DETECTION OF FIG MOSAIC VIRUS, FIG LEAF MOTTLE-ASSOCIATED VIRUS 1 AND FIG MILD MOTTLE-ASSOCIATED VIRUS IN BOSNIA AND HERZEGOVINA

During May 2015, a small scale survey was conducted on 27 fig trees situated in three locations (Mostar, Trebinje and Ljubuski) in Bosnia and Herzegovina, to investigate the presence of Fig mosaic virus (FMV), Fig leaf mottle-associated virus 1 (FLMaV-1), Fig leaf mottle-associated virus 2 (FLMaV- 2) and Fig mild mottle-associated virus (FMMaV). Samples consisted of leaves singularly taken from trees with symptoms of mosaic, vein yellowing, ring spots, necroses and leaf malformations (23 plants) and one asymptomatic tree, all situated in a fig germplasm collection plot. Additional three samples were taken from symptomatic fig plants of three outdoor gardens. Total RNAs extracted from the leaf midrib using RNeasy Plant Mini Kit (QIAGEN), were used in RT-PCR assays with specific primer pairs for each virus following protocols described in Elbeaino et al. (2006, 2009, 2010). RT-PCR results revealed 19 FMV- (70%), 25 FLMaV-1- (92%) and 4 FMMaV- (15%) positive samples. Double infections with FLMaV-1 and FMV were detected in 74% samples, while triple infections with FLMaV- 1, FMV and FMMaV affected 15% of tested figs. After cloning and bi-directional sequencing of 2 clones from random PCR amplicons for each virus, nucleotide BLASTn sequence analyses for FLMaV- 1 (350 bp) (GenBank Accession Nos. KU198378-KU198382), FMMaV (311 bp) (KU198388) and FMV (302 bp) (KU198367-KU198373) showed identity levels of 84-95%, 89-92% and 83-97%, respectively, with homologues available in GenBank. To our knowledge, this is the first report of three fig-infecting viruses (FLMaV-1, FMMaV and FMV) in Bosnia and Herzegovina and the results obtained, even if limited, reflect a precarious sanitary status of fig that needs a clean stock program in this country.