Genotypic Diversity among Agrobacterium tumefaciens and A. vitis Isolates From Different hosts in Some of Western and Southern Provinces of Iran using RAPD Marker

To assess the genetic diversity of Agrobacterium tumefaciens and A.vitis , during 2013 and 2014, samples of grapevine, rose and sugar beet with gall symptoms on crown and roots were collected from different areas of southern (Fars and kohgiluye and Boyerahmad) and western (Kurdistan and Kermanshah) provinces of Iran. Based on phenotypic characteristics, biochemical and pathogenicity tests as well as using the generic primers virD2A/virD2C the isolates were identified as Agrobacterium. Furthermore, species-specific primer pairs VCF/VCR for A. tumefaciens, as well as PGF/PGR for A. vitis were used to determine the identity of the causal agent.For detection of opine type among A. vitis isolates, a PCR assay was carried out using primer pairs VisF/VisR and VisFF1/ R2. Genetic diversity of A. tumefaciens and A.vitis isolates were investigated by random amplified polymorphic DNA technique using 11 random primers. The A.tumefaciens isolates clustered in three groups at 73% level of similarity and A.vitis clustered in two groups. Based on the results, A. tumefaciens and A.vitis isolates showed high genetic diversity even in a same geographical region. In conclusion, no correlation was observed between genetic diversity and geographical origins or their host plants. For detection of opine genes among A. vitis isolates, a PCR assay was carried out using primer pairs VisF/VisR and VisFF1/R2. Our results revealed that 57.9% of A. vitis isolates produced vitopine and 26.3% of them produced octopine or nopaline, whereas 15.8% of isolates were unable to amplify specific fragment.