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Detection of WWE2-related Lentisphaerae by 16S rRNA gene sequencing and fluorescence in situ hybridization in landfill leachate

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Abstract:

We collected samples of anaerobic landfill leachate from municipal solid waste landfill (Vert-le-Grand, France) and constructed 16S rRNA clone libraries using primers targeting Planctomycetes and relatives (Pla46F and 1390R). Analyses of 16S rRNA gene sequences resulted in the abundant representation of WWE2-related Lentisphaerae, members of the phylum Lentisphaerae, in the clone library (98% of the retrieved sequences). Although the sequences that are phylogenetically affiliated with the cultured isolate Victivallis vadensis were identified (WWE2 subgroup II), the majority of the sequences were affiliated with an uncultured Lentisphaerae lineage (WWE2 subgroup I). We designed oligonucleotides probes targeting the specific 16S rRNA gene regions of those 2 subgroups. Fluorescence in situ hybridization confirmed the abundance of the uncultivated WWE2 subgroup I in our leachate samples.

Nous avons récolté des échantillons anaérobies du lixiviat d’une décharge municipale de déchets solides (Vert-le-Grand, France), et nous avons construit des banques de clones d’ARNr 16S à l’aide d’amorces ciblant Planctomycetes et les bactéries apparentées (Pla46F et 1390R). Les analyses des séquences géniques de l’ARNr 16S ont révélé que les séquences du groupe WWE2 relié aux Lentisphaerae, appartenant au phylum Lentisphaerae étaient abondamment représentées dans la banque de clones (98 % des séquences retrouvées). Même si des séquences affiliées d’un point de vue phylogénique à l’isolat cultivé Victivallis ont été identifiées (sous-groupe II de WWE2), la majorité des séquences étaient affiliées à un lignage non cultivé de Lentisphaerae (sous groupe I de WWE2). Nous avons conçu des sondes d’oligonucléotides qui ciblaient des régions spécifiques du gène de l’ARNr 16S de ces 2 sous-groupes. L’hybridation in situ fluorescente a confirmé l’abondance du sous-groupe I de WWE2 non cultivé dans les échantillons de lixiviat.

Document Type: Research Article

Publication date: October 1, 2010

More about this publication?
  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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