Abstract
The overall goal of this study was to examine the role of quorum-sensing (QS) signals in a multispecies microbial community. Toward this aim, we studied QS signals produced by an indigenous member and an invading pathogen of the microbial community of the cabbage white butterfly (CWB) larval midgut (Pieris rapae). As an initial step, we characterized the QS system in Pantoea CWB304, which was isolated from the larval midgut. A luxI homolog, designated panI, is necessary for the production of N-acyl-L-homoserine lactones (AHLs) by Pantoea CWB304. To determine whether AHL signals are exchanged in the alkaline environment of the midgut, we constructed AHL-sensing bioluminescent reporter strains in Pantoea CWB304 and a panI mutant of this strain. In the gut of the CWB larvae, the reporter in an AHL-deficient Pantoea CWB304 detected AHLs when coinoculated with the wild type. To study the role of AHL signals produced by a community invader, we examined pathogenesis of Pseudomonas aeruginosa PAO1 in CWB larvae. Mortality induced by P. aeruginosa PAO1 was significantly reduced when signaling was interrupted by either a potent chemical inhibitor of QS or mutations in the lasI and rhlI AHL synthases of P. aeruginosa PAO1. These results show that AHLs are exchanged among bacteria in the alkaline gut of CWB larvae and contribute to disease caused by P. aeruginosa PAO1.
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Acknowledgements
We are grateful to Ronald Binder for generously providing synthetic AHLs; to Professors Leo Eberl, Michael Givskov, Max Teplitski and Barbara Iglewski for providing plasmids and strains; and to Gary Splitter and Gireesh Rajashekara for the assistance with and use of the IVIS BLI 100 CCD camera. We are also grateful to Professor Kenneth Raffa for guidance and use of his insect-rearing facilities and Anders Boyd for sharing data on the midgut pH. BRB was supported by an Advanced Opportunity Fellowship from the University of Wisconsin–Madison Graduate School. This work was supported by the Howard Hughes Medical Institute, NIH Grant no. 5RO1GM75830-2 and the University of Wisconsin–Madison College of Agricultural and Life Sciences Hatch Project no. 4038. Research in the Blackwell lab is supported in part by the NSF (CHE-0449959), Burroughs Welcome Foundation, Johnson & Johnson and Greater Milwaukee Foundation Shaw Scientist Program. HEB is an Alfred P Sloan Foundation Fellow and a Research Corporation Cottrell Scholar. GDG was supported by an American Chemical Society Division of Medicinal Chemistry predoctoral fellowship. CJR was supported by the Biotechnology Training Program (NIH 5 T32 GM08349) and an Advanced Opportunity Fellowship from the University of Wisconsin–Madison Graduate School.
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Borlee, B., Geske, G., Robinson, C. et al. Quorum-sensing signals in the microbial community of the cabbage white butterfly larval midgut. ISME J 2, 1101–1111 (2008). https://doi.org/10.1038/ismej.2008.70
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DOI: https://doi.org/10.1038/ismej.2008.70
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