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Area of Science:

  • Microbial Ecology
  • Evolutionary Biology
  • Genetics

Background:

  • Model microbial systems like Pseudomonas fluorescens are crucial for understanding ecological trait evolution.
  • Experimental evolution in structured environments drives rapid divergence in microbial populations.
  • Wrinkly spreader (WS) variants of P. fluorescens overproduce cellulose for mat formation at the air-liquid interface.

Purpose of the Study:

  • To investigate alternative evolutionary pathways for mat formation in P. fluorescens when cellulose production is disabled.
  • To identify new traits and genetic mechanisms enabling surface colonization.
  • To explore the role of diguanylate cyclases and cyclic di-GMP (c-di-GMP) networks in adaptation.

Main Methods:

  • Genetic manipulation: deletion of cellulose-encoding genes in the ancestral P. fluorescens genotype.
  • Phenotypic analysis: characterization of mat formation and cell-cell adhesion.
  • Competition experiments: assessing the fitness of evolved variants against cellulose-based WS types.
  • Genetic analysis: investigating the roles of diguanylate cyclases (WspR, AwsR, MwsR) in relation to cellulose and poly-beta-1,6-N-acetyl-d-glucosamine (PGA) production.

Main Results:

  • Deletion of cellulose genes led to the evolution of two new mat-forming strategies: one utilizing poly-beta-1,6-N-acetyl-d-glucosamine (PGA) via pgaABCD, and another involving cell chaining due to nlpD defects.
  • Both novel mat types exhibited reduced fitness compared to cellulose-based WS types.
  • Diguanylate cyclases previously linked to cellulose also regulated the evolution of PGA-based mats, suggesting conserved regulatory mechanisms.
  • Evidence suggests c-di-GMP networks are adaptable, accommodating the loss and gain of exopolysaccharide production pathways.

Conclusions:

  • Evolution can generate diverse phenotypic solutions for niche adaptation, even with genetic constraints.
  • The poly-beta-1,6-N-acetyl-d-glucosamine (PGA) pathway and cell chaining represent alternative strategies for mat formation in P. fluorescens.
  • Cyclic di-GMP (c-di-GMP) regulatory networks play a pivotal role in microbial adaptation by integrating signals for exopolysaccharide production and cell behavior.
  • These findings highlight the plasticity of microbial regulatory networks in response to environmental pressures and genetic perturbations.