Schmid M, Twachtmann U, Klein M, Strous M, Juretschko S, Jetten M, Metzger J W, Schleifer K H, Wagner M
Lehrstuhl für Mikrobiologie, Technische Universität München, Freising, Germany.
Syst Appl Microbiol. 2000 Apr;23(1):93-106. doi: 10.1016/S0723-2020(00)80050-8.
Recently, a bacterium capable to oxidize ammonium anaerobically at a high rate was identified as novel member of the Planctomycetales (Strous, M., Fuersi, J. A., Kramer, E. H. M., Logemann, S., Muyzer, G., van de Pas-Schoonen, K. T., Webb, R. I., Kufnen, J. G., and Jetten, M. S. M.: Nature 400, 446-449, 1999). Here we investigated the microbial community structure of a trickling filter biofilm with a high anaerobic ammonium oxidation activity. Fluorescence in situ hybridization (FISH) with a set of nine probes designed for specific identification of the recently described anaerobic ammonium oxidizer demonstrated that only one probe hybridized to bacteria within the biofilm. For phylogenetic characterization of putative biofilm anaerobic ammonium oxidizers a full-cycle 16S rDNA approach was performed by using a Planctomycetales-specific forward primer for PCR amplification. Of the twenty-five 16S rDNA fragments (1364 bp in length) amplified from the biofilm, nine were affiliated to the Planctomycetales. Comparative analysis showed that these sequences were more than 98.9% similar to each other but only distantly related to the previously recognized anaerobic ammonium oxidizer (below 91% similarity) and all other organisms represented in public 16S rRNA databases (similarities of below 79%). The retrieved sequences and the previously recognized anaerobic ammonium oxidizer represent two well-separated groups of a deep-branching lineage within the Planctomycetales. Quantitative FISH analysis with a newly designed specific probe showed that the novel bacterium, provisionally classified as "Candidatus Kuenenia stuttgartiensis" constituted the dominant fraction of the biofilm bacteria. In situ probing revealed that ammonia-oxidizing bacteria of the beta-subclass of Proteobacteria were also present, albeit in significant smaller amounts, within the anoxic biofilm. Comparative sequence analysis of a stretch of the gene encoding ammonia-monooxygenase (amoA) demonstrated the occurrence of the DNA of at least three different populations of beta-subclass ammonia oxidizers within the biofilm.
最近，一种能够在厌氧条件下高速氧化铵的细菌被鉴定为浮霉菌目（Planctomycetales）的一个新成员（斯特劳斯，M.，富尔西，J. A.，克莱默，E. H. M.，洛格曼，S.，穆伊泽尔，G.，范德帕斯 - 斯科嫩，K. T.，韦伯，R. I.，库夫嫩，J. G.，和耶滕，M. S. M.：《自然》400，446 - 至449，1999年）。在此，我们研究了具有高厌氧铵氧化活性的滴滤池生物膜的微生物群落结构。使用一组为特异性鉴定最近描述的厌氧铵氧化菌而设计的九个探针进行荧光原位杂交（FISH），结果表明只有一个探针与生物膜内的细菌杂交。为了对假定的生物膜厌氧铵氧化菌进行系统发育特征分析，通过使用浮霉菌目特异性正向引物进行PCR扩增，采用了全循环16S rDNA方法。从生物膜中扩增出的25个16S rDNA片段（长度为1364 bp）中，有9个属于浮霉菌目。比较分析表明，这些序列彼此之间的相似度超过98.9%，但与先前确认的厌氧铵氧化菌的亲缘关系较远（相似度低于91%），并且与公共16S rRNA数据库中代表的所有其他生物的相似度也低于79%。检索到的序列和先前确认的厌氧铵氧化菌代表了浮霉菌目内一个深分支谱系的两个明显不同的类群。使用新设计的特异性探针进行定量FISH分析表明，这种暂定为“斯图加特考氏菌（Candidatus Kuenenia stuttgartiensis）”的新型细菌构成了生物膜细菌的主要部分。原位探测显示，变形菌β亚类的氨氧化细菌在缺氧生物膜中也有存在，尽管数量明显较少。对编码氨单加氧酶（amoA）的一段基因进行比较序列分析表明，生物膜内至少存在三个不同群体的β亚类氨氧化菌的DNA。
