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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 謝文陽 | |
| dc.contributor.author | Yu-Te Lin | en |
| dc.contributor.author | 林育德 | zh_TW |
| dc.date.accessioned | 2021-06-13T04:27:29Z | - |
| dc.date.available | 2006-07-25 | |
| dc.date.copyright | 2006-07-25 | |
| dc.date.issued | 2006 | |
| dc.date.submitted | 2006-07-20 | |
| dc.identifier.citation | Altabet, M. A., Francois, R., Murray, D. W. & Prell, W. L. (1995). Climate-related variations in denitrification in the Arabian Sea from sediment 15N/14N ratios. Nature 373, 506-509.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/33164 | - |
| dc.description.abstract | 脫氮作用為地球生態系氮循環的重要過程。具脫氮能力之細菌則種類繁多，並不侷限於單一菌屬或菌種。本研究針對台灣北部河口紅樹林，包括挖仔尾、竹圍、關渡、紅毛港、客雅溪口和中港溪口等處沉積物中的脫氮細菌菌相進行研究。使用 PYN (polypepton-yeast-nitrate) 液體培養基，配合最可能數計數法 (most-probable-number counts，略稱 MPN counts)，估得各地紅樹林沉積物中，脫氮菌數約介於 2.6×104 至 1.2×105 cells/g wet wt. 間。自沉積物樣本中共分得 101 株脫氮分離株，經 AluI 和 RsaI 兩種限制內切酶，進行其 16S rRNA 基因之限制酶片段長度多型性 (restriction fragment length polymorphisms，略稱 RFLP) 圖譜之分析後，共可分為 21 個菌群。在進行多項生理生化特性檢測後發現，其中 66 株 (65.3 %) 具鈉鹽生長需求，應屬海洋原生種細菌。所有分離株中有 28 株 (27.7 %) 能行發酵作用，屬於兼性嫌氣性脫氮細菌。分析各菌群內各地代表株 16S rRNA 基因序列之親緣關係後顯示，自各地區紅樹林沉積物中所得之脫氮分離株分屬於 11 個菌屬，其中以 Marinobacter、Shewanella 和 Oceanimonas 等菌屬為主要組成。
所分得脫氮分離株中，編號為 CK1和K12，分別分離自中港溪口和關渡的菌株，除脫氮能力外，亦同時具發酵能力，屬於異營性、兼性嫌氣性，藉單根極鞭毛進行運動之革蘭氏陰性桿菌。在嫌氣狀態下，它們都能利用硝酸根或氧化亞氮作為電子接受者，進行脫氮生長；亦能利用葡萄糖或蔗糖等碳水化合物作為基質，進行發酵生長。CK1 和 K12 最適合生長於 30 ºC 至 35 ºC 和 pH 7。它們皆無鈉鹽生長需求，但最適合生長於氯化鈉濃度為 1 % 至 3 % 之環境；兩分離株中只有 K12 能在13 % 至 14 % 之氯化鈉濃度下生長。16S rRNA 基因序列分析顯示，CK1和 K12 的序列相似度為 96.8%；且與已知最相近菌種 Oceanimonas doudoroffii、Oceanimonas baumannii、Oceanimonas smirnovii 和 Oceanisphaera litoralis 在親緣樹上明顯分歧，彼此之間只具有94.1 % 至 96.8 % 的序列相似度。在細胞脂肪酸方面，CK1 和 K12 皆以 C18:1ω7c (32.6-35.7%)、C16:1ω7c (27.5-29.4%) 和 C16:0 (20.1-22.0%) 為主要組成，不同於 Oceanimonas 和 Oceanisphaera 所含菌種皆以 C16:1ω7c 為主。以G + C含量而言，CK1和K12的62-64 mol%，亦與 Oceanimonas 和 Oceanisphaera 屬所含菌種之54-59 mol% 明顯有別。多元分類數據顯示，CK1和K12 兩脫氮分離株應為不同種，且可歸類為屬於γ-Proteobacteria 之新屬細菌。目前已將其歸屬於 Zobellella 此新的菌屬，分別命名為 Zobellella denitrificans 和 Zobellella taiwanensis，並以 Zobellella denitrificans 為此菌屬之標準種。相關論文並已發表於 International Journal of Systematic and Evolutionary Microbiology 期刊 (Lin, Y.-T. & Shieh, W. Y. Int J Syst Evol Microbiol (2006). 56, 1209-1215)。 除了針對紅樹林沉積物中脫氮細菌的多樣性，及分類地位進行研究外，本研究亦利用屬於亞硝酸根還原酶 (nitrite reductase) 基因 (nirK 和 nirS) 中保守區域的引子對，透過聚合酶連鎖反應 (polymerase chain reaction，略稱 PCR) 分析脫氮分離株具有的 nir 形式。所得 101 個脫氮分離株中，有 64 株 (63.4 %) 含有 nirS 基因，僅 16 個 (15.8 %) 分離株帶有 nirK 基因，其餘 21 株則尚無法以此些引子對確定其亞硝酸根還原酶形式。分析部分分離株 nir 基因片段序列之親緣關係顯示，此些分離株具有之 nirK 基因與已知者具高度差異，序列相似度皆不及 80 %，而 nirS 基因與目前已知者則較為相近，序列相似度皆高於 93 %。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2021-06-13T04:27:29Z (GMT). No. of bitstreams: 1 ntu-95-D87241004-1.pdf: 2330234 bytes, checksum: c4cdb8790d05d2d42f7b0562cbbe1909 (MD5) Previous issue date: 2006 | en |
| dc.description.tableofcontents | 摘要 ..……………....……………………………………………… I
Abstract ...………………………………………………………….. III 第 1 章 前言 ..…………………………………………………… 1 1.1 脫氮作用之重要性 ..………………………………………... 1 1.2 紅樹林與脫氮作用 ..……………………………………….. 2 1.3 脫氮細菌之多樣性 ………………………………………… 3 1.4 兼性嫌氣性，能同時進行發酵與脫氮作用之海洋細菌： Pseudovibrio denitrificans ………………………………….. 4 1.5 好氣性，但能進行脫氮作用之脫氮細菌： Bowmanella denitrificans …………………………………... 4 1.6 研究目的 …………………………………………………… 5 第 2 章 紅樹林沉積物中脫氮細菌之菌相組成 ……………..... 6 2.1 前言 ………………………………………………………… 6 2.2 材料與方法 ……………………………………………….... 7 2.2.1 培養基 ………………………………………………….. 7 2.2.2 紅樹林沉積物樣本採集 ……………………………….. 7 2.2.3 紅樹林沉積物中脫氮細菌菌數之估算 ……………….. 8 2.2.4 沉積物內脫氮細菌之分離、純化與保存 …………….. 9 2.2.5 分離株之 DNA 萃取與 PCR 反應 ………………….. 10 2.2.6 16S rRNA 基因RFLP 圖譜之分析 …………………… 11 2.2.7 脫氮分離株各項生理生化特性之分析 ……………….. 11 2.2.8 脫氮分離株親緣關係之分析 ………………………….. 12 2.3 結果與討論 ……………………………………………….... 13 2.3.1 各地紅樹林沉積物中脫氮細菌菌數 ……...…………... 13 2.3.2 脫氮細菌之分離與其 16S rRNA 基因 RFLP 圖譜之分析 …………………………………………….. 14 2.3.3 脫氮分離株之生理生化活性 ………………………….. 15 2.3.4 各菌群各地代表株親緣關係之分析 ………………….. 15 圖 ……………………………………………………………….. 22 表 ……………………………………………………………….. 26 第 3 章 能兼行發酵作用與脫氮作用之新屬新種脫氮細菌： Zobellella denitrificans 和 Zobellella taiwanensis ……….. 33 3.1 前言 ………………………………………………………... 33 3.2 材料與方法 ………………………………………………... 34 3.2.1 培養基 …………………………………………………. 34 3.2.2 菌株來源 ………………………………………………. 35 3.2.3 DNA 萃取、16S rRNA 基因 PCR 反應與 親緣關係之分析 ……………………………………….... 35 3.2.4 菌株 G + C 含量分析 ………………………………… 35 3.2.5 DNA-DNA 相關性 (DNA-DNA relatedness) 之分析 .. 36 3.2.6 脂肪酸與極性脂質組成之分析 ………………………. 36 3.2.7 不同因子對分離株生長之影響 ……............................. 37 3.2.8 分離株生理生化特性之檢測 …………..……………... 37 3.2.9 分離株之抗生素藥劑感受性試驗 …............................. 38 3.2.10 分離株 quinone 之萃取與檢測 …………………….. 38 3.3 結果與討論 ………………………………………………... 39 3.3.1 分離株16S rRNA 基因序列之分析 ………………….. 39 3.3.2 分離株DNA 之 G + C 含量 …………………………. 40 3.3.3 分離株之 DNA-DNA 相關性 ....................................... 40 3.3.4 分離株之脂肪酸與極性脂肪組成 …………………….. 40 3.3.5 不同因子對分離株生長之影響 …….............................. 41 3.3.6 分離株之各項生理生化特性 …………………...……... 42 3.3.7 分離株對抗生素藥劑之感受性 ……………………….. 44 3.3.8 分離株之 quinone 組成 ………………………………. 44 圖 ……………………………………………………………….. 46 表 ……………………………………………………………….. 51 第 4 章 紅樹林沉積物中脫氮基因之多樣性 ………….…….... 58 4.1 前言 ………………………………………………………... 58 4.2 材料與方法 ………………………………………………... 59 4.2.1 培養基 …………………………………………………. 60 4.2.2 培養條件 ………………………………………………. 60 4.2.3 分離株之 DNA 萃取 …………………………………. 60 4.2.4 nir 基因引子對與 PCR 反應 ………………………… 60 4.2.5 分離株之 nir 基因定序與序列分析 ……………….... 62 4.3 結果與討論 ……………………………………………….. 62 4.3.1 nir 基因之 PCR 反應和沉積物脫氮分離株之 nir 基因形式 ………………………………………………... 62 4.3.2 紅樹林沉積物脫氮分離株 nir 基因之親緣關係 …… 64 圖 ………………………………………………………………. 66 表 ………………………………………………………………. 68 參考文獻 ………………………………………………………… 69 附錄一 …………………………………………………………… A1 附錄二 …………………………………………………………… A2 附錄三 …………………………………………………………… A3 | |
| dc.language.iso | zh-TW | |
| dc.title | 紅樹林沉積物中脫氮細菌之研究 | zh_TW |
| dc.title | The Study of Denitrifying Bacteria from Mangrove Sediments | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 94-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 蔡懷楨,蔡珊珊,李佳音,陳卓昇,劉玫英,李重義,李永安 | |
| dc.subject.keyword | 脫氮細菌,沉積物,紅樹林, | zh_TW |
| dc.subject.keyword | Denitrifying bacteria,Sediments,Mangrove, | en |
| dc.relation.page | 78 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2006-07-22 | |
| dc.contributor.author-college | 理學院 | zh_TW |
| dc.contributor.author-dept | 海洋研究所 | zh_TW |
| 顯示於系所單位： | 海洋研究所 | |
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