本研究嘗試以此技術處理受柴油污染濕地土壤，並對香蒲（Typha orientalis Presl）、鹹草（Cyperus malaccensis）、雲林莞草（Bolbos choenus planieulmis）及蘆葦（Phragmites communis）等四種本土常見水生植物進行耐油污能力測試，結果顯示香蒲在含有柴油污染土壤中，葉綠素削減率較低，生長旺盛、存活率最高。最後，本研究以香蒲為初步實驗對象。
經過240天的溫室實驗，比較栽種香蒲與未栽種香蒲的處理組，於人工配製的柴油污染濕地土壤試驗上，結果顯示香蒲的栽種有助於柴油污染濕地土壤中的真菌、柴油利用菌（DUB）生長及去氫酵素活性的增加，而營養鹽的添加對於香蒲對抗土壤柴油污染的逆境具有正面的幫助。
實驗最後，並利用分子生物技術PCR/DGGE方法，分析土壤中生物的多樣性，由DGGE剖面圖中顯示，香蒲的栽種對於柴油污染土壤可有較高的微生物多樣性。至於土壤TPH-D的降解分析方面，栽種香蒲並施加營養鹽實驗組，對TPH-D的生物分解效率大為提高。實驗至第240天時，土壤的柴油污染的去除約可達80 %，土壤中的TPH-D可由原先高濃度的16000 mg kg-1降解至3500 mg kg-1。

In this study we used the phytoremediation techniques to treatment diesel contaminated wetland soil. At first, we compared the four common wetland plants, Typha orientalis Presl, Cyperus malaccensis, Bolbos choenus planieulmis and Phragmites communis, on the treatment efficiency of the diesel contaminated wetland soils. From the results, we find out that the Typha orientalis Presl has highest growth rate and activity on rhizosphere among the four species.
The Typha orientalis Presl was planted on artifical diesel contaminated wetland soil and incubated inside a greenhouse, while a control system without vegetation is compared. After 240 days, the result shows that soil planted with Typha orientalis Presl can enhance the microbial and dehydrogenase activity. And adding with nutrients can help plants to prevent the diesel stress. Finally, we utilized the PCR/DGGE methods to analyze soil microbial diversity. According to the DGGE profiles, presence of Typha orientalis Presl can augment microbial diversity .
So far as degradation of TPH-d to be concerned, because of the period was too short, it doesn’t have significant difference between treatments. However, presence of Typha orientalis Presl and addition of nutrients, the TPH-D degradation rate was measured to be approximately 80 % and concentration of TPH-D could degrade from 16000 mg kg-1 to 3500 mg kg-1 after 240 days.

目錄
中文摘要………………………………………………………….…………I
英文摘要……………………………………………………………………II
表目錄…………………………………………………………..…………VI
圖目錄…………………………………………………………….………VII
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 溢油污染與環境的影響……………………………………………..3
2.2 柴油化學組成及性質………………………………………………..4
2.3 濕地植物介紹………………………………………………………..8
2.4 植物復育……………………………………………………………..9
2.4.1 植物選擇………………………………………………………...13
2.4.2 根圈微生物……………………………………………………...15
2.4.3 根部分泌液……………………………………………………...16
2.4.4 污染物生物可利用性…………………………………………...19
2.4.5 植物與微生物交互作用………………………………………...19
2.4.6 去氫酵素活性…………………………………………………...20
2.4.7 植物復育之優缺點…...…………………………………………22
2.5 處理成本比較………………………………………………………23
2.6 分子生物技術應用於土壤微生物分析……………………………23
2.6.1 聚合酵素鏈鎖反應……………………………………………...23
2.6.2 變性梯度凝膠電泳……………………………………………...24
2.7 國外案例…………….…………………………………………….24
第三章 研究方法與步驟………………………………………….…..…..26
3.1 實驗流程……………………………………………………………27
3.2 植物採集……………………………………………………………28
3.3 土壤採樣與樣品保存………………………………………………30
3.4 植物初步篩選世試驗………………………………………………30
3.5 柴油土壤配製……………………………………………………....30
3.6 土壤分析方法………………………………………………………32
3.6.1 土壤含水率………………………………………………….…32
3.6.2 土壤中酸鹼值………………………………………………….32
3.6.3 土壤有機碳…………………………………………………….33
3.6.4 土壤總凱氏氮………………………………………………….33
3.6.5 土壤氨氮……………………………………………………….34
3.6.6 總石油碳氫化合物………………………………………….…34
3.6.7 去氫酵素活性………………………………………………….35
3.6.8 土壤真菌……………………………………………………….35
3.6.9 土壤柴油利用菌……………………………………………….36
3.7 根圈中菌相分析……………………………………………………38
3.7.1土壤DNA萃取………………………………………………...38
3.7.2 聚合酶連鎖反應分析………………………………………….39
3.7.3 變性梯度膠凝電泳分析……………………………………….42
3.8 植體分析……………………………………………………………43
3.8.1 植體葉綠素…………………………………………………….43
3.8.2 植體生物質量………………………………………………….43
3.9 實驗之QA/QC……………………………………………………...44
3.10 實驗相關器材……………………………………………………..44
第四章 結果與討論……………………………………………………….46
4.1 土壤基本性質…………………………………………………..…..46
4.2 耐油濕地植物初步篩選…………………………………………....47
4.3 植物生長受柴油污染的影響………………………………………49
（1）實驗初期植物生長狀況………………………………………..49
（2）實驗最終植物高度比較………………………………………..49
（3）實驗最終植物葉片質量比較………………………………….50
4.4 土壤去氫酵素活性…………………………………………………54
4.5 土壤中微生物的含量………………………………………………57
（1）土壤中柴油利用菌的含量………………………………….57
（2）土壤中真菌的含量………………………………………….58
4.6 分子生物技術比較土壤微生物多樣性……………………………61
4.6.1 DNA抽取比較…………………………………………………61
4.6.2 PCR/DGGE分析土壤微生物多樣性………………………….63
4.7 TPH-D分解…………………………………………………………66
第五章 結論與建議……………………………………………………….72
5.1 結論…………………………………………………………………72
5.2 建議………………………………………………………………....73
參考文獻…………………………………………………………...……75
附錄……………………………………………………………………...80

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