Cultivation-independent examination of horizontal transfer and host range of an IncP-1 plasmid among gram-positive and gram-negative bacteria indigenous to the barley rhizosphere.
The host range and transfer frequency of an IncP-1 plasmid (pKJK10) among indigenous bacteria in the barley rhizosphere was investigated. A new flow cytometry-based cultivation-independent method for enumeration and sorting of transconjugants for subsequent 16S rRNA gene classification was used. Indigenous transconjugant rhizosphere bacteria were collected by fluorescence-activated cell sorting and identified by cloning and sequencing of 16S rRNA genes from the sorted cells. The host range of the pKJK10 plasmid was exceptionally broad, as it included not only bacteria belonging to the alpha, beta, and gamma subclasses of the Proteobacteria, but also Arthrobacter sp., a gram-positive member of the Actinobacteria. The transfer frequency (transconjugants per donor) from the Pseudomonas putida donor to the indigenous bacteria was 7.03 x 10(-2) +/- 3.84 x 10(-2). This is the first direct documentation of conjugal transfer between gram-negative donor and gram-positive recipient bacteria in situ.
Musovic
Sanin
S
Dept. of Microbiology, University of Copenhagen, Institute of Biology, Sølvgade 83H, 1307K Copenhagen K, Denmark.
Oregaard
Gunnar
G
Kroer
Niels
N
Sørensen
Søren J
SJ
D003227:Conjugation, Genetic / Q000235:genetics*; D022761:Gene Transfer, Horizontal; D006090:Gram-Negative Bacteria / Q000235:genetics; D006094:Gram-Positive Bacteria / Q000235:genetics; D001467:Hordeum / Q000382:microbiology*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000032:analysis / Q000235:genetics; D013559:Symbiosis / Q000502:physiology; D014169:Transformation, Bacterial
D012336:RNA, Ribosomal, 16S