ارزیابی بیان ژنهای TaWRKY10 ، TaWRKY53،NAC2 و برخی صفات بیوشیمیایی و آنزیمی تحت تنش شوری در ارقام گندم نان

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد ژنتیک و به نژادی گیاهی ، گروه اصلاح نباتات و بیوتکنولوژی دانشگاه منابع طبیعی و علوم کشاورزی گرگان، گرگان، گلستان، ایران.

2 استاد گروه اصلاح نباتات و بیوتکنولوژی دانشگاه منابع طبیعی و علوم کشاورزی گرگان، گرگان، گلستان، ایران.

3 دانش‌آموخته دکتری، گروه اصلاح نباتات و بیوتکنولوژی دانشگاه منابع طبیعی و علوم کشاورزی گرگان، گرگان، گلستان، ایران.

چکیده

مقدمه: تنش شوری یکی از محدودکننده­ترین عامل غیرزنده در تولید گندم است، برای غلبه بر این مشکل با شناخت نحوه پاسخ و عمل ژن­ها در تنش می­توان اطلاعات مفیدی برای اصلاح گیاهان به­منظور تحمل تنش­های محیطی فراهم کرد. واکنش بیوشیمیایی و مولکولی گندم به تنش شوری بسیار متنوع است. با توجه به نقش مؤثرخانواده مهم عوامل رونویسیNAC و WRKY در تقابل با تنش­ها، در این پژوهش میزان بیان سه ژن مهم این خانواده­ها شامل TaWRKY10 و TaWRKY53  وNAC2  و هم­چنین میزان محتوای کلروفیل، مالون­دی­آلدئید، آنزیم­های کاتالاز و پلی­فنل ­اکسیداز در ارقام گندم نان (کلاته، بهاران و گنبد) مورد ارزیابی قرار گرفتند.
مواد و روش‌ها: آزمایش به‌صورت کرت­های خرد شده در قالب طرح بلوک کامل تصادفی با چهار تکرار در شرایط گلخانه انجام شد. عامل اصلی، تیمار شوری (شاهد، 9 و 12 دسی­زیمنس) با اعمال آب آبیاری پس از جوانه زنی و استقرار گیاهان اعمال شد. عامل فرعی سه رقم گندم (کلاته، گنبد و بهاران ) بود. نمونه­بردای جهت ارزیابی بیان ژن و صفات بیوشیمیایی انجام شد. جهت بررسی بیان ژن‌ها تکنیک Real Time PCR مورد استفاده قرار گرفت.
یافته‌ها: در هر سه رقم با افزایش میزان شوری از میزان محتوای کلروفیل کاسته شد، بیشترین میزان کلروفیل a و  b به ترتیب با (5/14 و 2/18میلی­گرم بر گرم وزن تر) در تیمار شاهد رقم کلاته مشاهده گردید. در هر سه رقم با افزایش میزان شوری نسبت به شاهد میزان آنزیم­های کاتالاز و پلی­فنل­اکسیداز و هم­چنین بیان ژن­های مورد بررسی (TaWRKY10 ، TaWRKY53  وNAC2) افزایش یافت. بیشترین میزان آنزیم­ها در تیمار شوری 12 دسی­زیمنس شوری رقم کلاته مشاهده شد. بیشترین میزان مالون دی­آلدئید 31 میکرومول بر گرم وزن تر در رقم بهاران تحت تنش شوری 12 دسی­زیمنس مشاهده شد که تخریب بیشتر غشای سلولی این رقم را نشان می­دهد. بر اساس نتایج حاصل از رقم کلاته با این پژوهش متحمل بودن بیشتر رقم کلاته به تنش شوری نسبت به دو رقم بهاران و گنبد را تأیید کرد.
نتیجه‌گیری: در بهبود و اصلاح گیاهان در مقاومت به شوری فاکتورهای رونویسی نقش مهمی دارند. نتایج این تحقیق در راستای تأیید نقش فاکتورهای رونویسی در مقاومت به شوری مورد توجه است و در ایجاد و معرفی ارقام گندم متحمل می­تواند به کار گرفته شود.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Evaluation of the expression of TaWRKY10, TaWRKY53, NAC2 genes and some biochemical and enzymatic traits under salt stress in bread wheat cultivars

نویسندگان [English]

  • hanifeh bahlakeh 1
  • saeid navabpour 2
  • mostafah hamidi 3
1 Master's student in genetics and plant breeding, Department of Plant Breeding and Biotechnology, Gorgan University of Natural Resources and Agricultural Sciences, Gorgan, Golestan, Iran.
2 Professor of Plant Breeding and Biotechnology Department of Gorgan University of Natural Resources and Agricultural Sciences, Gorgan, Golestan, Iran.
3 PhD student, Department of Plant Breeding and Biotechnology, Gorgan University of Natural Resources and Agricultural Sciences, Gorgan, Golestan, Iran.
چکیده [English]

Introduction: Salinity stress is one of the most limiting non-living factors in wheat production. To overcome this problem, helpful information can be provided by understanding how genes respond and act in stress, to help plants withstand environmental stress. The biochemical and molecular response of wheat to salinity stress is diverse. Considering the effective role of the important family of transcription factors NAC and WRKY in dealing with stresses, this study evaluated the expression levels of three important genes from these families, TaWRKY10, TaWRKY53, and NAC2, as well as the content of chlorophyll, malondialdehyde, catalase, and polyphenol oxidase enzymes in bread wheat cultivars (Kalateh, Baharan, and Gonbad).
Materials and methods: The experiment was conducted in a randomized complete block design with four replications in greenhouse conditions. The main factor was salinity treatment (control, 9 and 12 deci-siemens) applied through irrigation after germination and plant establishment. The secondary factor was wheat varieties (Kalate, Gonbad, and Baharan). Sampling was performed to evaluate gene expression and biochemical traits. Real-Time PCR technique was used to check gene expression.
Results: In all tested cultivars, the chlorophyll content decreased with increasing salinity. The highest amounts of chlorophyll a and b were observed in the control treatment of the Kalate variety (14.5 and 18.2 mg/g fresh weight, respectively). Catalase and polyphenol oxidase enzymes, as well as the expression of TaWRKY10, TaWRKY53, and NAC2 genes, increased with salinity compared to the control in all three cultivars. The highest enzyme levels were observed in 12 deci-siemens salinity treatment of the Kalateh variety. The highest malondialdehyde amount (31 μmol/g fresh weight) was observed in the Baharan cultivar under 12 deci-siemens salt stress, indicating greater cell membrane damage in this cultivar. Based on the results, the Kalateh variety showed greater tolerance to salt stress compared to Baharan and Gonbad varieties.
Conclusion: Transcription factors play a crucial role in enhancing plant resistance to salinity. The results of this study confirm the role of transcription factors in salinity resistance and could be utilized in developing and introducing tolerant wheat cultivars.

کلیدواژه‌ها [English]

  • gene expression
  • catalase
  • chlorophyll
  • bread wheat
  • salt

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