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تهیۀ گیاه توتون تراژن مصون به طیف گسترده‌ای از سویه‌های ویروس وای سیب‌زمینی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه گیاه‌پزشکی پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

2 بخش ویروس‌های گیاهی DSMZ آلمان، برانشوایگ

چکیده

ویروس وای سیب‌زمینی (Potato virus Y, PVY) از مهم‌ترین عوامل خسارت‌زا در مزارع توتون مناطق مختلف استان‌های گلستان، مازندران و گیلان است. با توجه به بی‌تأثیری حشره‌کش‌ها در کاهش آلودگی به PVY، شکسته شدن منابع طبیعی مقاومت توسط سویه‌های خاص و مشکلات روش‌های سنتی اصلاحی، به‌کارگیری روش‌های جایگزین ایجاد مقاومت به PVY در توتون با به‌کارگیری مقاومت مشتق‌شده از بیمارگر می‌تواند راهکاری برای کاهش خسارت در نظر گرفته شود. در این پژوهش گیاهان توتون تراژنی تولید شدند که در برابر طیف گسترده‌ای از جدایه‌های PVY از سویه‌های مختلف این ویروس مقاوم‌اند. ناحیه‌ای به اندازۀ ۴۷۲ جفت‌باز از ژنوم یک جدایۀ ایرانی PVY شامل توالی نوکلئوتیدی بخش‌هایی از CP و 3'UTR به منظور تهیۀ یک سازۀ سنجاق‌سری برای مقاومت به PVY به‌کار گرفته شد و تراژن‌سازی توتون رقم Wisconsin 38 با این سازه به کمک اگروباکتریوم انجام گرفت. ۶۱ درصد از گیاهان تراژن T0 حاصل به PVY مقاوم بودند و انتقال مقاومت به نسل بعد در 9 لاین مختلف با استفاده از روش داس الایزا تأیید شد. یکی از این لاین‌ها با ۱۲ جدایۀ مختلف PVY شامل چهار جدایۀ ایرانی و هشت جدایه از کشورهای دیگر تحت آزمایش قرار گرفت و مصونیت آن بر اساس عدم ظهور علائم و نتایج آزمون‌‌های داس الایزا و RT-PCR به تأیید رسید. این پژوهش نشان داد که سازۀ سنجاق‌سری به‌کاررفته دارای کارایی بسیار زیادی برای ایجاد توتون تراژن مصون در مقابل سویه‌های مختلف PVY بوده است.

کلیدواژه‌ها


عنوان مقاله [English]

Generation of transgenic tobacco plants with immunity against a broad spectrum of Potato virus Y strains

نویسندگان [English]

  • Hadi Khateri 1
  • Gholam-Hosein Mosahebi-Mohammadi 1
  • Stephan Winter 2
  • Mina Koohi-Habibi 1
  • Akbar Dizadji 1
1 Department of Plant Protection, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran.
2 Plant virus Department, German Collection of Microorganisms and Cell Cultures (DSMZ), Braunschweig, Germany.
چکیده [English]

Potato virus Y (PVY) is one of the most important plant viruses affecting tobacco fields in Golestan, Mazandaran and Guilan provinces (Iran). Given the lack of insecticide impact in reducing infection to PVY, breakdown of the natural resistance resources by certain strains and the problems with traditional breeding methods, the use of alternative methods to generate PVY resistant tobacco by pathogen-derived resistance could be considered as a method for reducing the damages. The objective of this study was to produce transgenic tobacco plants with resistance against a diverse spectrum of PVY isolates from different strains. A 472 bp fragment of the genome of an Iranian PVY isolate including the partial nucleotide sequences of CP and 3'UTR regions was used for constructing a hairpin structure for PVY resistance. This construct was used for Agrobacterium-mediated transformation of tobacco variety Wisconsin 38. Following transformation, 61% of the resulting transgenic plants in T0 were resistant to PVY and the inheritance of resistance to the next generation of  nine different lines was confirmed using DAS-ELISA which confirmed the immunity in all tested plants. One of these lines was tested against 12 different PVY isolates, including four Iranian and eight foreign ones, and the immunity against PVY was confirmed based on the lack of symptoms and the results of DAS-ELISA and RT-PCR tests. The results showed that the hairpin construct had a high-performance for generating transgenic tobacco plants with resistance against different PVY strains

کلیدواژه‌ها [English]

  • hairpin construct
  • broad-spectrum resistance
  • Agrobacterium-mediated transformation
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