Characterization of agrobacteria from weeping fig ( Ficus benjamina )

Ficus benjamina plants, galled both at epi‐ and hypogeous parts, were observed in Italy and in The Netherlands, and these were the first records of the appearance of weeping fig crown gall in Europe. A total of 241 Agrobacterium isolates was obtained from 41 tumours and studied for their morphological, physiological and phytopathological characters. Two main groups of agrobacteria were distinguished by their colony morphology and through classical biovarietal tests that allowed allocation of 86 isolates into biovar 1 and 155 into an intermediate biovar rather different from any of the three biovars defined for agrobacteria. Most of the isolates were unable to utilize mannopine, nopaline or octopine as C and N sources; only 62 strains utilized nopaline. However, when nonopine‐utilizing strains were inoculated into F. benjamina , only nopaline was detected in the developing tumours. BIOLOG ML 1™ system analysis applied to 50 representative strains allowed identification of the biovar 1 isolates as Agrobacterium tumefaciens and most of the intermediate biovar isolates as the newly proposed species Agrobacterium fici . Analysis of sensitivity to a set of 14 antibiotics confirmed the allocation of the 50 strains into two well defined main clusters matching the BIOLOG identification. Out of 141 tumorigenic isolates, 66 were sensitive in vitro to agrocine 84, but four of these strains showed scarce or no sensitivity to the antagonist A . radiobacter K84 when tested in fig plants. The two types of agrobacteria could usually be isolated from the same tumours. Tumorigenic strains were able to induce tumours in six herbaceous plant species, in eight to 10 out of 12 woody plants and in six to eight out of nine Ficus species, indicating a wide host‐range Ti plasmid. Agrobacteria were able to survive and move in the vascular system of galled ficus plants and to induce tumour growth in stem‐cutting propagated plants. Moreover, agrobacteria were detected in many healthy F. benjamina plants as part of the endophytic microflora. These findings suggest potential for spread of the disease through latently infected plant propagation material produced as cuttings or by tissue culture.