Molecular characterization of community structures and sulfur metabolism within microbial streamers in Japanese hot springs
Molecular characterization of community structures and sulfur metabolism within microbial streamers in Japanese hot springs
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DOI：
10.1128/aem.69.12.7044-7057.2003
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发表时间：
2003-12-01
影响因子：
4.4
通讯作者：
Fukui, M
中科院分区：
文献类型：
--
作者：
Nakagawa, T;Fukui, M
采用基于16S rRNA基因的分子分析方法，研究了日本Nakabusa和Yumata温度为72~80℃的硫化物温泉中沉水微生物黏液流(SMSS)的群落结构。温泉硫化物含量较低(小于0.1 mm)时，出现第一个联合体；当硫化物含量较高(大于0.1 mm)时，第二个联合体占主导地位。联合体I的细胞形态以丝状和小杆状为主。丝状细胞与领域细菌、领域古生菌和水族植物的荧光寡核苷酸探针杂交。用Eub341-F的GC钳和Univ907R和Eub341-F的GC钳和Arch915R的两套引物对变性梯度凝胶电泳(DGGE)条带的分析表明，优势带与Aquifex属的微生物系统发育有关。另一方面，联合体H以长而小的杆状细胞为主，与本研究开发的寡核苷酸探针S-*-TDES-0830a-A-20杂交，用于耐热杆菌类中大多数尚未培养的微生物。经聚合酶链式反应和逆转录-聚合酶链式反应得到的DGGE显性条带属于硫氢化钠属。此外，我们对从这两个联合体中检索到的异化亚硫酸盐还原酶(DSR)基因序列的分析表明，DSR基因与类高温杆菌的DSR基因相对应的频率很高。利用硫化物监测和(SO_4~(2-)-S-35示踪实验)，我们观察到硫化物的产生和消耗，表明在Nakabusa和Yumata温泉的SMSS中，尚未培养的类热脱硫杆菌类微生物原位产生硫化物，类硫氢菌原位消耗硫化物。
Community structures of submerged microbial slime streamers (SMSS) in sulfide-containing hot springs at 72 to 80degreesC at Nakabusa and Yumata, Japan, were investigated by molecular analysis based on the 16S rRNA gene. The SMSS were classified into two consortia; consortium I occurred at lower levels of sulfide in the hot springs (less than 0.1 mM), and consortium II dominated when the sulfide levels were higher (more than 0.1 mM). The dominant cell morphotypes in consortium I were filamentous and small rod-shaped cells. The filamentous cells hybridized with fluorescent oligonucleotide probes for the domain Bacteria, the domain Archaea, and the family Aquificaceae. Our analysis of the denaturing gradient gel electrophoresis (DGGE) bands by using reverse transcription (RT)-PCR amplification with two primer sets (Eub341-F with the GC clamp and Univ907R for the Bacteria and Eub341-F with the GC clamp and Arch915R) indicated that dominant bands were phylogenetically related to microbes in the genus Aquifex. On the other hand, consortium H was dominated by long, small, rod-shaped cells, which hybridized with the oligonucleotide probe S-*-Tdes-0830a-A-20 developed in this study for the majority of as-yet-uncultivated microbes in the class Thermodesuffibbacteria. The dominant DGGE band obtained by PCR and RT-PCR was affiliated with the genus Sulfurihydrogenibium. Moreover, our analysis of dissimilatory sulfite reductase (DSR) gene sequences retrieved from both consortia revealed a high frequency of DSR genes corresponding to the DSR of Thermodesuffibbacteria-like microorganisms. Using both sulfide monitoring and (SO42-)-S-35 tracer experiments, we observed microbial sulfide production and consumption by SMSS, suggesting that there is in situ sulfide production by as-yet-uncultivated Thermodesulfobacteria-like microbes and there is in situ sulfide consumption by Sulfurihydrogenibium-like microbes within the SMSS in the Nakabusa and Yumata hot springs.