Cyclic Lipopeptide Surfactant Production by Pseudomonas fluorescens SS101 Is Not Required for Suppression of Complex Pythium spp. Populations

ABSTRACT Previously, the zoosporicidal activity and control of Pythium root rot of flower bulbs by Pseudomonas fluorescens SS101 was attributed, in part, to the production of the cyclic lipopeptide surfactant massetolide A. The capacity of strain SS101 and its surfactant-deficient massA mutant 10.24 to suppress populations and root infection by complex Pythium spp. communities resident in orchard soils was assessed on apple and wheat seedlings and on apple rootstocks. Both strains initially became established in soil and persisted in the rhizosphere at similar population densities; however, massA mutant 10.24 typically was detected at higher populations in the wheat rhizosphere and soil at the end of each experiment. Both strains effectively suppressed resident Pythium populations to an equivalent level in the presence or absence of plant roots, and ultimately suppressed Pythium root infection to the same degree on all host plants. When split-root plant assays were employed, neither strain suppressed Pythium spp. infection of the component of the root system physically separated from the bacterium, suggesting that induced systemic resistance did not play a role in Pythium control. Strain SS101 only marginally suppressed in vitro growth of Pythium spp. and growth was not inhibited in the presence of mutant 10.24. When incorporated into the growth medium, the cyclic lipopeptide massetolide A significantly slowed the rate of hyphal expansion for all Pythium spp. examined. Differences in sensitivity were observed among species, with Pythium heterothallicum, P. rostratum, and P. ultimum var. ultimum exhibiting significantly greater tolerance. Pythium spp. populations indigenous to the two soils employed were composed primarily of P. irregulare, P. sylvaticum, and P. ultimum var. ultimum. These Pythium spp. either do not or rarely produce zoospores, which could account for the observation that both SS101 and mutant 10.24 were equally effective in disease control. Collectively, the results showed that (i) Pseudomonas fluorescens SS101 is very effective in controlling diverse Pythium populations on different crops grown in different soils and (ii) production of the cyclic lipopeptide massetolide A does not play a significant role in disease suppression. Other, as yet undefined mechanisms appear to play a significant role in the interaction between P. fluorescens SS101 and soilborne Pythium spp. communities.