Turunçgil Bitkilerinde Görülen Citlodavirus citri (CCDaV) ve Potexvirus citriflavivenae (CYVCV) Etmenlerinin Nitroselüloz ve Kaba Filtre Kağıtları Üzerindeki Kalıcılık Sürelerinin Araştırılması

Öz

Bu araştırmada, turunçgil yetiştiriciliğinde ekonomik kayıplara yol açan önemli iki viral patojen olan Citlodavirus citri (CCDaV) ve Potexvirus citriflavivenae (CYVCV)'nin, nitroselüloz ve kaba filtre kağıtları üzerindeki canlılık ve tespit edilebilirlik süreleri karşılaştırmalı olarak incelenmiştir. Çalışmada virüsler, üç farklı yöntemle (CTAB protokolü ile ekstrakte edilmiş total nükleik asit/TNA, enfekteli bitki dokusunun direkt baskısı ve bitki öz suyunun damlatılması) membranlara aktarılmıştır. Beş hafta boyunca 7., 14., 21., 28. ve 35. günlerde RT-PCR/PCR ile yapılan analizlerde, CCDaV'nin TNA formunda her iki kağıt türünde de 35 gün boyunca stabil kaldığı tespit edilmiştir. CYVCV ise yalnızca TNA formunda benzer dayanıklılık göstermiş, diğer yöntemlerde özellikle kaba filtre kağıdında tespit süresi kısalmış veya hiç sonuç alınamamıştır. Mekanik inokulasyon testlerinde, CCDaV 21. güne kadar enfektivitesini korurken CYVCV hiçbir örnekte enfeksiyon oluşturamamıştır. Sonuçlar, özellikle nitroselüloz filtre kağıtlarının her iki virüsün de nükleik asit düzeyinde uzun süre saklanması için güvenilir bir ortam sağladığını göstermektedir. Bu bulgular filtre kağıtlarının, uzak mesafelere viral materyal gönderimi ve moleküler tanı laboratuvarları için sürekli kontrol materyali temininde pratik, ekonomik ve etkili bir araç olarak kullanılabileceğini ortaya koymaktadır.

Anahtar Kelimeler

• CCDaV
• CYVCV
• PCR
• Nitroselüloz filtre kağıdı
• Kaba filtre kağıdı

Investigation of the Persistence Periods of Citlodavirus citri (CCDaV) and Potexvirus citriflavivenae (CYVCV) Agents Present in Citrus Plants on Nitrocellulose and Coarse Filter Papers

Öz

This study comparatively investigated the stability and detection periods of two significant viral pathogens in citrus cultivation, Citlodavirus citri (CCDaV) and Potexvirus citriflavivenae (CYVCV), on nitrocellulose and coarse filter papers. The viruses were transferred to the membranes using three different methods: total nucleic acid (TNA) extracted via the CTAB protocol, direct tissue imprinting from infected plant material, and application of crude plant sap. Analysis via RT-PCR/PCR at 7, 14, 21, 28, and 35 days post-application revealed that CCDaV remained stable for 35 days on both paper types when applied as TNA. CYVCV showed similar durability only in TNA form, while its detection period shortened or yielded no results with other methods, particularly on coarse filter paper. In mechanical inoculation tests, CCDaV retained its infectivity up to 21 days, whereas CYVCV failed to induce infection in any sample. The results demonstrate that nitrocellulose filter papers provide a reliable medium for long-term preservation of both viruses at the nucleic acid level. These findings suggest that filter papers can serve as a practical, cost-effective, and efficient tool for shipping viral material over long distances and supplying consistent control materials for molecular diagnostics laboratories.

Anahtar Kelimeler

• CCDaV
• CYVCV
• PCR
• Nitrocellulose filter paper
• Coarse filter paper

Kaynakça

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