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Titolo:
Analysis of broad-scale differences in microbial community composition of two pristine forest soils
Autore:
Chatzinotas, A; Sandaa, RA; Schonhuber, W; Amann, R; Daae, FL; Torsvik, V; Zeyer, J; Hahn, D;
Indirizzi:
ETH Zurich, Inst Terr Ecol, CH-8952 Schlieren, Switzerland ETH Zurich Schlieren Switzerland CH-8952 CH-8952 Schlieren, Switzerland
Titolo Testata:
SYSTEMATIC AND APPLIED MICROBIOLOGY
fascicolo: 4, volume: 21, anno: 1998,
pagine: 579 - 587
SICI:
0723-2020(199812)21:4<579:AOBDIM>2.0.ZU;2-O
Fonte:
ISI
Lingua:
ENG
Soggetto:
WHOLE-CELL HYBRIDIZATION; TARGETED OLIGONUCLEOTIDE PROBES; 16S RIBOSOMAL-RNA; BACTERIAL COMMUNITY; LABELED OLIGONUCLEOTIDES; BACILLUS-MEGATERIUM; OXIDIZING BACTERIA; DNA HYBRIDIZATION; GRASSLAND SOILS; DIVERSITY;
Keywords:
DNA reassociation; in situ hybridization; % G+C-content; probes; rRNA;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
56
Recensione:
Indirizzi per estratti:
Indirizzo: Hahn, D ETH Zurich, Inst Terr Ecol, Grabenstr 3, CH-8952 Schlieren, Switzerland ETH Zurich Grabenstr 3 Schlieren Switzerland CH-8952 Switzerland
Citazione:
A. Chatzinotas et al., "Analysis of broad-scale differences in microbial community composition of two pristine forest soils", SYST APPL M, 21(4), 1998, pp. 579-587

Abstract

Broad-scale differences in soil microbial community composition were analyzed in two contrasting soils using DNA reassociation and % G+C profiles foranalysis on the community-level, and filter- and whole cell hybridization techniques for a coarse-level characterization of larger phylogenetic groups of bacteria. Reassociation analysis of DNA from bacterial fractions extracted from the organic soil Seim and the mineral soil Hau revealed similar complexity of the communities with 5700 and 4900 different bacterial genomes(g soil [dry wt])(-1), respectively. Thermal denaturation studies showed wide % G+C distributions in DNA from bacteria of both soils. Differences in the median % G+C with 55 to 61% for the bacterial community in soil Seim and 61 to 66% for that in soil Hall indicated a higher proportion of bacteriawith a high DNA G+C content in soil Hau. In situ hybridization with fluorescent (Cy3-labeled) probes targeting larger phylogenetic groups showed minor differences between both soils, and between direct detection of bacteria in dispersed soil slurries and in bacterial fractions extracted from soils though about 90% of the total bacteria were lost during extraction. In dispersed slurries of both soils, only probes ALF1b, SRB385, and PLA46 hybridized to cells accounting for more than 1% of the DAPI-stained cells, while numbers obtained after hybridization with probes ARCH915, BET42a, GAM42a, HGC69a, and CF319a were below the detection limit set at <1%. These results were confirmed by in situ hybridization with horseradish peroxidase (HRP)-labeled probes and subsequent Cy3-tyramide signal amplification. In contrast, dot blot hybridization with probe HGC69a indicated significant amounts of Gram-positive bacteria with a high DNA G+C content in both soils. These could subsequently be visualized in non-dispersed soil slurries by in situ hybridization with HRP-labeled probe HGC69a and Cy3-tyramide signal amplification. Filamentous Gram-positive bacteria with a high DNA G+C content, likely actinomycetes, which are present in soil Hau in significant numbers are obviously destroyed by procedures used for soil dispersion.

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Documento generato il 28/09/20 alle ore 19:29:46