Effect of Incubation Conditions on the Enrichment of Pyrene-degrading Bacteria Identified by Stable-isotope Probing in an Aged, PAH-contaminated Soil
| Autor: | Sabrina N. Powell, Michael D. Aitken, David R. Singleton, Darryl P. Carstensen, Julie S. Swanson, Frederick K. Pfaender, Maiysha D. Jones |
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| Rok vydání: | 2007 |
| Předmět: |
DNA
Bacterial Nitrogen Molecular Sequence Data Stable-isotope probing Soil Science Soil chemistry.chemical_compound RNA Ribosomal 16S Soil Pollutants Food science Polycyclic Aromatic Hydrocarbons Incubation Phylogeny Soil Microbiology Ecology Evolution Behavior and Systematics Carbon Isotopes Pyrenes Bacteria Ecology biology Genes rRNA Sequence Analysis DNA Mineralization (soil science) Ribosomal RNA biology.organism_classification chemistry Biochemistry Pyrene Soil microbiology Temperature gradient gel electrophoresis |
| Zdroj: | Microbial Ecology. 56:341-349 |
| ISSN: | 1432-184X 0095-3628 |
| DOI: | 10.1007/s00248-007-9352-9 |
| Popis: | To determine whether the diversity of pyrene-degrading bacteria in an aged polycyclic aromatic hydrocarbon-contaminated soil is affected by the addition of inorganic nutrients or by slurrying the soil, various incubation conditions (all including phosphate buffer) were examined by mineralization studies and stable-isotope probing (SIP). The addition of nitrogen to either continuously mixed slurry or static field-wet soil incubations increased the rate and extent of mineralization of [(14)C]pyrene, with the most rapid mineralization observed in slurried, nitrogen-amended soil. Microcosms of slurry and static field-wet soil amended with nitrogen were also examined by SIP with [U-(13)C]pyrene. Recovered (13)C-enriched deoxyribonucleic acid (DNA) was analyzed by denaturing-gradient gel electrophoresis (DGGE) and 16S ribosomal ribonucleic acid (rRNA) gene clone libraries. DGGE profiles of (13)C-enriched DNA fractions from both incubation conditions were similar, suggesting that pyrene-degrading bacterial community diversity may be independent of treatment method. The vast majority (67 of 71) of the partial sequences recovered from clone libraries were greater than or equal to 97% similar to one another, 98% similar to sequences of pyrene-degrading bacteria previously detected by SIP with pyrene in different soil, and only 89% similar to the closest cultivated genus. All of the sequences recovered from the field-wet incubation and most of the sequences recovered from the slurry incubation were in this clade. Of the four sequences from slurry incubations not within this clade, three possessed greater than 99% similarity to the 16S rRNA gene sequences of phylogenetically dissimilar Caulobacter spp. |
| Databáze: | OpenAIRE |
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