石油組成成分中具有許多的碳氫化合物，會對自然環境造成傷害。其中多環芳香族碳氫化合物（Polycyclic aromatic hydrocarbons, PAHs）因其不易自然降解，且生物毒性較高，成為許多學者研究的主題，近來已有許多文獻在探討其代謝菌群以及其代謝路徑。而在此一類研究中，culture-independent approach利用16S rRNA gene序列的比對，不須經由培養過程，即可快速獲得菌群的分類資訊並建立PAHs相關分解菌群的資料庫，但其所能提供的有關生理代謝方面的資訊卻相當少。為了能得到直接證據回答”Who eat what”，穩定同位素標定法（Stable-isotope probing, SIP）技術開始廣泛應用在環境微生物研究上，其可以同時得到有關微生物基質代謝以及分類學上的資訊。本實驗中，利用優厚培養的方法建立PAHs分解菌群的16S rRNA gene的選殖資料庫，並利用SIP的方法，探討naphthalene分解菌群的結構。
在選殖資料庫建立的研究中，Naphthalene的主要分解菌群皆屬於Gammaproteobacteria中的Pseudomonas菌屬（100%）。Anthracene分解菌群主要分布於Alphaproteobacteria（90%）。Phenanthrene的分解菌群則主要分布在Betaproteobacteria（70%）。而Pyrene分解菌群分布較廣，但主要集中在Betaproteobacteria以及Xanthomonadales（64%）。在13C-naphthalene的SIP實驗中，Pseudomonas aeruginosa為其主要菌群，顯示其具有很強的Naphthalene代謝能力，與先前clone library 結果比較，Pseudomonas aeruginosa可能在分解初期有很好的表現量，但隨培養時間的延長，其可能在族群競爭上較不具優勢，以至在優厚培養的clone library中，所佔比例較少。由實驗結果可知，clone library的建立可有效的迅速掌握PAHs分解菌群的資料，但其無法提供有關代謝速率優勢上的資訊。而利用SIP技術，可正確回答”Who eat what”的問題。

Crude oil is a complex mixture of hydrocarbons, and it cause lot of environment pollution. Polycyclic aromatic hydrocarbons, PAHs, are one class of hazardous organic chemicals in crude oil mixture. They are difficult to be degraded in the natural environment and have more toxicity. In this study, we used the method of enrich culture to build 16S rRNA gene database of PAHs degrading bacteria community and also used the SIP to discuss the community structure of naphthalene in environmental soil samples.
In the 16S rRNA gene database of PAHs degrading bacteria, all of the dominant naphthalene degrading bacteria belong to the genus of Pseudomonas of γ-proteobacteria. Anthracene degrading bacteria and Phenanthrene degrading bacteria belong to the α-proteobacteria (90%) and β-proteobacteria (70%), individually. Others are pyrene degrading bacteria, which are the Xanthomonadales and the class of β-proteobacteria (64%). In the result of 13C-naphthalene SIP, Pseudomonas aeruginosa is the dominant bacterium, and show it has the stroungly ablity to degrade naphthalene. Compare with the 16S rRNA gene clone library of naphthalene enrich culture (there is only 5% belong to Pseudomonas aeruginosa), Pseudomonas aeruginosa may has better growth rate in primary stage, but it has little competitive advantage in later stage. The result presented here provide that the building of 16S rRNA gene clone library database can fast get the information of PAHs degrading bacteria community structure, but it can’t provide the information about the metabolism capabilities. And using the tecology of SIP, we can get this information and answer the question “who eat what”.

鄭竹逸。萘分解菌對於芳香族碳氫化合物的分解與菌種基本特性之研究。國立成功大學生物學研究所碩士論文。1997。
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邱憲明。利用傳統培養方法和分子生物方法探討油污染土壤的微生物社會結構。國立成功大學生物學研究所碩士論文。2004。
林徽鳴。利用變性梯度凝膠電泳與末端螢光標定限制酵素片段長度多型性分析法監測微生物社會之分子指紋變化。國立成功大學生物學研究所碩士論文。2005。
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