臺灣博碩士論文加值系統

植物微生物燃料電池(Plant microbial fuel cell, PMFC)作為一種新興的綠色能源，具有透過植物將太陽能轉換為電力之能力，藉由系統內部產電菌與甲烷菌、脫硝菌的競爭可能有溫室氣體減量的效果。本研究之目的在於探討植物微生物燃料電池技術應用於濕地對溫室氣體(甲烷和氧化亞氮)的減量以及產電效能，並比較PMFC應用於無鹽害土和鹽害土的差異性。同時對土壤中甲烷、氧化亞氮相關功能性基因進行定量，以了解隨著PMFC系統操作土壤中的功能性菌群變化，以及功能性基因與溫室氣體排放之間的相互關係。
結果顯示無鹽害土PMFC閉路及SMFC (Sediment microbial fuel cell)閉路產生之平均電壓分別為0.21 ± 0.07 V、0.34 ± 0.08 V，PMFC電壓輸出低於SMFC，與文獻中之結果有所出入，可能與研究時期為冬季有關；鹽害土PMFC閉路、SMFC閉路產生之平均電壓分別為0.06 ± 0.04 V、0.02 ± 0.02 V，推測是高鹽度對植物和微生物造成抑制，進而影響電壓輸出。
PMFC系統在產電良好的情況下有甲烷減量的效果，減少氧化亞氮排放的能力則不明確。無鹽害土PMFC甲烷通量由閉路低於開路轉變為閉路高於開路是由於甲烷菌增加，SMFC開路的甲烷菌豐度高於閉路，與SMFC甲烷通量結果相符。鹽害土的甲烷菌豐度隨著時間有上升但幅度不大，鹽害土的甲烷菌豐度遠低於無鹽害土，驗證了鹽害土甲烷通量明顯低於無鹽害土的結果。無鹽害土與鹽害土norB、nosZ、nirK和nirS基因的豐度數量級相當，與兩者氧化亞氮通量差異不大的結果一致。本研究結果尚無法清楚解釋脫硝菌與產電菌的競爭，若要探討脫硝菌與產電菌的關係需要更進一步研究。

Plant microbial fuel cell (PMFC), as a emerging green energy source, has the ability to convert solar energy into electricity through plants. It is hypothesized that through competition between electricity-generating bacteria and methanogens and denitrifying bacteria, greenhouse gas emissions could be reduced. The aim of this study is to explore the reduction of greenhouse gases (methane and nitrous oxide) and the power generation efficiency of PMFC technology applied to wetlands, and to compare the difference between PMFC applied to fresh and salt-affected soil. Meanwhile, by quantifying the functional genes related to methane and nitrous oxide in the soil, we can understand the functional consortium shifts as the PMFC systems operate and the correlation between functional genes and greenhouse gas emissions.
The results showed that the average voltages generated by fresh soil close circuit PMFC and close circuit SMFC (Sediment microbial fuel cell) were 0.21 ± 0.07 V and 0.34 ± 0.08 V, respectively. PMFC voltage output was lower than SMFC, which is inconsistent with the literature and may be related to the study period being winter. In salt-affected soil, the average voltages of close circuit PMFC and close circuit SMFC were 0.06 ± 0.04 V and 0.02 ± 0.02 V, respectively. The inhibition of plant and microorganism growth by high salinity may cause the low voltage output.
The PMFC system may have an effect on methane reduction under optimal operation, but the ability to reduce nitrous oxide emissions is unclear. In fresh soil, the PMFC methane flux change from close circuit lower than open circuit to the opposite was due to increased methanogens. In addition, the abundance of methanogens in open circuit SMFC is higher than that in close circuit, which is consistent with the results of SMFC methane flux. The abundance of methanogens in salt-affected soil increased slightly with time, and is much lower than that in fresh soil, which verifies that the methane flux of salt-affected soil is significantly lower than that of fresh soil. The abundances of the norB, nosZ, nirK and nirS genes of fresh soil and salt-affected soil were of the same order of magnitude, which is consistent with the little difference in nitrous oxide flux between the two. From the results of this study, the competition between denitrifying bacteria and electricity-generating bacteria cannot be clearly explained. Further research is required to provide evidence of the relationship between denitrifying bacteria and electricity-generating bacteria.

論文口試委員審定書 I
致謝 II
摘要 III
目錄 VI
圖目錄 IX
表目錄 XI
第一章 緒論 1
1.1 研究背景 1
1.2 研究動機與目的 2
第二章 文獻回顧 3
2.1 微生物燃料電池發展與原理 3
2.1.1微生物燃料電池發展 3
2.1.2微生物燃料電池的發電原理 4
2.1.3電子傳遞機制 4
2.2 植物微生物燃料電池 7
2.2.1植物微生物燃料電池原理 7
2.2.2 植物微生物燃料電池應用 8
2.2.3土壤鹽份對植物微生物燃料電池之影響 9
2.3 濕地溫室氣體 12
2.3.1濕地溫室氣體排放 12
2.3.2甲烷及氧化亞氮相關功能性基因 13
2.3.3濕地溫室氣體減量 14
2.4 數位核酸偵測系統(digital PCR) 16
第三章 材料與方法 17
3.1 研究架構 17
3.2 植物微生物燃料電池結構與運行 19
3.2.1實驗土壤來源 19
3.2.2植物選擇 19
3.2.3植物微生物燃料電池架設 20
3.2.4植物微生物燃料電池運行 21
3.3 土壤性質分析 22
3.3.1採樣及保存 22
3.3.2土壤含水率 22
3.3.3土壤元素分析 23
3.3.4土壤酸鹼值 23
3.3.5土壤導電度 24
3.4 溫室氣體通量分析 25
3.4.1氣體採樣罩設計 25
3.4.2氧化亞氮與甲烷 25
3.5 微生物分析方法 28
3.5.1土壤微生物樣本保存 28
3.5.2土壤DNA萃取 28
3.5.3微滴式數位核酸偵測系統(droplet digital PCR, ddPCR) 30
3.5.4次世代定序(Next generation sequencing, NGC) 33
3.6 統計分析 34
第四章 結果與討論 35
4.1 植物微生物燃料電池之產電 35
4.2 土壤酸鹼值變化 40
4.3 土壤導電度變化 44
4.4 植物微生物燃料電池對溫室氣體通量之影響 47
4.2.1甲烷通量 47
4.2.2氧化亞氮通量 49
4.5 植物微生物燃料電池對基因豐度之影響 51
4.5.1微滴式數位核酸偵測系統品保品管 51
4.5.2細菌與古菌16S rDNA基因 52
4.5.3產甲烷功能性基因 55
4.5.4氧化亞氮相關功能性基因 57
4.5.5菌群結構 64
第五章 結論與建議 67
5.1 結論 67
5.2 建議 69
第六章 參考文獻 71
附錄 81

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