با همکاری مشترک دانشگاه پیام نور و انجمن بیوتکنولوژی جمهوری اسلامی ایران

نوع مقاله : علمی پژوهشی

نویسندگان

1 دانشجوی دکتری فیزیولوژی گیاهی، دانشگاه پیام نور، تهران، ایران

2 استادیار، گروه زیست‌شناسی دانشگاه پیام نور، تهران، ایران

3 استادیار، پژوهشکده زیست فناوری کشاورزی، پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری، تهران، ایران.

4 استاد، گروه زیست‌شناسی دانشگاه پیام نور، تهران، ایران.

چکیده

تنش‌های محیطی اثرات جبران‌ناپذیری بر تولید گندم نان (Triticum aestivum L.) که از مهمترین محصولات زراعی است می‌گذارند. از طرفی اعضا خانواده AP2/ERF از مهمترین تنظیم‌کنندگان رونویسی هستند که بر رشد و پاسخ گیاه به تنش‌های زنده و غیرزنده موثرند. برای ارزیابی سازوکار تحمل تنش شوری در گندم فعالیت آنزیم‌های سوپر اکسیددیسموتاز، آسکوربات‌پراکسیداز و کاتالاز در دو لندریس متحمل گندم (3623 و 3625) تحت شوری با آزمایش فاکتوریل در قالب طرح کاملا تصادفی در دو سطح شاهد و 250 میلی مولار شوری در سه تکرار انجام شد. از گیاهچه‌ها در زمان‌های صفر، 1، 3، 6، 12 و 24 ساعت و 10 روز پس از تنش نمونه‌برداری شد. فعالیت آنزیم‌ها در ریشه و اندام هوایی گیاهان اندازه‌گیری شد. توالی نوکلئوتیدیAP2-21 از پایگاه‌ NCBI استخراج و آغازگرها طراحی و قطعه ژن از گندم جدا، همسانه-سازی و توالی‌یابی و با حضور دومین حفاظت شده AP2 تأیید شد. تغییرات بیان TaAP2-21 با روش PCR کمی و با استفاده از آغازگرهای ویژه و ژن بتا اکتین بررسی شد. نتایج نشان‌دهنده اختلاف معنی‌دار فعالیت آنزیم‌ها در زمان‌های مختلف نسبت به شاهد در هر دو بافت هر دو رقم بود و بیشترین تفاوت در تنش‌های کوتاه و میان‌مدت مشاهده شد با این‌حال ظاهرا" در تنش بلندمدت سازوکار آنتی‌اکسیدانی آنزیم‌ها در 3623 فعالتر از 3625 عمل می کنند. بیان ژن تحت شوری در بافت‌های هر دو لندریس کاهش معنی‌داری داشت. احتمالا ژن TaAP2-21 یکی از عوامل بازدارنده رونویسی از ژن‌های پاسخ‌دهنده به شوری بوده و باعث ایجاد حساسیت به شوری در گندم می‌شود.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Biochemical and molecular responses of two native landraces of bread wheat to salt stress at different time periods

نویسندگان [English]

  • Narjes Fatahi 1
  • Hamid Sobhanian 2
  • Khadijeh Razavi 3
  • Tahmineh Lohrasebi 3
  • Gholamreza BakhshiKhaniki 4

1 Assistant Professor, Department of Biotechnology, Payame Noor University, Tehran, Iran.

2 Assistant Professor, Department of Biotechnology, Payame Noor University, Tehran, Iran.

3 Assistant Professor, Department of Agriculture Biotechnology, National Institute of Genetic engineering and Biotechnology (NIGEB), Tehran, Iran.

4 Professor, Department of Biology, Payame Noor University, Tehran, Iran.

چکیده [English]

Environmental stresses have an irreversible effect on the production of bread wheat (Triticum aestivum L.), one of the most important crop plants. On the other hand, AP2/ERF members are the most important transcriptional regulators that influence plant growth and response to biotic and abiotic stresses. To evaluate the mechanism of salt stress tolerance in wheat the activities of superoxide dismutase, ascorbate peroxidase and catalase in two tolerant wheat landraces (3623 and 3625) under salinity were investigated by completely randomized factorial experiment at control and 250 mM salinity in three replications. Seedlings were sampled at 0, 1, 3, 6, 12 and 24 h and 10 days after stress. The activity of enzymes was measured in the root and shoot of plants. The AP2-21 nucleotide sequence was extracted from the NCBI database and primers were designed and the gene fragment was isolated from wheat and then cloned and sequenced and confirmed by the presence of AP2 conserved domain. TaAP2-21 expression was evaluated by qPCR using specific primers and β-actin housekeeping genes. The results showed a significant difference in enzyme activity at different times compared to control in both tissues of both landraces and the highest was observed in short and medium-term stresses, however, apparently in long term stress the antioxidant mechanism of the enzymes is more active in 3623 than in 3625. Gene expression decreased significantly under salinity in both tissues. The TaAP2-21 gene is probably one of the inhibitors of the transcription of saline responsive genes and causes salt sensitivity in wheat.

کلیدواژه‌ها [English]

  • bread wheat
  • salinity
  • AP2 transcription factor
  • oxidative stress
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