بهینه‌سازی استخراج آران‌ای دو رشته‌ای ویروسیاز برخی گونه‌های Fusarium
محورهای موضوعی : گیاه پزشکیداود کولیوند 1 , مهدی داوری 2 , نعمت سخندان بشیر 3 , مهدی ارزنلو 4
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کلید واژه: Biological control, کنترل بیولوژیک, استخراج, Hypersensitive, Fusarium head blight, dsRNA Extraction, کم‌آزاری, سوختگی فوزاریومی سنبله گندم, dsRNA,
چکیده مقاله :
اغلب ویروس های آلوده کننده ی قارچ های بیمارگر گیاهی که تا کنون گزارش شده اند، دارای ژنوم آران ای دو رشته ای (Double Strand RNA,dsRNA) می باشند. این نوع ویروس ها با توجه به خاصیت کم آزاری به عنوان یکی از عوامل کنترل بیولوژیکی قارچ ها مطرح هستند. در این تحقیق، هشت جدایه متعلق به پنج گونه Fusarium شامل F. graminearum s. str. (lin. 7)، F. culmorum، F. tricinctum، F. proloferatum و F. incarnatum برای وجود dsRNA مورد بررسی قرار گرفتند. با توجه به وجود پروتکل های متفاوت برای استخراج dsRNA که اغلب دارای مراحل پیچیده و استفاده از مواد خطرناک و سرطان زا می باشند، در روش مذکور از فنول و کلروفورم استفاده نشد و ضمن استفاده از میزان بافت کمتر، مدت زمان استخراج آران ای دورشته ای نسبت به سایر روش ها کاهش پیدا کرد. نهایتاً در چهار جدایه Fusarium پس از استخراج و الکتروفورز، وجود آران ای دو رشته ای مشخص شد و نتایج حاصله نشان داد روش مذکور یک روش مناسب و کارآمد برای استخراج dsRNA از بافت قارچی می باشد. همچنین در این تحقیق وجود آر.ان.ای دو رشته ای در گونه F. incarnatum (F. semitectum) برای اولین بار گزارش می شود.
The majority of viruses infected fungi which have been reported contain double-strand RNA (dsRNA) genomes. Mycoviruses are introducing as agent of the biological control of fungi according to hypersensitive properties. In this research, eight isolates belong to Fusariumgenus include ofF. graminearum, F. culmorum, F. tricinctum, F. proliferatum and F. incarnatum survived to dsRNA infection. Due to using of hazardous material such as phenol and complicated procedure in different dsRNA extraction protocols in mentioned research phenol and chlorophorm was removed , using less tissue and dsRNA extraction time was reduced compared to other methods. Result showed that four isolates infected by dsRNA also, mentioned method has sufficient efficiency and convenient for dsRNA extraction from fungi tissue. Also, dsRNA was reported fromF. incarnatum (F. semitectum) for first time.
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