Increasing survival and e⁄cacyofa bacterial biocontrol agentof ¢re blightof rosaceousplants bymeansofosmoadaptation

The efficacy of Pseudomonas fluorescens EPS62e in the biocontrol of Erwinia amylovora, the causal agent of fire blight of apple and pear, depends on the colonization of plant surfaces after spray application. A procedure to increase cell survival in the phyllosphere was developed consisting of saline stress and osmolyte amendment to the growth medium during inoculum preparation. Hyperosmotic stress induced the synthesis of the osmolytes trehalose, N-acetylglutaminylglutamine amide and glucosyl-glycerol, but decreasing growth rate. Amendment of the growth medium with glycine betaine increased growth rate and cell yield and promoted its intracellular accumulation. Under controlled environment conditions, osmoadaptation increased by 10to 100-fold cell survival to desiccation and to low relative humidity conditions on plant surfaces, in comparison with the nonosmoadapted controls. In the field, cell survival increased 100–1000 times in immature fruit upon osmoadaptation but was not significantly affected in flowers where active colonization occurs. The efficacy in the control of fire blight infections was increased 30–50% upon osmoadaptation on immature fruits but was not affected in blossoms. The method of osmoadaptation may be useful for improving the fitness and efficacy of biological control agents of phyllosphere pathogens under limiting humidity conditions.