Detection and phylogenetic analyses of fig-infecting viruses in Bosnia and Herzegovina and Montenegro

During spring 2016, a survey was carried out in Bosnian-Herzegovinian (BiH) and Montenegrin (MNE) fig orchards, germplasm collection plots and outdoor gardens, to investigate the presence of unreported fig viruses possibly present in both countries, i.e. Fig leaf mottle-associated virus 2 (FLMaV-2), Fig latent virus 1 (FLV-1), Fig cryptic virus 1 (FCV-1), Fig fleck-associated virus (FFkaV) and Fig badnavirus 1 (FBV-1); as well as those previously reported, i.e. Fig leaf mottle-associated virus 1 (FLMaV-1), Fig mild mottle-associated virus (FMMaV) and Fig mosaic emaravirus (FMV). A total of 84 fig samples (49 from BIH and 35 from MNE) were collected and tested by PCR/RT-PCR using sets of virus-specific primers. Results showed that FBV-1 was the prevailing virus with all samples (100%) infected, followed by FLMaV-1 (54%), FMV (35%), FMMaV (7%), FFkaV (6%) and FLMaV-2 (1%); whereas FLV-1 and FCV-1 were not detected. Excluding the FBV-1 detection, 35% of tested trees were infected with at least one other virus. Sequence analyses of PCR/RT-PCR fragments obtained from different viruses showed that FBV-1 was the least variable (0.9% of nucleotides divergent) compared with FLMaV-1 (15.7% sequence variation), FLMaV-2 (17.4%), FMMaV (14.9%), FMV (16.9%) and FFkaV (14.3%). Phylogenetic trees constructed with obtained sequences, together with their homologues retrieved from the Genbank database, showed distinct separation of the BiH and MNE isolates from those of different origins, in particular for FFkaV and FMV; whereas for closteroviruses (FLMaV-1, FLMaV-2 and FMMaV), there was no distinction between the isolates. This is the first report on sequence analyses of fig viruses in this geographical region, and of the presence of FBV-1 in BiH and MNE, and of FLMaV-2 and FFkaV.