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Phasins are important proteins controlling poly-3-hydroxybutyrate (PHB) granules formation, their number into the cell and stability. The genome sequencing of the endophytic and diazotrophic bacterium  Herbaspirillum seropedicae  SmR1 revealed two homologous phasin genes. To verify the role of the phasins on PHB accumulation in the parental strain  H. seropedicae  SmR1, isogenic strains defective in the expression of  phaP1, phaP2  or both genes were obtained by gene deletion and characterized in this work. Despite of the high sequence similarity between PhaP1 and PhaP2, PhaP1 is the major phasin in  H. seropedicae , since its deletion reduced PHB accumulation by ≈50% in comparison to the parental and Δ phaP2 . Upon deletion of  phaP1 , the expression of  phaP2  was sixfold enhanced in the Δ phaP1  strain. The responsive backup expression of  phaP2  partially rescued the Δ phaP1  mutant, maintaining about 50% of the parental PHB level. The double mutant Δ phaP1.2  did not accumulate PHB in any growth stage and showed a severe reduction of growth when glucose was the carbon source, a clear demonstration of negative impact in the fitness. The co-occurrence of  phaP1  and  phaP2  homologous in bacteria relatives of  H. seropedicae , including other endophytes, indicates that the mechanism of phasin compensation by  phaP2  expression may be operating in other organisms, showing that PHB metabolism is a key factor to adaptation and efficiency of endophytic bacteria.

Backup Expression of the PhaP2 Phasin Compensates for phaP1 Deletion in Herbaspirillum seropedicae, Maintaining Fitness and PHB Accumulation