Kloep Frank, Manz Werner, Röske Isolde
Institute of Microbiology, Dresden University of Technology, Dresden, Germany.
FEMS Microbiol Ecol. 2006 Apr;56(1):79-94. doi: 10.1111/j.1574-6941.2006.00049.x.
The microbial communities of three different habitat types and from two sediment depths in the River Elbe were investigated by fluorescence in situ hybridization at various levels of complexity. Differences in the microbial community composition of free-flowing river water, water within the hyporheic interstitial and sediment-associated bacteria were quantitatively analyzed using domain- and group-specific oligonucleotide probes. Qualitative data on the presence/absence of specific bacterial taxa were gathered using genus- and species-specific probes. The complete data set was statistically processed by univariate statistical approaches, and two-dimensional ordinations of nonmetric multidimensional scaling. The analysis showed: (1) that the resolution of microbial community structures at microenvironments, habitats and locations can be regulated by targeted application of oligonucleotides on phylogenetic levels ranging from domains to species, and (2) that an extensive qualitative presence/absence analysis of multiparallel hybridization assays enables a fine-scale apportionment of spatial differences in microbial community structures that is robust against apparent limitations of fluorescence in situ hybridization such as false positive hybridization signals or inaccessibility of in situ oligonucleotide probes. A general model for the correlation of the phylogenetic depth of focus and the relative spatial resolution of microbial communities by fluorescence in situ hybridization is presented.
通过不同复杂程度的荧光原位杂交技术,对易北河三种不同栖息地类型以及两个沉积物深度的微生物群落进行了研究。使用针对结构域和类群的特异性寡核苷酸探针,定量分析了自由流动河水、潜流间隙水以及与沉积物相关细菌的微生物群落组成差异。使用属特异性和种特异性探针收集特定细菌类群存在与否的定性数据。完整数据集通过单变量统计方法和非度量多维尺度的二维排序进行统计处理。分析表明:(1)通过在从结构域到物种的系统发育水平上有针对性地应用寡核苷酸,可以调节微生物群落结构在微环境、栖息地和位置上的分辨率;(2)对多平行杂交试验进行广泛的定性存在/不存在分析,能够对微生物群落结构的空间差异进行精细划分,这种划分对荧光原位杂交的明显局限性(如假阳性杂交信号或原位寡核苷酸探针无法到达)具有稳健性。本文提出了一个关于荧光原位杂交聚焦的系统发育深度与微生物群落相对空间分辨率相关性的通用模型。