Identification of the genes controlling biofilm formation in the plant commensal Pseudomonas protegens Pf-5
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ORIGINAL PAPER
Identification of the genes controlling biofilm formation in the plant commensal Pseudomonas protegens Pf‑5 Akihiro Ueda1,2 · Shinta Ogasawara2 · Keishi Horiuchi3 Received: 14 February 2020 / Revised: 20 June 2020 / Accepted: 24 June 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Determinant genes controlling biofilm formation in a plant commensal bacterium, Pseudomonas protegens Pf-5, were identified by transposon mutagenesis. Comprehensive screening of 7500 transposon-inserted mutants led to the isolation of four mutants exhibiting decreased and five mutants exhibiting increased biofilm formation. Mutations in the genes encoding MFS drug resistance transporter, LapA adhesive protein, RetS sensor histidine kinase/response regulator, and HecA adhesin/ hemagglutinin led to decreased biofilm formation, indicating that these genes are necessary for biofilm formation in Pf-5. The mutants exhibiting increased biofilm formation had transposon insertions in the genes coding for an outer membrane protein, a GGDEF domain-containing protein, AraC transcriptional regulator, non-ribosomal peptide synthetase OfaB, and the intergenic region of a DNA-binding protein and the Aer aerotaxis receptor, suggesting that these genes are negative regulators of biofilm formation. Some of these mutants also showed altered swimming and swarming motilities, and a negative correlation between biofilm formation and swarming motility was observed. Thus, sessile-motile lifestyle is regulated by divergent regulatory genes in Pf-5. Keywords Biofilm · Pseudomonas protegens Pf-5 · Swimming motility · Swarming motility
Introduction Successful bacterial colonization on the surface and the inside of plants is the first step in plant-bacterial interactions. Biofilm formation is one strategy by which bacteria attach to surfaces (Ryder et al. 2007). Biofilm-forming bacteria produce extracellular polysaccharides (EPS), proteins, and nucleic acids to establish a developed structure of biofilm matrix (Friedman and Kolter 2004b; Ueda and Wood 2010; Whitchurch et al. 2002). Bacteria living in biofilms can adhere to surfaces and increase their population. Slow penetration of antimicrobial compounds into biofilms affects Communicated by Erko Stackebrandt. * Akihiro Ueda akiueda@hiroshima‑u.ac.jp 1
Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi‑Hiroshima 739‑8528, Japan
2
Graduate School of Biosphere Science, Hiroshima University, Higashi‑Hiroshima 739‑8528, Japan
3
School of Applied Biological Science, Hiroshima University, Higashi‑Hiroshima 739‑8528, Japan
the physiological status of bacterial cells, especially antibiotic resistance (Stewart and Costerton 2001). Therefore, pathogenic bacterial infections accompanied by biofilm formation cause chronic diseases (Davies 2003). However, little is known about the influence of biofilm formation by beneficial bacteria on plants. Mechanisms of biofilm formation have been extensively studied using the model bacteria Pseudomonas
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