J. Zindović, M. Viršček Marn, I. Mavrič Pleško
doi: 10.4454/JPP.V97I2.024
Raspberry leaf blotch virus (RLBV, genus Emaravirus) has recently been reported to be associated with red raspberry cv. Glen Ample showing severe symptoms of raspberry leaf blotch disorder in Scotland (McGavin et al., 2012) and in several samples of raspberry in England, Serbia, Finland and Bulgaria (Bi et al., 2012; McGavin et al., 2012; Mavrič Pleško et al., 2014). In June 2011, 57 red raspberry plants were sampled in three municipalities in Montenegro to test for raspberry infecting viruses. Many samples showed virus-like symptoms, including chlorotic mottling and yellow blotches. Total RNA was extracted from all samples using RNeasy Plant Mini kit (Qiagen, Germany) and served as a template in conventional RT-PCR using primers 1571 and 1286 (McGavin et al., 2012) for P5 gene. Products of the expected size (377 bp) were obtained from 38 samples. Amplified products from two positive samples were cloned into p-GEM T-Easy Vector (Promega, USA) and sequenced in both directions using SP6/T7 primers (Macrogen, Korea). Both sequences were identical and were deposited in GenBank (accession No. KP709057). BLAST analysis of Montenegrin RLBV nucleotide sequence showed 93% nucleotide identity with three available RLBV RNA5 sequences (HG738849, HG738846 and FR823303) from Bulgaria and the UK. The deduced protein sequence was 96.1% identical to sequences from Bulgaria (CDJ26745) and showed 100% similarity to P5 sequence from the UK (CBZ42028). To our knowledge, this is the first report of RLBV occurrence in red raspberry in Montenegro. Due to the increasing importance of red raspberry production in Montenegro in the last decade, sanitary measures to prevent further virus spread and additional sanitary control of domestic and imported plant material should be adopted.