Characterization of a strain of Pepino mosaic virus found in Sicily.

During winter 2005, an unusual virus-like yellow leaf disorder associated to interveinal leaf yellowing and marbling on fruits, was observed in some greenhouse tomato crops in Sicily, exactly in province of Ragusa. Two leaves from 250 symptomatic tomato plants were serologically tested by DAS-ELISA technique using the sera to Tomato spotted wilt virus (TSWV), Impatiens necrotic spot virus (INSV), Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV) and Pepino mosaic virus (PepMV). Of the 250 samples tested, 215 were positive to PepMV presence. This virus was mechanically transmitted to Cucumis sativus “Cubit”, Datura metel, Nicotiana benthamiana and tomato “Rio Grande”. The host range of PepMV-Ragusa differed from that of PepMV found infecting in 2002 tomato “Camone” in Sardinia. To amplify PepMV were utilized primers PepMV-TGB and PepMV-UTR that amplify a fragment of 844 bp containing CP gene, part of TGB gene and UTR3 (14) for PepMV PCR fragments were cloned in apCR2.1 – TOPO vector. Purified recombinant plasmids were sequenced on an ABI PRISM DNA 377 sequencer using standard M13 forward and reverse primers. To compare the PepMV-Ragusa (genbank acc. N. DQ517884) isolate, 14 nucleotide sequences were retrieved from the GenBank entries: nucleotide sequences alignments of CP-PepMV-Ragusa showed homology of 99% with both isolates US2 and Spain-Murcia. From an epidemiological point of view, aspects of the long-distance dissemination of PepMV should be considered, like the trade of living plantlets, seeds, contaminated pots in Southern Italy, occur together, indifferently in the open and greenhouse crops. Restrictive measures are required to avoid the PepMV spreading in others Italian regions