Detection of XIIA phytoplasma group on cultivar Župljanka in Župa vineyard region by RFLP analysis of 16s rDNA sequences

Abstract

'Bois noir' (BN) is an important grapevine disease associated with phytoplasmas belonging to ribosomal subgroup 16SrXII-A. Phytoplasmas cause diseases in several hundred plant species. The number of infected cultivars is growing each year and it is important to follow the spreading of the phytoplasma in the different regions and identify which strains are present in specific regions on specific cultivars. Phytoplasmas are identified and classified based on direct sequencing of phytoplasma 16S rDNA or the 16S to 23S intergenic spacer region, but this approach is not always practical when a large number of unknown phytoplasmas is to be analyzed. Classification by RFLP analysis has provided a simple and rapid method that can be used to differentiate and identify a large number of unclarified phytoplasmas. Our objective was to investigate presence of phytoplasmas of 16SrXII-A group (Stolbur) in Zupa vineyard region. Detection was based on RFLP analysis of 16s rDNA sequences using four restriction enzymes: Tru1I, AluI, KpnI and TaqI. We identified phytoplasmas of XIIA group on two of three investigated cultivars (Zupljanka and Frankovka, but not on Plovdina) in the Zupa vineyard regions (Gornje Rataje and Tules locality). This is the first report of Stolbur phytoplasma on cv. Zupljanka in Zupa region.

'Bois noir' (BN) je značjna bolest vinove loze izazavana fitoplazmama 16SrXII-A grupe. Fitoplazme izazaivaju bolesti kod nekoliko stotina biljnih vrsta. Ovaj broj se povećeva svake godine, pa je veoma značajno pratiti širenje fitoplazmi u različitim regionima i utvrditi koji izazivač je prisutan u nekom regionu i na određenoj sorti. Identifikacija i klasifikacija fitoplazmi bazirana je na sekvenciranju 16S rDNA ili intergenskog regiona 16S - 23S, ali ovaj metod se retko primenjuje na analizu velikog broja uzoraka. Primena RFLP analize omogućava brzo i jednostavno detektovanje, diferenciranje i klasifikaciju velikog broja fitoplazmi. Cilj ovog rada je detektovanje fitoplazmi 16SrXII-A grupe (Stolbur) u Župskom vinogorju. Detekcija je izvršena RFLP analizom 16s rDNA sekvenci korišćenjem restrikcionih enzima: Tru1I, AluI, KpnI and TaqI. Identifikovane su fitoplazme XIIA grupe na dve od tri ispitivane sorte (na Župljanki i Frankovki, ali ne na Plovdini) u Župskom vinogorju (lokaliteti Gornje Rataje i Tuleš ). U ovim istraživanjima je prvi put dokazano prisustvo Stolbur fitoplazme na sorti Župljanka u Župskom regionu.