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Ispanak önemli bir kışlık sebze olup ülkemizin her bölgesinde üretimi yapılmaktadır. Ispanak üretiminde verim ve kaliteyi etkileyen birçok hastalık etmeni vardır ve bunlar içerisinden virüsler önemli bir yer tutmaktadır. Bu virüs hastalıklarından bir tanesi “Rhizomonia” olarak ta adlandırılan Pancar nekrotik sarı damar virüsü (Beet necrotic yellow vein virus; BNYVV)’dür. BNYVV’nin ülkemizde ıspanak bitkisinde varlığı serolojik yöntemlerle belirlenmiş ancak moleküler düzeyde bir çalışma yapılmamıştır. Bu amaçla Çanakkale’de il ve ilçe düzeyinde ıspanak yetiştiriciliği yapılan alanlara BNYVV’nin varlığını belirlemek ve tanısını yapmak amacı ile 2017-2018 üretim yılında bir arazi çalışması gerçekleştirilmiştir. Bu arazi çalışması kapsamında tipik virüs simptomları gösteren ıspanak bitkilerinden 66 örnek alınmıştır. Alınan örneklerdeki BNYVV varlığı virüs proteinine dayalı bir yöntem olan DAS-ELISA testi ile belirlenmiştir. DAS-ELISA testi sonuçlarına göre 66 örneğin 12’si BNYVV ile enfekteli bulunmuştur. Enfekteli bulunan örnekler içerisinden bir tanesi seçilerek (CNKSpinach39) örtü protein geni (coat protein; CP) RT-PCR ile çoğaltılmıştır. CP geni çoğaltılan izolatın RT-PCR ürünleri saflaştırılmış ve nükleotid dizileri belirlenmiştir. Elde edilen nükleotit (nt) dizileri kullanılarak seçilen izolata ait CP geninin amino asit (aa) dizileri çıkarılmıştır. Bu nükleotid ve amino asit dizileri dünya BNYVV izolatlarının CP genlerinin nükleotid ve amino asit dizileri ile karşılaştırılarak, göstermiş oldukları benzerlik oranları ve filogenetik ilişkileri saptanmıştır. Gerçekleştirilen çoklu nt ve aa temelli dizi karşılaştırmaları sonucunda CNKSpinach39 izolatının dünya izolatları ile %95-99 oranında benzerlikler gösterdiği belirlenmiştir. Gerçekleştirilen filogenetik analizler sonucunda ise CNKSpinach39 izolatının B patotipinde yer aldığı tespit edilmiştir. Bu çalışma ile ülkemizde ıspanakta ilk defa BNYVV’nin B patotipi bulunmuştur.
Spanish is an important winter vegetable and is produced in every region of our country. There are many diseases that affect efficiency and quality in the production of spinach, and the viruses hold an important place in them. One of the diseases of this virus is the Pancar necrotic yellow vein virus (BNYVV), which is called "Rhizomonia". The presence of BNYVV in the spinach plant in our country has been determined by serological methods but no molecular study has been done. For this purpose, a land work in the production year 2017-2018 was carried out in Çanakkale with the aim of determining and diagnosing the presence of BNYVV in the areas of spinach cultivation at the province and district level. In this land study, 66 samples of spinach plants with typical viral symptoms were taken. The presence of BNYVV in the taken samples was determined by the DAS-ELISA test, a method based on viral protein. According to the results of the DAS-ELISA test, 66 of the 12 were infected with BNYV. One of the infected samples was selected (CNKSpinach39) covered protein gene (coat protein; CP) multiplied with RT-PCR. The RT-PCR products of the CP gene multiplied isolat are purified and the nucleotid series are determined. Amino acids (aa) of the CP gene belonging to the selected isolate with the use of the derived nucleotites (nt) series are removed. These series of nucleotides and amino acids, compared with the series of nucleotides and amino acids of the CP genes of the world’s BNYVV isolates, have identified their similarity rates and philogenetic relationships. The multi-nt and aa-based series comparisons have found that the CNKSpinach39 isolat has a 95%-99% similarity to the world isolat. The philogenetic analysis found that the CNKSpinach39 is located in the B pathotype. For the first time in our country, BNYVV’s B patotype was found in Spain.
Spinach is an important winter vegetable and it is produced in all regions in Turkey. There are many diseases affecting the yield and quality in its production. Viruses have an important role among them. One of these virus diseases is Beet necrotic yellow vein virus (BNYVV), called Rhizomania disease. The presence of BNYVV in the spinach plant was determined by serological methods in Turkey, but no studies were conducted at the molecular level. For this purpose, field studies were carried out in 2017-2018 in order to determine the presence of BNYVV in the fields of spinach cultivation at the level of Çanakkale province and its districts. In these field studies, 66 samples were collected from spinach plants showing typical virus symptoms. The presence of BNYVV in the samples was determined by DAS-ELISA test, a method based on virus protein. According to the DAS-ELISA test, 12 out of 66 samples were infected with BNYVV. One of the infected samples was selected (CNKSpinach39) and the coat protein gene (CP) was amplified by RT-PCR. Its RT-PCR product was purified and sequenced. The amino acid (aa) sequences of this isolate was deduced based on its nt sequence. These nt and aa sequences were compared with the related sequences of the world BNYVV isolates and their similarities and phylogenetic relationships were determined. As a result of multiple sequence comparisons based on nt and aa, it was determined that CNKSpinach39 isolates showed similarities with 95-99% of the world isolates. As a result of the phylogenetic analysis, it was determined that the isolate CNKSpinach39 was found in the B pathotype. In this study, the first BNYVV B pathotype was found in spinach in our country.
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