Detection and Identification of Tomato Ring Spot Virus in Ornamental Plants in North Khorasan Province
محورهای موضوعی : مجله گیاهان زینتیJavad Mahmoodi Safa 1 , Saeed Nasrolahnejad 2 , Farveh Sadat Mostafavi Neishaburi 3
1 - Department of Plant Protection, Faculty of Plant Production, Gorgan University of Agriculture Sciences and Natural Resources, Gorgan, Iran.
2 - Department of Plant Protection, Faculty of Plant Production, Gorgan University of Agriculture Sciences and Natural Resources, Gorgan, Iran
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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