Semih ERKAN, İsmail Can PAYLAN, Müge ERGÜN

Comparison of RNA extraction methods for the molecular detection of Artichoke latent virus in globe artichoke

Enginar, içeriğindeki vitamin ve mineraller ile insan sağlığı için oldukça yararlı bir sebzedir. Yoğun olarak Akdeniz havzasında yetiştirilen bu ürün kendine has tadıyla Akdeniz mutfağında, çorba, salata ve yemek şeklinde tüketilmektedir. Son yıllarda yetiştiriciliği artmakta ve dünyaya yayılmaktadır. Tarımı yapılan her bitki gibi enginar da birçok patojenden etkilenmektedir. Bu çalışmada enginarlarda verim düşüşlerine neden olan Artichoke latent virus tespitinde önemli bir aşama olan RNA ekstraksiyon yöntemleri araştırılmıştır. Virüslerin tespiti ve tanılanmasında yaygın olarak kullanılan RT-PCR’ın temelini oluşturan hastalıklı bitki dokularından RNA ekstraksiyonu testin güvenilirliği açısından en önemli aşamadır. Enginar fenolik asit içeriği yüksek bir bitkidir. Bu maddeler RNA ekstraksiyonunu zorlaştırmakta dolayısıyla RT-PCR sonuçlarını etkilemektedir. Bu çalışmada yaygın kullanılan üç ekstraksiyon yöntemi ve ticari RNA izolasyon kitinin fenolik madde içeriği yüksek olan enginar bitkilerinde RNA ekstraksiyon etkinliklerinin araştırılması amaçlanmıştır. Yapılan çalışma sonuçları büyük ölçüde literatürle paralellik göstermiş, silica capture yöntemi ve ticari kitin yüksek saflıkta ve yoğunlukta RNA elde edilmesinde etkili olduğu saptanmıştır.

Enginarda artichoke latent virus (ArLV)’ün moleküler tanısında kullanılan RNA ekstraksiyon yöntemlerinin karşılaştırılması

Globe artichoke is very beneficial vegetable for human health because of involve vitamins and minerals within its structure. This vegetable which is intensively cultivated in the Mediterranean basin, are consumed in the form of meals, soups and salads with its own flavor in Mediterranean cuisine. In recent years, its production and consuming are increasing and spreading all over the world. As other cultivated plants, globe artichoke has been affected by many plant pathogens. In this study, the success of some RNA extraction methods that is very important step for detecting Artichoke latent potyvirus which cause yield and quality loss were investigated. RT-PCR is a reliable method and commonly used for detection and identification of viruses. Globe artichoke is a plant with high content of phenolic acids which make difficult isolation of RNA. Thereby the results of RT-PCR are affected. In this study, our aim is investigation of three widely used extraction methods and a commercial isolation kit’s effectiveness on artichoke with high content of phenolic compounds. The results of this study showed substantially parallel to the literature and also the commercial kit and silica capture methods were found effective to obtain high purity RNA with high concentration.

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