Epiphytic bacteria were isolated from strawberry plants cultivated in the field or in the greenhouse in order to investigate their interaction with leaf-surface transport properties. Colonization of lower leaf sides was higher on field-grown plants, whereas upper leaf sides were more densely colonized on plants cultivated in the greenhouse. Fungal isolates significantly contributed to total microbial biomass on leaf surfaces of greenhouse-grown strawberry plants, whereas these organisms were rarely abundant on field-grown plants. Microscopic investigations of bacteria in the phyllosphere revealed that the highest densities of bacteria were observed on living trichomes, which obviously provide a source of nutrients. Isolated strains were characterized by colony morphology, microscopy and histochemistry. About 324 isolated bacterial strains were grouped into 38 morphotypes. Of the morphotypes, 12 were identified by 16S rRNA gene sequencing. Dominating bacteria belonged to the genus Pseudomonas, Stenotrophomonas, Bacillus and Arthrobacter. Cuticular water permeability of isolated cuticular membranes and intact leaf disks was measured before and after treatment with one of the most prominent epiphytic bacteria, Pseudomonas rhizosphaerae. Results showed that cuticular transpiration was significantly increased by P. rhizosphaerae. This shows that leaf-surface properties, such as cuticular water permeability, can be influenced by bacteria, leading to improved habitable conditions in the phyllosphere.