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  • 學位論文

台灣西南部關子嶺泥泉之微生物代謝多樣性研究

Microbial Diversity Study of the Kuantzuling Mud Spring in the Southwestern Taiwan

指導教授 : 王珮玲

摘要


台灣西南部泥質溫泉中常富含有機碳、硫酸根、甲烷及氫氣等化學成分,可提供許多微生物族群進行多樣化的代謝活動。關子嶺地區噴出的泥泉溫度相對西南部其它泥火山水溫高,水溫可達 80℃ 左右,適合做為研究嗜熱菌與陸域熱泉生態系統的題材,本研究即是運用微生物富化培養方法,探討關子嶺泥泉中實際存在的微生物代謝,並透過分離泥泉中的微生物,及其生理測試,了解泥泉中各種微生物在碳循環中所扮演的角色,討論各種微生物之間競爭或互利的關係。 本研究根據關子嶺泥泉的水化學及氣體化學分析資料,設計泥泉中可能存在的各種代謝反應之培養基。使用的培養基種類包括厭氧條件下使用醋酸、甲醇、甲胺、氫氣與二氧化碳等四種甲烷菌培養基、使用葡萄糖的硫代硫酸鹽還原菌培養基、發酵菌培養基,及好氧條件下異營菌和甲烷消耗菌等培養基。確認使用特定代謝微生物能夠適應培養基後,再進行純菌分離的工作,以得到單一菌株,並測試生理特性。 上述八類培養基均可富化出使用該代謝反應的微生物群落,但並非所有培養溫度均可見微生物群落的持續生長,厭氧條件下使用醋酸鹽、甲醇、氫氣和二氧化碳的產甲烷菌、使用葡萄糖之硫代硫酸鹽還原菌、發酵菌、好氧異營菌在50℃以上培養中可被富化,其中大部分的代謝在 80℃ 仍能被富化而持續進行,只有使用甲胺之產甲烷菌及好氧甲烷消耗菌僅在50℃以下的培養中被富化而生長。研究證實關子嶺泥泉中微生物具有多樣化的代謝方式,加上前人研究可富化培養之 使用乳酸鹽之硫酸鹽還原菌共為九種代謝反應,與前人之代謝通量計算和分子生物分析結果一致。利用富化培養結果顯示關子嶺泥泉中微生物參與環境中的有機碳降解和碳循環,彼此之間產生互相競爭或是互利的關係。 由好氧異營菌培養基中,純化出二株嗜熱好氧異營性細菌,編號分別為KTL01-S-121106-I1 與 KTL01-S-121106-I2。I1 菌株與Meiothermus ruber strain SPS242 之 16S rDNA 序列的相似度達 99%,可生長在酸鹼值 5~9 之間,可生長溫度範圍在 20~70℃ 之間,最適合生長溫度則為 60℃,其最適生長條件與關子嶺泥泉的地化條件相符; I2 菌株與 Rhodobacter vinaykumarii JA123 之16S rDNA序列的相似度達 95%,但此菌株並非光合作用菌,該菌株生長在酸鹼值 6~9 之間,可生長的溫度範圍在 20~60℃ 之間,最適合生長溫度則為 50℃,為一新菌種。

並列摘要


Organic carbon, sulfate, methane, and hydrogen are abundant in mud springs,southwestern Taiwan. These compounds can provide the microbial community to carry on diverse metabolisms. The temperature of the Kuantzuling mud spring could be as high as 80℃, thus the mud spring may offer a great opportunity to study thermophilic microorganisms and terrestrial hot spring ecosystems. This study performed a set of enrichment experiments, stain isolations and physiological tests to understand the interaction among different microorganisms. According to the aqueous and gas chemistry data of the Kuantzuling mud spring,we designed eight types of cultural media to enrich different kinds of microbial metabolism in the slurry. They are 6 types of anaerobic media for methanogens using acetate, methanol, methylamine or hydrogen and carbon dioxide to produce methane, thiosulfate reducers using glucose, fermenters and 2 types of aerobic media for heterotrophs and methanotrophs. Once the microorganisms could adapt to the certain types of media, pure stains were tend to be isolated and tested their physiological characteristics. Microorganisms can be enriched in all types of media, but not in all cultivated temperatures. Methanogens using acetate, methanol or hydrogen and carbon dioxide , thiosulfate reducter, fermenter, aerobic heterotroph could be enriched at temperatures higher than 50℃. Mostly of them can also be enriched at 80℃. Methanogen using methylamine and methanotroph can only be enriched at temperature of lower than 50 ℃. Total nine types of microbial metabolisms, including sulfate reducer using lactate confirmed by previous studies, can be enriched in the Kuantzuling mud spring. This result is consistent with previous energetic evaluation of microbial metabolisms and molecular analysis of the slurry. It also inferred that the microorganisms exhibited a great diversity of metabolic pathways and were either competitive or beneficial to each others while processing organic carbon degradation or carbon cycling. Strain KTL01-S-121106-I1 and KTL01-S-121106-I2 were isolated from the Kuantzuling mud spring with heterotrophy media. The 16S rDNA gene sequence of strain I1 exhibited a very close affiliation (at a similarity of 99%) with Meiothermus ruber strain SPS242. It can grow at the pH between 5 and 9, in the temperature ranging from 20 to 70℃ with the optimal growth temperature at 60℃. The 16S rDNA gene sequence of strain I2 was close to that of Rhodobacter vinaykumarii JA123 at the similarity of 95%, but stain I2 is not phototroph and probably is a new species. Strain I2 can grow at the pH between 6 and 9, in the temperature ranging from 20 to 60℃ with the optimal growth temperature at 50℃.

參考文獻


Academy of Sciences of the United States of America, v. 87, p. 4576-4579.
王宏霖, 2006, 以反射震測法研究木屐寮斷層及其附近構造, 國立中央大學地球物理研
朱寶萱, 2008, 厭氧型甲烷氧化作用在台灣西南部陸域泥火山的可能性評估, 國立台灣
陳冠樵, 2009, 台灣熱泉生態系統中微生物代謝之生物能量評估, 國立台灣大學理學院海洋研究所碩士論文, p. 160.
James G. Cappuccino, 2008, Natalie Sherman, Microbiology-a laboratory manual 8th edition,

被引用紀錄


陳韻如(2012)。台灣西南部關仔嶺溫泉微生物生成甲烷作用造成之同位素分化與族群結構關係〔碩士論文,國立臺灣大學〕。華藝線上圖書館。https://doi.org/10.6342%2fNTU.2012.02016
Lu, G. S. (2011). 台灣與菲律賓地熱溫泉中可培養嗜熱砷轉化菌之多樣性與生理代謝特性 [master's thesis, National Taiwan University]. Airiti Library. https://doi.org/10.6342%2fNTU.2011.02921

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