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Phenotypic characterizations demonstrated that diversity among 19 naturalized North American and 5 commercial Kura clover ( Trifolium ambiguum M.B.) rhizobial strains was limited. Growth chamber and field evaluations indicated the superiority of North American isolates, increasing foliage accumulation by 30% when compared to commercial inoculant strains. Nitrogen fertilization, however, produced greater accumulations in all evaluations. Genetic diversity among 128 isolates from the lower Caucasus was significant. Nodulation specificity of rhizobia from the lower Caucasus was demonstrated to be more complex than what was reported in the literature, as plants of different ploidy levels and even of different species were, in some cases, nodulated by the same isolate. Specificity of a given rhizobial strain varied depending on the isolate. This study identified naturalized North American rhizobial isolates that are more efficient than currently used commercial strains and increased the genetic diversity of Kura clover rhizobia currently available.