1st International Molecular Plant Protection Congress, Adana, Turkey, 10 - 13 April 2019, pp.70
Grapevine trunk diseases associated with Botryosphaeriaceae fungi are common and important diseases in Turkey vineyards. The members of Botryosphaeriaceae family cause Botryosphaeria Dieback and the disease are described by symptoms such as wedge-shaped dark brown cankers, xylem necrosis, and lack of spring growth. Up to date 24 species of this family have been reported to cause these symptoms all over the World. These species can easily be isolated from a symptomatic vine in a laboratory but species discrimination doing based on morphological and microscopic examinations is very complicated and time consuming. Moreover, misidentification can change achievement of disease control due to different species have a different susceptibility to various control methods (hot-water treatments, protective fungicide applications etc.). For a long time, molecular tools have been using to confirm classical identification of Botryosphaeriaceae fungi and to get fast and accurate results in all studies. In this study, symptomatic vines (showing Botryosphaeria Dieback) were received to the laboratory (by extension service) from Adana, Mersin and Kahramanmaraş vineyards (18) for disease diagnosis and 21 Botryosphaeriaceous isolates were obtained in 2015-2018 years. Their identification and phylogenetic discrimination were done by molecular and phylogenetic analysis. DNA of the isolates was extracted using CTAB protocol and DNA samples were used in PCR reactions as template. ITS and translation elongation factor 1-alpha (TEF1-α) gene regions were amplified with PCR using primers ITS4/ITS5, EF728F/EF986R and EF688F/EF1251R. PCR products were subsequently sequenced and the sequences were compared with those deposited in the NCBI GenBank database using the BLASTn program. Two sets of sequences were aligned using MEGA7 software and a combined (ITS and TEF1-α) phylogenetic tree was made. After BLASTn and phylogenetic analysis 7 Botryosphaeriaceous fungi (Botryosphaeria dothidea, Diplodia seriata, Lasiodiplodia theobromae, L. pseudoteobromae, Neofusicoccum parvum, N. vitifusiforme, Neoscytalidium novaehollandiae) were identified and pathogenicity tests were performed on one-year old grapevine plants in greenhouse conditions (for two months). According to lesion occurrence, their pathogenicity was proven (measuring lesion lengths) and variance analysis was performed for statistical separation of the isolates’ virulence. This study is important for the new studies aiming at determination of Botryosphaeriaceae diversity in Turkey viticulture.