Plant-mediated restriction of Salmonella enterica on tomato and spinach leaves colonized with Pseudomonas plant growth-promoting rhizobacteria

- Pseudomonas S2, S4 act as plant growth-promoting rhizobacteria (PGPR) on produce.
- Rhizosphere S2 and S4 promoted biomass accumulation in spinach, lettuce and tomato.
- S2 or S4 root inoculation boosted chlorophyll content in lettuce and tomato leaves.
- Spinach and tomato colonized with S4 restricted Salmonella populations on leaves.
- PGPR-induced effects could prime plants against human enteric pathogens.

Reducing Salmonella enterica association with plants during crop production could reduce risks of fresh produce-borne salmonellosis. Plant growth-promoting rhizobacteria (PGPR) colonizing plant roots are capable of promoting plant growth and boosting resistance to disease, but the effects of PGPR on human pathogen-plant associations are not known. Two root-colonizing Pseudomonas strains S2 and S4 were investigated in spinach, lettuce and tomato for their plant growth-promoting properties and their influence on leaf populations of S. enterica serovar Newport. Plant roots were inoculated with Pseudomonas in the seedling stage. At four (tomato) and six (spinach and lettuce) weeks post-germination, plant growth promotion was assessed by shoot dry weight (SDW) and leaf chlorophyll content measurements. Leaf populations of S. Newport were measured after 24 h of leaf inoculation with this pathogen by direct plate counts on Tryptic Soy Agar. Root inoculation of spinach cv. 'Tyee', with Pseudomonas strain S2 or S4 resulted in a 69% and 63% increase in SDW compared to non-inoculated controls (p < 0.005 and p < 0.01, respectively). Similarly, Romaine lettuce cv. 'Parris Island Cos' responded positively to S2 and S4 inoculation (53% and 48% SDW increase, respectively; p < 0.05), and an increase in leaf chlorophyll content (p < 0.001), compared to controls. Tomato cv. 'Nyagous' yielded significantly greater SDW (74%, p < 0.01 and 54%, p < 0.05 for S2 and S4, respectively), and also higher leaf chlorophyll content (19% and 29%, p < 0.001, respectively) relative to controls. Leaf chlorophyll content only increased in S4-inoculated tomato cv. 'Moneymaker' plants (27%, p < 0.001), although both S2 and S4 promoted plant growth by over 40% compared to controls (p < 0.01 and p < 0.05, respectively). No significant growth promotion was detected in tomato cv. 'BHN602', but S2-inoculated plants had elevated leaf chlorophyll content (13%, p < 0.01). Root inoculation with Pseudomonas S4 restricted S. Newport populations inoculated on leaves of spinach (p < 0.001) and all three tomato cultivars (p < 0.05), compared to controls, 24 h post Salmonella inoculation. Impairment of S. Newport leaf populations was also observed on spinach when plant roots were inoculated with S2 (p < 0.01). With an initial leaf inoculum of approximately 6.0 log CFU of S. Newport/plant, the significantly greater reduction of S. Newport populations on Pseudomonas-treated plants than those on non-inoculated control plants after 24 h was modest with differences of one log or less. By contrast, the survival of S. Newport on the leaves of Romaine lettuce was not influenced by Pseudomonas root colonization. These findings provide evidence that root inoculation of certain specialty crops with beneficial Pseudomonas strains exhibiting PGPR properties may not only promote plant growth, but also reduce the fitness of epiphytic S. enterica in the phyllosphere. Plant-mediated effects induced by PGPR may be an effective strategy to minimize contamination of crops with S. enterica during cultivation.