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Occurrence of Agrobacterium Vitis Carrying Two Opine-Type Plasmids in Tunisian Vineyards

Samir Chebil, Rabeb Fersi, Synda Chenenaoui, Emna Abdellatif, Giuseppe Durante, Elena Zacchi, Ali Rhouma and Ahmed Mliki

Fifty DNA samples were recovered from infected wood tissue of 1 and 2-year-old grapevine (Vitis vinifera L.) cultivars collected from several vineyards located in the Center of Tunisia. The strains were differentiated using a Multiplex PCR assay with combination of pehA, virF and virD2 gene-specific primers which amplified the corresponding fragments from nearly all strains and distinguished clearly between Agrobacterium vitis and A. tumefaciens strains carrying octopine or nopaline pTis and A. vitis vitopine strains. The isolates were segregated into three main groups, the first group carries octopine type Ti plasmids; the second vitopine type Ti plasmids and the third group both octopine and vitopine type Ti plasmids. The polygalacturonase gene sequence from 10 isolates showed 94-97% identity to the sequences of A. vitis previously deposited in NCBI GenBank database. Obtained nucleotide sequences were submitted to Genbank under accession numbers from JX946285 to JX946294.