1887

Abstract

The ability to monitor the spatial and temporal distribution of signals in complex environments is necessary for an understanding of the function of bacteria in the wild. To this end, an existing recombinase-based transcriptional reporter strategy (recombinase-based expression technology, RIVET) has been extended and applied to the plant-colonizing bacterium SBW25. Central to the project was a rhizosphere-inducible locus, , which functional analyses show is , a histidine-inducible gene that is required for histidine utilization. A transcriptional fusion between and a promoterless site-specific recombinase ( ) results in excision of a chromosomally integrated tetracycline-resistance cassette in a histidine-dependent manner. The dose- and time-responsiveness of the promoterless recombinase to histidine closely mirrored the histidine responsiveness of an identical fusion to promoterless . To demonstrate the effectiveness of the strategy, the activity of was monitored on sugar beet seedlings. Low levels of transcriptional activity were detected in the phyllosphere, rhizosphere and in plant extract, but not in vermiculite devoid of seedlings. The histidine concentration in the rhizosphere was estimated to be 0.6 μg ml. The ecological significance of the locus was examined by competing a deletion mutant against the wild-type during colonization of sugar beet seedlings. No impact on competitive fitness was detected, suggesting that the ability to utilize plant-derived histidine is not essential for bacterial colonization.

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2006-06-01
2019-09-24
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