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Applied and Environmental Microbiology, September 2002, p. 4509-4516, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4509-4516.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Lipopeptide Production in Pseudomonas sp. Strain DSS73 Is Regulated by Components of Sugar Beet Seed Exudate via the Gac Two-Component Regulatory System

Birgit Koch,1 Tommy H. Nielsen,1 Dan Sørensen,2 Jens Bo Andersen,3 Carsten Christophersen,2 Søren Molin,3 Michael Givskov,3 Jan Sørensen,1 and Ole Nybroe1*

Section of Genetics and Microbiology, Department of Ecology, Royal Veterinary and Agricultural University, DK-1871 Frederiksberg C,1 Marine Chemistry Section, Department of Chemistry, University of Copenhagen, DK-2100 Copenhagen,2 Section of Molecular Microbiology, BioCentrum-DTU, Technical University of Denmark, DK-2800 Lyngby, Denmark3

Received 12 February 2002/ Accepted 20 June 2002

Pseudomonas sp. strain DSS73 isolated from the sugar beet rhizosphere produces the cyclic lipopeptide amphisin, which inhibits the growth of plant-pathogenic fungi. By Tn5::luxAB mutagenesis, we obtained two nonproducing mutant strains, DSS73-15C2 and DSS73-12H8. The gene interrupted by the transposon in strain DSS73-15C2 (amsY) encoded a protein with homology to peptide synthetases that was designated amphisin synthetase. DSS73-12H8 carried the transposon in a regulatory gene encoding a protein with homology to the sensor kinase GacS. Growth of strain DSS73-15C2 (amsY) was impaired during the transition to stationary phase in a minimal medium amended with an exudate of sugar beet seeds. This growth phenotype could be complemented by purified amphisin. Seed exudate further induced expression of bioluminescence from the amsY::luxAB reporter during the transition to stationary phase. This agreed with an increase in amphisin production by the DSS73 wild-type strain during early stationary phase. Amphisin synthesis in DSS73 was strictly dependent on GacS, and even induction by seed exudate depended on a functional gacS locus. Hence, a signal triggering the GacS/GacA two-component system appeared to be present in the seed exudate.

* Corresponding author. Mailing address: Section of Genetics and Microbiology, Department of Ecology, Royal Veterinary and Agricultural University, Thorvaldsenvej 40, DK-1871 Frederiksberg C, Denmark. Phone: 45 3538 2629. Fax: 45 3528 2606. E-mail: oln{at}kvl.dk.

Applied and Environmental Microbiology, September 2002, p. 4509-4516, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4509-4516.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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