اثر سرکوبگر‌های خاموشی ویروس موزائیک زرد راه راه جو (Barley yellow striate mosaic virus) بر میزان بیان ژن‌های مرتبط با اتوفاژی در گیاهNicotiana benthamiana 16c

نوع مقاله: مقاله کامل پژوهشی

نویسندگان

چکیده

اتوفاژی یک فرایند محافظت شده در یوکاریوت‌ها برای حذف اجزای سلولی آسیب دیده یا نا خواسته است. این مسیر در مقاومت به بیماری‌های گیاهی نیز دخیل می‌باشد اما مکانیزم آن دقیقا مشخص نیست. در این مطالعه تاثیر دو پروتیین سرکوبگر (:Pفسفوپروتیین و :P3پروتیین فرعی 3) متعلق به ویروس موزائیک زرد راه راه جو (BYSMV) بر بیان چهار ژن مهم دخیل در اتوفاژی شامل ATG2، ATG6، ATG7 و AGO1 در گیاه N. benthamiana مورد بررسی قرار گرفت. ژن‌های P و P3 BYSMV در ناقل pCAMBIA-1302 تحت پروموتور 2×35S و برچسبHemagglutinin در انتهای آمینی به منظور ردیابی پروتیین‌‌ها، همسانه‌سازی شدند. سازه‌ هر ژن با استفاده از اگروباکتریوم در سطح پشت برگ گیاهان N. benthamiana تزریق گردید. پنج روز پس از تزریق، میزان بیان این ژن‌ها با استفاده از پی‌سی‌آر در زمان واقعی اندازه‌گیری شد. نتایج نشان داد که هر سه تیمار(P, P3, P+P3) منجر به افزایش بیان ژن‌های ATG2، ATG6 و ATG7 گردیدند. این افزایش بیان در ژن ATG2 به ترتیب 57/5، 15/6 و 26/5 برابر در تیمار‌هایP/GFP ، P3/GFP و P/P3/GFP بود. در حالی که هر سه تیمار بر بیان ژن AGO1 اثر منفی داشتند بنحوی که بیان آن را تقریبا 5/1 برابر، نسبت به تیمار کنترل، کاهش دادند. این نتایج حاکی از نقش ژن‌های AGO1، ATG6،ATG7 و ATG2 در پاسخ دفاعی گیاه N. benthamiana در مقابل سرکوبگر BYSMV می‌باشد که می‌تواند به درک بهتر مکانیز‌م‌های دفاعی پیچیده گیاه در مقابل بیمارگر‌های ویروسی و دست یابی به روش‌های جدید کنترل ویروس‌های گیاهی کمک نماید.

کلیدواژه‌ها


عنوان مقاله [English]

Effect of RNA silencing suppressors of Barley yellow striate mosaic virus on expression levels of autophagy-related genes in Nicotiana benthamiana16c

نویسندگان [English]

  • S. Rabiee
  • A. Afsharifar
  • K. Izadpanah
چکیده [English]

Autophagy is a degradation process in eukaryotes through which damaged or unwanted intracellular components are degraded. This process is also involved in plant disease resistance, although its mechanisms are not precisely known. Autophagy is regulated by multiple autophagy-related proteins (ATGs).In this study, we investigated the possible impact of two Barley yellow striate mosaic virus (BYSMV) proteins, i.e., phosphoprotein (P) and ancillary protein 3 (P3), on four important genes involved in autophagy (ATG2, ATG6, ATG7, and AGO1) in N. benthamiana. P and P3 genes were cloned in pCAMBIA-1302 vector under the control of the 2 × 35S promoter and Hemagglutinin tag. Constructs of each gene were agroinfiltrated in the abaxial side of the N. benthamiana leaves. Five days after agroinfiltration, the expression level of these genes was measured using RT- qPCR. The results showed that expression of ATG2, ATG6 and ATG7 genes increased in all treatments (P, P3, P+P3).The level of ATG2 expression was 5.57, 15.6 and 5.6 fold, in P/GFP, P3/GFP and P/P3/GFP treatments, respectively, while a 1.5-fold reduction was obtained in expression of AGO1 in all treatments. These results implied that P and P3 proteins of BYSMV can modify the expression of autophagy related genes in N. benthamiana plant. These findings suggest the involvement of AGO1, ATG6, ATG7 and ATG2 in immune responses of N. benthamiana against BYSMV, which provide a better understanding of plant host defense mechanisms against virus infections and might be an opportunity to exploit a novel antivirus approach.

کلیدواژه‌ها [English]

  • autophagy
  • Phosphoprotein
  • RNA silencing suppressor
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