Evolution of the IncP-7 carbazole-degradative plasmid pCAR1 improves survival of its host Pseudomonas fluorescens Pf0-1 in artificial water microcosms

Microbiology (Reading). 2011 Aug;157(Pt 8):2276-2286. doi: 10.1099/mic.0.049064-0. Epub 2011 May 12.

Abstract

In our previous study, Pseudomonas fluorescens Pf0-1L, harbouring the IncP-7 carbazole-degradative plasmid pCAR1 : : rfp, was shown to be undetectable within 5 days post-inoculation in carbazole-contaminated artificial freshwater microcosms containing several plasmid-free bacteria in addition to Pf0-1L(pCAR1 : : rfp). Fourteen days after the inoculation, carbazole degraders become detectable. Here, we revealed that these isolates were not pCAR1 transconjugants, but Pf0-1L(pCAR1 : : rfp) mutants, based on RFLP and BOX-A1R-based repetitive extragenic palindromic-PCR (BOX-PCR) analysis. Notably, the mutants displayed more rapid initiation of carbazole degradation than the parent strain Pf0-1L(pCAR1 : : rfp). The mutants were unable to degrade anthranilate due to a 163 bp deletion in the antA gene, which was overcome by their transformation with a wild-type antABC-expressing plasmid. Quantitative RT-PCR analysis indicated that the transcriptional induction of carbazole-, anthranilate- and catechol-degradative genes was comparable in both parent and mutant strains. The deletion mutants became dominant in the artificial water microcosm. The mutation caused anthranilate to accumulate instead of catechol, a toxic compound for the parent strain, and may be beneficial to host survival in artificial microcosms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotransformation
  • Carbazoles / metabolism*
  • Carbazoles / toxicity
  • Carcinogens / metabolism*
  • Carcinogens / toxicity
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Evolution, Molecular*
  • Gene Deletion
  • Gene Expression Profiling
  • Microbial Viability / drug effects*
  • Molecular Sequence Data
  • Molecular Typing
  • Mutation
  • Plasmids*
  • Polymorphism, Restriction Fragment Length
  • Pseudomonas fluorescens / drug effects*
  • Pseudomonas fluorescens / genetics
  • Pseudomonas fluorescens / physiology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Water Microbiology
  • ortho-Aminobenzoates / metabolism

Substances

  • Carbazoles
  • Carcinogens
  • DNA, Bacterial
  • ortho-Aminobenzoates
  • carbazole
  • anthranilic acid

Associated data

  • GENBANK/AB571236
  • GENBANK/AB571237
  • GENBANK/AB571238
  • GENBANK/AB571239