Yoshida Nobuyuki, Yagi Kazuhiro, Sato Daisuke, Watanabe Noriko, Kuroishi Takeshi, Nishimoto Kana, Yanagida Akira, Katsuragi Tohoru, Kanagawa Takahiro, Kurane Ryuichiro, Tani Yoshiki
Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma 630-0192, Japan.
J Biosci Bioeng. 2005 Feb;99(2):143-9. doi: 10.1263/jbb.99.143.
Bacterial communities in crude-oil samples from Japanese oil stockpiles were investigated by 16S rRNA gene cloning, followed by denaturing gradient gel electrophoresis (DGGE) analysis. 16S rRNA genes were successfully amplified by PCR after isooctane treatment from three kinds of crude-oil sample collected at four oil stockpiles in Japan. DGGE profiles showed that bacteria related to Ochrobactrum anthropi, Burkholderia cepacia, Stenotrophomonas maltophilia, Propionibacterium acnes, and Brevundimonas diminuta were frequently detected in most crude-oil samples. The bacterial communities differed in the sampling time and layer. Among the predominant bacteria detected in the crude oil, only three species were found for bacteria isolated on agar plates and were related to Burkholderia, Stenotrophomonas, and Propionibacterium, while Ochrobactrum sp. could not be isolated although this species seemed to be the most abundant bacterium in crude oil from the DGGE profiles. Using an archaea-specific primer set, methanogens were found in crude-oil sludge but not in crude-oil samples, indicating that methanogens might be involved in sludge formation in oil stockpiles.
通过16S rRNA基因克隆，随后进行变性梯度凝胶电泳（DGGE）分析，对来自日本石油储备的原油样品中的细菌群落进行了研究。在对从日本四个石油储备库采集的三种原油样品进行异辛烷处理后，通过PCR成功扩增出了16S rRNA基因。DGGE图谱显示，在大多数原油样品中经常检测到与嗜油 Ochrobactrum、洋葱伯克霍尔德菌、嗜麦芽窄食单胞菌、痤疮丙酸杆菌和短小鞘氨醇单胞菌相关的细菌。细菌群落在采样时间和层面上存在差异。在原油中检测到的主要细菌中，在琼脂平板上分离出的细菌只有三种，分别与伯克霍尔德菌、嗜麦芽窄食单胞菌和丙酸杆菌有关，而尽管从DGGE图谱来看Ochrobactrum sp.似乎是原油中最丰富的细菌，但却无法分离出来。使用古菌特异性引物对，在原油污泥中发现了产甲烷菌，但在原油样品中未发现，这表明产甲烷菌可能参与了石油储备库中污泥的形成。
