Isparta İli Biber Üretim Alanlarında Patates Y Virüsü’nün Belirlenmesi

Öz

Bu çalışmanın amacı indikatör bitkilerinin mekanik inokulasyonu, Double Antibody Sandwich Enzyme Linked Immunosorbent Assay (DAS-ELISA) ve Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) yöntemleri kullanılarak Isparta'daki biber yetiştirme alanlarında Patates Y Virüsü (PVY) varlığının belirlenmesidir. Tarlalardan toplam 184 biber bitkisi toplanmış ve tüm örnekler DAS-ELISA'da spesifik bir antiserum kullanılarak PVY açısından test edilmiştir. PVY, 33 örnekte tespit edilmiş ve tarla örneklerinin %17,93'ünde enfeksiyon oranı saptanmıştır. En yüksek PVY enfeksiyon oranı Senirkent'te (%63,63) gözlemlenirken, bunu Kuleönü (%57,14) izlemiştir. DAS-ELISA testinde PVY virüsü için pozitif çıkan yaprak örneklerinde elde edilen PVY izolatları, daha sonra mekanik aşılama, toplam RNA izolasyonu ve RT-PCR çalışmalarında kullanılmıştır. Hassas test bitkilerinin mekanik inokülasyonunu takiben PVY'nin tipik simptomları gözlenmiştir. RT-PCR çalışmalarında, PVY için spesifik primer çiftleri kullanılmış ve PVY'nin kılıf protein bölgesine ait 801 bp'lik bölge çoğaltılmış ve agaroz jel elektroforezinde gözlemlenmiştir. RT-PCR testi, DAS-ELISA pozitif dokuz izolatın ve simptomatik on bir bitkinin tümünün PVY ile enfekte olduğunu doğrulamıştır. Negatif kontrol olarak kullanılan biber bitkilerinde, N. tabacum L. ‘White Burley’, N. tabacum L. ‘Samsun NN’, C. amaranticolor ve C. quinoa'da virüse özgü PCR bantları gözlemlenmedi ve bu bitkilerin tümü mekanik inokülasyon testlerinde simptomsuz kaldı. Bu çalışmada, Türkiye'nin Isparta ilindeki biber üretim alanlarında PVY'nin varlığı ilk kez tespit edilmiştir. Bu durum gelecekte PVY izolatlarının sekans analizinin yapılması, dayanıklılık çalışmalarının sürdürülmesi ve yetiştiricilerin bilgilendirilmeye devam edilmesi gerektiğini göstermektedir.

Anahtar Kelimeler

• Biber
• Virüs
• Patates Y virüsü
• Teşhis

Detection of Potato Y Virus in Pepper Growing Areas in Isparta Province

Öz

The aim of the study was to determine the presence of Potato Y Virus (PVY) in pepper-growing areas in Isparta, using mechanical inoculation of indicator plants, Double Antibody Sandwich Enzyme Linked Immunosorbent Assay (DAS-ELISA) and Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). A total of 184 pepper plants were collected from pepper fields and all samples were tested for PVY using a specific antiserum in a DAS-ELISA. PVY was detected in 33 samples, amounting to an infection rate of 17.93% of the field samples. The highest PVY infection rate was observed in Senirkent (63.63%), followed by Kuleönü (57.14%). Leaf samples that tested positive for the PVY in the DAS-ELISA test were then used in mechanical inoculation, total RNA isolation, and RT-PCR studies. Following mechanical inoculation of sensitive test plants, typical symptoms of PVY were observed. In RT-PCR studies, primer pairs specific to PVY were used to amplify 801 bp fragments representing the coat protein region of PVY, which were then observed using agarose gel electrophoresis. RT-PCR testing confirmed that all nine DAS-ELISA-positive isolates and all eleven symptomatic plants were infected with PVY. No virus-specific PCR bands were observed in the pepper plants used as negative controls, nor in Nicotiana tabacum L. ‘White Burley’, N. tabacum L. ‘Samsun NN’, Chenopodium amaranticolor and C. quinoa, all of which remained symptomless in the mechanical inoculation assays. In this study, the presence of PVY was detected for the first time in pepper production areas in the province of Isparta, Turkey. It indicates that sequence analysis of PVY isolates should be carried out in future, that resistance studies should continue and that growers should be kept informed.

Anahtar Kelimeler

• Pepper
• Virus
• Potato Y virus
• Detection

Kaynakça

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