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Applied and Environmental Microbiology, September 2005, p. 5309-5317, Vol. 71, No. 9
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.9.5309-5317.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Plasmid Donor Affects Host Range of Promiscuous IncP-1ß Plasmid pB10 in an Activated-Sludge Microbial Community

Leen De Gelder,1 Frederik P. J. Vandecasteele,2,3 Celeste J. Brown,1 Larry J. Forney,1 and Eva M. Top1*

Department of Biological Sciences, Initiative for Bioinformatics and Evolutionary Studies, University of Idaho, Moscow, Idaho 83844-3051,1 Department of Biological and Agricultural Engineering, University of Idaho, Moscow, Idaho 83844-0904,2 Environmental Biotechnology Institute, University of Idaho, Moscow, Idaho 83844-10523

Received 18 February 2005/ Accepted 21 April 2005

Horizontal transfer of multiresistance plasmids in the environment contributes to the growing problem of drug-resistant pathogens. Even though the plasmid host cell is the primary environment in which the plasmid functions, possible effects of the plasmid donor on the range of bacteria to which plasmids spread in microbial communities have not been investigated. In this study we show that the host range of a broad-host-range plasmid within an activated-sludge microbial community was influenced by the donor strain and that various mating conditions and isolation strategies increased the diversity of transconjugants detected. To detect transconjugants, the plasmid pB10 was marked with lacp-rfp, while rfp expression was repressed in the donors by chromosomal lacIq. The phylogeny of 306 transconjugants obtained was determined by analysis of partial 16S rRNA gene sequences. The transconjugants belonged to 15 genera of the {alpha}- and {gamma}-Proteobacteria. The phylogenetic diversity of transconjugants obtained in separate matings with donors Pseudomonas putida SM1443, Ralstonia eutropha JMP228, and Sinorhizobium meliloti RM1021 was significantly different. For example, the transconjugants obtained after matings in sludge with S. meliloti RM1021 included eight genera that were not represented among the transconjugants obtained with the other two donors. Our results indicate that the spectrum of hosts to which a promiscuous plasmid transfers in a microbial community can be strongly influenced by the donor from which it transfers.

* Corresponding author. Mailing address: Department of Biological Sciences, 252 Life Sciences South, University of Idaho, Moscow, ID 83844-3051. Phone: (208) 885-5015. Fax: (208) 885-7905. E-mail: evatop{at}uidaho.edu.

Applied and Environmental Microbiology, September 2005, p. 5309-5317, Vol. 71, No. 9
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.9.5309-5317.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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