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bioRxiv · 10.1101/499731

Bacterial analogs of plant piperidine alkaloids mediate microbial interactions in a rhizosphere model system

Lozano, G. L.·Park, H. B.·Bravo, J. I.·Armstrong, E. A.·Denu, J. M.·Stabb, E. V.·Broderick, N. A.·Crawford, J. M.·Handelsman, J.
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Abstract

Plants expend significant resources to select and maintain rhizosphere communities that benefit their growth and protect them from pathogens. A better understanding of assembly and function of rhizosphere microbial communities will provide new avenues for improving crop production. Secretion of antibiotics is one means by which bacteria interact with neighboring microbes and sometimes change community composition. In our analysis of a taxonomically diverse consortium from the soybean rhizosphere, we found that Pseudomonas koreensis selectively inhibits growth of Flavobacterium johnsoniae and other members of the Bacteroidetes grown in soybean root exudate. A genetic screen in P. koreensis identified a previously uncharacterized biosynthetic gene cluster responsible for the inhibitory activity. The metabolites were isolated based on biological activity and were characterized using tandem-mass spectrometry, multidimensional NMR, and Mosher ester analysis, leading to the discovery of a new family of bacterial piperidine alkaloids, koreenceine A-D (1-4). Three of these metabolites are analogs of the plant alkaloid {gamma}-coniceine. Comparative analysis of the koreenceine cluster with the {gamma}-coniceine pathway revealed distinct polyketide synthase (PKS) routes to the defining piperidine scaffold, suggesting convergent evolution. Koreenceine-type pathways are widely distributed among Pseudomonas species, and koreenceine C was detected in another Pseudomonas sp. from a distantly related cluster. This work suggests that Pseudomonas and plants convergently evolved the ability to produce similar alkaloid metabolites that can mediate inter-bacterial competition in the rhizosphere.\n\nIMPORTANCEThe microbiomes of plants are critical to host physiology and development. Microbes are attracted to the rhizosphere due to massive secretion of plant photosynthates from roots. Microorganisms that successfully join the rhizosphere community from bulk soil have access to more abundant and diverse molecules, producing a highly competitive and selective environment. In the rhizosphere, as in other microbiomes, there is little known about the genetic basis for individual species behaviors within the community. In this study, we characterized competition between Pseudomonas koreensis and Flavobacterium johnsoniae, two common rhizosphere inhabitants. We identified a widespread gene cluster in several Pseudomonas spp., which is necessary for the production of a novel family of piperidine alkaloids that are structural analogs of plant alkaloids. We expand the known repertoire of antibiotics produced from Pseudomonas in the rhizosphere and demonstrate the role of the metabolites in interactions with other bacteria of the rhizosphere.

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Lozano, G. L., Park, H. B., Bravo, J. I., Armstrong, E. A., Denu, J. M., Stabb, E. V., Broderick, N. A., Crawford, J. M., Handelsman, J.. 2018-12-19. Bacterial analogs of plant piperidine alkaloids mediate microbial interactions in a rhizosphere model system. https://doi.org/10.1101/499731

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