本研究之目的主要是利用植物復育技術處理被pyrene污染之土壤。試驗時選用蘆葦、香蒲、培地茅、萬年青、鹹草、雲林莞草及大花咸豐草等七種植物，進行土壤復育，並分析土壤中pyrene之降解效率及微生物之變化，植物組織中之pyrene濃度。
結果顯示，實驗二十二週後，種植培地茅、萬年青及香蒲等植物可提高土壤中pyrene降解效率，尤其是當實驗進行十四週後，種植培地茅、萬年青及香蒲等植物土壤中pyrene降解百分率分別為86％、84％及77％，比未種植物之66％高出10％到20％。實驗分別在夏天或在冬天進行，實驗經過時間相同時，夏天測得土壤中pyrene降解效率皆大於冬天所測的數值。當pyrene污染土壤經過滅菌後，種植香蒲進行植物復育時，結果發現土壤中之pyrene降解效率皆比未經滅菌的土壤為低，實驗進行十四週後，不論是在有種植或無種植香蒲的試驗中，經滅菌過的污染土壤中pyrene的降解效率分別為59％及55％，比未經過滅菌土壤之77％及66％為低。由此可知微生物對土壤中pyrene之降解具有極大的影響性。種植有香蒲之土壤，若添加肥料(花寶2號)，則土壤中pyrene之降解效率具有明顯提高的作用，實驗進行十四週後。有種植及無種植香蒲在施肥之污染土壤中，pyrene降解效率分別為84％及73％，此數據顯示，比未施肥之77％及66％高出7％。研究結果發現，當土壤中加入界面活性劑Triton X-100及混合界面活性劑(Triton X-100、Tween 20及十二烷苯磺酸鈉)，對pyrene之降解效率確實有提升之作用，例如實驗十週後，種植培地茅且在土壤中添加界面活性劑Triton X-100及混合界面活性劑(Triton X-100、Tween 20及十二烷苯磺酸鈉)時，pyrene之降解效率分別為85％及87％，比未加入界面活性劑之81％分別高出4％及6％。實驗二十二週後，種植培地茅且在土壤中添加界面活性劑Triton X-100及混合界面活性劑(Triton X-100、Tween 20及十二烷苯磺酸鈉)時，pyrene之降解效率分別為96％及96％，同樣也較未加入界面活性劑土壤之94％為高。在種植萬年青之相同實驗中，也有相似之結果。因此選擇適當之界面活性劑對植物復育污染之土壤具有正面之效果。
上層土壤及下層土壤中，由pyrene之降解研究結果得知，由於上層土壤之含氧量較高故好氧微生物之活性較大，因此上層土壤較下層土壤對pyrene之降解效率為高。當實驗進行二十二週後，土壤中種植培地茅時，上層及下層土壤pyrene的濃度則分別為5.7 mg/㎏及10.8 mg/㎏，而上層濃度只有下層之ㄧ半，由此可推測pyrene降解應為好氧反應。另外，土壤中種植萬年青時也有相似之結果。
植物復育機制分析時，本研究分別對植物之根、莖及葉組織進行濃度分析，結果發現植物莖及葉中並無pyrene之存在，pyrene沒有被植物吸入至植物體中，因此植物萃取(phytoextraction)及植物蒸散(phytovolatilization)作用等機制，應並不是本研究中植物復育的機制，而本研究中，對pyrene的降解主要應為根區效應所造成。
當土壤中種植培地茅時，對乳酸菌屬的微生物似乎有抑制的作用，而在土壤中種植萬年青及香蒲時，則會對根黴菌屬微生物造成不利之影響。當實驗進行4週之後原分別存在於土壤中之乳酸菌屬及根黴菌屬已不存在。而土壤總生菌數分析，也發現當土壤中種植培地茅、萬年青及香蒲時，微生物之總生菌數及活性因而增大，所以土壤中之pyrene降解效率也提高。

The purpose of this study was to treat soils contaminated by pyrene through phytoremediation. The plant species selected were Phragmites communis Trin., Typha orientalis Presl, Vetiveria zizanioides, Rohdea japonica (Thunb.) Roth et Kunth, Cyperus malaccensis. Lam. subsp. monophyllus (Vahl) T. Koyama, Bolboschoenus planiculmis (F. Schmidt) T. Koyama and Bidens pilosa respectively. The degradation efficiencies of pyrene in soils and concentration of pyrene in the plant tissues were evaluated in this study. In addition, the change of microbial biota in soils was investigated in the tests of this study.
The experimental results indicated that after twenty-two weeks, soils planted with V. zizanioides, R. japonica and T. orientalis have better pyrene degradation efficiencies. Especially, after fourteen weeks the pyrene degradation efficiencies were 86%, 84% and 77% respectively, which showed that the efficiencies 10% to 20% higher than those unplanted control experiments, which was 66%. In addition, the pyrene degradation efficiencies in summer were found to be higher than those in winter. The degradation efficiencies of pyrene in sterilized soil with and without T. orientalis were found equal to 59% and 55%, respectively. These values were found lower than those in the experiment without sterilization, in which the pyrene degradation efficiencies with and without T. orientalis were 77% and 66%, respectively, after the fourteen weeks experiment. Hence the rhizospheric microorganisms had a significant effect on the degradation of pyrene in soils.
The pyrene degradation efficiencies were improved with application of fertilizer (HYPONeX No.2, HYPONeX Co., USA). After fourteen weeks, it was found that the experiment with fertilizer and with or without T. orientalis planted were 7% higher, which were 84% and 73% respectively, compared to 77% and 66% with no application of fertilizer.
Proper surfactants have positive effect on phytoremediation. In this study, we found that addition of the surfactant Triton X-100 or combined surfactants (Triton X-100, Tween 20 and sodium dodecylbenzene sulfonate) both presented better pyrene degradation efficiencies than the system without adding surfactant. After ten weeks, soils planted with V. zizanioides and added with surfactants showed the pyrene degradation efficiencies equal to 85% and 87% (combined) respectively, which showed that 4% and 6% higher rate than the system without adding surfactants (81% ). After twenty two weeks, soils planted with V. zizanioides and added with surfactants showed that the pyrene degradation efficiencies were 96% and 96% (combined) respectively. They were all higher than the system without adding surfactants (94%). Soils planted with R. japonica also showed the same results.
In this study, it was also found that the degradation efficiencies were higher at the surface layer of the soil than subsurface layer due to better oxygen content there. Hence the activities of microorganisms in the surface layer were higher than those in the subsurface layer of soils. After twenty two weeks, soils planted with V. zizanioides showed the residual concentration of pyrene were 5.7mg/Kg (surface layer) and 10.8 mg/Kg (subsurface layer). The difference between them was about 50%. Soils planted with R. japonica, T. orientalis or unvegetation also showed the same results.
The pyrene concentrations of the roots, stems and leaves were analyzed and the results showed that pyrene did not exist in the plant stems or its leaves. Since pyrene could not be absorbed into plant’s tissues by plants, the phytoextraction and phytovolatilization did not occur in this study. It was concluded that the degradation of pyrene in soils was mainly in rhizoremediation.
The soils planted with V. zizanioides showed that the inhibition of Lactobacillus sp, while the soils planted with R. japonica and T. orientalis showed unfavorable conditions to Rhibopus sp.. Four weeks after this experiment, both Lactobacillus sp. and Rhibopus sp. were not existent. The soils planted with V. zizanioides, R. japonica and T. orientalis showed an increase of the number of bacteria (CFU), and thus the pyrene degradation efficiency was increased.

中文摘要-------------------------------------------------Ⅰ
英文摘要-------------------------------------------------Ⅲ
目錄-----------------------------------------------------Ⅵ
圖目錄---------------------------------------------------XI
表目錄--------------------------------------------------XIV
第一章 緒論-----------------------------------------------1
1-1 研究動機-------------------------------------------1
1-2 植物復育之趨勢-------------------------------------1
1-3 研究目的-------------------------------------------3
第二章 文獻回顧-------------------------------------------4
2-1多環芳香烴碳氫化合物(PAHs)--------------------------4
2-1-1多環芳香烴碳氫化合物(PAHs)之特性--------------5
2-1-2多環芳香烴碳氫化合物(PAHs)之降解反應機構-----10
2-2土壤復育技術---------------------------------------14
2-2-1工程處理及化學處理技術-----------------------15
2-2-2 自然處理------------------------------------19
2-2-3 生物復育------------------------------------20
2-2-4 植物復育------------------------------------22
2-2-4-1 植物復育機制------------------------22
2-2-4-2 植物復育之優缺點--------------------25
2-2-4-3 植物篩選----------------------------25
2-2-4-4 根區效應----------------------------28
2-2-4-5 界面活性劑之分類特性及作用機制------31
2-2-4-6 植物復育技術之回顧------------------35
2-2-4-7 植物復育技術與其他復育法之成本比較--38
第三章 實驗及分析方法------------------------------------39
3-1 實驗藥品及設備材料--------------------------------39
3-1-1 實驗藥品------------------------------------39
3-1-2 儀器設備及材料------------------------------40
3-1-3 植物種類------------------------------------41
3-2 實驗方法------------------------------------------42
3-2-1 Pyrene污染土壤之製備------------------------42
3-2-2 土壤及植物滅菌------------------------------43
3-2-3 植物植入------------------------------------43
3-2-4 菌種植入------------------------------------46
3-2-5 界面活性劑之添加----------------------------46
3-2-6 有機酸之添加--------------------------------46
3-2-7 肥料之添加----------------------------------46
3-2-8 溫室培養------------------------------------47
3-3 分析方法------------------------------------------51
3-3-1 土壤樣品中pyrene之萃取----------------------51
3-3-1-1 土壤之取樣--------------------------51
3-3-1-2 樣品中Pyrene之萃取------------------52
3-3-2 植物組織中pyrene之萃取----------------------54
3-3-3 高效液相層析儀(HPLC)分析--------------------56
3-3-4 總生菌數(CFU)之分析-------------------------58
3-3-5 土壤中之菌種分析----------------------------59
第四章 植物復育技術土壤中pyrene降解之研究---------------62
4-1 前言----------------------------------------------62
4-2 結果與討論----------------------------------------63
4-2-1 植物種類對pyrene降解之影響-----------------63
4-2-2 季節(冬天及夏天)對pyrene降解之影響---------65
4-2-3 施肥對pyrene降解之影響---------------------70
4-2-4 土壤滅菌對pyrene降解之影響-----------------72
4-2-5 界面活性劑對pyrene降解之影響---------------74
4-2-5-1 培地茅植物復育系統------------------74
4-2-5-2 萬年青植物復育系統------------------82
4-2-6 有機酸對pyrene降解之影響-------------------91
4-2-7 植物種類及添加微生物對pyrene降解之影響-----95
4-2-8 土壤上層及土壤下層之pyrene降解分析---------97
4-3 小結---------------------------------------------100
第五章 植物組織中pyrene之分析--------------------------102
5-1 前言---------------------------------------------102
5-2 結果與討論---------------------------------------102
5-3 小結---------------------------------------------104
第六章 植物復育技術對土壤菌相變化及pyrene降解之影響----105
6-1 前言---------------------------------------------105
6-2 結果與討論---------------------------------------105
6-2-1 植物種類對土壤中總生菌數之影響-------------105
6-2-2 土壤滅菌對土壤中總生菌數之影響-------------107
6-2-3 界面活性劑對土壤中總生菌數之影響-----------109
6-2-4 土壤中菌種分析-----------------------------112
6-4 小結---------------------------------------------117
第七章 統計分析-----------------------------------------119
7-1 前言---------------------------------------------119
7-2 統計分析方法-------------------------------------119
7-3 結果與討論---------------------------------------121
7-3-1 植物種類對pyrene降解之影響----------------121
7-3-2 植物種植及土壤滅菌對pyrene降解之影響------124
7-3-3 溫度(冬天及夏天)對pyrene降解之影響--------127
7-3-4 施肥對pyrene降解之影響--------------------129
7-3-5 植物種類對土壤中總生菌數之影響-------------130
7-3-6 植物種植及土壤滅菌對土壤中總生菌數之影響---131
第八章 結論與建議---------------------------------------132
8-1 結論---------------------------------------------132
8-2 建議---------------------------------------------136
第九章 參考文獻-----------------------------------------138
附錄 Pyrene 回收率之影響之研究--------------------------153
附錄-1 前言------------------------------------------153
附錄-2 結果與討論------------------------------------154
附錄-2-1 萃取溶劑對污染土壤中pyrene萃取回收率之
影響------------------------------------154
附錄-2-2 萃取溶劑二氯甲烷量對污染土壤中pyrene回
收率之影響------------------------------156
附錄-2-3 污染土壤中pyrene濃度對pyrene 回收率之
影響------------------------------------157
附錄-2-4 細胞粉碎機之萃取時間對污染土壤中pyrene
回收率之影響----------------------------158
附錄-2-5 細胞粉碎機之萃取次數對污染土壤中pyrene 回
收率之影響------------------------------159
附錄-3 小結------------------------------------------161

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