The path to re-evolve cooperation is constrained inPseudomonas aeruginosa

ABSTRACT Background A common form of cooperation in bacteria is based on the secretion of beneficial metabolites, shareable as public good among cells at the group level. Because cooperation can be exploited by “cheat” mutants, which contribute less or nothing to the public good, there has been great interest in understanding the conditions required for cooperation to remain evolutionarily stable. In contrast, much less is known about whether cheats, once fixed in the population, are able to revert back to cooperation when conditions change. Here, we tackle this question by subjecting experimentally evolved cheats of Pseudomonas aeruginosa , partly deficient for the production of the iron-scavenging public good pyoverdine, to conditions previously shown to favor cooperation. Results Following approximately 200 generations of experimental evolution, we screened 720 evolved clones for changes in their pyoverdine production levels. We found no evidence for the re-evolution of full cooperation, even in environments with increased spatial structure, and reduced costs of cooperation – two conditions that have previously been shown to maintain cooperation. In contrast, we observed selection for complete abolishment of pyoverdine production. The patterns of complete trait degradation were likely driven by “cheating on cheats” in unstructured, iron-limited environments where pyoverdine is important for growth, and selection against a maladaptive trait in iron-rich environments where pyoverdine is superfluous. Conclusions Our study shows that the path to re-evolve cooperation seems constrained. One reason might be that the number of mutational targets potentially leading to reversion is limited. Alternatively, it could be that the selective conditions required for revertants to spread from rare are much more stringent than those needed to maintain cooperation.