Detection and Identification of Tomato Spotted Wilt Virus in Ornamental Plants in North Khorasan Province
محورهای موضوعی : مجله گیاهان زینتیJavad Mahmodi Safa 1 , Saeed Nasrolahnejad 2 , Mohamad Rezaei 3 , Farve Sadat Mostafavi Neishaburi 4
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
2 - Associate Professor, Department of Plant Protection, Gorgan University of Agriculture Sciences and Natural Resources, Gorgan, Iran
3 - M.Sc Graduated in Plant Pathology, Management of Plant Protection of North Khorasan
4 - Ph.D. Graduated in Plant Pathology, Zabol University
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.
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.
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