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کارایی بخشی از ژنوم ویروس موزاییک شلغم برای ایجاد مقاومت در کلزا

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی سابق دکتری، گروه بیماری شناسی گیاهی، دانشکده کشاورزی، دانشگاه تربیت مدرس، تهران، ایران

2 دانشیار، گروه اصلاح نباتات، دانشکده کشاورزی، دانشگاه تربیت مدرس، تهران، ایران

3 استاد، دانشکده کشاورزی، دانشگاه تربیت مدرس، تهران، ایران

چکیده

ویروس موزاییک شلغم (Turnip mosaic virus, TuMV) متعلق به جنس Potyvirus و خانواده Potyviridae، یکی از ویروس‌های‏ شایع مزرعه‏های کلزا در ایران است و تاکنون لاین یا رقم تجاری مقاومی به آن در کشور گزارش نشده است. در این پژوهش، امکان ایجاد مقاومت به TuMV در لاین کلزای تراریختۀ حاوی قطعه کوتاهی از ژن رمزکنندۀ‏ پروتئین پوششی ویروس بررسی شد. بدین منظور توالی 130 نوکلئوتیدی از ژن رمزکنندۀ‏ پروتئین پوششی در حامل pFGC5941 در جهت‏های سنس و آنتی‌سنس‏ به‌طور مستقل همسانه­سازی‏ شد و با استفاده از Agrobacterium tumefaciens سویۀ LBA4404 به دم‏برگ کوتیلدونی لاین کلزای Hayola R-Line 401 منتقل شد. گزینش و تأیید گیاهان تراریختۀ T0 و T1 با استفاده از آزمون‌های‏ PCR و مقاومت به علف‌کش‏ گلوفوسینت آمونیوم (نشانگر) در شرایط گلخانه انجام شد. پس از مایه‌زنی‏ ویروس به گیاهان لاین T1 ارزیابی واکنش با استفاده از سامانۀ نمره‌دهی‏ انجام شد. نتایج نشان داد که تمام گیاهان تراریخته حاوی سازۀ سنس هیچ‌گونه مقاومتی را نشان ندادند درحالی‌که تعدادی از گیاهان تراریختۀ حاوی سازۀ آنتی‏سنس سطوح متفاوتی از مقاومت شامل تأخیر در ظهور علائم به همراه کاهش شدت علائم (0 تا 30 درصد نتاج نسل T1) و یا بهبودی (10 تا 66 درصد نتاج نسل T1) را نشان دادند. این اولین گزارش از افزایش مقاومت به TuMV از راه انتقال یک توالی کوتاه ویروسی در جهت آنتی‌سنس‏ در کلزا است.

کلیدواژه‌ها


عنوان مقاله [English]

The efficiency of the partial genome of Turnip mosaic virus (TuMV) on resistance induction in canola

نویسندگان [English]

  • Majid Jafari 1
  • Ahmad Moieni 2
  • Masoud Shams-bakhsh 3
1 Former Ph. D. Student, Plant Pathology Department, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
2 Associate Professor, Plant Breeding Department, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
3 Professor, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
چکیده [English]

Turnip mosaic virus (TuMV) a species of the genus Potyvirus in the family Potyviridae, is one of the most prevalent viruses of canola fields in Iran. So far, commercial resistant lines or varieties against TuMV have not been known in Iran. In this research, the possibility of inducing resistant to TuMV in transgenic canola lines carrying a short fragment of the coat protein gene of the virus was evaluated. Therefore, a 130 nucleotide fragment of TuMV coat protein gene, as sense or antisense orientations, were cloned in the pFGC5941 vector independently and then were transformed to canola cotyledonary (Hayola R-Line 401 variety) explants using Agrobacterium tumefaciens strain LBA4404. Selection and confirmation of transgenic T0 and T1 lines were carried out via PCR and glufosinate ammonium herbicide (selective marker) resistance assay under greenhouse conditions. After virus inoculation to T1 lines, resistance assay was done by the scoring system. The obtained results confirmed that transgenic lines with sense construction did not show any resistant. However, several transgenic lines with antisense construction showed a different level of resistant to TuMV including delay in symptoms appearance and a decrease in symptoms severity (0-30% of T1 progeny) or symptoms recovery (10-66% of T1 progeny). This is the first report of increased resistant against TuMV via transformation of a short viral fragment sequence with the antisense orientation in canola.

کلیدواژه‌ها [English]

  • Potyvirus
  • recovery
  • short sequence
  • scoring system
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