First Report of Broad bean wilt virus 2 and Ranunculus mild mosaic virus in Ranunculus asiaticus in Southern Italy

During 2013 to 2014, about 20% of Ranunculus asiaticus cv. Elegance plants in a commercial greenhouse in Campania, Southern Italy, were found showing leaf deformation and mosaic. Leaves from twenty-five symptomatic plants were tested by double-antibody sandwich (DAS)-ELISA using commercial diagnostic kits (Loewe, Munich, Germany) against Broad bean wilt virus 1 (BBWV-1), Broad bean wilt virus 2 (BBWV-2), Cucumber mosaic virus (CMV), Impatiens necrotic spot virus (INSV), Potato virus Y (PVY), Turnip mosaic virus (TuMV), and Tomato spotted wilt virus (TSWV). Commercial positive and negative controls (Loewe) were included in each ELISA. The same samples were also analyzed using fast lateral flow kits provided by Istituto per la Protezione Sostenibile delle Piante (IPSP), Torino, Italy, using polyclonal antibodies against Ranunculus leaf distortion virus (RanLDV-RN122), Ranunculus mild mosaic virus (RanMMV-RN129), and Ranunculus mosaic virus (RanMV-RN136). Using BLASTn, the Ranunculus BBWV-2 isolate sequence (GenBank Accession No. KJ715961) showed 94 to 97% nucleotide identity with more than 28 BBWV-2 isolates in GenBank, including a Korean isolate from pepper (JX183225) and an isolate from tomato in Xinjiang (FN985164).