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Mutualisms are common in nature, though these symbioses can be quite permeable to cheaters in situations where one individual parasitizes the other by discontinuing cooperation yet still exploits the benefits of the partnership. In the  Rhizobium -legume system, there are two separate contexts, namely nodulation and nitrogen fixation processes, by which resident  Rhizobium  individuals can benefit by cheating. Here, we constructed reversible and irreversible mutations in key nodulation and nitrogen-fixation pathways of  Rhizobium etli  and compared their interaction with plant hosts  Phaseolus vulgaris  to that of wild type. We show that  R. etli reversible  mutants deficient in nodulation factor production are capable of  intra- specific cheating, wherein mutants exploit other  Rhizobium  individuals capable of producing these factors. Similarly, we show that  R. etli  mutants are also capable of cheating  inter- specifically, colonizing the host legume yet contributing nothing to the partnership in terms of nitrogen fixation. Our findings indicate that cheating is possible in both of these frameworks, seemingly without damaging the stability of the mutualism itself. These results may potentially help explain observations suggesting that legume plants are commonly infected by multiple bacterial lineages during the nodulation process.

Applying Reversible Mutations of Nodulation and Nitrogen-Fixation Genes to Study Social Cheating in Rhizobium etli-Legume Interaction