Abstract
The potential for real-time PCR (RTm-PCR) detection of the genetically engineered strain Pseudomonas putida GN2 was studied during 2-chlorobenzoate (2-CB) degradation in three different soils. The strain contained the constructed plasmid pGN2 which encoded genes for 2-CB oxidation (cbdA) and the green fluorescent protein (gfp). P. putida GN2 numbers were assessed by plating onto 2-CB minimal media and also by RTm-PCR detection of cbdA and gfp. Addition of P. putida GN2 decreased the time required to degrade 2-CB in all tested soils by more than 7 days. The RTm-PCR estimations of P. putida GN2 numbers strongly correlated with those obtained from plate count methods during active 2-CB degradation. However, after 2-CB degradation in the soils had ceased, RTm-PCR estimations of cbdA and gfp genes were generally one order of magnitude lower than those from plate counts. These results indicate the potential for RTm-PCR to rapidly determine degrader numbers in soil following bioaugmentation but also the need to exercise caution when attempting to determine cell numbers of degraders from the RTm-PCR quantification of plasmid encoded genes after substrate is depleted.
Original language | English (US) |
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Pages (from-to) | 307-314 |
Number of pages | 8 |
Journal | FEMS Microbiology Letters |
Volume | 233 |
Issue number | 2 |
DOIs | |
State | Published - Apr 15 2004 |
Keywords
- Bioaugmentation
- Chlorobenzoate
- GFP
- Plasmid
- Real-time PCR
- cbdA
ASJC Scopus subject areas
- Microbiology
- Molecular Biology
- Genetics