Na Hyunsoo, Lever Mark Alexander, Kjeldsen Kasper Urup, Schulz Frederik, Jørgensen Bo Barker
Center for Geomicrobiology, Department of Bioscience, Aarhus University, Ny Munkegade 114, DK-8000, Aarhus, Denmark.
Division of Microbial Ecology, Department of Microbiology and Ecosystem Science, University of Vienna, Althanstraße 14, 1090, Vienna, Austria.
Environ Microbiol Rep. 2015 Aug;7(4):614-22. doi: 10.1111/1758-2229.12296. Epub 2015 Jun 18.
Stable isotope probing (SIP) of deoxyribonucleic acid (DNA) was used to identify microbes incorporating (13) C-labeled acetate in sulfate-reducing sediment from Aarhus Bay, Denmark. Sediment was incubated in medium containing 10 mM sulfate and different (13) C-acetate (10, 1, 0.1 mM) concentrations. The resultant changes in microbial community composition were monitored in total and SIP-fractionated DNA during long-term incubations. Chemical analyses demonstrated metabolic activity in all sediment slurries, with sulfate-reducing activity largely determined by initial acetate concentrations. Sequencing of 16S rRNA gene PCR amplicons showed that the incubations shifted the bacterial but not the archaeal community composition. After 3 months of incubation, only sediment slurries incubated with 10 mM (13) C-acetate showed detectable (13) C-DNA labeling. Based on 16S rRNA and dsrB gene PCR amplicon sequencing, the (13) C-labeled DNA pool was dominated by a single type of sulfate reducer representing a novel genus in the family Desulfobacteraceae. In addition, members of the uncultivated Crenarchaeotal group C3 were enriched in the (13) C-labeled DNA. Our results were reproducible across biological replicate experiments and provide new information about the identities of uncultured acetate-consuming bacteria and archaea in marine sediments.
利用脱氧核糖核酸(DNA)的稳定同位素探测(SIP)技术,鉴定了丹麦奥胡斯湾硫酸盐还原沉积物中摄取¹³C标记乙酸盐的微生物。沉积物在含有10 mM硫酸盐和不同浓度¹³C-乙酸盐(10、1、0.1 mM)的培养基中进行培养。在长期培养过程中,对总DNA和SIP分级分离的DNA中微生物群落组成的变化进行了监测。化学分析表明,所有沉积物悬浮液均具有代谢活性,硫酸盐还原活性主要由初始乙酸盐浓度决定。16S rRNA基因PCR扩增子测序显示,培养使细菌群落组成发生了变化,但古菌群落组成未变。培养3个月后,只有用10 mM¹³C-乙酸盐培养的沉积物悬浮液显示出可检测到的¹³C-DNA标记。基于16S rRNA和dsrB基因PCR扩增子测序,¹³C标记的DNA库主要由一种硫酸盐还原菌主导,该菌代表脱硫杆菌科中的一个新属。此外,未培养的泉古菌C3组成员在¹³C标记的DNA中富集。我们的结果在生物学重复实验中具有可重复性,并为海洋沉积物中未培养的消耗乙酸盐细菌和古菌的身份提供了新信息。