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Identification and characterization of Agrobacterium spp. isolated from apricot in Serbia

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Crown gall, caused by tumorigenic strains of Agrobacterium spp., is considered one of the most important diseases in stone fruit nurseries throughout the world. Since the crown gall disease has not been studied extensively in Serbia for more than 30 years, the objective of this study was to isolate, identify and characterize the bacterium associated with crown gall symptoms on one-year-old apricot trees. Samples were collected from a nursery in central Serbia and subjected to laboratory analysis. Bacteria were isolated from tumour tissue on yeast mannitol agar (YMA) and six gram-negative isolates were selected for further study. PCR assay using primers specific for virD2, virC, ipt and tms2 pathogenicity-associated plasmid genes revealed that isolates harbour Ti plasmids. All studied strains carrying Ti plasmids were classified as nopaline-type based on further molecular analysis. Using a multiplex PCR assay, targeting 23S rRNA gene sequences, and physiological and biochemical tests, five strains were assigned as A. rhizogenes/biovar 2 and the remaining one as A. tumefaciens/biovar 1. Identity of the strains was confirmed by sequence analysis of the 16S rRNA gene. In pathogenicity assay, all six strains caused tumour formation on inoculated carrot root discs, young tomato and sunflower plants.

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This research was supported by the project III46008 financed by Ministry of Education, Science and Technological Development, Republic of Serbia, COST Action 873 and by Polish Ministry of Science and Higher Education Grant 118/N-COST/2008/0

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Correspondence to Nemanja Kuzmanović.

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Nucleotide sequence data are available in GenBank database under the accession numbers: KC466565 and KC466566.

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Kuzmanović, N., Ivanović, M., Prokić, A. et al. Identification and characterization of Agrobacterium spp. isolated from apricot in Serbia. Eur J Plant Pathol 137, 11–16 (2013).

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