A collection of 31 strains received as Agrobacterium tumefaciens and A. radiobacter was subjected to detailed phenotypic and genomic studies. These strains were recovered from plants, soil, water and clinical specimens. Type strains of A. tumefaciens, A. radiobacter, A. rhizogenes and A. rubi were also included. The strains were tested for their ability to use 169 organic compounds as sources of carbon and energy. In addition, 11 conventional characters were studied for each strain. Relatedness among the strains was assessed by determining the extent of reassociation in heterologous DNA preparations. S1 nuclease and diethylaminoethyl-cellulose filters were used to separate reassociated from non-reassociated nucleotide sequences, and to determine the thermal stability of related nucleotide sequences. The resultant data revealed the following points: regardless of their phytopathogenic effects, 3-ketolactose-producing A. tumefaciens and A. radiobacter strains group into one species; this species contains 9 taxa which can be differentiated from each other by phenotypic and genomic characters; clinical isolates did not induce tumours on plants and clustered in three taxa of this species; the clinical and ecological significance of these organisms is not known; the present classification of the genus Agrobacterium is based on phytopathogenicity and does not reflect the phylogenetic relationships amongst these bacteria; as proposed previously by several workers, the genus Agrobacterium should be divided into 3 species on the basis of phenotypic and genomic characteristics. Different aspects of the classification and nomenclature of Agrobacterium are discussed.

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