The role of rpoS on the survival of a p-nitrophenol degrading Pseudomonas putida strain in planktonic and biofilm phases

The survival of and interactions between a Pseudomonas putida strain labelled with a red fluorescent protein gene (WT-rfp) and its green fluorescent protein gene-labelled rpoS– mutant (KO-gfp) were examined. The generation times of the planktonic WT-rfp and KO-gfp in trypticase soy broth were not significantly different (i.e., p > 0.05) from each other at 30 °C. However, the biovolume of the KO-gfp biofilm was about 7 times larger than its WT-rfp counterpart after 48 h of growth. Furthermore, the presence of WT-rfp suppressed the biofilm development of KO-gfp significantly in co-culture biofilms. In planktonic conditions, the pre-carbon-starved WT-rfp achieved a 3-fold greater survival than the pre-carbon-starved KO-gfp in 0.85% saline after a 13-day incubation. In a 1:1 ratio co-culture, the pre-carbon-starved WT-rfp outcompeted the pre-carbon-starved KO-gfp by 20-fold. However, the survival of WT-rfp and KO-gfp were not significantly different from each other in biofilm conditions. Additionally, 11.4% and 61.2% of the WT-rfp and KO-gfp biofilms, respectively, remained intact after washing in 0.2% SDS for 60 min. In conclusion, the rpoS had a significant impact on survival and competitiveness of planktonic P. putida, and on biofilm development, being implicated in competitive suppression of biofilm development in co-culture biofilms and decreased biofilm cohesiveness.