RRC ID |
34061
|
著者 |
Shintani M, Matsui K, Inoue J, Hosoyama A, Ohji S, Yamazoe A, Nojiri H, Kimbara K, Ohkuma M.
|
タイトル |
Single-cell analyses revealed transfer ranges of IncP-1, IncP-7, and IncP-9 plasmids in a soil bacterial community.
|
ジャーナル |
Appl Environ Microbiol
|
Abstract |
The conjugative transfer ranges of three different plasmids of the incompatibility groups IncP-1 (pBP136), IncP-7 (pCAR1), and IncP-9 (NAH7) were investigated in soil bacterial communities by culture-dependent and culture-independent methods. Pseudomonas putida, a donor of each plasmid, was mated with soil bacteria, and green fluorescent protein (GFP), encoded on the plasmid, was used as a reporter protein for successful transfer. GFP-expressing transconjugants were detected and separated at the single-cell level by flow cytometry. Each cell was then analyzed by PCR and sequencing of its 16S rRNA gene following either whole-genome amplification or cultivation. A large number of bacteria within the phylum Proteobacteria was identified as transconjugants for pBP136 by both culture-dependent and culture-independent methods. Transconjugants belonging to the phyla Actinobacteria, Bacteroidetes, and Firmicutes were detected only by the culture-independent method. Members of the genus Pseudomonas (class Gammaproteobacteria) were identified as major transconjugants of pCAR1 and NAH7 by both methods, whereas Delftia species (class Betaproteobacteria) were detected only by the culture-independent method. The transconjugants represented a minority of the soil bacteria. Although pCAR1-containing Delftia strains could not be cultivated after a one-to-one filter mating assay between the donor and cultivable Delftia strains as recipients, fluorescence in situ hybridization detected pCAR1-containing Delftia cells, suggesting that Delftia was a "transient" host of pCAR1.
|
巻・号 |
80(1)
|
ページ |
138-45
|
公開日 |
2014-1-1
|
DOI |
10.1128/AEM.02571-13
|
PII |
AEM.02571-13
|
PMID |
24141122
|
PMC |
PMC3911017
|
MeSH |
Bacteria / classification*
Bacteria / genetics*
Conjugation, Genetic*
DNA, Bacterial / chemistry
DNA, Bacterial / genetics
DNA, Ribosomal / chemistry
DNA, Ribosomal / genetics
Flow Cytometry
Gene Transfer, Horizontal*
Genes, Reporter
Green Fluorescent Proteins / analysis
Molecular Sequence Data
Plasmids*
Polymerase Chain Reaction
RNA, Ribosomal, 16S / genetics
Sequence Analysis, DNA
Single-Cell Analysis
Soil Microbiology*
|
IF |
4.016
|
引用数 |
40
|
WOS 分野
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
MICROBIOLOGY
|
リソース情報 |
一般微生物 |
JCM 1090
JCM 1236
JCM 1237
JCM 1677
JCM 5833
JCM 6203
JCM 13063
JCM 13333
JCM 20371
JCM 20682 |