Abstract
An underutilized experimental design was used to isolate adapted mutants
of the model bacterium Pseudomonas putida KT2440. The
experimental design consisted of subjecting a random pool of
mini-Tn5 mutants of P. putida KT2440 to several rounds of
selection in the rhizosphere of soybean irrigated with NaCl solution.
Isolated adapted mutants (MutAd) showed a mutation in a gene encoding
the membrane-binding protein LapA, which is involved in the early stages
of biofilm formation on abiotic surfaces. Two MutAd bacteria (MutAd160
and MutAd185) and a lapA deletion mutant were tested to study the
effect of this gene on salt tolerance, rhizosphere fitness,
extracellular polymeric substances (EPS) production, and plant growth
promotion. The inability of the mutants to form biofilm did not hinder
attachment to soybean seeds and roots. MutAd bacteria showed an
overproduction of EPS when grown under saline conditions, which would
compensate for the lack of biofilm formation. MutAd185 bacteria showed
increased root attachment and growth promotion of soybean in slightly
saline soils. The proposed experimental design would be useful to
accelerate bacterial adaptation to the rhizosphere of plants under a
given environmental condition, identify genetic mutations that benefit
bacterial fitness in that condition, and thus increase their ability to
promote plant growth.