Analysis of Microbial Ecological Structure of Anaerobic Granular Sludge Consortia by In-situ Hybridization of 16S rRNA targeted oligonucleotides
Analysis of Microbial Ecological Structure of Anaerobic Granular Sludge Consortia by In-situ Hybridization of 16S rRNA targeted oligonucleotides
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16S rRNA 靶向寡核苷酸原位杂交分析厌氧颗粒污泥群落的微生物生态结构
DOI：
10.11532/proes1992.34.51
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
N. Araki
中科院分区：
文献类型：
--
作者：
K. Syutsubo;Y. Sekiguchi;H. Harada;A. Ohashi;T. Tagawa;Hirokazu Oseki;N. Araki
采用荧光寡核苷酸全细胞原位杂交技术，研究了上流式厌氧污泥床（UASB）反应器中厌氧颗粒污泥的微生物群落结构。所用的分子探针与结构域的16 S rRNA的保守区域互补，所述结构域为甲烷菌属（Methanosaeta）（甲烷丝菌属（Methanothrix））和甲烷菌组，所述甲烷菌组包括甲烷杆菌属（Methanobacterium）、甲烷短杆菌属（Methanobrevibacter）、甲烷菌属（Methanosaera）的所有物种。结果表明，在相应的颗粒污泥中，DAPI（6-diamidino-2-phenylindole）染色细胞（相当于总细胞数）中约有40%至60%为嗜甲烷菌属，DAPI染色细胞中约有15%至35%为甲烷菌属。一个很强的正相关性之间的存在比例属Methanosaeta确定的FISH和乙酸裂解各自的污泥产甲烷活性。通过将颗粒分为外层和内层两部分，研究了两种类型颗粒（嗜温生长颗粒和嗜热生长颗粒）的微生物结构拓扑。内层部分比外层部分更显著地含有产甲烷菌。与中温颗粒相比，高温颗粒具有更高的甲烷化基团的倾向，甲烷化基团利用氢或甲酸进行甲烷化。这支持了我们先前的发现，即互养乙酸盐通过氢（或甲酸盐）形成氧化甲烷在嗜热条件下变得更加突出。
Whole cell in-situ hybridization with fluorescent oligonucleotides was used to study the microbial community structure of anaerobic granular sludges developed in upflow anaerobic sludge blanket (UASB) reactors receiving various types of wastewaters. The molecular probes used were complementary to conserved region of the 16S rRNA of the domain Archaea, genus Methanosaeta (Methanothrix) , and Methanobacter group which includes all species of Methanobacterium, Methanobrevibacter, Methanoshaera. The result indicated that Archaea accounted for approximately 40 to 60% of DAPI (6-diamidino-2phenylindole) stained cells (corresponding to the total cell number) in the respective granular sludges and Methanosaeta species constituted 15 to 35% of DAPI stained cells. A strong positive correlation was observed between the presence ratio of genus Methanosaeta determined by FISH and acetoclastic methanogenic activity of the respective sludges. Topology of microbial structure of two types of granules, mesophilically-grown and thermophilicallygrown granules, was investigated by dividing the individual whole-granules into the outer-layer and the innerlayer portions. The inner-layer portion contained methanogens more significantly than the outer-layer portion. Thermophilic granules had a tendency to contain higher percentage of Methanobacter group, which utilize hydrogen or formate for methanogenesis, compared with mesophilic granule. This supports our previous finding that syntrophic acetate oxidizing methanogenesis through hydrogen (or formate) formation became more prominent under thermophilic conditions.