Importance of biofilm formation for promoting plant growth under salt stress in Pseudomonas putida KT2440

An underutilized experimental design was employed to isolate adapted mutants of the model bacterium Pseudomonas putida KT2440. The design involved subjecting a random pool of mini-Tn5 mutants of P. putida KT2440 to multiple rounds of selection in the rhizosphere of soybean plants irrigated with a NaCl solution. The isolated adapted mutants, referred to as MutAd, exhibited a mutation in the gene responsible for encoding the membrane-binding protein LapA, which plays a role in the initial stages of biofilm formation on abiotic surfaces. Two MutAd bacteria, MutAd160 and MutAd185, along with a lapA deletion mutant, were selected for further investigation to examine the impact of this gene on salt tolerance, rhizosphere fitness, production of extracellular polymeric substances (EPS), and promotion of plant growth. Despite the mutants' inability to form biofilms, they were able to attach to soybean seeds and roots. The MutAd bacteria demonstrated an elevated production of EPS when cultivated under saline conditions, which likely compensated for the absence of biofilm formation. MutAd185 bacteria exhibited enhanced root attachment and promoted the growth of soybean plants in slightly saline soils. The proposed experimental design holds promise for expediting bacterial adaptation to the rhizosphere of plants under specific environmental conditions, identifying genetic mutations that enhance bacterial fitness in those conditions, and thereby increasing their capacity to promote plant growth.