Serological and Molecular Detection of Single and Mixed Infections Symptomatology Associated with Potato Viruses Infecting Commercial Potato in Punjab Province, Pakistan

Öz

Potato is an important source of food in developing countries, and potato viruses have been a major constraint on sustainable production of the potato crop in Sialkot, Punjab Province, Pakistan. During February 2021, a survey of nine commercial potato fields was conducted to determine the incidence of PVY, PVX, and PLRV, respectively. The results of serological and RT-PCR assays indicated that PVX was not detected in any of the fields. PVY and PLRV were dominant in all fields with infection levels up to 63.15% and 45.83% respectively. The incidence of co-infection of potato with PVY and PLRV was 8.3%. The RT-PCR was the most reliable method for the detection and identification of viruses. The coat protein sequences of PVY-SKT1, PVY-SKT2, PLRV-SKT1 and PLRV-SKT2 were deposited in the Genbank database and assigned the accession numbers MW881191, MW881192 MW881193, and MW881194. The BLASTn search indicated PVY sequence shared the greatest similar score, at 99% similarity, with PVY isolates from China (HM036202.1) and Kashmir (KY851109.1). The PLRV sequence was most closely related to PLRV-Pakistan (MF276872.1) (99.8%) and China (KR051180.1) (99%) isolates. The results of co-inoculation experiments corroborated previous reports that infection of potato by multiple viruses dramatically increased disease severity (50-70%), compared to single virus infections (45-65%). The ELISA and RT-PCR results were confirmatory in that virus was detected in plants by serological and molecular methods, respectively. The prevalence of single and mixed PVY and PLRV infections in commercial potato fields underscores the need for routine virus indexing to foster routine planting of virus-free seed tubers.

Anahtar Kelimeler

• Molecular detection
• Potato leafroll virus
• Potato virus Y
• Plant virus co-infection
• Serology

Pakistan'ın Pencap Eyaletinde Ticari Patatesleri Etkileyen Patates Virüsleriyle İlişkili Tek ve Karışık Enfeksiyonların Semptomatolojisinin Serolojik ve Moleküler Tespiti

Öz

Patates, gelişmekte olan ülkelerde önemli bir gıda kaynağıdır ve patates virüsleri, Pakistan’ın Pencap Eyaleti, Sialkot bölgesindeki patates üretiminin sürdürülebilirliğini ciddi şekilde kısıtlamaktadır. Şubat 2021’de, PVY, PVX ve PLRV virüslerinin görülme sıklığını belirlemek amacıyla dokuz ticari patates tarlasında bir tarama yapılmıştır. Serolojik ve RT-PCR analiz sonuçları, PVX’in hiçbir tarlada tespit edilmediğini göstermiştir. PVY ve PLRV’nin ise tüm tarlalarda baskın olduğu ve enfeksiyon oranlarının sırasıyla %63,15 ve %45,83’e ulaştığı tespit edilmiştir. PVY ve PLRV’nin birlikte enfekte ettiği patateslerin oranı %8,3 olarak belirlenmiştir. RT-PCR, virüslerin tespiti ve tanımlanmasında en güvenilir yöntem olarak öne çıkmıştır. PVY-SKT1, PVY-SKT2, PLRV-SKT1 ve PLRV-SKT2’ye ait dış kılıf proteini dizileri GenBank veri tabanına kaydedilmiş ve sırasıyla MW881191, MW881192, MW881193 ve MW881194 erişim numaraları atanmıştır. BLASTn analizi, PVY dizisinin Çin’den (HM036202.1) ve Keşmir’den (KY851109.1) izolatlarla %99 oranında benzerlik gösterdiğini ortaya koymuştur. PLRV dizisinin ise Pakistan’dan (MF276872.1) %99,8 ve Çin’den (KR051180.1) %99 oranında benzerlik gösterdiği belirlenmiştir. Ortak enfeksiyon deneylerinin sonuçları, önceki raporlarla uyumlu olarak, birden fazla virüsün patatese bulaşmasının, tek bir virüsle enfeksiyona kıyasla hastalık şiddetini (%50-70) önemli ölçüde artırdığını doğrulamıştır (tek virüs enfeksiyonlarında %45-65). ELISA ve RT-PCR sonuçları, bitkilerde virüslerin serolojik ve moleküler yöntemlerle tespit edildiğini doğrulamıştır. Ticari patates tarlalarındaki tekli ve karışık PVY ve PLRV enfeksiyonlarının yaygınlığı, virüssüz tohum yumrularının düzenli olarak dikimini teşvik etmek için rutin virüs indeksleme uygulamalarının önemini vurgulamaktadır.

Anahtar Kelimeler

• Moleküler tespit
• Patates yaprak kıvırcıklığı virüsü
• Patates virüsü Y
• Bitki virüsü ko-enfeksiyonu
• Seroloji

Kaynakça

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