Resumo. Non-symbiotic diazotrophic bacteria (NSDB) influence plant growth mainly through biological nitrogenfixation and production and release of plant growth-regulating substances. The effect of these bacteria on rooting of olive is not described in the international literature, which has been provided by the use of hormonal treatment based on indole butyric acid (IBA). The purpose of this study was to evaluate the physiological diversity of isolates obtained from soil samples of the Biological Reserve Serra dos Toledos, Itajubá, MG state, and of reference strains of NSDB, and their potential of substitution of IBA on rooting of olive semi-hardwood cuttings from the experimental farm ‘Maria da Fé’ (FEMF-EPAMIG). The isolates were divided into five groups (G1 to G5) according with cultural phenotypic characteristics based on color, diameter and consistence of colonies, and reference strains used were Azospirillum brasilense (BR11001 t ), Azospirillum amazonense (BR11040 t ), Herbaspirillum seropedicae (BR11175 t ) and Burkholderia brasilensis (BR11340 t ). The diversity of physiological was performed using phosphate solubilization, tolerance to different conditions of temperature and pH and production of indole acetic acid (IAA). To evaluate the potential of semi-hardwood cuttings rooting in olive cultivars were used the cultivars Ascolano 315, Arbequina and Grappolo 541, from the germoplasm blank of the FEMF-EPAMIG. There were 33 treatments, and the cuttings were inoculated with the bacterial isolate groups, with the reference strains, without inoculation (control) and only IBA, commercially used in rooting. Only two isolates showed ability to solubilize phosphate. Groups of isolates and of reference strains showed very similar environmental requirements, with wide ranges of optimum conditions of pH (5,0-9,0) and temperature (15-35ºC) for maximum growth. The isolates and reference strains showed a production IAA ranging from 110.53 to 383.58 μg mL -1 , and the groups of isolates generally did not differ significantly from the values obtained by reference strains. All the groups of isolates present potential of rooting of olive cuttings, and those of groups 1 and 4 are more indicated to Arbequina, group 2 to Grappolo 541 and groups 1 and 5 to Ascolano 315, with performances similar to those obtained from IBA.