title: Single-cell analyses revealed transfer ranges of IncP-1, IncP-7, and IncP-9 plasmids in a soil bacterial community.
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.
authors.affiliation: Japan Collection of Microorganisms, RIKEN BioResource Center, Tsukuba, Ibaraki, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Shintani
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Matsui
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Inoue
authors.forename: Akira
authors.initials: A
authors.lastname: Hosoyama
authors.forename: Shoko
authors.initials: S
authors.lastname: Ohji
authors.forename: Atsushi
authors.initials: A
authors.lastname: Yamazoe
authors.forename: Hideaki
authors.initials: H
authors.lastname: Nojiri
authors.forename: Kazuhide
authors.initials: K
authors.lastname: Kimbara
authors.forename: Moriya
authors.initials: M
authors.lastname: Ohkuma
mesh_terms: D001419:Bacteria / Q000145:classification* / Q000235:genetics*; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D005434:Flow Cytometry; D022761:Gene Transfer, Horizontal; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D059010:Single-Cell Analysis; D012988:Soil Microbiology
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D049452:Green Fluorescent Proteins