بررسی الگوی بیانی برخی ژن‌های خانواده NAC در عدس(Lens Culinaris M.) تحت تنش سرما

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد، گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌‌آباد، ایران

2 نویسنده مسئول، دانشیار، گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد، ایران.

3 دانشیار، گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد، ایران

4 دانش‌آموخته دکتری، گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد، ایران

چکیده

مقدمه: سرما یکی از تنش‌های مهم مناطق سردسیر و معتدل است و در برنامه‌های اصلاحی عدس، بهبود تحمل به سرما در اولویت قرار دارد. فاکتور‌های رونویسی NAC در واکنش به تنش‌های مختلف غیرزیستی نقش مهمی دارند.
مواد و روش‌ها: در این پژوهش به‌ منظور ارزیابی بیان ژن‌های MfNAC، MtNAC57 و GmNAC2، گیاهان رقم گچساران عدس تحت پنج تیمار مدت زمان تنش سرما در قالب طرح تصادفی کامل با سه تکرار قرار گرفتند. تیمارهای مدت زمان تنش سرمایی °C4-2 شامل h6، h12، h24 و h48 بود که روی گیاهان 29 روزه اعمال شد و گیاهان رشد یافته در شرایط گلخانه (دمای °C23) به عنوان شاهد در نظر گرفته شدند. استخراج RNA از بافت‌ها، با استفاده از روش لیتیم‌کلراید و ساخت cDNA با استفاده از کیت TaKara انجام شد. واکنش Real-time PCR برای ژن‌های مورد نظر در هر دو نوع اندام هوایی و زمینی در سه تکرار بیولوژیک و دو تکرار تکنیکی انجام شد. از ژن‌ Actin به عنوان کنترل داخلی استفاده شد.
نتایج: نتایج تجزیه واریانس بیان ژن‌ها نشان داد که بین سطوح مختلف تیمارهای مدت زمان تنش دمایی در هر دو اندام اختلاف معنی‌داری وجود داشت. بیان ژن GmNAC2 در اندام‌ هوایی، 6 ساعت پس از تنش کاهش معنی‌داری یافت در حالی‌که در اندام زمینی در تیمار 24 ساعت پس از تنش بیان این ژن کاهش نشان داد. بیان ژن MtNAC57 در تیمارهای h6، h12 و h24 نسبت به شاهد کاهش نشان داد ولی در اندام زمینی تیمار h12 افزایش معنی‌دار مشاهده شد. ژن mfNAC در اندام هوایی در همه تیمارها نسبت به شاهد کاهش بیان معنی‌دار نشان داد در حالی‌که در اندام زمینی فقط در تیمار بلند مدت h48 افزایش بیان معنی‌دار مشاهده شد.
نتیجه‌گیری: در این تحقیق، بیان برخی از ژن‌های خانواده NAC تحت تنش سرما در عدس برای اولین بررسی شد و نتایج حاصل می‌تواند برای مطالعات به‌نژادی این گیاه و سایر حبوبات مفید واقع شود.

کلیدواژه‌ها


عنوان مقاله [English]

Analysis of Gene Expression Pattern of Some Members of NAC Gene Family in Lentil (Lens Culinaris M.) Under Cold Stress

نویسندگان [English]

  • Bentolhoda Farokhpour 1
  • Ahmad Ismaili 2
  • Hamid Reza Eisvand 3
  • Seyyed Mohsen Sohrabi 4
1 Former M.Sc. Student, Agronomy and Plant Breeding Department, Faculty of Agriculture, Lorestan University, Khorramabad, Iran
2 Corresponding author, Associate Professor, Agronomy and Plant Breeding Department, Faculty of Agriculture, Lorestan University, Khorramabad, Iran
3 Associate Professor, Agronomy and Plant Breeding Department, Faculty of Agriculture, Lorestan University, Khorramabad, Iran.
4 Former Ph.D. Student, Agronomy and Plant Breeding Department, Faculty of Agriculture, Lorestan University, Khorramabad, Iran
چکیده [English]

Objective
 Cold is one of the important stresses in cold and temperate regions and improving cold tolerance is in the top priority of the lentil breeding programs. NAC transcription factors play an important role in response to various biological stresses, including cold stress.
 
Materials and Methods: In this study, to evaluate the expression of MfNAC, MtNAC57 and GmNAC2 genes, the plants of Gachsaran lentil cultivar were subjected to five temprature treatments in a completely randomized design with three replications. Twenty-nine old plants were subjected to duration of cold stress treatments including 6h, 12h, 24h and 48h at 2°-4°c and plants grown under greenhouse conditions (23°C) were considered as controls. Total RNA extraction from tissues was performed using lithium chloride method and cDNA was synthesized using TaKara kit. For each treatment, real-time PCR was performed in both shoot and root in three biological and two technical replications. Actin gene was used as internal control.
 
Results
The results of analysis of variance of all studied genes showed that there was a significant difference between the different duration of cold treatments in both shoot and root. GmNAC2 gene expression decreased significantly in the shoot, 6 hours after stress, while in the root, the expression decreased 24 hours after stress. The expression of MtNAC57 gene in 6h, 12h, and 24h treatments decreased compared to control, but in the root in 12h treatment, a significant expression was observed. MfNAC gene in shoot showed a significant reduction in expression in all treatments, while in root a significant increase was only observed during long-term treatment of 48h.
 
Conclusions
In the present study, for first time, the expression pattern of some members of NAC gene family was examined in lentils under cold stress and the results could be useful for studies on this plant and other legumes.

کلیدواژه‌ها [English]

  • Cold
  • lentil
  • real time PCR
  • root
  • shoot
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