Molecular identification of Grapevine fanleaf Virus and Grapevine leafroll-associated virus-1,2,3,4,9 in vineyards in Tokat province

In this present research, during the 2018 summer season, samples were collected from fresh leaf and a one year old shoots showing virus-like symptoms from vineyards from Centrum, Erbaa, Niksar and Pazar districts of Tokat. Total 197 grapevine samples were tested molecularly and two step polymerase chain reaction (PCR) was performed with virus-specific primers to Grapevine fanleaf virus (GFLV), Grapevine leafroll-associated virus-1 (GLRaV-1), Grapevine leafroll-associated virus-2 (GLRaV-2), Grapevine leafroll-associated virus-3 (GLRaV-3), Grapevine leafroll-associated virus-4 (GLRaV-4) and Grapevine leafroll-associated virus-9 (GLRaV-9). Detection single or dual infections of these viruses have been determined in 106 (53.80%) samples. 103 (52.28%) out of 197 samples, tested resulted in positive with GLRaV-1, 10 out of 197 samples with GLRaV-3 (%5.18), 5 of samples with GLRaV-4 (%2.53), 2 of samples with GLRaV-2 (%1.03), 2 of samples with GLRaV-9 (%1.01) and 2 of samples with GFLV (%1.01). Positive PCR products of GLRaV-1 and GLRaV-3 were sequenced in two directions, aligned with MEGA10 and analysed in BLAST for multiple comparison of other isolates in Genebank. According to phlogenetic analaysis of GLRaV-1 and GLRaV-3 isolates, isolates infected with GLRaV-1 showed 94-96% of identity with others in NCBI Genebank. UO3-3 isolate infected with GLRaV-3 showed 99-100% of identity with those from Genebank