Characterisation of mixed virus infections in <i>Ribes</i> species in Switzerland

Abstract

Various virus disease-like symptoms are frequently observed in Ribes sp. in Switzerland but the aetiology remains poorly documented, although a number of viruses infecting Ribes sp. were described elsewhere. Therefore, symptomatic and apparently healthy plants from diverse origins were analysed by electron microscopy (EM), immunoprecipitation electron microscopy (IPEM), Western blot and (RT-)PCR. By EM, at least four different particle types, often in combination, were observed. (1) Bacilliform particles were typical for the Badnavirus genus with dimensions of 145 x 28 nm. This virus was identified by PCR as the Gooseberry vein banding associated virus (GVBaV). (2) Filamentous particles were mainly observed on black currants with downward rolling of leaves with interveinal reddening during summer and fall. We tentatively named this unknown virus Blackcurrant leafroll-associated virus 1 (BCLRaV-1). In phylogenetic analysis of HSP70h nucleotide sequences, BCLRaV-1 felt in the Closterovirus genus. In Western blot analysis, one dominant protein with an estimated molecular weight of about 28 kDa was detectable.

The virus was shown to be different from the Raspberry mottle closterovirus (RMoV) by IPEM and RT-PCR. (3) RTPCR and sequencing of products also clearly demonstrated the presence in our Ribes samples of Rubus chlorotic mottle virus (RuCMV), a Sobemovirus recently described in Scotland. This finding correlates with the presence of the 30 nm diameter particles observed by EM. (4) A further structure with isometrical particles of 60 nm could not yet be attributed to a particular genus. Altogether, our data suggest the presence of multiple virus infections in sp. in Switzerland and emphasize the need for an efficient sanitary selection process.

Keywords: Ribes sp., Gooseberry vein banding associated virus (GVBaV), Blackcurrant leafroll-associated virus 1 (BCLRaV-1), Rubus chlorotic mottle virus (RuCMV)