Cultivation-independent screening revealed hot spots of IncP-1, IncP-7 and IncP-9 plasmid occurrence in different environmental habitats

Simone Dealtry, Guo-chun Ding, Viola Weichelt, Vincent Dunon, Andreas Schlüter, María Carla Martini, María Florencia Del Papa, Antonio Lagares, Gregory Charles Auton Amos, Elizabeth Margaret Helen Wellington, William Hugo Gaze, Detmer Sipkema, Sara Sjöling, Dirk Springael, Holger Heuer, Jan Dirk Van Elsas, Christopher Thomas, Kornelia Smalla

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Abstract

IncP-1, IncP-7 and IncP-9 plasmids often carry genes encoding enzymes involved in the degradation of man-made and natural contaminants, thus contributing to bacterial survival in polluted environments. However, the lack of suitable molecular tools often limits the detection of these plasmids in the environment. In this study, PCR followed by Southern blot hybridization detected the presence of plasmid-specific sequences in total community (TC-) DNA or fosmid DNA from samples originating from different environments and geographic regions. A novel primer system targeting IncP-9 plasmids was developed and applied along with established primers for IncP-1 and IncP-7. Screening TC-DNA from biopurification systems (BPS) which are used on farms for the purification of pesticide-contaminated water revealed high abundances of IncP-1 plasmids belonging to different subgroups as well as IncP-7 and IncP-9. The novel IncP-9 primer-system targeting the rep gene of nine IncP-9 subgroups allowed the detection of a high diversity of IncP-9 plasmid specific sequences in environments with different sources of pollution. Thus polluted sites are “hot spots” of plasmids potentially carrying catabolic genes.
Original languageEnglish
Article numbere89922
JournalPLoS ONE
Volume9
Issue number2
DOIs
Publication statusPublished - 24 Feb 2014

Keywords

  • Marine geology
  • Pesticides
  • Plant signaling
  • Polymerase chain reaction
  • Probe hybridization
  • Sediment
  • Sequence analysis
  • Surface water

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