Zhu Xiang Y, Lubeck John, Kilbane John J
Environmental Science and Technology Center, Gas Technology Institute, Des Plaines, Illinois 60018, USA.
Appl Environ Microbiol. 2003 Sep;69(9):5354-63. doi: 10.1128/AEM.69.9.5354-5363.2003.
Culture-independent techniques, denaturing gradient gel electrophoresis (DGGE) analysis, and random cloning of 16S rRNA gene sequences amplified from community DNA were used to determine the diversity of microbial communities in gas industry pipelines. Samples obtained from natural gas pipelines were used directly for DNA extraction, inoculated into sulfate-reducing bacterium medium, or used to inoculate a reactor that simulated a natural gas pipeline environment. The variable V2-V3 (average size, 384 bp) and V3-V6 (average size, 648 bp) regions of bacterial and archaeal 16S rRNA genes, respectively, were amplified from genomic DNA isolated from nine natural gas pipeline samples and analyzed. A total of 106 bacterial 16S rDNA sequences were derived from DGGE bands, and these formed three major clusters: beta and gamma subdivisions of Proteobacteria and gram-positive bacteria. The most frequently encountered bacterial species was Comamonas denitrificans, which was not previously reported to be associated with microbial communities found in gas pipelines or with microbially influenced corrosion. The 31 archaeal 16S rDNA sequences obtained in this study were all related to those of methanogens and phylogenetically fall into three clusters: order I, Methanobacteriales; order III, Methanomicrobiales; and order IV, Methanosarcinales: Further microbial ecology studies are needed to better understand the relationship among bacterial and archaeal groups and the involvement of these groups in the process of microbially influenced corrosion in order to develop improved ways of monitoring and controlling microbially influenced corrosion.
采用非培养技术、变性梯度凝胶电泳（DGGE）分析以及从群落DNA中扩增的16S rRNA基因序列的随机克隆，来确定天然气工业管道中微生物群落的多样性。从天然气管道获取的样本直接用于DNA提取，接种到硫酸盐还原菌培养基中，或用于接种模拟天然气管道环境的反应器。分别从9个天然气管道样本分离的基因组DNA中扩增细菌和古菌16S rRNA基因的可变V2-V3区域（平均大小384 bp）和V3-V6区域（平均大小648 bp）并进行分析。总共从DGGE条带中获得了106个细菌16S rDNA序列，这些序列形成了三个主要簇：变形菌门的β和γ亚群以及革兰氏阳性菌。最常遇到的细菌种类是反硝化丛毛单胞菌，此前未报道其与天然气管道中的微生物群落或微生物影响的腐蚀有关。本研究中获得的31个古菌16S rDNA序列均与产甲烷菌的序列相关，并且在系统发育上分为三个簇：第一类，甲烷杆菌目；第三类，甲烷微菌目；第四类，甲烷八叠球菌目。需要进一步开展微生物生态学研究，以更好地了解细菌和古菌类群之间的关系以及这些类群在微生物影响的腐蚀过程中的参与情况，从而开发出改进的监测和控制微生物影响腐蚀的方法。
