Detection and Identification of Tomato Ring Spot Virus in Ornamental Plants in North Khorasan Province
محورهای موضوعی : مجله گیاهان زینتی
Javad Mahmoodi Safa
1
(Department of Plant Protection, Faculty of Plant Production, Gorgan University of Agriculture Sciences and Natural Resources, Gorgan, Iran.)
Saeed Nasrolahnejad
2
(Department of Plant Protection, Faculty of Plant Production, Gorgan University of Agriculture Sciences and Natural Resources, Gorgan, Iran)
Farveh Sadat Mostafavi Neishaburi
3
(Department of Plant Protection, Faculty of Agriculture, University of Zabol, Iran.)
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 ring spot virus (ToRSV) belongs to the Nepovirus genus of Secoviridae family. 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 ToRSV, in the early autumn of 2014, 400 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 Cucumis sativus L. (Cucumber), Vigna unguiculata L. (Cowpea) and Nicotiana glutinosa L. (Tobacco). 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 449bp. 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 لکه حلقوی گوجه­فرنگی Tomato ring spot virus(ToRSV) متعلق به جنس Nepovirusو خانوادهSecoviridaeمی­باشد. این عامل ویروسی یکی از عوامل اصلی آلوده کننده گل­های زینتی در جهان محسوب می­شود. در این تحقیق جهت بررسی درصد آلودگی ویروس ToRSV، در اوایل پاییز سال 1393 مجموعا 400 نمونه گیاه زینتی به صورت کاملا تصادفی و بدون در نظر گرفتن علائم از پارک­ها، بوستان­ها و معابر استان خراسان شمالی (شهرستان­های بجنورد و شیروان) جمع­آوری شد و در شرایط خنک به آزمایشگاه منتقل گردید. در آزمایشگاه به منظور ارزیابی وجود ویروس در نمونه­های مشکوک،­ آزمون سرولوژیکی DAS-ELISA انجام شد و نمونه­های آلوده به ویروس که در آزمون الایزا مشخص شدند به گیاهان محک Cucumis sativus، Vigna unguiculata،Nicotiana glutinousa وNicotiana rusticaمایه­زنی گردیدند. پس از ظهور علائم، گیاهان محک مجددا برای تایید وجود آلودگی توسط آزمون DAS-ELISA تست شدند. شناسایی مولکولی گیاهان آلوده توسط کیت استخراج RNA (کیاژن) انجام شد و با استفاده از آغازگرهای اختصاصی ویروس در واکنش RT-PCR، یک قطعه 449bp تکثیر شد. نتایج آزمون الایزا و واکنش RT-PCR حضور ویروس را در گیاهان زینتی استان خراسان شمالی تایید کرد. در مایه­زنی مکانیکی ویروس هر سه گیاه محک فوق علائم بیماری را نشان دادند.
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