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The application of the biocontrol agent Pantoea agglomerans EPS125 to unwounded fruits was practically ineffective for control of postharvest blue mould caused by Penicillium expansum when the treatment and subsequent wounding and pathogen inoculation were separated by periods of unfavourable conditions. This was due to a rapid decrease in viability of the alocthonous introduced biocontrol agent in the intact peel surface. A system for osmoadaptation of the biocontrol agent was developed by combining saline osmotic stress and osmolyte amendment to the growth medium. Osmoadapted cells accumulated trehalose and glycine betaine (GB) intracellularly and showed a higher tolerance to desiccation than non-osmoadapted cells. Osmoadaptation in NaCl plus GB during inoculum preparation increased considerably survival on the peel surface of apple fruits. This effect was significant under low relative humidity (RH) and fluctuating RH conditions, but was not significant at high RH. Osmoadaptation significantly improved blue mould control under conditions where the standard biological control treatments were ineffective. The rot diameter was significantly reduced in apple fruits which were treated with EPS125 and incubated for several days under low, high or fluctuating RH, followed by wounding and inoculation of P. expansum. Growth of EPS125 with NaCl, either with or without the addition of GB, was an effective osmoadaptation treatment for improving blue mould rot control. However, the addition of GB to the NaCl amended growth medium increased 4-5-fold growth rate and OD of the cultures. This is an advantage for mass production of P. agglomerans EPS125 in a NaCl amended growth medium.

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 10- to 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.

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