Use of a Whole-Cell Biosensor and Flow Cytometry to Detect AHL Production by an Indigenous Soil Community During Decomposition of Litter |
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Authors: | Mette Burmølle Lars Hestbjerg Hansen Søren Johannes Sørensen |
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Institution: | 1. Department of Microbiology, University of Copenhagen, S?lvgade 83H, 1307, Copenhagen K, Denmark
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Abstract: | Quorum sensing, mediated by acylated homoserine lactones (AHLs), is well described for pure culture bacteria, but few studies
report detection of AHL compounds in natural bacterial habitats. In this study, we detect AHL production during a degradation
process in soil by use of whole-cell biosensor technology and flow cytometry analysis. An indigenous soil bacterium, belonging
to the family of Enterobacteriaceae, was isolated and transformed with a low-copy plasmid harboring a gene encoding an unstable
variant of the green fluorescent protein (gfpASV) fused to the AHL-regulated PluxI promoter originating from Vibrio fischeri. This resulted in a whole-cell biosensor, responding to the presence of AHL compounds. The biosensor was introduced to compost
soil microcosms amended with nettle leaves. After 3 days of incubation, cells were extracted and analyzed by flow cytometry.
All microcosms contained induced biosensors. From these microcosms, AHL producers were isolated and further identified as
species previously shown to produce AHLs. The results demonstrate that AHL compounds are produced during degradation of litter
in soil, indicating the presence of AHL-mediated quorum sensing in this environment. |
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