Marmara Bölgesi domates üretim alanlarında Cucumber mosaic virus izolatlarının yaygınlığı ve moleküler karakterizasyonu

Öz

Türkiye’de domates virüsleri ile ilgili olarak gerçekleştirilmiş çok sayıda çalışma vardır. Ancak domatesin en önemli virüs hastalıklarından bir tanesi olan Cucumber mosaic virus (CMV)’nin geniş alanlarda yaygınlığı ve genetik çeşitliliği üzerine gerçekleştirilmiş bir çalışma bulunmamaktadır. Bu bağlamda Marmara Bölgesi domates üretim alanlarında sürveyler düzenlenerek virüs ve virüs benzeri simptom gösteren 113 domates bitkisinden örnekler alınarak DAS-ELISA testi ile CMV enfeksiyonunun belirlenmesi amacı ile testlemeler gerçekleştirilmiştir. Gerçekleştirilen testlemeler sonucunda 34 bitkide CMV enfeksiyonu tespit edilmiştir. Enfekteli örnekler içerisinden elde edildikleri iller temel alınarak 10 CMV izolatı moleküler karakterizasyon çalışmaları için seçilmiştir. Seçilen bu izolatların kılıf protein (CP) ve hareket protein (MP) genleri RT-PCR ile amplifiye edilmiş ve sekanslanmıştır. Elde edilen sekanslar kullanılarak gerçekleştirilen çoklu dizi analiz çalışmaları sonucunda, Marmara Bölgesi CMV izolatlarının CP genine göre kendi içlerinde nükleotid ve amino asit düzeyinde %97-100, dünya izolatları ile nükleotid düzeyinde %77-100, amino asit düzeyinde ise %79-100 oranlarında benzerlikler gösterdiği tespit edilmiştir. MP gen bölgesine göre ise CMV izolatlarının kendi içlerinde nükleotid düzeyinde %97-100, amino asit düzeyinde ise %96-100 oranlarında benzerlikler gösterdiği belirlenmiştir, dünya izolatları ile gösterdikleri benzerlik oranları ise nükleotid düzeyinde %79-100, amino asit düzeyinde ise %81-100 olarak belirlenmiştir. Gerçekleştirilen filogenetik analizler sonucunda Marmara Bölgesi CMV izolatlarının hem CP hem de MP gen bölgesine göre birbirleri ile yakın ilişkili olduğu ve Ia altgrubunda olduğu belirlenmiştir.

Anahtar Kelimeler

• CMV
• DAS-ELISA
• RT-PCR
• sekanslama

Prevalence and molecular characterization of Cucumber mosaic virus isolates infecting tomato plants in Marmara region of Turkey

Abstract

There are many studies carried out on tomato viruses in Turkey. However, there is no study on the prevalence and genetic diversity of Cucumber mosaic virus (CMV), one of the most important viral diseases of tomatoes. In this context, surveys were conducted in the tomato production areas of the Marmara region, and samples were taken from 113 tomato plants showing virus and virus-like symptoms, and tests were carried out to determine CMV infection by DAS-ELISA. As a result of the tests, CMV infection was detected in 34 plants. And, 10 CMV isolates were selected for further studies. Coat protein (CP) and movement protein (MP) genes of selected isolates were amplified by RT-PCR and sequenced. As a result of multiple sequence analysis, CMV isolates from the Marmara region showed 97-100% similarity in nucleotide and amino acid levels within themselves, 77-100% in nucleotide level, and 79-100% in amino acid level in the world isolates according to the CP gene. According to the MP gene region, it was determined that the CMV isolates showed 97-100% and 96-100% similarities at the nucleotide and amino acid levels in each other, respectively. The similarity rates with world isolates were determined as 79-100% at the nucleotide level and 81-100% at the amino acid level. As a result of the phylogenetic analyses performed, tomato CMV isolates were closely related to each other according to both gene regions and were in the Ia subgroup.

Keywords

• CMV
• DAS-ELISA
• RT-PCR
• sequencing

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