Mikrobiol. Z. 2019; 81(2):65-72.
Grapevine Trunk Diseases Pathogens Identification on Grapevine Rootstocks in Ukraine
Muljukina N.А.1, Pecenka J.2, Geretskij R.V.1, Eichmeier А.2
1Tairov Research Institute of Viticulture and Wine-Making, NAAS of Ukraine
27 40 let Pobedi Str., Odesa, 65496, Ukraine
2Mendel University in Brno
1/1665 Zemědělská Str., Brno, 61300, Czech Republic
The objective of this study was isolation of fungi from the grapevine trunks, DNA extraction and identification of potential esca pathogens in Ukraine. Methods. Isolation and identification of fungal cultures based on morphology and DNA sequencing. Total DNA of fungal colonies was extracted by NucleoSpin Tissue kit (Macherey-Nagel, Düren, Germany) according to manufacturer’s instructions. To amplify ITS region, ITS1 and ITS4 primers were used. The PCR products corresponding to the size of approx. 550 bp. were sequenced and the obtained nucleotide sequences were analysed using CLC Main Workbench 5.0 (CLC bio, Aarhus, Denmark). Results. Determination of potential pathogens complex showed the presence of species associated with “trunk diseases”. Among specific esca pathogens 3 isolates of Cadophora luteo-olivacea were found out. Esca-affected samples also showed the presence of Eutypa lata (causal agent of Eutypa dieback) and Botryosphaeria dothidea (causal agent of Botrioshaeria dieback). Conclusions. DNA identification of fungal pathogens showed a discrepancy in species composition between asymptomatic plants and esca-affected plants. Esca-affected cultivars showed the presence of Eutypa lata and Botryosphaeria dothidea, which were absent in asymptomatic plants.
Keywords: grapevine trunk diseases, esca, Cadophora luteo-olivacea, PCR, DNA sequencing.
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