Microbial diversity in Shengli petroleum reservoirs analyzed by T-RFLP

Abstract:

Recent investigations on the microbial ecology of oil reservoirs in a variety of locales indicated that these habitats harbor various assemblages. In this study, a cultured-independent molecular technique, Terminal Restriction Fragment Length Polymorphism (T-RFLP), was used to analyze the microbial diversity of an injection well (S12-ZHU) and three related production wells (S12-4、S12-5 and S12-19) in the ShengLi oilfield (Shandong province, China). The 16S rRNA genes were amplified by PCR with the 5′carboxy-fluorescein (5-FAM)-labelled universal forward primers (27F for bacteria and 21F for archaea) and a universal reverse primer (1495R). Then the 16S rRNA genes were digested with restriction enzymes (HaeⅢ and HhaⅠ) and analyzed by using an automated DNA sequencer. The Shannon-Wiener Diversity index, based on the T-RFLP profiles, indicated that the bacterial and archaeal species richness in the injection well was higher than those of the production ones. The similarity coefficient showed the microbial community similarity among the four samples was 22.4%～30.8% (Bacteria) and 20.8%～34.5％ (Archaea), respectively. According to the analysis by TAP T-RFLP program, species belonging to Pseudomonas, Marinobacter and Methanosarcina as well as some uncultured archaeon were supposed to be the dominant bacteria in all four samples. Thus, this study indicates that T-RFLP is useful for analysis of the microbial diversity in petroleum reservoirs.