Erzincan İlinde Bal Arısı (Apis mellifera L.) Kolonilerinde Tespit Edilen Deforme Kanat Virüslerünün Moleküler Karakterizasyonu

Öz

Iflaviridae ailesine ait olan deforme kanat virüsü (DWV), tüm dünyada bal arısı kolonilerinde ciddi maddi kayıplara neden olan yıkıcı bir virüstür. 2020'de (Kasım) Erzincan ilinden toplanan virüs şüphesi olan bal arısı örnekleri, DWV enfeksiyonuna karşı moleküler testlerle tarandı. Test edilen 12 örneğin 6'sında muhtemel virüs varlığını gösteren yaklaşık 700 bp DNA bandı elde edildi. Virüsün varlığını gösteren bantlardan rastgele seçilen iki amplifiye edilmiş DNA bandı pGEM-T Easy vektörüne klonlanmış ve elektriksel olarak kompetan E. coli bakterisine aktarılmıştır. Klonlamadan elde edilen rekombinant plazmitler, E. coli'den saflaştırılmış ve yeni nesil dizileme (NGS) ile dizilenmiştir. DWV'nin kısmi nükleotid dizilerinin 711 nükleotid içerdiği tespit edilmiş ve bu diziler MW962981 ve MW962982 erişim numarası ile NCBI GenBank'a kaydedilmiştir. Her iki izolatın en yüksek dizi benzerliği, nükleotid düzeyinde %98.03 ile %98.17 arasında değişmiştir. Ayrıca, Erzincan DWV dizilerinin filogenetik ilişkileri, GenBank'ta kayıtlı farklı ekolojik bölgelerden 15 benzersiz dizi kullanılarak ortaya çıkarılmıştır. Filogenetik ağaç, DWV Erzincan izolatlarının İngiltere (HM067438), İspanya (MK262743) ve İsveç (MN746311) izolatları ile kümelendiğini göstermiştir. Literatür bilgilerimize göre, bu çalışma ile DWV (Deforme kanat virüsü) Türkiye'nin Erzincan ili bal arılarında ilk kez rapor edilmiştir.

Anahtar Kelimeler

• Deforme kanat virüsü
• bal arısı
• RT-PCR
• klonlama

Molecular characterization of Deformed wing viruses identified in honeybee (Apis mellifera L.) colonies in Erzincan province of Turkey

Öz

Deformed wing virus (DWV), which belongs to the Iflaviridae family, is a devastating virus causing severe financial losses in honey bee colonies all over the world. In 2020 (November), virus-suspected honey bee specimens collected from Erzincan province were screened by molecular assays against DWV infection. In 6 of 12 samples, an expected approximately 700 bp DNA bands were achieved, indicating probably virus-positive. Two randomly selected amplified DNA band suggesting the presence of the virus was further cloned into a pGEM-T Easy vector and electrically transformed into E. coli competent bacteria. Recombinant plasmids obtained from cloning were purified from E. coli and sequenced by next-generation sequencing (NGS). The partial nucleotide sequences of DWV were ascertained to contain 711 nucleotides and these sequences were recorded in the NCBI GenBank with MW962981 and MW962982 accession number. The highest sequence similarity of both isolates ranged from 98.03% to 98.17%, at the nucleotide level. Furthermore, phylogenetic relationships of Erzincan DWV sequences were revealed using 15 unique sequences from different ecological regions registered in the GenBank. Phylogenetic tree displayed that DWV Erzincan isolates are clustered with UK (HM067438), Spain (MK262743), and Sweden (MN746311) isolates. According to our best knowledge, DWV has been reported for the first time in the honey bees from Erzincan province of Turkey by this study.

Anahtar Kelimeler

• Deformed wing virus
• honey bee
• RT-PCR
• cloning

Kaynakça

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