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PCR amplification of the fas-1 gene for the detection of virulent strains of Rhodococcus fascians






Plant Pathology (Oxford) 45(3): 407-417

PCR amplification of the fas-1 gene for the detection of virulent strains of Rhodococcus fascians

Thirty-six isolates of the fasciation-inducing bacterium Rhodococcus fascians were evaluated for the presence and location of the fas-1 gene, which codes for an isopentenyl transferase, the committed step in cytokinin biosynthesis. The presence of fas-1 was determined by PCR using a set of primers to the most conserved regions of the gene and by Southern hybridization to genomic digests using the PCR fragment as a probe. Both methods revealed the presence of the gene in 18 virulent isolates and the absence of the gene in 18 avirulent isolates. Thus, there is a strong relationship between the presence of the gene and virulence of the organism. The location of fas-1 was determined by probing blots of linear and circular DNA. For most of the virulent isolates, the gene was localized to a 200 +- 10 kb linear plasmid. Three virulent isolates lacked a plasmid of this size, but contained fas-1 either on a linear plasmid of 130 kb or on a large circular plasmid.

Accession: 002914391

DOI: 10.1046/j.1365-3059.1996.d01-154.x

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