Detection and Identification of Tomato Spotted Wilt Virus in Ornamental Plants in North Khorasan Province

محورهای موضوعی : مجله گیاهان زینتی

Javad Mahmodi Safa ¹ (Ph.D. Student, Department of Entomology and Plant Pathology, University of Tehran, College of Agriculture and Natural Resources, Faculty of Agricultural Technology, Tehran, Iran)
Saeed Nasrolahnejad ² (Associate Professor, Department of Plant Protection, Gorgan University of Agriculture Sciences and Natural Resources, Gorgan, Iran)
Mohamad Rezaei ³ (M.Sc Graduated in Plant Pathology, Management of Plant Protection of North Khorasan)
Farve Sadat Mostafavi Neishaburi ⁴ (Ph.D. Graduated in Plant Pathology, Zabol University)

تاریخ دریافت : 1402/07/08 تاریخ پذیرش : 1402/08/30 تاریخ انتشار : 1402/09/10

کلید واژه: ornamental plants, RT-PCR, DAS-ELISA, Mechanical inoculation, Viral diseases,

چکیده مقاله :

In recent years, the symptoms of viral diseases such as dwarfism, mosaic, discoloration, necrosis, and circular spots have been prevalent in ornamental plants in parks, gardens, and streets in North Khorasan Province. Tomato spotted wilt virus (TSWV), the type species of the genus Orthotospovirus, the family Tospoviridae, and the order Bunyavirales, is a tripartite negative/ambisense ssRNA virus. This virus is a major cause of the infection of ornamental plants in the world. In this study, to evaluate the percentage of infection to TSWV, in the early autumn of 2020, 350 samples of ornamental plants were collected based on the suspicious viral symptoms from parks, gardens, and streets of North Khorasan province (Bojnord and Shirvan cities) and were transported to the laboratory in cold conditions. In the laboratory, DAS-ELISA serological test was performed to evaluate the presence of virus in the suspected samples. Then, the infected samples identified by ELISA test were inoculated to the test plants of Chenopodium album L. (lamb's quarters), Vigna unguiculata L. (cowpea) and Datura stramonium L. (jimsonweed). After the appearance of symptoms, in order to verify the infection, they were tested again using DAS-ELISA. The molecular identification of the infected samples was done by Qiagene RNA extraction kit. Using specific primers in RT-PCR reaction, a fragment was amplified in the band of 276 bp. The results of DAS-ELISA and RT-PCR tests proved the presence of the virus on ornamental plants in North Khorasan province. In the mechanical inoculation of the virus, three above-mentioned plants showed the symptoms of the disease.

چکیده انگلیسی:

در سال‌های اخیر علائم بیماری‌های ویروسی مانند کوتولگی، موزائیک، رنگ پریدگی، بافت مردگی و لکه‌های حلقوی روی گیاهان زینتی در پارک‌ها، بوستان‌ها و معابر استان خراسان شمالی شیوع پیدا کرده است. ویروس پژمردگی لکه-ای گوجه‌فرنگی (TSWV) ، گونه‌ای از جنس Orthotospovirus، خانواده Tospoviridae ، راسته Bunyavirales، با ژنوم سه قسمتی (negative/ambisense) و ویروس دارای ژنوم تک رشته‌ای RNA است. این ویروس عامل اصلی آلودگی گیاهان زینتی در جهان است. در این مطالعه به منظور بررسی درصد آلودگی به TSWV، در اوایل پاییز ۱۳۹۹، ۳۵۰ نمونه گیاه زینتی بر اساس نشانه‌های مشکوک به علائم ویروسی از پارک‌ها، بوستان‌ها و معابر استان خراسان شمالی (شهرستان‌های بجنورد و شیروان) جمع آوری شد و در شرایط خنک به آزمایشگاه منتقل گردید. در آزمایشگاه به منظور ارزیابی وجود ویروس در نمونه‌های مشکوک،‌ آزمایش سرولوژیکی DAS-ELISA انجام شد و نمونه‌های آلوده به ویروس که در آزمون الایزا مشخص شدند به گیاهان محک Chenopodium album ، Vigna unguiculata و Datura stramonium مایه‌زنی گردیدند و پس از ظهور علائم به منظور تایید آلودگی مجددا با آزمون DAS-ELISA آزمایش شدند. در شناسایی مولکولی نمونه های آلوده به ویروس با استفاده از کیت کیاژن استخراج RNA صورت گرفت و با استفاده از جفت آغازگرهای اختصاصی در واکنش RT-PCR قطعه ای در محدوده باندی bp ۲۷۶ تکثیر یافت. نتایج حاصل از آزمون DAS-ELISA و RT-PCR وجود ویروس را در گیاهان زینتی استان خراسان شمالی به اثبات رساند. در مایه‌زنی مکانیکی ویروس مورد مطالعه هر سه گیاه محک فوق علائم مشخص بیماری را نشان دادند.

منابع و مأخذ:

Anonymous. 2018. Agricultural statistics 2017. Agriculture Jihad Publication, Tehran, Iran. (In Persian)

Anonymous. 2021. International statistics flowers and plants. Edited by A.                           Schubach. Voorhout, Netherlands. www.aiph.org

Ayazpour, K. 2014. Alphabetic list of plant viruses and viroids reported from Iran. Islamic Azad University Press, Jahrom, Iran. (In Persian)

Clark, M.F. and Adams, A.N. 1977. Characteristics of the micro plate method of enzyme-linked immunosrobent assay for the detection of plant viruses. Journal of General Virology, 34: 475- 483.

Farzadfar, Sh., Golnaraghi, A.R. and Pourrahim, R. 2002. Plant viruses of Iran. Saman Company, Tehran, Iran, p. 203. (In Persian)

Ghotbi, T. and Shahraeen, N. 2012. Incidence and distribution of viruses infecting propagated ornamentals in Northern Iran. International Research Journal of Microbiology, 3(11): 373-381.

Golnaraghi, A.R., Pourrahim, R., Ahoonmanesh, A., Zamani-Zadeh, H. R. and Farzadfar, Sh. 2008. Detection and characterization of a distinct isolate of tomato yellow fruit ring virus from potato. Plant Disease, 92(9): 1280-1287.

Gumus, M. and Paylan, I.C. 2013. Detection of viruses in seeds of some vegetables by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). African Journal of Biotechnology, 12 (25): 3891-3897.

Loebenstein, G., Lawson, R.H. and Brunt, A.A. 1995. Virus and virus-like disease of bulb and flower crops. Wiley and Sons Publisher. 543 pp.

Mahmoodi Safa, J., Nasrolahnejad, S. and Mostafavi Neishabouri, F. 2016. Detection and identification of tomato ring spot virus in ornamental plants in North Khorasan province. Journal of Ornamental Plants, 6(3): 141-150.

Mahmoodi Safa, J., Nasrolahnejad, S., Rezai, M. and Mostafavy Neyshabouri, F.  2015. Serological survey tomato ring spot virus (ToRSV) on ornamental plants in North Khorasan. Proc. 1st Annual Iranian Agriculture Research Conference, Shiraz, Iran, 22 July. pp. 726-729. (In Persian)

Massumi, H., Shaabanian, M., Hosseini Pour, A., Heydarnejad, J. and Rahimian, H. 2009. Incidence of viruses infecting tomato and their natural hosts in the south east and central regions of Iran. Plant Disease, 93: 67-72.

Mathews, D.M. 2010. Optimizing detection and management of virus diseases of plants (1) and Emerging tree diseases in Southern California (2). In: Proceedings of the Landscape Disease Symposium, Camarillo, CA, 10-20 pp.

Morris, J. 2004. Protocol for the diagnosis of quarantine organisms tomato spotted wilt virus, impatiens necrotic spot virus and watermelon silver mottle virus. OEPP/EPPO Bulletin, 34: 271-279.

Mostashar Nezami, I., Mira, A. and Nikookar, Gh. 2013. Explanation of factors affecting the export of flowers in Iran: A case study in Alborz province. Journal of Natural and Social Sciences, 2 (3): 3217- 3227.

Oliver, J.E. and Whitfield, A.E. 2016. The genus tospovirus: Emerging bunyaviruses that threaten food security. Annual Review of Virology, 3: 101–124.

OPRC. 2023. Flower and plant industry of Iran: Production of flowers and ornamental plants. Mahallat. www.oprc.hsri.ac.ir

Pappu, H.R., Jones, R.A. and Jain, R.K. 2009. Global status of tospovirus epidemics in diverse cropping systems: Successes achieved and challenges ahead. Virus Research, 141: 219–236.

Shoshtari, S., Jafarpour, B. and Falahati-Rastegar, M. 2013. Molecular and serological detection of tomato spotted wilt virus from Khorasan Razavi. Plant Protection, 26: 348-356. (In Persian)

Wangai, A. W., Mandal, B., Pappu, H. R. and Kilonzo, S. 2001. Outbreak of Tomato spotted wilt virus in Tomato in Kenya. Plant Disease, 85 (10): 1123. doi:10.1094/PDIS. 2001.85.10.1123B.

Zhang, W., Jiao, Y., Ding, C., Shen, L., Li, Y., Yu, Y., Huang, K., Li, B., Wang, F. and Yang, J. 2021. Rapid detection of tomato spotted wilt virus with Cas13a in tomato and Frankliniella occidentalis. Frontiers in Microbiology, 12: 745173.