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Loy, A.* ; Schulz, C.* ; Lücker, S.* ; Schöpfer-Wendels, A. ; Stöcker, K.* ; Lehner, A.* ; Wagner, M.*

16S rRNA gene-based oligonucleotide microarray for environmental monitoring of the betaproteobacterial order "Rhodocyclales".

Appl. Environ. Microbiol. 71, 1373-1386 (2005)
DOI
Open Access Green as soon as Postprint is submitted to ZB.
For simultaneous identification of members of the betaproteobacterial order “Rhodocyclales” in environmental samples, a 16S rRNA gene-targeted oligonucleotide microarray (RHC-PhyloChip) consisting of 79 probes was developed. Probe design was based on phylogenetic analysis of available 16S rRNA sequences from all cultured and as yet uncultured members of the “Rhodocyclales.” The multiple nested probe set was evaluated for microarray hybridization with 16S rRNA gene PCR amplicons from 29 reference organisms. Subsequently, the RHC-PhyloChip was successfully used for cultivation-independent “Rhodocyclales” diversity analysis in activated sludge from an industrial wastewater treatment plant. The implementation of a newly designed “Rhodocyclales”-selective PCR amplification system prior to microarray hybridization greatly enhanced the sensitivity of the RHC-PhyloChip and thus enabled the detection of “Rhodocyclales” populations with relative abundances of less than 1% of all bacteria (as determined by fluorescence in situ hybridization) in the activated sludge. The presence of as yet uncultured Zoogloea-, Ferribacterium/Dechloromonas-, and Sterolibacterium-related bacteria in the industrial activated sludge, as indicated by the RHC-PhyloChip analysis, was confirmed by retrieval of their 16S rRNA gene sequences and subsequent phylogenetic analysis, demonstrating the suitability of the RHC-PhyloChip as a novel monitoring tool for environmental microbiology.
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Publication type Article: Journal article
Document type Scientific Article
ISSN (print) / ISBN 0099-2240
e-ISSN 1098-5336
Journal Applied and Environmental Microbiology
Quellenangaben Volume: 71, Issue: 3, Pages: 1373-1386 Article Number: , Supplement: ,
Publisher American Society for Microbiology (ASM)
Reviewing status Peer reviewed
Institute(s) Institute of Epidemiology (EPI)