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Members of theRoseobacter lineage of marine bacteria are prolific surface colonizers in marine coastal environments, and antimicrobial secondary metabolite production has been hypothesized to provide a competitive advantage to colonizing roseobacters. Here, we report that the roseobacterPhaeobacter sp. strain Y4I produces the blue pigment indigoidine via a nonribosomal peptide synthase (NRPS)-based biosynthetic pathway encoded by a novel series of genetically linked genes:igiBCDFE . A Tn5 -based random mutagenesis library of Y4I showed a perfect correlation between indigoidine production by thePhaeobacter strain and inhibition ofVibrio fischeri on agar plates, revealing a previously unrecognized bioactivity of this molecule. In addition, igiD null mutants (igiD encoding the indigoidine NRPS) were more resistant to hydrogen peroxide, less motile, and faster to colonize an artificial surface than the wild-type strain. Collectively, these data provide evidence for pleiotropic effects of indigoidine production in this strain. Gene expression assays support phenotypic observations and demonstrate thatigiD gene expression is upregulated during growth on surfaces. Furthermore, competitive cocultures ofV. fischeri and Y4I show that the production of indigoidine by Y4I significantly inhibits colonization ofV. fischeri on surfaces. This study is the first to characterize a secondary metabolite produced by an NRPS in roseobacters.

applied and environmental microbiology

Production of the Antimicrobial Secondary Metabolite Indigoidine Contributes to Competitive Surface Colonization by the Marine Roseobacter Phaeobacter sp. Strain Y4I