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

Year 2025, Volume: 22 Issue: 2, 268 - 275, 26.05.2025

Samına Yasmın , Mukaddes Kayım

https://doi.org/10.33462/jotaf.1320130

Abstract

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.

Keywords

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

Ethical Statement

Bu çalışma için etik kuruldan izin alınmasına gerek yoktur.

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

Abstract

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.

Keywords

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

Ethical Statement

There is no need to obtain permission from the ethics committee for this study.

References

• Abbas, M. F., Hameed, S. and Rauf, C. A. (2014). Presence of new strain of potato virus Y in Pakistan. International Journal of Agriculture and Biology, 16: 941-945.
• Abbas, M. F., Hameed, S., Rauf, C. A, Nosheen, Q., Gilani, A., Qadir, A. and Zakia, S. (2012). Incidence of six viruses in potato growing areas of Pakistan. Pakistan Journal of Phytopathology 24: 44-47.
• Ali, M. C., Mariana, R. R., Kelsie, J. G., Dong, J. K., Sang, M. C., Joseph, C. K. and Alexander, V. K. (2019). Identification and molecular characterization of recombinant potato virus Y (PVY) in potato from South Korea, PVYNTN strain. Plant Disease, 103: 137-142.
• Clark, M. F. and Adam, A. W. (1977). Characteristics of microplate methods of enzyme-linked immune sorbent assay (ELISA) for detection of plant viruses. Journal of General Virology, 34:475-483.
• Devaux, A., Kromann, P. and Ortiz, O. (2014). Potato for sustainable global food security. Potato Research, 57:185-199.
• Dietgen, R. G. (2002). Application of PCR in plant virology. In: plant viruses as molecular pathogens: Khan, J.A. Dijkstra, J, (Eds.), The Haworth press, New York, NY, USA.
• Garcia-cano, E., Resen, D. R., Fernandez-Mounz, R. and Moriones, E. (2006). Synergistic between tomato chlorosis virus and tomato spotted wilt virus results in breakdown of resistance in tomato. Phytopathology, 96: 1263-1269.
• Gibbs, A. J. and Ohshima, K. (2010). Potyviruses and the digital revolution. Annual of Phytopathology, 8: 205-223.
• Hameed, A., Iqbal, Z., Asad, S. and Mansoor, S. (2014). Detection of multiple potato viruses in the field suggests synergistic interactions among potato viruses in Pakistan. Plant Pathology Journal, 30: 407-415.
• Harahagazwe, D., Condor, B. and Barreda, C. (2018). How big is the potato (Solanum tuberosum L.) yield gap in Sub-Saharan Africa and why? A participatory approach. Open Agriculture, 3: 180-189.
• İlbağı, H. and Geyik, S. (2014). Detection of virus diseases in corn (Zea mays L.) fields in Bursa Province of Turkey. Journal of Tekirdag Agricultural Faculty, 11(1): 122-125.
• Kreuze, J. F., Souza-Dias, J. A. C., Jeevalatha, A., Figueira, A. R, Valkonen, J. P. T. and Jones, R. A. C. (2020). Viral diseases in potato. In: Campos H., Ortiz O. (eds) The Potato Crop. Its agricultural, nutritional and social contribution to humankind. Cham (Switzerland). Springer, Cham. ISBN: 978-3-030-28683-5. pp. 389-430.
• Kumar, R., Jeevalatha, A., Baswaraj, R., Kumar, R., Sharma, S. and Nagesh, M. (2017). A multiplex RT-PCR assay for simultaneous detection of five viruses in potato. Journal of Plant Pathology, 99: 37-45.
• Kumar, S., Stecher, G. and Tamura, K. (2016). MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33: 1870-1874.
• Matthews, R. H. (2002). Plant Virology. 4th ed., Academic Press, UK.
• Mughal, S. M, Khalid, S. and Hussain, Z. (1981). Isolation, identification, and prevalence of tobacco viruses in Pakistan. Pak Tobacco. 10: 5-9.
• Önder, S. and Korkmaz, S. (2008) Determination of virus and viroid diseases on satsuma owari mandarins by biological and serological methods in Edremit Gulf Region. Journal of Tekirdag Agricultural Faculty, 5(2):205-214.
• Rahman, M., Akanda, A., Mian, I. H., Bhuian, M. and Karim, M. R. (2010). Growth and yield performance of different generations of seed potato affected by PVY and PLRV. Bangladesh Journal of Agriculture and Research, 35: 37-50.
• Shrestha, D., Erik, J.W., Pamela, J. S. H, Jonathan, L. W., Shaonpius, M., Sanford, D. E., Nilsa, A. B. P. (2014). Interactions among potato genotypes, growth stages, virus strains, and inoculation methods in the potato virus Y and green peach aphid pathosystem. Environmental Entomology, 43: 662-671.
• Thompson, J. D., Higgins, D. G. and Gibson, T. J. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research, 22: 4673-4680.
• Veerbek, M., Prion, P. G. M., Dollemans, A. M., Cuperus, C., Van, D. and Vlugt, R. A. A. (2010). Determination of aphid transmission efficiencies for N and NTN strains of potato viruses. Annual of Applied Biology, 56: 39-49. Wales, S., Platt, H. W. and Cattlin, N. (2008). Disease pest and disorder of potato. Color Handbook, 1st ed. Manson Publishing Ltd. London UK.
• Wang, B., Ma, Y. L., Zhang, Z. B., Wu, Z. M., Wu, Y. F., Wang, Q. C. and Li, M. F. (2011). Potato viruses in China. Crop Protection, 30: 1117-1123.
• Wright, G. C. and Bishop, G. W. (1981). Volunteer potatoes as source of potato leaf roll virus and potato virus X. American Journal of Potato Research, 58: 603-609.
• Wrobel, S. (2014). Modification of ELISA by replacing incubation of microtitre plate in an incubator with their shaking in PVY, PVM and PLRV detection. American Journal of Potato Research, 10: 1-7.
• Yardımcı, N., Handan, Ç. K. and Demir, Y. (2015). Detection of PVY, PVX, PVS, PVA, and PLRV on different potato varieties in Turkey using DAS-ELISA. Journal of Agriculture Science and Technology, 7: 757-764.