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.
Full text (PDF, in English)
- Fontaine F, Gramaje D, Armengol J, Smart R, Nagy ZA, Morgo M, Rego C, Corio-Costet MF. Grapevine trunk diseases. A review. OIV Publications; 2016.
- Shmatkovska KA. [Grapevine esca spread on Odessa and Mykolaiv regions vineyards]. Viticulture and Wine-Making. 2010; 47:209–212. Ukrainian.
- Bertsch C, Ramírez-Suero M, Magnin-Robert M, Larignon P, Chong J, Abou-Mansour E, Spagnolo A, Clement C, Fontaine F. Grapevine trunk diseases: complex and still poorly understood. Plant Pathol. 2013; 62:243–265. https://doi.org/10.1111/j.1365-3059.2012.02674.x
- Mugnai L, Graniti A, Surico G. Esca (black measles) and brown wood-streaking: two old and elusive diseases of grapevines. Plant Disease. 1999; 83:404–418. https://doi.org/10.1094/PDIS.19220.127.116.114
- Calzarano F, Di Marco S, D'Agostino V, Schiff, S, Mugnai L. Grapevine leaf stripe disease symptoms (esca complex) are reduced by a nutrients and seaweed mixture. Phytopathologia Mediterranea. 2014; 53(3):543–558.
- Di Marco S, Osti F, Mugnai L. First studies on the potential of a copper formulation for the control of of leaf stripe disease within esca complex in grapevine. Phytopathologia Mediterranea. 2011; 50:300–309.
- Surico G. The Esca Disease Complex. In: Ciancio A, Mukerji KG, eds. Integrated Management of diseases Caused by Fungi, Phytoplasma and Bacteria. 2008;119–136. https://doi.org/10.1007/978-1-4020-8571-0_6
- Vlasov V, Konup L, Muljukina N, Geretskij R. Sanitary certification in the production of grapevine planting material biological categories: European experience and Ukrainian realities. Bulletin of Georgian Academy of Agricultural Sciences. 2018; 1(39):83–86.
- White TJ, Bruns T, Lee SB, Taylor JL. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR protocols: a guide to methods and applications. 1990; 18(1):315–322. https://doi.org/10.1016/B978-0-12-372180-8.50042-1
- Eichmeier A, Peňázová E, Pečenka J, Baránková K, Mynarzová Z. First report on molecular detection of fungal trunk pathogens in grapevine wood focusing on Moravian certified rootstock mother plants. Mitteilungen Klosterneuburg. 2016; 66:153–163.
- Eichmeier A, Baranek M, Pidra M. Analysis of genetic diversity and phylogeny of partial coat protein domain in Czech and Italian GFLV isolates. Plant Protection Science. 2010; 46:145–148. https://doi.org/10.17221/10/2010-PPS
- Díaz GA, Latorre BA. Infection caused by Phaeomoniella chlamydospora associated with esca-like symptoms in grapevine in Chile. Plant Dis. 2014; 98:351–360. https://doi.org/10.1094/PDIS-12-12-1180-RE
- Fisher M. A new wood-decay basidiomycete species associated with esca of grapevine: Fomitiporia mediterranea (Hymenochaetales). Mycological Progress. 2002; 1(3):315–324. https://doi.org/10.1007/s11557-006-0029-4
- Armengol J, Vicent A, Torne L, Garcia-Figueres F, Garcia-Jimenes J. Fungi associated with esca and grapevine declines in Spain: a three-year survey. Phytopathol. Mediterr. 2001; 40:325–329.
- Gramajje D, Armengdol J. Fungal trunk pathogens in the grapevine propagation process: potential inoculum sourses, detection, identification and management strategies. Plant Disease. 2011; 95(9):1040–1055. https://doi.org/10.1094/PDIS-01-11-0025
- Travadon R., Lawrence, D, Rooney-Latham S, Gubler W, Rolshausen PE, Baumgartner K. Cadophora species as trunk pathogens and wood-infecting fungi of grapevine in North America. Fungal Biology. 2015; 119:53–66. https://doi.org/10.1016/j.funbio.2014.11.002
- Hofstetter V. What if esca disease of grapevine were not a fungal disease? Fungal Diversity. 2012; 54(1):51–67. https://doi.org/10.1007/s13225-012-0171-z