謝文陽臺灣大學:海洋研究所廖建勛Liao, Jian HsunJian HsunLiao2010-05-062018-06-282010-05-062018-06-282008U0001-1707200815404800http://ntur.lib.ntu.edu.tw//handle/246246/181190耐冷細菌 (psychrotolerant bacteria) 一般的定義，是指最適生長溫度位於20 - 40 ℃之間，而在低於20 ℃、甚至更低溫的環境下仍能生長良好的細菌。本研究於2006至2007年間，在台灣東北角海岸 (潮境公園、磅磅子、南雅)、宜蘭石城與墾丁南灣等地採集水樣，進行序列稀釋後將稀釋海水塗佈於PY培養基，並放置於4 ℃恒溫箱中培養，藉此篩選出具耐冷特性的分離株。研究一共篩選出101株具耐冷特性的分離株，由菌落型態與觸酶、鈉鹽需求、可生長鹽度等表型特性，初步將分離菌株區分為10群；將分離株 16S rDNA 以4種限制酶 (DdeI、RsaI、AluI和HhaI) 進行限制片段長度多樣性 (restriction fragment length polymorphism, RFLP) 分析，共可分為13種基因型。3種基因型代表株經16S rDNA定序，並與標準菌株序列比對，大致可歸類為Cobetia、Enterobacter、Pseudomonas、Pseudoalteromonas、Psychrobacter、Shewanella、Vibrio等七個菌屬，其中以Pseudoalteromonas屬佔最多數，而Vibrio屬涵蓋了4種基因型。本研究顯示，雖然台灣位處亞熱帶地區，仍有多種耐冷細菌存在沿岸海水中。Psychrotolerant bacteria are those grow with optima of 20 - 40 ℃, but can grow below 20 ℃, even lower to 0 ℃. These bacteria are called. Attempting to know the diversity of psychrotolerant bacteria in coastal seawater of Taiwan, the sea water samples were collected from northeastern coast of Taiwan (Chaojing Park, Bangbangzih and Nanya), Shihcheng and South Bay during 2006 to 2007. These sea water samples were diluted to 101 - 103 solutions, then plated on PY agar and placed at 4 ℃ to screen out psychrotolernat bacteria.n this study, there were 101 psychrotolernat bacteria strains isolated from the sea water samples. They could be divided into ten groups according to the results of the following tests: colors of the colony, requirement of Na+ for growth, and the ability to grow in 10 % NaCl. These strains were divided into thirteen genotypes within 16S rDNA RFLP after the response by four kinds of restrictive enzyme (AluI, DdeI, HhaI and RsaI).he 16S rDNA of the representative strains of thirteen genotypes were sequenced, and they were belonged to seven genus: Cobetia, Enterobacter, Pseudomonas, Pseudoalteromonas, Psychrobacter, Shewanella and Vibrio. This study indicates that there are many species of psychrotolerant bacteria in coastal seawaters in Taiwan.目錄文摘要 i文摘要 ii一章 前言 1二章 材料與方法 5、培養基配置 5、樣本採集 5、菌株分離與篩選 6、分離株之形態及生理、生化特性檢測 6、分離株之16S rDNA分析 11三章 結果 15、耐冷細菌的分離與篩選 15、分離株形態與生理特性分群 15、分離株16S rDNA RFLP 分析 17、各基因型代表株之細部生理生化特性分析 17、各基因型代表株16S rDNA序列分析 18四章 討論 20、海洋細菌的定義與判別 20、表型與基因型分群比較 21、各基因型代表株與相近菌種的比較 23、台灣沿岸的耐冷細菌多樣性 29考文獻 30 42一、各群分離株之生理生化特性 42二、分離株16S rDNA RFLP分析結果 43三、13種基因型代表株之生理生化特性 44四、13種基因型代表株之單一碳源利用與醣類發酵測試 46五、各基因型代表株與已知菌種的16S rDNA相似度 48六、表型與基因型分群差異比較 49 50一、採樣地點 50二、根據表型特性差異進行分離株初步分群 51三、RFLP分析酶切片段模擬圖 (限制酶AluI、DdeI) 52四、RFLP分析酶切片段模擬圖 (限制酶HhaI、RsaI) 53五、以限制酶AluI、DdeI進行酶切反應所得的酶切片段電泳圖 54六、以限制酶HhaI、RsaI進行酶切反應所得的酶切片段電泳圖 55七、12種基因型代表株前段序列比對之親緣關係圖 56錄 57application/pdf1055413 bytesapplication/pdfen-US海洋細菌耐冷多樣性限制片段長度多樣性分析sea waterbacteriapsychrotolerantdiversityRFLPthesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/181190/1/ntu-97-R95241213-1.pdf