生物復育是近年備受關注的一門環境復育技術，微生物社會結構之分析則是了解其作用機制不可或缺的部份。本研究樣本取自台灣南部LY、YK、FD、KS和KT等石油污染場址，以傳統微生物培養與分子生物學的方法，來探討油污染場址的微生物社會結構。從各場址建立的16S rRNA基因選殖資料庫，可發現彼此間微生物社會結構有極大的差異，各場址均有特定的菌群存在，其中β-proteobacteria又可依五處場址分成五組不同的小群。LY場址是一個污染較久場址，其微生物社會組成和目前已知的油污染分解菌均不同，大多屬於uncultured α-、β-和γ-proteobacteria，而YK場址則以β-proteobacteria的Burlkhoderia菌屬較多。在油污染場址16S rRNA基因選殖資料庫中，與油分解相關的OTUs(operational taxonomic units)共有21個，其中YK廠址以Pseudomonas與Burlkhoderia菌屬所佔比例最大，分別為28.0%和24.5%。本研究以T-RFLP來監測KS(柴油含量較多)與KT(重油含量較多)兩場址在生物復育過程中微生物社會結構的變化，並以nonmetric multidimensional scaling (NMS)統計分析兩場址間微生物社會結構之相似性，可看出兩場址第0天的微生物社會結構差異性大，可能出自於兩者所含油的組成成分不同所致。至於同場址之不同時間點相似性的差異，則顯示出微生物社會結構在生物復育過程中有明顯變化。
關於PCR造成模板與產物菌群比例的偏差方面，本篇發現一般一階段的PCR會造成模板與產物菌群比例的偏差，且當引子與模板有一個鹼基無法配對時，造成的偏差則越大。然而，利用兩階段PCR策略則可減小上述造成的偏差。因此，在以T-RFLP分析微生物社會結構上，此方法可以改善因引子與模板不完全配對造成的模板與產物菌群比例之偏差。

Bioremediation is the most concern technology of environmental remediation in recent year. Analysis of microbial community is indispensable to understand mechanism of bioremediation. The soil samples of this study were from petroleum-contaminated sites, including LY, YK, FD, KS, and KT. We used traditional culture and molecular biology methods to investigate the microbial communities of these sites. In 16S rRNA gene clone libraries of each site, large diversity was found between these microbial communities. Microbes at each site had specific locus in phylogenetic tree of 16S rRNA gene clone libraries. Interestingly, β-proteobacteria could be divided five groups by these five sites. LY site is contaminated longer time. As we known, it had different microbial community from others and most of the microbes here was uncultured α-, β-, and γ-proteobacteria. Meanwhile, most of the microbes in YK was Burlkhoderia. In 16S rRNA gene clone libraries of petroleum-contaminated sites, there were 21 OTUs (operational taxonomic units) related to degradation of petroleum, and YK site had a large fraction of Pseudomonas and Burlkhoderia, 28.0% and 24.5% respectively. This study used T-RFLP to monitor the change of microbial community at KS (more diesel oil) and KT (more heavy oil) sites during bioremediation, and used NMS (nonmetric multidimensional scaling) to analyze the similarity of microbial community. As the result, similarity between the two sites was small in 0 day. It may be caused by the different component of petroleum at two sites. As for the dissimilarity at different time, it suggests that microbial community change obviously in bioremediation.
This study also found that one-stage PCR amplification produced bias of bacterial ratio between templates and products, and the bias increased if there was a mismatch site between primers and templates. Two-stage PCR amplification could diminish the bias. Therefore, in T-RFLP, this method could ameliorate the bias caused by mismatch between primers and templates.

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。
林裕家。利用變性梯度凝膠電泳與即時定量聚合酵素連鎖反應監測油污染場址在復育過程中微生物社會結構之變化。2006。
趙奎驊。利用傳統培養方法及分子生物方法建立分解多環芳香族碳氫化合物的微生物社會結構並探討其重要分解菌群的角色。2007。
潘柏岑。應用土耕法配合生物添加促進法整治柴油污染土壤之研究。2006。

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