engleski

Detection and partial molecular characterization of Grapevine rupestris stem pitting-associated virus in Croatia

Grapevine rupestris stem pitting-associated virus (GRSPaV) is one of the most widespread grapevine viruses. It causes pits and grooves on woody cylinder of indicator plant Vitis rupestris "St. George" and it is transmitted through infected planting material and by using buds and cuttings from infected mother plants. The aim of this study was to determine occurrence of mentioned virus within Croatian autochthonous grapevine cultivars. Investigation was conducted on 12 autochthonous cultivars (Babica, Babić, Dobričić, Glavinuša, Ljutun, Maraština, Mladenka, Ninčuša, Plavac mali, Pošip, Vlaška, Vugava) in their main grape-growing areas. Totally 26 randomly selected samples in form of well wooded cuttings were taken from 26 different vineyards. Samples were tested by RT-PCR using primers 13 and 14 and by Western blot using As7-276 antibody. Virus was detected in all samples by RT-PCR and in 25 samples by Western blot. RT-PCR products from 5 different samples were sequenced in both directions and aligned with each other and with RSPaV-1 isolate as one of the fully sequenced GRSPaV genomes. It was determined that Croatian isolates shared between 87.63% and 98.99% identity on the nucleotide level and between 94.95% and 100% identity on the amino acid level. In comparison with RSPaV-1 variant, analyzed isolates showed the nucleotide-level identities ranging from 87.29% to 99.67%, while the amino acid-level identities varied from 95.96% to 100%.

autochthonous grapevine cultivars; RT-PCR; sequencing; Western blot

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