Patateste patates Y virüsü ırklarının tanılanması ve virüs miktarının kantitatif real time RT-PCR ile belirlenmesi
Yükleniyor...
Tarih
2017
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Niğde Ömer Halisdemir Üniversitesi / Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Patates bitkisi yumrularıyla vejetatif olarak üretildiği için virüsler gibi yumru ile taşınan hastalıklardan önemli ölçüde etkilenmektedir. Tohumluk yumru üretiminde en önemli virüs etmeni Potato virus Y (PVY)’dir. PVY ırklarına bağlı olarak, tohumlukların özelliklerini yitirmesine ve % 50–80 arasında verim kaybına neden olmaktadır. Virüs ile mücadelede virüsün ırklarının bilinmesi, özellikle tohumluk yumru üretiminde damızlıklıların doğru, hassas ve hızlı teşhis yöntemleri ile test edilmesi oldukça önem taşımaktadır. Bu amaçla, PVY’nin ırklarını belirlemede Immunocapture- revers trancription- multipleks polimeraz zincir reaksiyonu (IC-RT-multipleks PCR) sistemi Niğde ilinden temin edilen toplam 54 PVY izolatına uygulanmıştır. Analiz sonucunda 17 adet PVYNW (B), 7 adet PVYNW (B), 1 adet PVYNTN (B) ırkları tespit edilmiştir. Ayrıca, 2 adet PVYNTN+PVYNW karışık ırk enfeksiyonu belirlenmiştir. İzolatların büyük bir çoğunluğunda (30 adet) 278 bp seviyesinde atipik amplikonlar elde edilmiştir. PVY’nin farklı ırklarında başarı ile uygulanabilen real-time PCR sistemi geliştirmek amacıyla ICRT- SyberGreen (SYBR) real time PCR sistemi PVY F/R primerleri kullanılarak uygulanmıştır. IC-RT-SYBR real time PCR Niğde ilinden temin edilen farklı PVY ırklarına uygulandığında, tüm örneklerde PVY teşhisinde başarılı olmuştur. Ayrıca ICRT-SYBR kantitatif real time PCR sistemi kurulmuştur. En düşük ve en yüksek ölçüm sonuçları alınmıştır. Desire ve Nectar çeşidi tohumluk yumrularda PVY taraması ve virüs miktarı tayini çalışmarında kantitatif real time PCR başarılı bir şekilde kullanılabilmiştir.
Since the potato plant is vegetatively produced by the tubers, it is significantly affected by tuber-borne diseases such as viruses. The most important virus in the production of seed tubers is Potato virus Y (PVY). According to PVY strains, it causes loss of seed characteristics and yield loss of 50-80%. In management of virus diseases, detection of virus strains and availability of accurate, sensitive and rapid diagnostic methods, has an imporant role in poducing virus free seed tuber. For this purpose, Immunocapture-reverse trancription-multiplex polymerase chain reaction (IC-RT-multiplex PCR) system for determining strains of PVY has been applied to a total of 54 PVY isolates from Niğde. As a result, 17 PVYNW (B), 7 PVYNW (B) and 1 PVYNTN (B) strains were detected. In addition, two PVYNTN + PVYNW mixed race infections were identified. In a large majority of isolates (30), 278 bp atypical amplicons were obtained. In order to develop a real-time PCR system that can be successfully applied in different strains of PVY, the IC-RTSyberGreen (SYBR) real time PCR system was applied using PVY F / R primers. When IC-RT-SYBR real time PCR was applied to different PVY strains from Niğde province, PVY was successfully diagnosed in all cases. In addition, the IC-RT-SYBR quantitative real-time PCR system was established and the minimum and maximum titer quanities were measured. Quantitative real time PCR was successfully used in the detection of PVY screening and virus quantitation in seeds of Desire and Nectar species.
Since the potato plant is vegetatively produced by the tubers, it is significantly affected by tuber-borne diseases such as viruses. The most important virus in the production of seed tubers is Potato virus Y (PVY). According to PVY strains, it causes loss of seed characteristics and yield loss of 50-80%. In management of virus diseases, detection of virus strains and availability of accurate, sensitive and rapid diagnostic methods, has an imporant role in poducing virus free seed tuber. For this purpose, Immunocapture-reverse trancription-multiplex polymerase chain reaction (IC-RT-multiplex PCR) system for determining strains of PVY has been applied to a total of 54 PVY isolates from Niğde. As a result, 17 PVYNW (B), 7 PVYNW (B) and 1 PVYNTN (B) strains were detected. In addition, two PVYNTN + PVYNW mixed race infections were identified. In a large majority of isolates (30), 278 bp atypical amplicons were obtained. In order to develop a real-time PCR system that can be successfully applied in different strains of PVY, the IC-RTSyberGreen (SYBR) real time PCR system was applied using PVY F / R primers. When IC-RT-SYBR real time PCR was applied to different PVY strains from Niğde province, PVY was successfully diagnosed in all cases. In addition, the IC-RT-SYBR quantitative real-time PCR system was established and the minimum and maximum titer quanities were measured. Quantitative real time PCR was successfully used in the detection of PVY screening and virus quantitation in seeds of Desire and Nectar species.
Açıklama
Anahtar Kelimeler
Immunocapture, Real Time PCR, Kantitatif PCR, Patates Yumrusu, Quantitaive PCR, Potato Tuber
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Bolat, V. (2017). Patateste patates Y virüsü ırklarının tanılanması ve virüs miktarının kantitatif real time RT-PCR ile belirlenmesi. (Yüksek Lisans Tezi) Niğde Ömer Halisdemir Üniversitesi, Fen Bilimleri Enstitüsü, Niğde
