Leloup Julie, Fossing Henrik, Kohls Katharina, Holmkvist Lars, Borowski Christian, Jørgensen Bo Barker
Laboratoire BioEmco CNRS 7618, site de l'Ecole Normale Supérieure, Paris, France.
Environ Microbiol. 2009 May;11(5):1278-91. doi: 10.1111/j.1462-2920.2008.01855.x. Epub 2009 Feb 10.
In order to better understand the main factors that influence the distribution of sulfate-reducing bacteria (SRB), their population size and their metabolic activity in high- and low-sulfate zones, we studied the SRB diversity in 3- to 5-m-deep sediment cores, which comprised the entire sulfate reduction zone and the upper methanogenic zone. By combining EMA (ethidium monoazide that can only enter damaged/dead cells and may also bind to free DNA) treatment with real-time PCR, we determined the distributions of total intact bacteria (16S rDNA genes) and intact SRB (dsrAB gene), their relative population sizes, and the proportion of dead cells or free DNA with depth. The abundance of SRB corresponded in average to 13% of the total bacterial community in the sulfate zone, 22% in the sulfate-methane transition zone and 8% in the methane zone. Compared with the total bacterial community, there were relatively less dead/damaged cells and free DNA present than among the SRB and this fraction did not change systematically with depth. By DGGE analysis, based on the amplification of the dsrA gene (400 bp), we found that the richness of SRB did not change with depth through the geochemical zones; but the clustering was related to the chemical zonation. A full-length clone library of the dsrAB gene (1900 bp) was constructed from four different depths (20, 110, 280 and 500 cm), and showed that the dsrAB genes in the near-surface sediment (20 cm) was mainly composed of sequences close to the Desulfobacteraceae, including marine complete and incomplete oxidizers such as Desulfosarcina, Desulfobacterium and Desulfococcus. The three other libraries were predominantly composed of Gram-positive SRB.
为了更好地了解影响硫酸盐还原菌(SRB)分布、其种群大小及其在高硫酸盐区和低硫酸盐区代谢活性的主要因素,我们研究了3至5米深沉积物岩心中的SRB多样性,这些岩心包括整个硫酸盐还原区和上部产甲烷区。通过将EMA(仅能进入受损/死亡细胞且也可能与游离DNA结合的单叠氮溴化乙锭)处理与实时PCR相结合,我们确定了总完整细菌(16S rDNA基因)和完整SRB(dsrAB基因)的分布、它们的相对种群大小以及死细胞或游离DNA随深度的比例。SRB的丰度平均相当于硫酸盐区总细菌群落的13%、硫酸盐-甲烷过渡区的22%和甲烷区的8%。与总细菌群落相比,SRB中存在的死/受损细胞和游离DNA相对较少,并且这一比例不会随深度系统性变化。通过基于dsrA基因(400 bp)扩增的DGGE分析,我们发现SRB的丰富度在地球化学区域中不会随深度变化;但聚类与化学分区有关。从四个不同深度(20、110、280和500厘米)构建了dsrAB基因(1900 bp)的全长克隆文库,结果表明近地表沉积物(20厘米)中的dsrAB基因主要由与脱硫杆菌科相近的序列组成,包括海洋完全和不完全氧化菌,如脱硫八叠球菌、脱硫杆菌和脱硫球菌属。其他三个文库主要由革兰氏阳性SRB组成。