A negative regulator mediates quorum-sensing control of exopolysaccharide production in Pantoea stewartii subsp. stewartii.

Classical quorum-sensing (autoinduction) regulation, as exemplified by the lux system of Vibrio fischeri, requires N-acyl homoserine lactone (AHL) signals to stimulate cognate transcriptional activators for the cell density-dependent expression of specific target gene systems. For Pantoea stewartii subsp. stewartii, a bacterial pathogen of sweet corn and maize, the extracellular polysaccharide (EPS) stewartan is a major virulence factor, and its production is controlled by quorum sensing in a population density-dependent manner. Two genes, esaI and esaR, encode essential regulatory proteins for quorum sensing. EsaI is the AHL signal synthase, and EsaR is the cognate gene regulator. esaI, DeltaesaR, and DeltaesaI-esaR mutations were constructed to establish the regulatory role of EsaR. We report here that strains containing an esaR mutation produce high levels of EPS independently of cell density and in the absence of the AHL signal. Our data indicate that quorum-sensing regulation in P. s. subsp. stewartii, in contrast to most other described systems, uses EsaR to repress EPS synthesis at low cell density, and that derepression requires micromolar amounts of AHL. In addition, derepressed esaR strains, which synthesize EPS constitutively at low cell densities, were significantly less virulent than the wild-type parent. This finding suggests that quorum sensing in P. s. subsp. stewartii may be a mechanism to delay the expression of EPS during the early stages of infection so that it does not interfere with other mechanisms of pathogenesis.

Relationships between soil nitrate desorbed from anion-exchange membranes, canopy reflectance and nitrate leaching losses from cool-season lawn turf.

Nutrient leaching studies are expensive and require expertise in water collection and analyses. Less expensive or easier methods that estimate leaching losses would be desirable. The objective of this study was to determine if anion-exchange membranes (AEMs) and reflectance meters could predict nitrate (NO3-N) leaching losses from a cool-season lawn turf. A two-year field study used an established 90% Kentucky bluegrass (Poa pratensis L.)-10% creeping red fescue (Festuca rubra L.) turf that received 0 to 98 kg N ha-1 month-1, from May through November. Soil monolith lysimeters collected leachate that was analyzed for NO3-N concentration. Soil NO3-N was estimated with AEMs. Spectral reflectance measurements of the turf were obtained with chlorophyll and chroma meters. No significant (p > 0.05) increase in percolate flow-weighted NO3-N concentration (FWC) or mass loss occurred when AEM desorbed soil NO3-N was below 0.84 µg cm-2 d-1. A linear increase in FWC and mass loss (p < 0.0001) occurred, however, when AEM soil NO3-N was above this value. The maximum contaminant level (MCL) for drinking water (10 mg L-1 NO3-N) was reached with an AEM soil NO3-N value of 1.6 µg cm-2 d-1. Maximum meter readings were obtained when AEM soil NO3 N reached or exceeded 2.3 µg cm-2 d-1. As chlorophyll index and hue angle (greenness) increased, there was an increased probability of exceeding the NO3-N MCL. These data suggest that AEMs and reflectance meters can serve as tools to predict NO3-N leaching losses from cool-season lawn turf, and to provide objective guides for N fertilization.