Chouari Rakia, Le Paslier Denis, Daegelen Patrick, Ginestet Philippe, Weissenbach Jean, Sghir Abdelghani
CNRS UMR 8030, Genoscope and Université d'Evry Val d'Essonnes, 91057 Evry, France.
Appl Environ Microbiol. 2003 Dec;69(12):7354-63. doi: 10.1128/AEM.69.12.7354-7363.2003.
We examined anoxic and aerobic basins and an anaerobic digestor of a municipal wastewater treatment plant for the presence of novel planctomycete-like diversity. Three 16S rRNA gene libraries were constructed by using a 16S rRNA-targeted universal reverse primer and a forward PCR primer specific for Planctomyces: Phylogenetic analysis of 234 16S rRNA gene sequences defined 110 operational taxonomic units. The majority of these sequences clustered with the four known genera, Pirellula (32%), Planctomyces (18.4%), Gemmata (3.8%), and Isosphaera (0.4%). More interestingly, 42.3% of the sequences appeared to define two distantly separated monophyletic groups. The first group, represented by 35.5% of the sequences, was related to the Planctomyces group and branched as a monophyletic cluster. It exhibited between 11.9 and 20.3% 16S rRNA gene sequence dissimilarity in comparisons with cultivated planctomycetes. The second group, represented by 6.8% of the sequences, was deeply rooted within the Planctomycetales tree. It was distantly related to the anammox sequences (level of dissimilarity, 20.3 to 24.4%) and was a monophyletic cluster. The retrieved sequences extended the intralineage phylogenetic depth of the Plantomycetales from 23 to 30.6%. The lineages described here may have a broad diversity of undiscovered biochemical and metabolic novelty. We developed a new 16S rRNA-targeted oligonucleotide probe and localized members of one of the phylogenetic groups using the fluorescent in situ hybridization technique. Our results indicate that activated sludge contains very diverse representatives of this group, which grow under aerobic and anoxic conditions and even under anaerobic conditions. The majority of species in this group remain poorly characterized.
我们对一座城市污水处理厂的缺氧池、好氧池和一个厌氧消化器进行了检测，以确定是否存在新型浮霉菌样多样性。使用靶向16S rRNA的通用反向引物和针对浮霉菌属的正向PCR引物构建了三个16S rRNA基因文库：对234个16S rRNA基因序列进行系统发育分析，确定了110个操作分类单元。这些序列中的大多数与四个已知属聚类，即皮氏菌属（32%）、浮霉菌属（18.4%）、芽单胞菌属（3.8%）和等球藻属（0.4%）。更有趣的是，42.3%的序列似乎定义了两个相距甚远的单系群。第一组由35.5%的序列代表，与浮霉菌属组相关，并作为一个单系簇分支。与培养的浮霉菌相比，其16S rRNA基因序列差异在11.9%至20.3%之间。第二组由6.8%的序列代表，深深扎根于浮霉菌目树中。它与厌氧氨氧化序列关系较远（差异水平为20.3%至24.4%），是一个单系簇。检索到的序列将浮霉菌目的谱系内系统发育深度从23%扩展到30.6%。这里描述的谱系可能具有广泛的未被发现的生化和代谢新奇性。我们开发了一种新的靶向16S rRNA的寡核苷酸探针，并使用荧光原位杂交技术定位了其中一个系统发育组的成员。我们的结果表明，活性污泥含有该组非常多样的代表，它们在好氧、缺氧甚至厌氧条件下生长。该组中的大多数物种仍未得到充分表征。
