Molecular Characterization of Polyprotein genes of Two BCMV (Bean common mosaic potyvirus) isolates in Antalya (Turkey) and Their Genomic Divergence

Year 2020, , 411 - 419, 24.04.2020

Mustafa Usta Abdullah Güller

https://doi.org/10.30910/turkjans.725819

Cited By: 2

Abstract

Common bean (Phaseolus vulgaris L.) is regarded as one of the most important crops of the Fabaceae family throughout the world. Diseases caused by viruses are the most important factor limiting the production of beans. Bean specimens with classic virus-like symptoms were collected from bean fields in Antalya (Turkey) in July and August, 2018. BCMV was examined by RT-PCR test (Reverse Transcriptase -Polymerase Chain Reaction) using appropriate primer pairs directed to the partial NIb and the capsid protein (CP) gene which was devised to identify and to characterize the viral agent. The PCR test produced approximately 850 bp amplicon of expected lengths in 11 out of 20 fresh leaf tissues, indicating the presence of BCMV. Two of them were randomly selected and molecularly cloned into a congruent plasmid vector to reveal the CP sequences of interested isolates. Obtained recombinant clones consisting of insert genes were bidirectionally sequenced and both of the sequences were registered in the GenBank with MN104839 and MN104840 accession number. The provided BCMV partial CP gene sequences comprised 823 bp coding for 274 amino acid residues. The CP gene of these isolates was aligned with those of 17 isolates deposited in the GenBank database from different geographical location and its phylogenetic relationships were determined. Molecular analysis of the CP gene sequences of Antalya isolates showed the highest identity rates between 91.22 % and 94.71 %, at the nucleotide level. Moreover, phylogenetic analyses revealed that BCMV-Antalya 1 and Antalya 10 are best clustered with the Turkish isolate (KT766179) and England isolate (AY112735), respectively. By this study, the genetic difference of BCMV isolates have been determined in the bean plant from Antalya province of Turkey.

Keywords

BCMV (Bean common mosaic potyvirus), RT-PCR, Characterization, Phylogenetic relationship

Antalya (Türkiye) 'da İki Fasulye Adi Mozaik Virüs (BCMV) İzolatlarının Polyprotein Genlerinin Moleküler Karakterizasyonu ve Genomik Farklılıkları

Abstract

Fasulye (Phaseolus vulgaris L.), dünya çapında Fabaceae familyasının en önemli ürünlerinden biri olarak kabul edilmektedir. Virüslerin sebep olduğu infeksiyonlar, fasulye üretimini sınırlayan en önemli faktördür. Antalya ilinde 2018 yılı Temmuz ve Ağustos ayında fasulye ekilen tarlalardan virüs benzeri simptomları gösteren fasulye örnekleri toplanmıştır. Toplanan örnekler, Bean common mosaic potyvirus (BCMV) virüsünü araştırmak ve viral ajanının kapsid proteinini (CP) karakterize etmek amacıyla dizayn edilen primerler yardımıyla RT-PCR testi ile testlenmiştir. 20 taze yaprak örneğinin 11’inde beklenen büyüklükte (850 bp) fragment elde edilmiştir. Rastgele iki pozitif izolat seçilmiş ve kısmi CP gen dizisinin ortaya çıkarılması amacıyla uygun bir plazmid vektöre klonlanmıştır. İnsert gen içeren rekombinant klonlar çift yönlü olarak dizilenmiş ve elde edilen her iki izolatın sekansı MN104839 ve MN104840 erişim numarası ile Gen Bankasına kaydedilmiştir. Antalya izolatlarının kısmi CP gen dizisinin 823 bp’den oluştuğu ve 274 amino asit parçasını kodladığı belirlenmiştir. BCMV Antalya izolatlarının CP geni, GenBankası veri tabanında ve farklı coğrafi bölgelerde belirlenen 17 izolat ile çoklu karşılaştırma yapılmış ve filogenetik ilişkileri belirlenmiştir. Antalya izolatlarının CP gen sekansları, nükleotit seviyesinde diğer izolatlar ile %91.22 ve % 94.71 arasındaki en yüksek benzerlik oranlarını göstermiştir. Ayrıca, filogenetik analizler, BCMV-Antalya izolatlarının en iyi şekilde Türkiye (KT766179) ve İngiltere izolatı (AY112735) ile kümelendiğini ortaya koymuştur. Bu çalışma ile Antalya (Türkiye) ili fasulye alanlarında belirlenen iki BCMV izolatının genetik farklılıkları ortaya konmuştur.

Keywords

Fasulye Adi Mozaik Virus (BCMV), RT-PCR, Karakterizasyon, Filogenetik ilişki

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