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Applied and Environmental Microbiology, October 2006, p. 6687-6692, Vol. 72, No. 10
0099-2240/06/$08.00+0 doi:10.1128/AEM.00013-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.
University of Copenhagen, Institute of Biology, Department of Microbiology, Sølvgade 83H, 1307K Copenhagen K, Denmark,1 National Environmental Research Institute, Department of Environmental Chemistry and Microbiology, Frederiksborgvej 399, 4000 Roskilde, Denmark2
Received 3 January 2006/ Accepted 24 July 2006
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 102 ± 3.84 x 102. This is the first direct documentation of conjugal transfer between gram-negative donor and gram-positive recipient bacteria in situ.
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