摘要

自然環境中，許多的菌種都具備有代謝或分解的能力，這些具代謝力以及分解能力的菌種常被用來解決環境污染的問題。硝化菌目前多半應用在廢水處理場的活性污泥或養殖環境中。本研究是針對高雄縣的養殖池進行研究，篩選異營性及自營性的氨氧化菌種。異營菌部分，共計挑出10株可培養的菌種進行硝化能力測試及16S rDNA的定序鑑種工作。由實驗結果，10株菌的16S rDNA的定序結果與最相似菌株的相似度皆達95%以上。其中有5株歸類於Pseudomonas屬的菌種、1株Alcaligenes屬及Serratia屬的菌種較具備氨的氧化分解能力。在自營菌的部分，採用富集化的方式培養，而後經PCR增幅出自營性菌的16S rDNA以及amoA基因，進行定序及序列分析的工作。由其16S rDNA的定序發現，所有的菌種皆歸類於Nitrosomonas屬的菌種，其中林園所篩出的菌種有別於其他兩地所篩出菌種，自成一個群落。而在amoA基因的定序結果所做成的親緣演化關係和16S rDNA類似，亦是全部皆落於Nitrosomonas屬的群落當中，且皆和Nitrosomonas nitrosa這株於氨氮有機物含量高的環境中，所分出來的自營性氨氧化菌有較為接近的親緣關係。分解能力較佳的異營性菌種及Nitrosomonas屬的菌種，皆可以提供研究發展本土性生物製劑的發展。而amoA基因PCR引子的專一性以及和amoA基因片段具有較高的變異度，亦是為研究自營性氨氧化菌的族群歧異度研究相當好方法。

Abstract

The process of nitrification is highly dependent on the microbial activities and transformation, which is carried out by autotrophic nitrifiers in general, however some heterotrophic nitrifiers also can carry out the process. The diversity of autotrophic and heterotrophic ammonia-oxidizing bacteria in aquaculture ponds in Kaohsiung county was investigated. Ten heterotrophic bacteria were isolated. The nitrification ability and 16S rDNA sequences were determined. Seven of the strains had higher nitrification ability, five of them are belong to the genus of Pseudomonas, and the other two belong to Alcaligenes and Serratia, respectively. Both 16S rDNA and amoA gene sequences results showed that all autotrophic ammonia-oxidizing strains in this study belong to Nitrosomonas genus. From the data of 16S rDNA sequences, the strains isolated from Linyuan Shiang were distinct to the other two sites. Besides, amoA gene represents a very powerful molecular tool for analyzing ammonia-oxidizing bacteria communities due to its specificity and fine-scale resolution of closely related populations.

目錄


章次 頁次

一、 前言……………………………….….1

二、 材料方法………………….…...….11

三、 結果……………………………..….26

四、 討論……………………………..….33

五、 參考文獻……………………….…..43

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