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研究生:陳怡文
研究生(外文):Chen, Yi-Wen
論文名稱:海洋兼性嫌氣性發酵產氣細菌之分離與特性研究
論文名稱(外文):Isolation and characterization of marine facultatively anaerobic bacteria that ferment carbohydrates with gas production
指導教授:謝文陽
指導教授(外文):Shieh, Wung Yang
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:海洋研究所
學門:自然科學學門
學類:海洋科學學類
論文種類:學術論文
論文出版年:2000
畢業學年度:88
語文別:中文
論文頁數:68
中文關鍵詞:好鹽性發酵產氣16S rDNA序列
外文關鍵詞:halophilicferment carbohydrates with gas production16S rDNA sequence
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本研究在台灣北部沿海設立11個採樣點,針對分布於此間的好鹽性兼性嫌氣性發酵產氣細菌之生菌數和種多樣性進行調查,並期待能發掘出前所未見的種類。調查結果發現在各採樣點中,對象菌的密度介於101至104 cells/100 ml之間。篩選出的四十三株好鹽性兼性嫌氣性發酵產氣菌中,有十二株在繼代培養過程中,失去於發酵醣類時產氣的性質,其餘分離株,皆為以極性鞭毛運動之革蘭氏陰性桿菌,它們都能以葡萄糖作為生長所需之唯一碳源,且都無法生長於不含鈉鹽的培養基中。這三十一株分離株依其表型特性,可區分成五群。第一群:僅有一株;菌落呈紅色,圓形;細胞為弧形桿狀;氧化反應為負;可還原硝酸鹽;能利用mannitol為唯一碳源。第二群:共六株;菌落呈白色,稍透明,圓形;細胞呈弧形桿狀;氧化反應為負;可還原硝酸鹽;能利用mannitol為唯一碳源。第三群:共三株;菌落呈白色,稍透明,圓形;細胞為弧形桿狀;氧化反應為正;無還原硝酸鹽能力;能利用mannitol為唯一碳源。第四群:共三株;菌落呈白色,不透明,圓形;細胞呈短直桿狀;氧化反應為正;可還原硝酸鹽;無法利用mannitol為唯一碳源。第五群:共十八株;菌落呈白色,稍透明,圓形;細胞呈弧形桿狀;氧化反應為正;可還原硝酸鹽;能利用mannitol為唯一碳源。這五群之代表株經16S rDNA定序與親緣分析後發現,它們都是屬於弧菌科( Vibrionaceae )中的弧菌屬( Vibrio )或發光菌屬 ( Photobacterium )。就16S rRNA序列之相似度而言,與第一群唯一菌株NFG1最相似之菌種為無產氣能力的Vibrio fluvialis (相似度93.7 % )及亦具有紅色色素和產氣能力的Vibrio gazogenes (相似度93.6 % )。第二群的代表株NFG2之最相似菌種為Vibrio furnissii (相似度97.8 % )。第三群代表株NFG3之最相似菌種為無產氣能力的Vibrio tubiashi (相似度96.6 % )。第四群代表株NFG4之最相似菌種為Photobacterium damselae subsp. damselae (相似度98.8 % )。第五群的代表株NFG5之最相似菌種為Vibrio furnissii (相似度96.7 % )。依照表型與16S rDNA序列分析的結果,NFG1與NFG3可確定為新種,NFG4應可歸類於Photobacterium damselae subsp. damselae,至於其他二株代表株之分類地位,則有待進一步確認。
A search for halophilic, facultatively anaerobic bacteria that ferment carbohydrates with gas production has been made in the present study. The most-probable-number ( MPN ) counting values of these bacteria in the coastal seawater collected from northern Taiwan were estimated to be in the range of 101 to 104 cells/100 ml. Forty-three bacterial strains that ferment carbohydrates with gas production were isolated from the MPN counting tubes. Twelve lost the gas-producing property after subculture among them. All the other isolates were Gram-negative, and motiled by polar flagella. They were able to grow in a mineral medium containing D-glucose, and could ferment carbohydrates with gas production. These isolates required Na+ for growth, too. According to phenotypes, thirty-one strains were divided into five groups. Group 1 contained only one strain, which produced red colonies. The cells were curved rods in shape and oxidase-negative. The strain was able to assimilate mannitol as sole carbon source, and reduced nitrate. Group 2 contained six strains, which produced white and transparent colonies. The cells were curved rods in shape and oxidase-negative. The strains were able to assimilate mannitol as sole carbon source, and reduced nitrate. Group 3 contained three strains, which also produced white and transparent colonies. The cells were curved rods in shape and oxidase-positive. The strains were able to assimilate mannitol as sole carbon source, and couldn’t reduce nitrate. Group 4 contained three strains, which produced white and opaque colonies. The cells were short and straight rods in shape and oxidase-positive. The strains weren’t able to assimilate mannitol as sole carbon source, and reduced nitrate. Group 5 contained eighteen strains, which produced white and transparent colonies. The cells were curved rods in shape and oxidase-positive. The strains were able to assimilate mannitol as sole carbon source, and reduced nitrate. The levels of 16S rDNA sequence similarity compared with any sequences available in the GeneBank database revealed that the representative strains NFG1 and NFG3 of Group 1 and Group 3 could be considered to represent various new species. The representative strain NFG4 of Group 4 was identified as Photobacterium damselae subsp. damselae. The taxonomic status of NFG2 and NFG5 that representative strains of Group 2 and Group 5 couldn’t be determined.
圖目錄…………………………………………………………Ⅱ
表目錄…………………………………………………………Ⅲ
中文摘要………………………………………………………Ⅳ
英文摘要………………………………………………………Ⅵ
前言……………………………………………………………1
材料與方法……………………………………………………5
結果……………………………………………………………16
討論……………………………………………………………22
參考文獻………………………………………………………29
圖表……………………………………………………………35
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