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通过结合微放射自显影和荧光原位杂交技术确定居住在有氧-缺氧污水生物膜中的硫酸盐还原菌的系统发育鉴定和底物摄取模式。

Phylogenetic identification and substrate uptake patterns of sulfate-reducing bacteria inhabiting an oxic-anoxic sewer biofilm determined by combining microautoradiography and fluorescent in situ hybridization.

作者信息

Ito Tsukasa, Nielsen Jeppe L, Okabe Satoshi, Watanabe Yoshimasa, Nielsen Per H

机构信息

Department of Urban and Environmental Engineering, Graduate School of Engineering, Hokkaido University, 060-8628, Sapporo, Japan.

出版信息

Appl Environ Microbiol. 2002 Jan;68(1):356-64. doi: 10.1128/AEM.68.1.356-364.2002.

Abstract

We simultaneously determined the phylogenetic identification and substrate uptake patterns of sulfate-reducing bacteria (SRB) inhabiting a sewer biofilm with oxygen, nitrate, or sulfate as an electron acceptor by combining microautoradiography and fluorescent in situ hybridization (MAR-FISH) with family- and genus-specific 16S rRNA probes. The MAR-FISH analysis revealed that Desulfobulbus hybridized with probe 660 was a dominant SRB subgroup in this sewer biofilm, accounting for 23% of the total SRB. Approximately 9 and 27% of Desulfobulbus cells detected with probe 660 could take up [(14)C]propionate with oxygen and nitrate, respectively, as an electron acceptor, which might explain the high abundance of this species in various oxic environments. Furthermore, more than 40% of Desulfobulbus cells incorporated acetate under anoxic conditions. SRB were also numerically important members of H(2)-utilizing and (14)CO(2)-fixing microbial populations in this sewer biofilm, accounting for roughly 42% of total H(2)-utilizing bacteria hybridized with probe EUB338. A comparative 16S ribosomal DNA analysis revealed that two SRB populations, related to the Desulfomicrobium hypogeium and the Desulfovibrio desulfuricans MB lineages, were found to be important H(2) utilizers in this biofilm. The substrate uptake characteristics of different phylogenetic SRB subgroups were compared with the characteristics described to date. These results provide further insight into the correlation between the 16S rRNA phylogenetic diversity and the physiological diversity of SRB populations inhabiting sewer biofilms.

摘要

我们通过将微自显影和荧光原位杂交(MAR-FISH)与家族和属特异性16S rRNA探针相结合,同时确定了以氧气、硝酸盐或硫酸盐作为电子受体的污水生物膜中硫酸盐还原菌(SRB)的系统发育鉴定和底物摄取模式。MAR-FISH分析表明,与探针660杂交的脱硫球菌是该污水生物膜中占主导地位的SRB亚群,占总SRB的23%。用探针660检测到的脱硫球菌细胞中,分别约有9%和27%能够以氧气和硝酸盐作为电子受体摄取[(14)C]丙酸盐,这可能解释了该物种在各种有氧环境中的高丰度。此外,超过40%的脱硫球菌细胞在缺氧条件下摄取乙酸盐。SRB也是该污水生物膜中利用H(2)和固定(14)CO(2)的微生物种群中的重要成员,约占与探针EUB338杂交的总H(2)利用细菌的42%。一项比较16S核糖体DNA分析表明,与地下脱硫微菌和脱硫脱硫弧菌MB谱系相关的两个SRB种群是该生物膜中重要的H(2)利用者。将不同系统发育SRB亚群的底物摄取特征与迄今描述的特征进行了比较。这些结果为深入了解污水生物膜中SRB种群的16S rRNA系统发育多样性与生理多样性之间的相关性提供了进一步的见解。

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