Savaş KORKMAZ, Bayram ÇEVİK, Serkan ÖNDER, N. Kemal KOÇ

Ülkemizin Farklı Turunçgil Üretim Bölgelerinden Elde Edilen Turunçgil Tristeza Virüsü İzolatlarının Biyolojik, Serolojik ve Moleküler Karakterizasyonu

Ülkemizin beş farklı turunçgil üretim bölgesinde 2005–2006 yılları arasında arazi çalışmaları yürütülmüş ve farklı turunçgil çeşitlerinden toplam 201 örnek toplanmıştır. Örnekler turunçgil tristeza virüsünün varlığını belirlemek amacıyla DAS-ELISA ve RTPCR yöntemleriyle test edilmiştir. ELISA testi sonucunda 41 örnek tristeza ile infekteli bulunurken, PCR çalışmalarında ilaveten 13 örnek daha tristeza ile infekteli bulunmuştur. Tristezanın şiddetli ırklarının belirlenmesi için geliştirilmiş olan MCA13 monoklonal antibadisi ile yapılan Western blot analizi çalışmalarında ise çoğunluğu Satsuma çeşitlerine ait olan 32 örnek pozitif sonuç vermiştir. MCA13 monoklonal antibadi sonuçlarını desteklemek ve aynı zamanda MCA 13 pozitif, negatif ya da karışık infeksiyonlarını belirlemek için geliştirilmiş olan iki yönlü PCR çalışmalarında ise MCA13 bulgularını destekler nitelikte sonuçlar elde edilmiştir. Ayrıca elde edildiği coğrafik bölgeler ve çeşitler göz önünde bulundurularak seçilen toplam 28 tristeza izolatının biyolojik özellikleri çalışılmıştır. Biyolojik indekslemeler sonucunda izolatların hiçbiri inokule edildiği portakal, turunç ve altıntop bitkilerinde simptom oluşturmazken, Meksika laymı bitkisinde ise damar açılması simptomu oluşturmuşlardır. İlaveten tüm Satsuma izolatları ve bir kamkat izolatı Meksika laymı bitkisinde gövde çukurlaşması simptomu oluşturmuştur. Bu sonuçlar farklı turunçgil üretim bölgelerinde turunçgil tristeza virüsünün şiddetli ırklarının bulunduğunu göstermiştir.

Anahtar Kelimeler:

Turuçgil tristeza virüsü, RT-PCR, Western blot analizi, Biyolojik indeksleme, ELISA, Türkiye

Biological, Serological, and Molecular Characterization of Citrus tristeza virus Isolates from Different Citrus Cultivation Regions of Turkey

Field surveys were carried out in 5 different citrus cultivation regions of Turkey in 2005 and 2006, and 201 samples were collected from different citrus species. Samples were tested for the presence of Citrus tristeza virus (CTV) by double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). While DAS-ELISA showed that 41 trees were infected with CTV, an additional 13 trees were found to be positive based on RT-PCR. When CTV-positive samples were tested with the Western blot method using the monoclonal antibody MCA13, which is specific to severe isolates of CTV, 32 isolates, mostly from satsuma, were found to be positive. These isolates were then verified by bidirectional/PCR (BD/PCR), allowing differentiation of the MCA13 positive and negative isolates, and detection of mixed infections. The BD/PCR results were generally in agreement with the results of the Western blot assay with MCA13. In total, 28 isolates representing different geographic locations and hosts were selected for biological indexing. Although none of these 28 isolates induced any symptoms in sour orange, grapefruit, or sweet orange, all isolates induced the vein clearing symptom in Mexican lime. Additionally, all the tested satsuma isolates and 1 kumquat isolate produced stem pitting in Mexican lime. The results revealed that potentially severe isolates of CTV are present in different citrus cultivation regions of Turkey.

Keywords:

Citrus tristeza virus, RT-PCR, Western blot, biological indexing, ELISA, Turkey,

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