Detection of Blueberry red ringspot virus in highbush blueberry cv. ‘Coville’ in Slovenia

Blueberry red ringspot virus (BRRSV) infects blueberries and is present in USA. It causes red ringspots or red blotches on one year old stems or older. In mid- to late summer reddish-brown spots develop on older leaves. In some blueberry cultivars also fruit symptoms, circular areas of light colour and/or fruit deformations, can be seen. In spring 2008 BRRSV was detected in symptomatic bark from blueberry cv. ‘Coville’ showing typical BRRSV symptoms. The obtained PCR product was sequenced and the identity of the virus confirmed. To our knowledge this was the first finding of BRRSV in Slovenia.Keywords: BRRSV, Vaccinium, PCR, detectio

Comparison of Raspberry bushy dwarf virus isolates from Hungary and Slovenia

In 2006 and 2007 samples of grapevine and Rubus species were collected and analysed by DAS-ELISA to survey the presence of Raspberry bushy dwarf virus (RBDV) in Slovenia and Hungary. Seven varieties of raspberry from one Hungarian collection orchard were found to be infected. In Slovenia the presence of RBDV was confirmed only in three samples of wild Rubus. None of the 133 samples from different locations in Hungary proved to be infected with RBDV, although this virus is found to be widely distributed in grapevine in neighbouring Slovenia. Serological characterisation with three monoclonal antibodies (R2, R5 and D1) was performed on positive samples. Selected positive samples were partially sequenced. The results of serological and molecular analyses were compared with the analyses of raspberry and grapevine isolates obtained in Slovenia from other projects and published RBDV sequences from the GeneBank database to study the variability among hosts and locations. Isolates from grapevine grouped separately from the black raspberry isolate and all the red raspberry isolates. RBDV isolates from Hungarian samples formed a subgroup within red and black raspberry group.Keywords: RBDV, variability, Rubus, raspberry, grapevine, sequences, monoclonal antibodie

Molecular characterization of the coat protein gene of prunus necrotic ringspot virus infecting peach in Montenegro

In 2011 and 2012, two major peach-growing areas in Montenegro were surveyed for the occurrence of eleven peach viruses. Samples collected from 105 peach and nectarine trees were tested by Reverse transcription-polymerase chain reaction (RT-PCR) using sets of primers specific for the detection of Apple chlorotic leaf spot virus (ACLSV), Apple mosaic virus (ApMV), Cherry mottle leaf virus (CMLV), Prunus necrotic ringspot virus (PNRSV), Prune dwarf virus (PDV), Peach mosaic virus (PMV), Peach rosette mosaic virus (PRMV), Tobacco ringspot virus (TRSV) and Tomato ringspot virus (ToRSV). The occurrence of Strawberry latent ringspot virus (SLRSV) was examined using nested RT-PCR, while a Real-time quantitative RT-PCR method was applied for detection of Plum pox virus (PPV). PPV, PNRSV and PDV were detected in 42.9, 24.8 and 0.9 % of samples. Mixed viral infections were recorded in 6.7 % of samples, while results showed the absence of ACLSV, ApMV, PMV, CMLV, SLRSV, PRMV, TRSV and ToRSV from all assayed samples. In particular, the PCR products of complete coat protein (CP) gene was cloned and sequenced from fourteen PNRSV isolates. A high identity (87.5–100 %) was found between CP nucleotide sequences of Montenegrin isolates and other PNRSV isolates from different hosts and geographic regions retrieved from GenBank. CP nucleotide sequence comparison, as well as phylogenetic reconstruction confirmed the clustering of the isolates into three molecular groups (PV-96, PV-32 and PE-5), whereby Montenegrin isolates were separated into two phylogroups (PV-96 and PE-5). Comparative sequence and phylogenetic analysis showed that clustering of various isolates was not associated with geographic and host origin