شناسایی و بررسی بیان برخی میکرو RNAهای مرتبط با تنش خشکی در گیاه جو

نوع مقاله : علمی پژوهشی

نویسندگان

1 دانشجوی دکتری اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم آباد، ایران

2 استاد گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم آباد، ایران

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

4 دانشیار، گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه شیراز، شیراز، ایران

چکیده

مطالعه سازوکار‏های تحمل تنش در گیاه جو، به درک بهتر اساس ژنتیکی تحمل تنش خشکی و در نهایت بهبود خصوصیات ژنتیکی مرتبط با تحمل تنش به کمک روش‏های نوین ژنتیک مولکولی می‌انجامد. در این پژوهش، به‌منظور شناسایی و بررسی بیان برخی میکرو RNAهای درگیر در تحمل تنش خشکی گیاه جو، واکاوی EST های برگ و ریشه در گیاه جو صورت گرفت. اطلاعات اولیه کتابخانه‌ها از پایگاه‌ NCBI دریافت گردید، سپس با کمک نرم افزارهای بیوانفورماتیکی، پیش‌پردازش داده‌ها و همچنین شناسایی ژن‌های با میزان بیان متفاوت در بین کتابخانه‌ها انجام گرفت. به منظور بررسی بیان ژن‌های منتخب با استفاده از روش Real time-PCR، یک آزمایش به صورت فاکتوریل اسپلیت پلات در قالب طرح کاملاً تصادفی در گلدان روی رقم زراعی نیمروز (متحمل در برابر تنش خشکی) و اکوتیپ جو وحشی اسپانتانئوم (Hordeum spontaneum) در سه سطح صفر، 24 و 72 ساعت پس از اعمال تنش خشکی صورت گرفت. با جستجو در بین کانتیگ‌های به‌دست‌آمده، سه miRNA با بیان بالا (ath-miR414، osa-miR2102-5p، osa-miR414) شناسایی شدند. بررسی بیان miR414 و miR2102 در نمونه‌های گیاهی جو، نشان داد که میزان بیان این دو miRNA در هر دو ژنوتیپ در پاسخ به تنش خشکی به‌طور معنی‌داری (05/0>P) افزایش یافت، به‌طوریکه بعد از 72 ساعت بیان ژن miR414 در ژنوتیپ نیمروز و اسپانتانئوم به ترتیب 61/2 و 2 برابر و بیان ژن miR2102 به ترتیب به میزان 4/2 و 8/2 برابر نسبت به اسپانتئوم (شاهد) در شرایط عدم تنش (صفر ساعت پس از اعمال تنش) افزایش یافتند.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Identification and investigation of microRNAs associated with drought stress in barley

نویسندگان [English]

  • Sajjad Zare 1
  • Farhad Nazarian-Firouzabadi 2
  • Ahmad Ismaili 3
  • Hassan Pakniyat 4
1 Ph.D.Candidate, Agronomy and Plant Breeding Department, Faculty of Agriculture, Lorestan University, Khorramabad, Iran.
2 Professor, Agronomy and Plant Breeding Department, Faculty of Agriculture, Lorestan University, Khorramabad, Iran.
3 Professor, Agronomy and Plant Breeding Department, Faculty of Agriculture, Lorestan University, Khorramabad, Iran.
4 Associate Professor, Agronomy and Plant Breeding Department, Faculty of Agriculture, Shiraz University, Shiraz, Iran.
چکیده [English]

Study of stress tolerance in barley can help for a better understanding of the genetic basis of drought stress tolerance mechanisms and lead to improve the genetic properties associated with drought tolerance through modern molecular genetic techniques. To this end, microRNAs associated with drought stress in barley leaf and root ESTs were analyzed in Nimruz and spantaneum barley genotypes. Bioinformatics databases were mined for relevant EST libraries and bioinformatics services were used for pre-processing and identify genes with different expressions among libraries. The expression profile of candidate genes was studied, by using Real time-PCR in a factorial-split plot design, including Nimruz as tolerant and Spontaneum as a drought sensitive in pots with three replications. Sampling time was also considered at 0, 24 and 72 hours after drought stress as sub factor. Results of this study led to identification of three highly-expressed miRNAs (ath-miR414, os-miR2102-5p and osa-miR414). The expression analysis showed that miR414 and miR2102 expression was significantly (P< 0.05) increased in both genotypes in response to drought stress. After 72h in Nimruz and Spontaneum, the expression of miR414 reached 2.61 and 2-fold and the expression of miR2102 was 2.4 and 2.8-fold of that of control (Spontaneum at control condition at 0 times), respectively.

کلیدواژه‌ها [English]

  • Bioinformatics
  • Drought stress
  • Gene expression
  • miRNA

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