Tokat ili Bağlarda Grapevine leafroll-associated virus -1,2,3,4'ün yaygınlığı ve Genetik Değişkenliği

Öz

Asma yaprak kıvırcıklığı hastalığı (GLRD), dünya çapında asmanın en önemli viral hastalıklarından biridir. Bu çalışma, 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), ve Grapevine leafroll-associated virus-9 (GLRaV-9) etmenlerinin yaygınlığının araştırılması amacıyla yapılmıştır. Bunun için Tokat ili Merkez, Erbaa, Niksar ve Pazar ilçelerindeki farklı bağ alanlarından virüs benzeri belirti gösteren asmalardan yaprak örnekleri ve bir yıllık bitki sürgünleri toplanmıştır. Surveyde elde edilen toplam 197 örnek GLRaV-1, GLRaV-2, GLRaV-3, GLRaV-4) ve GLRaV-9 etmenlerine özgü primerler ile iki aşamalı polimeraz zincir reaksiyonu (PCR) kullanılarak moleküler olarak test edilmiştir. 197 örnekten 106'sı (%53.80) bu virüslerle tekli veya çoklu enfeksiyon tespit edilmiştir. 103 (%52,28) örnek GLRaV-1 ile pozitif sonuçlanırken, enfeksiyon oranları 10 örnekle GLRaV-3 için %5,18, GLRaV-4 için 5 örnekle %2,53, GLRaV-2 ve GLRaV-9 için %1,03 olarak belirlenmiştir. GLRaV-1, GLRaV-2 ve GLRaV-3'ün pozitif PCR ürünleri çift yönlü sekans analizinden sonra MEGAX ile GenBank'taki referans diğer izolatlarla karşılaştırılması yapılmıştır. GLRaV-1 izolatlarının sekans analizine göre GenBank'taki referans izolatlarla %94-96, GLRaV-2 izolatları %96-99 ve GLRaV-3'ün UO3-3 izolatı %99-100 nükleotid benzerliği göstermiştir.

Anahtar Kelimeler

• Vitis vinifera
• virus
• RT-PCR
• Tokat
• Asma yaprak kıvırcıklığı hastalığı

Genetic variability of Grapevine Leafroll-Associated Virus-1,2,3,4 Infecting Vineyards in Tokat Province of Turkey

Abstract

Grapevine leafroll disease (GLRD) is one of the most important viral diseases of grapevine worldwide. The aim of this study was to investigate the prevelance of 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) in Tokat province of Turkey. In 2018, leaf samples and one-year-old shoots of plants, showing virus-like symptoms, were collected from different vineyards in Central, Erbaa, Niksar, and Pazar districts of Tokat province. Totally 197 samples were obtained and tested molecularly using two-step polymerase chain reaction (PCR) by using virus-specific primers for GLRaV-1, GLRaV-2, GLRaV-3, GLRaV-4) and GLRaV-9. At least 106 (53.80%) samples were found to be infected showed single or multiple infections. The 103 (52.28%) samples resulted in positive with GLRaV-1 while the infection rates were %5.18 for GLRaV-3 with 10 samples, %2.53 for GLRaV-4 with 5 samples, %1.03 for GLRaV-2 and GLRaV-9 with 2 samples, respectively. The amplified PCR products of GLRaV-1, GLRaV-2 and GLRaV-3 were sequenced, aligned with MEGAX, and analyzed in BLAST for multiple comparisons of other isolates in the GenBank. Sequence analysis of viruses showed 94-96 % , 96-99 % and, 99-100 % nucleotide identity with references isolates in the GenBank for GLRaV-1, GLRaV-2, and UO3-3 isolate of GLRaV-3 respectively.

Keywords

• Vitis vinifera
• virus
• RT-PCR
• Tokat
• Grapevine leafroll disease

Kaynakça

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