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Non-ribosomal Peptide Synthases from Pseudomonas aeruginosa Play a Role in Cyclodipeptide Biosynthesis, Quorum-Sensing Regulation, and Root Development in a Plant Host

  • Plant Microbe Interactions
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Abstract

Diverse molecules mediate cross-kingdom communication between bacteria and their eukaryotic partners and determine pathogenic or symbiotic relationships. N-acyl-L-homoserine lactone-dependent quorum-sensing signaling represses the biosynthesis of bacterial cyclodipeptides (CDPs) that act as auxin signal mimics in the host plant Arabidopsis thaliana. In this work, we performed bioinformatics, biochemical, and plant growth analyses to identify non-ribosomal peptide synthase (NRPS) proteins of Pseudomonas aeruginosa, which are involved in CDP synthesis. A reverse genetics strategy allowed the identification of the genes encoding putative multi-modular-NRPS (MM-NRPS). Mutations in these genes affected the synthesis of the CDPs cyclo(L-Pro-L-Val), cyclo(L-Pro-L-Leu), and cyclo(L-Pro-L-Tyr), while showing wild-type-like levels of virulence factors, such as violacein, elastase, and pyocyanin. When analyzing the bioactivity of purified, naturally produced CDPs, it was found that cyclo(L-Pro-L-Tyr) and cyclo(L-Pro-L-Val) were capable of antagonizing quorum-sensing-LasR (QS-LasR)-dependent signaling in a contrasting manner in the cell-free supernatants of the selected NRPS mutants, which showed QS induction. Using a bacteria-plant interaction system, we further show that the pvdJ, ambB, and pchE P. aeruginosa mutants failed to repress primary root growth, but improved root branching in A. thaliana seedlings. These results indicated that the CDP production in P. aeruginosa depended on the functional MM-NRPS, which influences quorum-sensing of bacteria and plays a role in root architecture remodeling.

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Acknowledgments

This study was funded by the Consejo Nacional de Ciencia y Tecnología (CONACYT) of México (grant number 256119), the Marcos Moshinsky Foundation, and Universidad Michoacana de San Nicolás de Hidalgo/C.I.C.2.14 grant. Omar Gonzalez received a scholarship from CONACYT.

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Authors and Affiliations

  1. Laboratorio de Biotecnología Microbiana, Instituto de Investigaciones Químico-Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Edif. B-3, Ciudad Universitaria, 58030, Morelia, Michoacán, México

    Omar González, Alma L. Díaz-Pérez & Jesús Campos-García

  2. Laboratorio de Biología del Desarrollo Vegetal, Instituto de Investigaciones Químico-Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Edif. A1´, Ciudad Universitaria, 58030, Morelia, Michoacán, México

    Randy Ortíz-Castro, Viridiana Magaña-Dueñas & José López-Bucio

  3. Instituto de Ecología A.C., Xalapa, Ver., México

    Randy Ortíz-Castro

  4. Depto. Ingeniería Agroindustrial, División de Ciencias de la Salud e Ingenierías, Universidad de Guanajuato, Salvatierra, Gto., México

    César Díaz-Pérez

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  1. Omar González
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Correspondence to Jesús Campos-García.

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González, O., Ortíz-Castro, R., Díaz-Pérez, C. et al. Non-ribosomal Peptide Synthases from Pseudomonas aeruginosa Play a Role in Cyclodipeptide Biosynthesis, Quorum-Sensing Regulation, and Root Development in a Plant Host. Microb Ecol 73, 616–629 (2017). https://doi.org/10.1007/s00248-016-0896-4

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