Detection of Cucumber Mosaic Virus (CMV) in Cucurbits by TaqMan Real-Time RT-PCR in Hakkari Province, Türkiye

Yıl 2024, Cilt: 11 Sayı: 3, 825 – 834, 24.07.2024

Ayşenur Biltekin , Nevin Akdura

https://doi.org/10.30910/turkjans.1466706

Öz

Cucumber mosaic virus (CMV) is among the important viruses that can infect many crops. In 2022, this study was conducted in the Kırıkdağ, Otluca, and Merzan areas of the Hakkari province of Türkiye to investigate the presence of CMV in cucurbits exhibiting viral disease symptoms (yellowing, necrosis, mosaic, and deformity). A total of 50 samples; 7 watermelons (Citrullus lanatus), 3 melons (Cucumis melo L.), 17 cucumbers (Cucumis sativus L.), 8 zucchinis (Cucurbita pepo L.), 9 pumpkins (Cucurbita moschata), 2 gourds (Lagenaria siceraria) and 4 gherkins (Cucumis melo var. flexuous) were collected. The presence of CMV in the samples was investigated by TaqMan real-time RT-PCR using probes and primers specific to the 105 bp region of partial the coat protein gene of the virus. CMV-specific viral RNA was detected in 6 (12%) of the 50 samples analyzed. CMV was detected most commonly in the Otluca area and melon (33.3%) among cucurbit varieties. CMV agent was not detected in the samples collected from the Merzan area. The TaqMan real-time RT-PCR assay used in this study demonstrated that the method can be widely used for the general detection of CMV in a wide range of hosts. This is the first report on the detection of CMV in cucurbits in the Hakkari province of Türkiye.

Anahtar Kelimeler

Cucumber mosaic virus, cucurbits, TaqMan real-time RT-PCR, Hakkari

Proje Numarası

FM22LTP15

Kaynakça

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  • Bashir, N. S., Kalhor, M. R., Zarghani, S. N. 2006. Detection, differentiation and phylogenetic analysis of cucumber mosaic virus isolates from cucurbits in the northwest region of Iran. Virus Genes, 32(3): 277-288. https://doi.org/10.1007/s11262-005-6912-2
  • Borah, M., Gowtham, K. R., Rupsanatan, M., Palash, D. N. 2023. Molecular characterization and Efficacy Study of Different Treatments Against Cucumber mosaic virus in Pumpkin (Cucurbita moschata). Biological Forum-An International Journal, 15(2): 1053-1063.
  • Brunt, A. A., Crabtree, K., Dallwitz, M. J., Gibbs, A. J., Watson, L., Zurcher, E. 1996. Plant Virus Online. Descriptions and Lists from the Vide Database. http://biology.anu.edu.au/Groups/MES/vide/.
  • Can, S. 2019. Akdeniz bölgesinde örtü altı biber üretim alanlarında hıyar mozaik virüsü (Cucumber Mosaic Virus, CMV)’nün saptanması ve karakterizasyonu, Çukurova Üniversitesi, Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi, Adana.
  • Chomicki, G., Kiers, E. T., Renner, S. S. 2020. The evolution of mutualistic dependence. Annual Review of Ecology, Evolution, and Systematics, 51, 409-432. https://doi.org/10.1146/annurev-ecolsys-110218-024629
  • Clementi, M., Menzo, S., Bagnarelli, P., Manzin, A., Valenza, A., Varaldo, P.E. 1993. Quantitative PCR and RT-PCR in virology. PCR methods and applications, 2, 191-191.
  • Çat, A., Yardımcı, N., Çulal Kılıç, H. 2016. Antalya İli ve İlçelerindeki Örtüaltı Hıyar (Cucumis sativus L.) ve Kabak (Cucurbita pepo L.) Üretim Alanlarında Viral Etmenlerin Saptanması. Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 20(1): 129-132. DOI: 10.19113/sdufbed.53635
  • Daş, F. 2020. Adana ve Mersin illeri domates alanlarında hıyar mozaik virüsü (Cucumber mosaic virus, CMV) nün karakterizasyonu. Çukurova Üniversitesi, Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi, Adana. Doolittle, S. P. 1916. A new infectious mosaic disease of cucumber. Phytopathology, 6(2), 145-147.
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  • Grube, R. C., Zhang, Y., Murphy, J. F., Loaıza-Fıgueara, F., Lackney, V. K., Provvidenti, R., Jahn, M. K. 2000. New Source of Resistance to Cucumber mosaic virus in Capsicum Frutescens. Plant Disease, 84: 885-891. https://doi.org/10.1094/PDIS.2000.84.8.885
  • Güller, A., Usta, M. 2020. Occurrence of Cucumber mosaic cucumovirus and Watermelon mosaic potyvirus on Melon exhibiting viral symptoms in Bingöl province of Turkey and Their Phylogenetic Affinities. Türk Tarım ve Doğa Bilimleri Dergisi, 7(4): 948-958. https://doi.org/10.30910/turkjans.740195
  • Güller, A., Demirel, S., Usta, M., Korkmaz, G. 2024. Genetic Diversity of Cucumber mosaic virus in Cucumber Plants Grown in Diyarbakır. Turkish Journal of Nature and Science, 13(1):1–6.
  • Günay, A., Usta, M. 2020. First Investigation of Five Tobacco Viruses using Pcr Based Methods In Tobacco Plants Grown In Adıyaman Turkey. Fresenius Environmental Bulletin, 29, 11624–11632.
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Hakkari İlinde Kabakgillerde TaqMan Real-Time RT-PCR ile Hıyar Mozaik Virüsü (CMV)‘nün Tespiti

Yıl 2024, Cilt: 11 Sayı: 3, 825 – 834, 24.07.2024

Ayşenur Biltekin , Nevin Akdura

https://doi.org/10.30910/turkjans.1466706

Öz

Hıyar mozaik virüsü (Cucumber mosaic virus; CMV) birçok ürünü enfekte edebilen önemli virüsler arasındadır. Bu çalışma, 2022 yılında Hakkari ili Kırıkdağ, Otluca ve Merzan alanlarında viral hastalık belirtileri (sararma, nekroz, mozaik ve şekil bozukluğu) gösteren kabakgillerde CMV varlığının araştırılması amacı ile yapılmıştır. Toplam 50 örnek; 7 karpuz (Citrullus lanatus), 3 kavun (Cucumis melo L.), 17 hıyar (Cucumis sativus L.), 8 kabak (Cucurbita pepo), 9 kabak (Cucurbita moschata), 2 su kabağı (Lagenaria siceraria) ve 4 acur (Cucumis melo var. flexuous) toplanmıştır. Örneklerde CMV varlığı, virüsün kısmi kılıf protein geninin 105 bç bölgesine spesifik problar ve primerler kullanılarak TaqMan real-time RT-PCR ile araştırılmıştır. Analizi yapılan 50 örneğin 6’sında (%12) CMV spesifik viral RNA tespit edilmiştir. CMV en sık Otluca alanında ve kabakgil çeşitleri arasında da kavunda (%33.3) tespit edilmiştir. Merzan’dan toplanan örneklerde CMV etmenine rastlanmamıştır. Bu çalışmada kullanılan TaqMan real-time RT-PCR testi, CMV’nin genel tespiti için bu yöntemin çok çeşitli konukçularda yaygın olarak kullanılabileceğini göstermiştir. Bu çalışma Türkiye’nin Hakkari ilinde kabakgillerde CMV’nin tespitine ilişkin ilk rapordur.

Anahtar Kelimeler

Hıyar mozaik virüsü, kabakgiller, TaqMan real-time RT-PCR, Hakkari

Destekleyen Kurum

Hakkari Üniversitesi

Proje Numarası

FM22LTP15

Teşekkür

This study was written from the Master's thesis. The study was supported by a grant from the Scientific Research Projects Unit of Hakkari University, Türkiye.

Kaynakça

  • Abdalla, O. A., Desjardins, P. R., Dodds, J. A. 1991. Identification, disease incidence and distribution of viruses infecting peppers in California. Plant Disease, 75: 1019-1023.
  • Ali, A. K., Ahmed, I. A., Al-Kuwaiti, N. A. 2023. Molecular of Cucumber Mosaic Virus Subgroup IB in Iraq. Plant Protection, 7(2), 289-294.
  • Andres, T.C. 2004. The Cucurbit Network. http://www.cucurbit.org. http://www.cucurbit.org/family.html.
  • Arlı-Sökmen, M., Şevik, M. A. 2004. Reverse transkriptaz-polimeraz zincir reaksiyon (RT-PCR) yöntemi kullanılarak hıyar mozayik virüsü enfeksiyonunun belirlenmesi. Ondokuz Mayıs Üniversitesi Ziraat Fakültesi Dergisi, 19(3): 14-18. https://doi/full/10.5555/20043205625
  • Ayo-John, E.I., Olorunmaiye, P. M., Odedara, O.O., Dada, O. B., Abiola, K. O., Oladokun, J. O. 2014. Assessment of field-grown cucurbit crops and weeds within farms in South West Nigeria for viral diseases. Academic Pres, Cluj-Napoca, Romania, Notulae Scientia Biologicae, 6(3): 321-325. https://doi.org/10.15835/nsb639315
  • Bashir, N. S., Kalhor, M. R., Zarghani, S. N. 2006. Detection, differentiation and phylogenetic analysis of cucumber mosaic virus isolates from cucurbits in the northwest region of Iran. Virus Genes, 32(3): 277-288. https://doi.org/10.1007/s11262-005-6912-2
  • Borah, M., Gowtham, K. R., Rupsanatan, M., Palash, D. N. 2023. Molecular characterization and Efficacy Study of Different Treatments Against Cucumber mosaic virus in Pumpkin (Cucurbita moschata). Biological Forum-An International Journal, 15(2): 1053-1063.
  • Brunt, A. A., Crabtree, K., Dallwitz, M. J., Gibbs, A. J., Watson, L., Zurcher, E. 1996. Plant Virus Online. Descriptions and Lists from the Vide Database. http://biology.anu.edu.au/Groups/MES/vide/.
  • Can, S. 2019. Akdeniz bölgesinde örtü altı biber üretim alanlarında hıyar mozaik virüsü (Cucumber Mosaic Virus, CMV)’nün saptanması ve karakterizasyonu, Çukurova Üniversitesi, Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi, Adana.
  • Chomicki, G., Kiers, E. T., Renner, S. S. 2020. The evolution of mutualistic dependence. Annual Review of Ecology, Evolution, and Systematics, 51, 409-432. https://doi.org/10.1146/annurev-ecolsys-110218-024629
  • Clementi, M., Menzo, S., Bagnarelli, P., Manzin, A., Valenza, A., Varaldo, P.E. 1993. Quantitative PCR and RT-PCR in virology. PCR methods and applications, 2, 191-191.
  • Çat, A., Yardımcı, N., Çulal Kılıç, H. 2016. Antalya İli ve İlçelerindeki Örtüaltı Hıyar (Cucumis sativus L.) ve Kabak (Cucurbita pepo L.) Üretim Alanlarında Viral Etmenlerin Saptanması. Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 20(1): 129-132. DOI: 10.19113/sdufbed.53635
  • Daş, F. 2020. Adana ve Mersin illeri domates alanlarında hıyar mozaik virüsü (Cucumber mosaic virus, CMV) nün karakterizasyonu. Çukurova Üniversitesi, Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi, Adana. Doolittle, S. P. 1916. A new infectious mosaic disease of cucumber. Phytopathology, 6(2), 145-147.
  • Francki, R. I. B., Mossop D. W., Hatta, T. 1979. Cucumber mosaic virus. CMI/AAB Description Viruses No. 213: 6p.
  • Gallitelli, D. 2000. The ecology of Cucumber mosaic virus and sustainable agriculture. Virus Research, 71: 9-21. https://doi.org/10.1016/S0168-1702(00)00184-2
  • Grube, R. C., Zhang, Y., Murphy, J. F., Loaıza-Fıgueara, F., Lackney, V. K., Provvidenti, R., Jahn, M. K. 2000. New Source of Resistance to Cucumber mosaic virus in Capsicum Frutescens. Plant Disease, 84: 885-891. https://doi.org/10.1094/PDIS.2000.84.8.885
  • Güller, A., Usta, M. 2020. Occurrence of Cucumber mosaic cucumovirus and Watermelon mosaic potyvirus on Melon exhibiting viral symptoms in Bingöl province of Turkey and Their Phylogenetic Affinities. Türk Tarım ve Doğa Bilimleri Dergisi, 7(4): 948-958. https://doi.org/10.30910/turkjans.740195
  • Güller, A., Demirel, S., Usta, M., Korkmaz, G. 2024. Genetic Diversity of Cucumber mosaic virus in Cucumber Plants Grown in Diyarbakır. Turkish Journal of Nature and Science, 13(1):1–6.
  • Günay, A., Usta, M. 2020. First Investigation of Five Tobacco Viruses using Pcr Based Methods In Tobacco Plants Grown In Adıyaman Turkey. Fresenius Environmental Bulletin, 29, 11624–11632.
  • Henson, J. M., French, R. 1993. The polymerase chain reaction and plant disease diagnosis. Annual Rev. Phytopathology, 31: 81-109.
  • Harth, J. E., Simmons, H. E., Stephenson, A. G. 2017. Vertical infection of Zucchini yellow mosaic virus via pollen transmission occurs at a lower frequency than ovule transmission. European Journal of Plant Pathology, 147(3): 717-720. https://doi.org/10.1007/s10658-016-1024-5
  • Hsu, H. T., Barzuna, L., Hsu, Y. H., Bliss, W., Perry, K. L. 2000. Identification and Subgrouping of Cucumber Mosaic Virus with Mouse Monoclonal Antibodies. Phytopathology, 90, 615- 620. https://doi.org/10.1094/PHYTO.2000.90.6.615
  • Jacquemond, M. 2012. Cucumber Mosaic Virus, Advances in Virus Research, 84, 439–504. https://doi.org/10.1016/B978-0-12-394314-9.00013-0
  • Jeffrey, C. 2005. A new system of Cucurbitaceae. Botanicheskii Zhurnal, 90: 332-335.
  • Kaper J. M., Waterworth H. M. 1981. Cucumoviruses. Hand Book of Plant Virus Infections, 257-263.
  • Karanfil, A., Korkmaz, S. 2021. Güney Marmara Bölgesi kabakgil üretim alanlarında cucumber virus enfeksiyonunun tespiti ve kılıf protein gen diziliminin filogenetik analizi. Ege Üniversitesi Ziraat Fakültesi Dergisi, 58(2), 239-246. https://doi.org/10.20289/zfdergi.682293
  • Karanfil, A. 2022. Marmara Bölgesi Kabak Üretim Alanlarında Tobacco Mosaic Virus İzolatlarının Yaygınlığı ve Moleküler Karakterizasyonu. Journal of Advanced Research in Natural and Applied Sciences, 8(2), 163-170. https://doi.org/10.28979/jarnas.981619
  • Kaya, A., Erkan, S. 2011. İzmir, Aydın, Manisa ve Balıkesir İllerinde üretilen kabakgillerdeki viral etmenlerin tanılanması ve yaygınlıklarının belirlenmesi. Bitki Koruma Bülteni, 51(4): 387-405.
  • Kurtoğlu, A., Korkmaz, S. 2018. Çanakkale İli Ispanak Üretim Alanlarındaki Hıyar Mozaik Virüsü (Cucumber Mosaic Virus; CMV) Enfeksiyonunun Belirlenmesi ve Moleküler Karakterizasyonu. J. Turk. Phytopath., 47(2): 43-51.
  • Lecoq, H., Desbiez, C. 2012. Virus of cucurbit crops in the Mediterranean Region: an ever-changing picture. In: Loebenstein, G., Lecoq, H. (Eds.), Viruses and Virus Diseases of Vegetables in the Mediterranean Basin. Adv. Virus Res., 84. Elsevier, USA, 67-126. https://doi.org/10.1016/B978-0-12-394314-9.00003-8
  • Li, S., Wang, H. Huo, Z. Pang, J. 2004. Detection of main casual virus in cucumber by RT-PCR. Acta Agriculturae Boreali-Sinica, 19(3): 100-102.
  • Lira-Saade, R., Montes-Hernandez, M. 1994. Cucurbits (Cucurbita spp.). In J. Hernando Bermejo, J. Leon (Eds.), Neglected crops: 1492 from a different perspective, plant production and protection 26 (pp. 63–77). Rome, Italy: FAO.
  • Mackay, I. M. 2004. Real-time PCR in the microbiology laboratory. Clinical microbiology and infection, 10(3): 190-212.
  • Mando, M. J., Kasem, A. A. H., Al-Chaabi, S., Kumari, S. G., Turina, M. 2011. Survey of some mosaic viruses on cucurbits in Syria and molecular detection of Zucchini yellow mosaic virus. Arab Journal of Plant Protection, 29(1): 14-20.
  • Martelli, G. P., Quacquarelli, A. 1988. The Present Status of Tomato and Pepper Viruses. ACTA Horticulture, 127: 39-64. DOI: 10.17660/ActaHortic.1983.127.3
  • Mnari-Hattab, M., Jebari, H., Zouba, A. 2008. Identification and distribution of viruses responsible for mosaic diseases affecting cucurbits in Tunisia. Bulletin OEPP/EPPO, 38(3): 497-506. DOI: 10.1111/j.1365-2338.2008.01269.x
  • Nagendran, K., Mohankumar, S., Aravintharaj, R., Balaji, C. G., Manorenjitham, S. K., Singh, A. K., Rai, A. B., Singh, B., Karthikeyan, G. 2017. The occurrence and distribution of major viruses infecting cucurbits in Tamil Nadu State, India. Crop Protection, 99, 10-16.
  • Neergaard, P. 1977. Seed-borne viruses. Chapter 3 in "seed Pathology, vol. I." Macmillan Press, London, and Madras, 839 p. https://doi.org/10.1016/j.cropro.2017.05.006
  • Nogay, A., Yorgancı, Ü. 1984. Investigations on the identification, seed transmission and host range of viruses infecting the culture plants in the Cucurbitaceae in Marmara region. 1. The identification of viruses infecting cucurbits in Marmara region. Journal of Turkish Phytopathology, 13(1): 9-27.
  • Nogay, A., Yorgancı, Ü. 1985. Investigations on the identification, seed transmission, and host range of viruses infecting the culture plants in the Cucurbitaceae in Marmara region. 2- The seed transmissibilities and cucurbit hosts of CMV and WMV-2 isolated from the culture plants in the Cucurbitaceae. Journal of Turkish Phytopathology, 14(1): 9-16.
  • Papan, A. N. 2022. Bandırma yöresinde salçalık domates yetiştirilen alanlarda Hıyar mozaik virüsü (Cucumber mosaic virus, CMV)’ nün saptanması ve karakterizasyonu. Çukurova Üniversitesi, Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi, 96s, Adana.
  • Omar, S. M., Fadel, S. F. 2022. Molecular Characterization of Cucumber mosaic virus (CMV) local isolates in Libya. Libyan Journal of Basic Sciences (LJBS), 18: 93-105.
  • Palukaitis, P., Rossinck, M. J., Dietzgen, R. G., Francki, R. I. B. 1992. Cucumber mosaic virus. Advances in Virus Research, 41: 281-348. DOI: 10.1016/s0065-3527(08)60039-1
  • Raccah, B., Galon, A., Eastop, V. F. 1985. The role of flying aphid vectors in the transmission of cucumber mosaic virus and potato virus to peppers in Israel. Ann. Appl. Biol., 106: 451-460. https://doi.org/10.1111/j.1744-7348.1985.tb03135.x
  • Roossinck, M. J. 2001. Cucumber mosaic virus, a model for RNA virus evolution. Molecular Plant Pathology, 22: 59-63. https://doi.org/10.1046/j.1364-3703.2001.00058.x
  • Sharma, S. 2023. Domateste Cucumber mosaic virus (CMV)’un Biyolojik ve Moleküler Karakterizasyonu. Ege Üniversitesi, Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi, İzmir.
  • Shehata, W. F., Iqbal, Z., Abdelbaset, T. E., Saker, K. I., El Shorbagy, A. E., Soliman, A. M., Sattar, M. N., El-Ganainy, S. M. 2023. Identification of a Cucumber Mosaic Virus from Cucurbita pepo on New Reclamation Land in Egypt and the Changes Induced in Pumpkin Plants. Sustainability, 15(12): 9751. https://doi.org/10.3390/su15129751
  • Silveira, L. M., Queiroz, M. A., Lima, J. A. A., Nascimento, A. K. Q., Lima Neto, I. S. 2009. Serological survey of virus in cucurbit species in the Lower Middle São Francisco River Basin, Brazil. Tropical Plant Pathology, 34(2): 123-126. https://doi.org/10.1590/S1982-56762009000200008
  • Simson, M. G. 2006. Plant Systematics. Elsevier Academic Pres., ISBN: 978-0-12-644460-9, 599. California, USA.
  • Soylu, B. 2014. Çanakkale ili Ev Bahçelerinde Yetiştirilen Soğanlı Süs Bitkilerinde Hıyar Mozaik Virüsü’nün Serolojik ve Moleküler teşhisi. Çannakkel Onsekiz Mart Üniversitesi, Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi, Çanakkale.
  • Suresh, L. M., Malathi, V. G., Shivanna, M. B. 2013. Serological diagnosis and host range studies of important viral diseases of a few cucurbitaceous crops in Maharashtra, India. Archives of Phytopathology and Plant Protection, 46(16): 1919-1930. https://doi.org/10.1080/03235408.2013.780841
  • Tomlinson, J. A., Carter, A. L. 1970. Studies on the seed transmission of cucumber mosaic virus in chickweed (Steleria media) in relation to the ecology of the virus. Annals of Applied Biology, 73: 293-298. https://doi.org/10.1111/j.1744-7348.1970.tb04617.x
  • Uzunoğulları, N., Gümüş, M. 2015. Marmara Bölgesi’nde bazı kültür bitkilerinde doğal enfeksiyona neden olan Hıyar mozaik virüsü (Cucumber mosaic virus, CMV)’nün tespiti. Trakya University Journal of Natural Sciences, 16(1): 9-15.
  • Vučurović, A., Bulajic,´A., Stankovic´, I., Ristic´, D., Berenji, J., Jovic´, J., Krstic´, B. 2012. Nonpersistently aphid-borne viruses infecting pumpkin and squash in Serbia and partial characterization of Zucchini yellow mosaic virus isolates. European Journal of Plant Pathology, 133(4): 935-947. https://doi.org/10.1007/s10658-012-9964-x
  • Yeşil, S. 2013. Konya, Karaman ve Aksaray illeri kabakgil ekim alanlarında görülen virüs hastalıklarının serolojik ve moleküler yöntemlerle tespiti ve enfeksiyon kaynaklarının belirlenmesi. Selçuk Üniversitesi, Fen Bilimleri Enstitüsü, Doktora Tezi, Konya.
  • Xu, L., Song, M., Ming, J. 2022. Application of Multiplex TaqMan Real-Time PCR Assay in Survey of Five Lily Viruses Infecting Lilium spp. Agronomy, 12, 47. https://doi.org/10.3390/agronomy12010047
  • Zitter, T. A., Murphy, J. F. 2009. Cucumber mosaic. The Plant Health Instructor. DOI: 10.1094/PHI-I-2009-0518-01.

Toplam 57 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Fitopatoloji
BölümAraştırma Makalesi
Yazarlar

Ayşenur Biltekin HAKKARİ ÜNİVERSİTESİ 0000-0003-3076-2463 Türkiye

Nevin Akdura Hakkari Üniversitesi 0000-0001-6162-0500 Türkiye

Proje NumarasıFM22LTP15
Yayımlanma Tarihi24 Temmuz 2024
Gönderilme Tarihi8 Nisan 2024
Kabul Tarihi24 Haziran 2024
Yayımlandığı Sayı Yıl 2024 Cilt: 11 Sayı: 3

Kaynak Göster

APABiltekin, A., & Akdura, N. (2024). Detection of Cucumber Mosaic Virus (CMV) in Cucurbits by TaqMan Real-Time RT-PCR in Hakkari Province, Türkiye. Türk Tarım Ve Doğa Bilimleri Dergisi, 11(3), 825-834. https://doi.org/10.30910/turkjans.1466706

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