Qualitative and quantitative detection of agricultural microorganisms expressing iturin and mop cyclase in soils

J Agric Food Chem. 2010 Dec 22;58(24):12657-63. doi: 10.1021/jf103216g. Epub 2010 Nov 15.

Abstract

The environmental release of genetically engineered microorganisms (GEMs) to improve agriculture or remediate environmental hazards has raised concern over the fate of the organisms and their engineered genes. To detect the microorganisms released into the environment at the molecular level, Bacillus subtilis KB producing iturin and Pseudomonas fluorescens MX1 carrying the moc (mannityl opine catabolism) region from the Agrobacterium tumefaciens were employed as model microorganisms. Using specific fusion primers and the TaqMan probes, qualitative and quantitative detections of the model organisms by PCR and real-time PCR were conducted employing a small-scale soil-core device and pots during the six month period. The data indicate that the model bacteria can be easily detected by qualitative and quantitative methods in the test systems employed, and they do not give significant impacts on the other bacteria in soils on the Southern blotting analysis, although long-term observation may be needed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium tumefaciens / enzymology
  • Agrobacterium tumefaciens / genetics
  • Bacillus subtilis / genetics*
  • Bacillus subtilis / isolation & purification
  • Bacillus subtilis / metabolism
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Base Sequence
  • Genetic Engineering*
  • Molecular Sequence Data
  • Pseudomonas fluorescens / genetics*
  • Pseudomonas fluorescens / isolation & purification
  • Pseudomonas fluorescens / metabolism
  • Soil / analysis*
  • Soil Microbiology*

Substances

  • Bacterial Proteins
  • Soil