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Analysis of a Pool of Small Plasmids from Soil Heterotrophic Cultivable Bacterial Communities.

Papaleo MC, Fondi M, Maida I, Perrin E, Bevivino A, Dalmastri C, Fani R - Open Microbiol J (2015)

Bottom Line: Bacterial communities were isolated from two different sites of a 13-year experimental field with a clay-silt texture.Plasmid molecules were detected at low frequency (27 isolates, 2%) with a size ranging between 2 Kb and 40 Kb.As it might be expected, even though the viable cells title did not differ significantly between the two samplings, the overall data disclosed an uneven distribution of both species and plasmid-harboring strains.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Microbial and Molecular Evolution, Department of Biology, University of Florence, Via Madonna del Piano 6, I-50019 Sesto Fiorentino (Florence), Italy.

ABSTRACT
In this work the analysis of the plasmid presence on soil aerobic cultivable heterotrophic bacterial communities was carried out checking a panel of 1,200 isolates, in order to establish the frequency of plasmid presence as well as the degree of plasmid flow between strains affiliated to the same or different taxon. Bacterial communities were isolated from two different sites of a 13-year experimental field with a clay-silt texture. Plasmid molecules were detected at low frequency (27 isolates, 2%) with a size ranging between 2 Kb and 40 Kb. The RAPD analysis performed on the plasmid-harboring isolates and the phylogenetic analysis of the whole community using the 16S rRNA gene sequences revealed the existence of transfer of the same plasmids between strains belonging to the same species and, in some cases, to different species of the same genus. As it might be expected, even though the viable cells title did not differ significantly between the two samplings, the overall data disclosed an uneven distribution of both species and plasmid-harboring strains.

No MeSH data available.


Agarose gel electrophoresis of RAPD profiles (upper) from 27 soil bacterial isolates harboring plasmids (lower). The last line of theright side was cut from another figure and pasted in Fig. (2), without changing neither the intensity nor the size of each band. Line M: (upper)DNA linear marker; (lower) reference plasmids of known size.
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Figure 2: Agarose gel electrophoresis of RAPD profiles (upper) from 27 soil bacterial isolates harboring plasmids (lower). The last line of theright side was cut from another figure and pasted in Fig. (2), without changing neither the intensity nor the size of each band. Line M: (upper)DNA linear marker; (lower) reference plasmids of known size.

Mentions: Two hundreds bacterial isolates from each of the six samples (2.2 A, 2.2 B, 2.2 C, 4.1 A, 4.1 B, and 4.1. C) were randomly selected for further characterization. The presence of plasmids was checked as described in Materials and Methods on each of the 1,200 bacterial isolates randomly selected and re-grown on 0.1 TSA medium (Tables 1 and 4). Data obtained are shown in Fig. (2) and revealed that only 27 out of the 1,200 bacterial isolates harbored plasmid molecules. Most of the strains exhibited only a single plasmid molecule, while in a few cases (i.e. isolates 308bis and 363) multiple plasmids were found in the same cell. Furthermore, some isolates showed plasmids with the same electrophoretic mobility. The size of plasmid molecules ranged between about 2 Kb and 40 Kb, as determined by comparing their eletrophoretic mobility with that of reference plasmids. However, we cannot a priori exclude the possibility that the genome of some of the bacterial isolates analyzed might contain large and/or low copy number plasmids, which might have not been revealed by the extraction methodology used in this work.


Analysis of a Pool of Small Plasmids from Soil Heterotrophic Cultivable Bacterial Communities.

Papaleo MC, Fondi M, Maida I, Perrin E, Bevivino A, Dalmastri C, Fani R - Open Microbiol J (2015)

Agarose gel electrophoresis of RAPD profiles (upper) from 27 soil bacterial isolates harboring plasmids (lower). The last line of theright side was cut from another figure and pasted in Fig. (2), without changing neither the intensity nor the size of each band. Line M: (upper)DNA linear marker; (lower) reference plasmids of known size.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4598422&req=5

Figure 2: Agarose gel electrophoresis of RAPD profiles (upper) from 27 soil bacterial isolates harboring plasmids (lower). The last line of theright side was cut from another figure and pasted in Fig. (2), without changing neither the intensity nor the size of each band. Line M: (upper)DNA linear marker; (lower) reference plasmids of known size.
Mentions: Two hundreds bacterial isolates from each of the six samples (2.2 A, 2.2 B, 2.2 C, 4.1 A, 4.1 B, and 4.1. C) were randomly selected for further characterization. The presence of plasmids was checked as described in Materials and Methods on each of the 1,200 bacterial isolates randomly selected and re-grown on 0.1 TSA medium (Tables 1 and 4). Data obtained are shown in Fig. (2) and revealed that only 27 out of the 1,200 bacterial isolates harbored plasmid molecules. Most of the strains exhibited only a single plasmid molecule, while in a few cases (i.e. isolates 308bis and 363) multiple plasmids were found in the same cell. Furthermore, some isolates showed plasmids with the same electrophoretic mobility. The size of plasmid molecules ranged between about 2 Kb and 40 Kb, as determined by comparing their eletrophoretic mobility with that of reference plasmids. However, we cannot a priori exclude the possibility that the genome of some of the bacterial isolates analyzed might contain large and/or low copy number plasmids, which might have not been revealed by the extraction methodology used in this work.

Bottom Line: Bacterial communities were isolated from two different sites of a 13-year experimental field with a clay-silt texture.Plasmid molecules were detected at low frequency (27 isolates, 2%) with a size ranging between 2 Kb and 40 Kb.As it might be expected, even though the viable cells title did not differ significantly between the two samplings, the overall data disclosed an uneven distribution of both species and plasmid-harboring strains.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Microbial and Molecular Evolution, Department of Biology, University of Florence, Via Madonna del Piano 6, I-50019 Sesto Fiorentino (Florence), Italy.

ABSTRACT
In this work the analysis of the plasmid presence on soil aerobic cultivable heterotrophic bacterial communities was carried out checking a panel of 1,200 isolates, in order to establish the frequency of plasmid presence as well as the degree of plasmid flow between strains affiliated to the same or different taxon. Bacterial communities were isolated from two different sites of a 13-year experimental field with a clay-silt texture. Plasmid molecules were detected at low frequency (27 isolates, 2%) with a size ranging between 2 Kb and 40 Kb. The RAPD analysis performed on the plasmid-harboring isolates and the phylogenetic analysis of the whole community using the 16S rRNA gene sequences revealed the existence of transfer of the same plasmids between strains belonging to the same species and, in some cases, to different species of the same genus. As it might be expected, even though the viable cells title did not differ significantly between the two samplings, the overall data disclosed an uneven distribution of both species and plasmid-harboring strains.

No MeSH data available.