Tokat İli Asma Alanında Bazı Asma Virüslerinin Moleküler Olarak Belirlenmesi ve Grapevine Virus A'nın Filogenetik Analizi

Türkiye bağcılığı sosyoekonomik anlamda en önemli ürün gruplarının biridir. Bağlarda birçok zararlı organizma grubu özellikle de virüs enfeksiyonları bitkiyi zayıflatıp, verim ve kalitesinde azalmalara yol açtığından karantina ve sertifikasyonda başı çekmektedir. Bu çalışma, bağcılığın çok önemli olduğu Tokat ilinde üretilen asmalarda verim kaybına neden olan bazı viral etmenleri tespit etmek amacıyla yapılmıştır. Tokat Merkez ve ilçelerindeki bazı bağ alanlarından virüs semptomu gösteren asmaların genç yapraklarından ve bir yaşındaki sürgünlerinden örnekler alınmıştır. Toplanan 189 asma örneğine Grapevine pinot gris virus (GPGV), Grapevine virus A (GVA) ve Strawberry latent ringspot virus (SLRSV) varlığını tespit etmek için virüse özgü primerler kullanılarak moleküler bir yöntem olan RT-PCR testine tabi tutulmuştur. Toplam 189 asma örneğinden 80 (%42.32) GVA, 3 (%1.58) GPGV enfeksiyonu tespit edilmiş ve hiçbir örnekte SLRSV (%0) tespit edilmemiştir. Test edilen 189 asma örneğinin 2'sinde (%1.05) birden fazla virüs tespit edilmiştir. Tokat Merkez ve ilçelerinden toplanan bitki örneklerinde en sık görülen virüsün GVA, en az tespit edilen virüsün ise GPGV olduğu belirlenmiştir. GVA ile enfekteli izolatların RT-PCR ürünlerin çift yönlü sekans analizi analizleri yapılmış ve MEGAX bilgisayar programı ile hizalandıktan sonra referans izolatlarla karşılaştırılarak filogenetik analizleri yapılmıştır. Filogenetik analiz çalışmalarına göre, GVA, GenBank'ta kayıtlı izolatlarla ve farklı çalışmada elde edilen izolatlarla farklı dallanma göstermiştir. GVA ile enfekte izolatlar referans izolatlarla %92-94 oranında benzerlik göstermiştir. Bu çalışmada Türk GVA izolatlarının moleküler analizi yapılmıştır. Bu moleküler bilgilerin ileride yapılacak çalışmalara ışık tutacağı öngörülmektedir.

Anahtar Kelimeler:

Filogenetik analiz, Grapevine virus A, Strawberry latent ringspot virus, Grapevine pinot gris virus, Tokat, Vitis vinifera, RT-PCR

Molecular Determination of the Presence of Some Grapevine Viruses and Phylogenetic analyses of Grapevine Virus A in Tokat Province Grapevine Area

Grapevine (Vitis spp.) is one of the major fruit crop with high socioeconomic importance for Turkey. In vineyards, many harmful organism, especially virus infections, weaken the plant and lead to decreases in yield and quality, so it takes the lead in quarantine and certification. This study was carried out to determine some viral agents that cause yield loss in vines produced in Tokat, where viticulture is very important. Samples were collected from young leaves and one-year-old shoots of grapevines showing virus symptoms from some vineyard areas in Tokat Center and its districts. Collected 189 grapevine samples were subjected to the RT-PCR test, which is a molecular method using virus-specific primers, to detect the presence of Grapevine pinot gris virus (GPGV), Grapevine virus A (GVA), Strawberry latent ringspot virus (SLRSV). Out of a total of 189 plant samples, 80 (42.32%) of GVA, 3 (1.58%) of GPGV were detected and SLRSV (0%) was not detected. More than one virus was found in 2 (1.05%) of 189 tested samples. It was determined that the most common virus was GVA, the least detected virus was GPGV in plant samples collected from Tokat Center and its districts. Bidirectional sequence analysis of RT-PCR products of GVA-infected isolates were performed and phylogenetic analyzes were done by comparing them with reference isolates after they were aligned with the MEGAX computer program. Based on phylogenetic analysis studies, GVA showed differential branching with isolates registered in GenBank and isolates obtained in the study. GVA-infected isolates showed similarity with reference isolates at rates of 92-94%. In this study, molecular analysis of Turkish GVA isolates was performed. This molecular information is important as it will shed light on future studies.

Keywords:

Phylogenetic analysis, Grapevine virus A, Strawberry latent ringspot virus, Grapevine pinot gris virus, Tokat, Vitis vinifera, RT-PCR,

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