Model bitki Nicotiana benthamiana’da Beet necrotic yellow vein virus izolatlarının simptom tipleri ve bazı viral proteinlerin araştırılması

Year 2023, Volume: 11 Issue: 2, 132 - 143, 21.12.2023

Murat Güngör Nazlı Dide Kutluk Yılmaz

https://doi.org/10.33409/tbbbd.1329616

Abstract

Beet necrotic yellow vein virus (BNYVV), dünyada şeker pancarı alanlarında ‘rhizomania’ olarak adlandırılan hastalığa neden olmaktadır. Bu çalışmada, Türkiye’nin farklı coğrafik bölgelerinde şeker pancarı üretim alanlarından elde edilmiş BNYVV ile bulaşık olduğu bilinen 15 toprak örneği kullanılmıştır. Öncelikle, tuzak bitki testi yöntemine göre BNYVV izolatlarının yeniden çoğaltımı sağlanmıştır. Daha sonra, 15 BNYVV izolatı virüsün sistemik konukçusu Nicotiana benthamiana’ya mekanik olarak inokule edilmiştir. Şiddetli (cücelik ve mozayik) ya da zayıf (mozayik) simptom oluşumu gösteren bitkilerin sistemik yapraklarından RNA izolasyonunu takiben, gen spesifik primerler kullanılarak RT-PCR çalışmaları gerçekleştirilmiştir. Bu çalışmalar sonucunda; 15 BNYVV izolatı ile enfekteli N. benthamiana’da P31 ile ilgili gen bölgesi saptanmasına ragmen, kılıf protein bölgesi 10 izolatta, P25 bölgesi bir izolatta, P26 bölgesi ise dört izolatta belirlenmiştir. Bu sonuç, N. benthamina’da BNYVV’nin simptom oluşumunda P31 geninin rol oynadığını desteklemiş, P25 ve P26 gen bölgelerinin ise bazı izolatlarda virüsün replikasyonu esnasında tespit edilemediğini göstermiştir.

Keywords

CP, ELISA, Rhizomania, P25, P26, P31, RT-PCR, sekans analizi

Supporting Institution

TÜBİTAK

Project Number

110O188

Thanks

Bu çalışmada kullanılan toprak örnekleri TÜBİTAK (TOVAG: 110O188) projesi kapsamında toplanmıştır.

Investigation of symptom types and viral proteins produced by Beet necrotic yellow vein virus isolates on the model plant Nicotiana benthamiana

Abstract

Beet necrotic yellow vein virus (BNYVV) causes a disease ‘rhizomania’ in sugar beet areas in the world. In this study, 15 BNYVV-infested soil samples were selected according to geographic locations. Firstly, BNYVV was propagated as bait plant technique. Then, systemic host Nicotiana benthamiana was mechanically inoculated by 15 BNYVV isolates. Some BNYVV isolates induced very strong symptom (dwarfing and mosaic) the others induced mild symptom (mosaic) on N. benthamiana. After RNA isolation, RT-PCR studies were conducted by using gene specific primers. This study showed that the P31 gene could be detected in N. benthamiana plants infected with 15 BNYVV isolates, the CP gene with only 10 isolates, the P25 gene with one isolate and the P26 gene with four isolates. This result supported that the P31 gene is involved symptom expression on N. benthamiana of BNYVV, the P25 and P26 genes were not possible to be detect with some BNYVV isolates during virus replication.

Keywords

CP, ELISA, P25, P26, P31, Rhizomania, RT-PCR, Sequencing

Project Number

110O188

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