Diversity, Abundance, and Activity of Archaeal Populations in Oil-Contaminated Groundwater Accumulated at the Bottom of an Underground Crude Oil Storage Cavity
Diversity, Abundance, and Activity of Archaeal Populations in Oil-Contaminated Groundwater Accumulated at the Bottom of an Underground Crude Oil Storage Cavity
复制标题
地下原油储存洞底部积聚的石油污染地下水中古菌种群的多样性、丰度和活性
DOI：
10.1128/aem.68.8.3899-3907.2002
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发表时间：
2002
影响因子：
4.4
通讯作者：
N. Kaku
中科院分区：
文献类型：
--
作者：
Kazuya Watanabe;Y. Kodama;N. Hamamura;N. Kaku
摘要荧光原位杂交显示，在地下原油储存腔底部积累的石油污染地下水中，标记有一种芽孢杆菌特异性探针（ARCH915）的细胞约占总细胞数的10%。虽然化学分析表明，洞地下水中的硝酸盐消耗旺盛，但本研究发现，甲烷的产生率高于硝酸盐的消耗率。为了表征可能的古生菌种群负责在这个系统中的甲烷生产，16S核糖体DNA（rDNA）的片段进行了扩增，使用8种不同的组合的通用和古生菌特异性引物的PCR。324个克隆的序列分析产生了23种不同的古菌序列类型，它们都隶属于广古菌界。其中，获得了5种丰富的层序类型（KuA1、KuA6、KuA12、KuA16和KuA22）。KuA1和KuA6分别与已知的产甲烷菌Methanosaeta concilii（99%相同）和Methanomethylovorans hollandica（98%）密切相关。虽然没有发现与KuA12密切相关的微生物，但它可能属于甲烷微生物科。KuA16和KuA22仅与部分环境克隆具有很高的同源性。这两个分支在广古菌中都有很深的分支，可能代表了新的目的。定量竞争PCR结果表明，KuA12的含量最高，约占总rDNA拷贝数的50%。KuA1和KuA16也构成了显着比例的总古细菌rDNA拷贝（7和17%，分别）。这些结果表明，有限的新古菌物种丰富的储油腔。比甲烷生产率的估计表明，他们是活跃的产甲烷菌。
ABSTRACT Fluorescence in situ hybridization has shown that cells labeled with an Archaea-specific probe (ARCH915) accounted for approximately 10% of the total cell count in oil-contaminated groundwater accumulated at the bottom of an underground crude oil storage cavity. Although chemical analyses have revealed vigorous consumption of nitrate in cavity groundwater, the present study found that the methane production rate was higher than the nitrate consumption rate. To characterize the likely archaeal populations responsible for methane production in this system, fragments of 16S ribosomal DNA (rDNA) were amplified by PCR using eight different combinations of universal and Archaea-specific primers. Sequence analysis of 324 clones produced 23 different archaeal sequence types, all of which were affiliated with the kingdom Euryarchaeota. Among them, five sequence types (KuA1, KuA6, KuA12, KuA16, and KuA22) were obtained in abundance. KuA1 and KuA6 were closely related to the known methanogens Methanosaeta concilii (99% identical) and Methanomethylovorans hollandica (98%), respectively. Although no closely related organism was found for KuA12, it could be affiliated with the family Methanomicrobiaceae. KuA16 and KuA22 showed substantial homology only to some environmental clones. Both of these branched deeply in the Euryarchaeota, and may represent novel orders. Quantitative competitive PCR showed that KuA12 was the most abundant, accounting for ∼50% of the total archaeal rDNA copies detected. KuA1 and KuA16 also constituted significant proportions of the total archaeal rDNA copies (7 and 17%, respectively). These results suggest that limited species of novel archaea were enriched in the oil storage cavity. An estimate of specific methane production rates suggests that they were active methanogens.
DOI：
10.1073/pnas.87.6.2264
发表时间：
1990-03-01
影响因子：
11.1
作者：
KARLIN, S;ALTSCHUL, SF
通讯作者：
ALTSCHUL, SF
影响因子：
14.9
作者：
GUTELL, RR
通讯作者：
GUTELL, RR
DOI：
10.1073/pnas.93.17.9188
发表时间：
1996-08-20
影响因子：
11.1
作者：
Barns, SM;Delwiche, CF;Pace, NR
通讯作者：
Pace, NR