تاثیر پروتئین ضد ویروسی گیاه لاله عباسی (MAP) در آلودگی های ناشی از ویروس موزاییک خیار (CMV) و ویروس Y سیب زمینی (PVY) در گیاه Nicotiana benthamiana در شرایط گلخانه

نوع مقاله : مقاله پژوهشی

نویسندگان

گروه گیاه پزشکی، دانشکده علوم کشاورزی و صنایع غذایی، واحد علوم و تحقیقات دانشگاه آزاد اسلامی، تهران

چکیده

چکیده
در این تحقیق تلاش شد تا با استفاده از پروتئین ضد ویروسی گیاه لاله عباسی Mirabilis Antiviral Protein (MAP) در شرایط گلخانه علاوه بر کنترل (CMV) Cucumber mosaic virus و (PVY) Potato virus Y ، مکانیسم کنترل کنندگی آن نیز مورد بررسی قرار گیرد. برای این منظور ابتدا MAP از عصاره­ های برگ و ریشه گیاه لاله عباسی جداسازی شد و سپس گیاهچه­ های Nicotiana benthamiana در گلدان­های 50 گرمی کاشته و در مرحله چهار تا شش برگی بصورت مکانیکی توسط ویروس­های CMV-Fny (Acc.no. D00356.1) و PVYº-UK (Acc.no. KY112748) مایه­ کوبی شدند. همچنین مایه­ کوبی PVYº با شته به‌­طور مجزا انجام شد. دو روز قبل و دو روز بعد از مایه ­کوبی گیاهچه ­های گیاه N. benthamiana، MAP به صورت مستقیم و به مقدار µg300 به محیط ریزوسفر در گلدان‌ها اضافه شد. آلودگی گیاه­ های N. benthamiana با هریک از ویروس­ها توسط آزمون Quantitative -ELISA و میزان نسخه‌های ژنی PVYº توسط آزمون Real-time RT-PCR تا 14 روز بعد از مایه­ کوبی مورد ارزیابی قرار گرفت. نتایج آزمون الایزا نشان داد تیمار گیاهچه‌های N. benthamiana با MAP باعث کاهش معنی ­دار تیتر هر یک از ویروس­ها تا 14 روز پس از تیمار شده است. نتایج Real-time PCR بیانگر کاهش میزان حضور نسخه‌های ژنوم PVYº تا ۲/۵ برابر در مقایسه با شاهد آلوده بود. همچنین مشخص شد پروتئینMAP به میزان بیشتری باعث کنترل PVYº نسبت به CMV-Fny می شود. نتایج آزمون انتقال با ناقل حاکی از آن بود که MAP موجب کاهش انتقال PVYº با شته نمی ­شود. همچنین وزن­های تر و خشک اندام‌های مختلف و نیز مقدار فنل در گیاهچه ­های N. benthamiana تیمار شده با MAP افزایش نشان دادند.

کلیدواژه‌ها


عنوان مقاله [English]

The effect of mirabilis antiviral protein (MAP) in infections by Cucumber mosaic virus (CMV) and Potato virus Y (PVY) in Nicotiana benthamiana plants under greenhouse condition

نویسندگان [English]

  • Farshad Rakhshandehroo
  • Zahra Pazooki
  • Saeed Rezaee
Department of Plant Protection, College of Agricultural Sciences and Food Industries, Science and Research Branch, Islamic Azad University, Tehran, Iran.
چکیده [English]

Abstract
In this study using with the Mirabilis antiviral protein (MAP), it was attempted to explore the MAP controlling effect mechanism as well its antiviral activity against the Cucumber mosaic virus-Fny (CMV) and Potato virus Y strain O (PVYº) under the greenhouse condition. Toward this aim, MAP was first isolated from different fractions of the crude extracts mixture of the leaves and roots of the four o'clock plant and then Nicotiana benthamiana seedlings were cultivated in 50 g pods and mechanically inoculated with the PVYO-UK (Acc.no. KY112748) and CMV-Fny (Acc.no. D00356.1) at 4 to 6 leaf stages. PVYO inoculation was also separately done by the aphid. MAP was administrated directly into the rhizosphere area of the pods at the concentration of 300 µg in two days after and two days before seedling inoculations. Infection with each of the inoculated viruses in MAP treated benthamiana plant seedlings were then assessed by DAS-ELISA, 14 days after inoculations. Using with the Real-time RT-PCR, PVYO transcript levels was assessed at this time. Results showed that soil treatments of the benthamiana plant seedling with the MAP significantly decreased the titre of the tested viruses up to 14 days after treatments. Results of the Real-time RT-PCR indicated to the significant decrease in PVYO transcript level compared with the infected control plant. It was also revealed that MAP is more effective in controlling PVYO compared to CMV-Fny. Results indicated that MAP has not inhibited the PVYO transmission by the aphid vector. The dry and wet weights and the phenol content were also increased in benthamiana plants treated with the MAP.

کلیدواژه‌ها [English]

  • Keywords: Antiviral Activity
  • Morphological Factor
  • MAP
  • Mirabilis jalapa
  • Ribosome Inactivating Protein
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