微生物 17 2.6.3重金屬與植物螯合素之關係 18 2.6.4 菌根菌與重金屬之關係 18 2.6.5 土壤去氫酵素活性 21 2.7 處理成本比較 22 2.8 植物修復之優缺點比較 23 第三章 研究方法與步驟 24 3.1 實驗設計流程 24 3.2 盆栽試驗 26 3.2.1 植物採集 26 3.2.2 重金屬鎘污染物土壤配製 28 3.2.2-1 第一階段實驗 28 3.2.2-2 第二階段實驗 30 3.3 土壤採樣及樣品保存 31 3.4 土壤分析方法 31 3.4.1 土壤中的含水率 31 3.4.2 土壤中酸鹼值 31 3.4.3 土壤有機碳 32 3.4.4 土壤總有機氮 32 3.4.5 土壤中有效性鎘之萃取 33 3.4.6 土壤中全量重金屬鎘之萃取 33 3.4.7 去氫酶素活性 34 3.5 土壤中微生物 34 3.5.1 總生菌 35 3.5.2 真菌 35 3.5.3 放線菌 35 3.5.4 菌根菌分析 37 3.6 植物生長及植體分析 37 3.6.1 植物生長高度 37 3.6.2 植物體重金屬分析 37 3.7 根圈菌相分析 38 3.7.1 土壤中DNA萃取 38 3.7.2 聚合酶連鎖反應 39 3.7.2-1 PCR-16SrDNA純化 41 3.7.2-2 PCR-16S rDNA片段瓊脂膠體電泳檢視 41 3.7.3 變性梯度膠凝電泳分析 41 3.8 實驗分析之QA�QC 42 3.9 實驗相關儀器及器材 43 第四章 結果與討論 45 4.1 土壤基本性質 45 4.2 耐重金屬鎘植物之初步篩選(第一階段實驗數據) 47 4.3 耐鎘植物之植物修復研究(第二階段實驗數據) 50 4.3.1 植物生長情形 50 4.3.2 土壤中酸鹼值 53 4.3.3 土壤有效性鎘 55 4.3.4 土壤去氫酶活性 58 4.3.5 土壤中微生物菌相 60 4.3.5-1 總菌落 60 4.3.5-2 真菌 60 4.3.5-3 放線菌 61 4.3.5-4 微生物菌相與有效性鎘之關係 65 (1)總菌與有效性鎘 65 (2)真菌與有效性鎘 65 (3)放線菌與有效性鎘 65 4.3.6 菌根菌 70 4.3.7 利用分子生物技術探討微生物之菌相 71 4.3.8 土壤中鎘之去除 73 4.3.9 植物體內累積鎘情形 74 第五章 結論與建議 76 5.1 結論 76 5.2 建議 77 參考文獻 78 附錄 一 86 附錄 二 87 附錄 三 89 |
參考文獻 References |
王裕文,2000,“培地茅(Vetiveria zizanioides)簡介”,中華民國雜草學會會刊,第二十一卷,第一期,第59-63頁。 行政院環境保護署,2002,土壤水分含量測定方法-重量法,NIEA S280.61C。 行政院環境保護署,1991,土壤中酸鹼值測定方法,NIEA S410.60T。 行政院環境保護署,2002,沈機物、污泥及油脂中重金屬元素總量之檢測方法-微波消化原子光譜法,NIEA R355.00C。 行政院環境保護署,1994,土壤中陽離子交換容量-醋酸鈉法,NIEA S202.60A。 吳昭慧,2002,“綠色草籬-培地茅”,台南區農業專訓,第42期第7-11頁。 吳繼光、林素禎,1998,叢囊枝內生菌根菌技術應用手冊,台灣省農業試驗所出版,台中縣。 林宏達,2004,“以植物復育技術處理遭受溢油污染濕地土壤之研究”,碩士論文,國立中山大學海洋環境及工程學系。 柯鴻慶,2004,“以植物萃取法處理銅鉛鎘污染之土壤”,碩士論文,嘉南藥理科技大學環境工程與科學系。 屏東縣環境保護局,2002,潮州鎮崙頂段916地號不明廢棄物場址清理作業細部調查規劃及監督技術服務工作期末報告。 袁又罡、郭書吟、沈家鳳,2003,“土壤內微生物對重金屬銅、鋅、鎘吸收及耐受程度之研究”,第二十八屆廢水處理研討會論文集。 陳尊賢,2001,“土壤整治技術國際會議”,科學發展月刊,第29卷,第3期,第207-210頁。 陳尊賢,2005,“污染土壤重金屬管制濃度之修正與評估”,研討會論文集,台灣土壤及地下水污染整治之回顧與展望研討會,第79-95頁。 陳慎德,2004,“重金屬污染整治技術與案例分析”,研習手冊,土壤及地下水應用技術講習會。 彭安、王文華,1993,環境生物無機化學,初版,淑馨出版社,台北市,第21-22頁。 曾顯雄,1985,“真菌學之最近研究發展”,專刊,行政院國家科學委員會生物科學研究中心-「真菌學之最近發展」專題演講論文集專刊,第12號,第45-70頁。 張尊國,2002,“台灣地區土壤污染現況與整治政策分析”,國政分析,永續(析)091-021號。 張維妮,2003,“不同營養特性之植物組織及部位對土壤中重金屬之累積吸附研究”,碩士論文,朝陽科技大學環境工程與管理系。 熊明彪、何建平等,2002,“根分泌物對根際微生物生態分佈地影響”,土壤通報,第33期,第二卷。 鄭秀珍,2006,“以植物修復技術處理受五氯酚污染土壤之研究”,碩士論文,國立中山大學海洋環境及工程學系。 簡志青、洪鈞煒,2005,“重金屬鎘污染環境微生物相與菌株之分離與探討”,第三屆土壤及地下水研討會論文集。 蕭茂讃,2004,“環保警察制度與商業倫理之探討:以台灣環保警察為例”,碩士論文,國立中央大學哲學研究所。 Ajwa, H.A., Banuelos, G.S. and Mayland, H.F. 1998,“Selenium uptake by plants fro- m soils amended with inorganic and organic materials”, Journal of Envirnment Quality, Vol. 27, pp.1218-1227. Banuelos, G.S., Ajwa, H.A., Mackey B., Wu, L., Cook, C., Akohoue, S. and Zambr- uzuski S. 1997, ”Evalution of different plant species used for phytoremediation of high soil selenium”, Journal of Environment Quality, Vol. 26, pp.639-646. Blake, R.C., Choate, D.M., Bardhan, S., Revis, N., Barton, L.L., Zocco, T.G. 1993. “Chemical transformation of toxic metals by Pseudomonas strain from a toxic waste site”, Environmental Toxicology and chemistry, Vol. 12, pp. 1365 - 1376. Blaylock, M.J., Salt, D.E., Dushenkov, S., Zakharova, O., Gussman, G., Kapulnik, Y., Ensley, B.D., Raskin, I., 1997,“Enhanced accumulation of Pb in Indian mustard by soil-applied chelating agents”, Environmental Science and Technology, Vol. 31, pp.860-865. Bremner, J.M., 1960, “Determination of nitrogen in soil by the Kjeldahl method”, Journal of Agricultural Science, Cambridge 55. pp. 11-33. Casida, L.E., Jr., Klein, D.A. and Santoro., 1994, “Soil dehydrogenase activity”, Soil Science, Vol. 98. pp. 371-376. Chen, H., Cutright, T., 2001,“EDTA and HEDTA effects on Cd, Cr, and Ni uptake by Helianthus annuus”, Chemosphere, Vol. 45, pp.21-28. Chen, Y., Shen, Z. and Li, X., 2004, “The use of vetiver grass (Vetiveria zizanioides) in the phytoremediation of soils contaminated with heavy metals”, Applied Ge- ochemistry, Vol. 19. pp. 1553-1565. Cunnigham, S.D. and Berti, W.R., 1993,“Remediation of contaminated soils with green plants: an overview”, In Vitro Cellular and Development Biology, Vol. 29, pp.207-212. Curl E. A. and Trueglove B., 1986, “The Rhizosphere”, Advanced Series in Agricul- tural Sciences, Vol. 15. Berlin Germany: Springer-Verlag. Ebbs, S.D., and Kochian, L.V., 1998,“Phytoextraction of Zinc by oat (Avena sativa), barley(Hordeum vulgare) and Indian mustard (Brassica juncea)”, Environment Science and Technology, Vol. 32, pp.802-806. Enkhtuya, B., Rydlová, J., Vosátka, M., 2000,“Effectiveness of indigenous and non- indigenous isolates or arbuscular mycorrhizal fungi in soils from degraded eco- systems and man-made habitats”, Applied Soil Ecology, Vol. 14, pp.201- 211. Fortina, N., Beaumiera, D., Leeb, K., and Greera, C.W., 2004,“Soil washing impro- ves the recovery of total community DNA from polluted and high organic cont- ent sediments”, Journal of Microbiological Methods, Vol. 56, pp. 181-191. Gadd, G.M., 1990, “Fungi and yeasts for metal accumulation”, In:Ehrlich. H.L., and Brierley C.L. (eds) Microbial mineral recovery, McGraw-Hill, New York, pp. 249–276. Glass D.J., 2000, “Economic potential of phytoremediation”, in “Phytoremediation of Toxic Metals: Using Plants to Clean up the Environment”, B. Ensley and I. Raskin, eds., John Wiley & Sons, pp.15-31. Hornburg, V. and Brümmer, G.W., 1993,“Behaviour of heavy metals in soil. 1. Inve- stigations on heavy metal mobility. (In German)”, Z. Pflanzenernähr Bodenkunde, Vol. 156, pp.467-477. Huang, J.W., and Cunningham, S.D., 1996,“Lead phyotextraction : Species variati- on in lead uptake and translocation”, New Phytologist, Vol. 134, pp.75-84. Huang, J.W., Chen, J.J., Berti, W.R. and Cunningham, S.D.,1997,“Phytoremediation of lead-contaminated soils: Role of synthetic chelates in lead phytoextraction”, Environment Science and Technology, Vol. 31, pp.800-805. Jones, D.L., Prabowo, A.M. and Kochian, L.V., 1996c,“Kinetics of malate transport and decomposition in acid soils and isolated bacterial populations – the effect of microorganisms on root exudation of malate under Al stress”, Plant and Soil, Vol. 182, pp.239-247. Khan, A.G., Kuek, C., Chaudhry, T.M., Khoo, C.S. and Hayes, W.J., 2000, “Role of plant, mycorrhizae and phytochelators in heavy metal contaminated land reme- diation”, Chemosphere Vol. 41, pp. 197-207. Keating, M.H., Mahaffey, K.R., Schoney, R., Rice, G.E., Bullock, O.R., Ambrose, R. B., Swartout, J., Nichols, J.W.,“Mercury Study Report to congress”, EPA-452/ R-97-003, Washington DC, Vol. I, Sec.3. p.6-7. Lai, H.Y. and Chen, Z.S., 2004, “Effects of EDTA on solubility of cadmium, Zinc, and lead and their uptake by rainbow pink and vetiver grass”, Chemosphere, Vol. 55. pp.421-430. Lasat, M.M., 2001, “The Use of Plant for the Removal of Toxic Metals from Conta- minated soil”, Grant No. CX824823/US EPA. Ma, L.Q., Kmar, K.M., Tu, C., Zhang, W., Cai, Y., Kennelley, E.D., 2001,“A fern th- at hyperaccumulates arsenic”, Nature, Vol. 409, pp. 579. Mejare, M. and Bulow, L., 2001, “Metal-binding protein and peptides in bioremedi- ation and phytoremediation of heavy metals”, Trends in Biotechnology, Vol. 19, No.2, pp. 67-73. Nelson, D. W., Sommers, L. E.,1986, in Methods of Soil Analysis: Part2 Chemical and Microbiological Properties, Page, A. L., Ed., American Society of Agrono- my/Soil Science Society of America: Madison, WI, pp. 539-580. Nicholas, DJ. D., 1965,“Influence of the rhizosphere on the mineral nutrition of the plant”, In K. F. Baker and Snyder W. C. (eds.), Ecology of soil-brone plant pa- thogen, University of California Press, Berkeley. pp.210-217. Nies, D.H., 2003, “Efflux- mediated heavy metal resistance in prokaryotes”, Annual Review of Microbology, Vol. 27, No.2-3, pp. 313-339. Page, A.L., Miller, R.H., and Keeney, D.R., 1982,“Methods of soil analysis”, Part II, 2nd ed., ASA-SSSA, Wesconsin. Park, C.H., Keyhan, M. and Matin, A., 1999, “Purification and characterization of chromate reductase in Pseudomonas putida”, Abs. Gen. Meet. American. Soc. Microbiol., Vol. 99, pp.536. Paul, E.A. and Clark, F.E., 1996, .“Soil Microbiology and Biochemistry ”, 2nd ed., ACADEMIC PRESS, pp.27. Raskin, I., Kumar, P.B.A.N., Dushenkoy, S. and Salt, D.E., 1994,“Bioconcentration of heavy metals by plants”, Current Opinion Biotechnology, Vol. 5, pp.285- 290. Reasoner, D.J. and Geldreich, E.E., 1985,“A new medium for the enumeration and subculture of bacteria from potable water."Applied and Environmental Microbiology, Vol. 49, pp. 1-7. Rowbotham, T. J. and Cross, T., 1977,“Ecology of Rhodococcus coprophilus and associated actinomycetes in fresh water and agricultural habitats.”Journal of General Microbiology, Vol. 100, pp. 231-240. Shen, Z.G., Li, X.D. Chen, H.M., Wang, C.C. and Chua, H., 2002, “Phytoextraction of Pb from a contaminated soil using high biomass species of plant”, Journal of Environmental Quality, Vol. 31, pp.1893-1900. Silver, S. and Phung, L.T., 1996, “Bacterial heavy metal resistence:new surprise”, Annual Review of Microbiology, Vol. 50, pp. 753-789. Teizel, G.M. and Paresk, M.R., 2003, “Heavy metal resistance of biofilm and plank- tonic Pseudomonas aeruginosa”, Applied and Environmental Microbiology, Vol. 69, No.4, pp. 2313-2320. Tüzen, M., 2003, “Determination of heavy metals in soil, mushroom and plant sam- ples by atomic absorption spectrometry”, Microchemical Journal, Vol. 74, pp. 289-297. US EPA, 2000.“Introduction to Phytoremediation.”EPA/600/R-99/107. Washing- ton DC, February. Vivas A., Barea, J.M. and Azcón, R., 2005,“Interactive effect of Brevibacillus brev- is and Glomus mosseae, both isolated from Cd contaminated soil, on plant gr- owth, physiological mycorrhizal fungal characteristics and soil enzymatic act- ivities in Cd polluted soil”, Environmental Pollution, Vol. 134, Issue 2, pp.257-266. Wang, C.L., Michels, P.C., Dawson, S.C., Kitisakkul, S., Baross, J.A., Keasling, J.D. and Clark, D.S., 1997, “Cadmium removal by a new strain of Pseudomo- nas aeruginosa in aerobic culture”, Applied and Environmental Microbiology, Vol. 63, No.10, pp. 4075-4078. Xia, H. and Ke.H, 2003, “Application vetiver and three other grasses to oil shale mine for revegetation and phytoremediation”, Third International Vetiver Conference Guangzhou, China, pp. 540. 台中縣中山國民小學,校園植物介紹。 (http://163.17.130.1/myweb/namaster/naneng/html/alph.htm) 全國法規資料庫,土壤及地下水污染整治法。 (http://law.moj.gov.tw/) 高職園藝科進修網頁,園藝植物小百科。 (http://140.127.10.49/hort/hort/research/weed.htm) University of Hawai`i at Manoa, Botany Department. (http://www.botany.hawaii.edu/faculty/webb/BOT410/Roots/Mycorrhizae.htm) |
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