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Communication dans un congrès

Improvement of RNA-SIP by pyrosequencing to identify putative 4-n-nonylphenol degraders in activated sludge

Abstract : Nonylphenols (NP) have estrogenic potential because of their phenolic ring, but the organisms involved in the degradation of this alkylated phenol remain unidentified. Using 16S ribosomal RNA (rRNA)-based stable isotope probing (SIP) and a new method based on pyrosequencing, we identified the bacteria involved in the degradation of the aromatic ring of [U-ring-13C] 4-n-NP in aerobic sludge. The first order degradation rate of 4-n-NP was 5.5 d-1. Single strand conformation polymorphism of densityseparated labeled and unlabeled 16S rRNA showed significant differences and enabled selection of four representative fractions for pyrosequencing. Nineteen phylotypes showed a significant enrichment in the heavy fraction in the labeled pulse. The relative abundances of these phylotypes were combined with the RNA concentration of each fraction to yield a simple model of the distribution of each phylotype across the gradient. This model was used to estimate the percentage of labeling for each phylotype. The sequences showing the highest labeling (11%) were closely related to Afipia sp. but represented only 2 % of the RNA in the heavy fraction of the labeled pulse. The sequences representing the largest fraction of the RNA in the heavy fraction were related to Propionibacterium acnes and Frateuria aurantia, which are known to possess enzymes of phenol degradation. The model shows that despite Afipia having the highest 13C enrichment, species encoding the phenol degradation pathway are responsible for more 13C incorporation. Last, we showed that some species represent 12% of the total RNA but contain only 1% 13C more than the natural abundance.
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Communication dans un congrès
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https://hal.inrae.fr/hal-02809518
Déposant : Migration Prodinra <>
Soumis le : samedi 6 juin 2020 - 07:04:33
Dernière modification le : jeudi 17 septembre 2020 - 08:24:02

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  • HAL Id : hal-02809518, version 1
  • PRODINRA : 306240

Citation

Olivier Zemb, Ml Guitierrez-Zammora, M. Lee, H. Chong, Jérôme Hamelin, et al.. Improvement of RNA-SIP by pyrosequencing to identify putative 4-n-nonylphenol degraders in activated sludge. Colloque Génomique Environnementale Lyon 2011, Nov 2011, Lyon, France. ⟨hal-02809518⟩

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