شناسایی miRNAهای حفاظت‌شده و ژن‌های هدف آنها در گل محمدی (Rosa damascena Mill)

نوع مقاله : مقاله علمی - پژوهشی

نویسندگان

1 دانشجوی دکتری، گروه زراعت و اصلاح نباتات، دانشکدگان کشاورزی و منابع طبیعی، دانشگاه تهران، کرج

2 استادیار، گروه زراعت و اصلاح نباتات، دانشکدگان کشاورزی و منابع طبیعی، دانشگاه تهران، کرج

3 استاد، گروه زراعت و اصلاح نباتات، دانشکدگان کشاورزی و منابع طبیعی، دانشگاه تهران، کرج

4 دانشیار، گروه زراعت و اصلاح نباتات، دانشکدگان کشاورزی و منابع طبیعی، دانشگاه تهران، کرج

5 استادیار، پژوهشکده توسعه صنایع شیمیایی، کرج

10.22092/ijrfpbgr.2023.360701.1428

چکیده

ژن­ های (miRNAs)، عناصر تنظیم‌کننده­ای هستند که نقش اصلی آنها کاهش بیان ژن در سطح mRNAها است. miRNA ها همچنین در چندین مسیر گیاهی مرتبط با فعالیت‌های مهم سلولی مانند رشد، تکثیر، تمایز، مورفوژنز، آپوپتوزیز و پاسخ به تنش‌های غیر زیستی و زیستی نقش‌های مهمی ایفا می‌کنند. این تحقیق به‌منظور شناسایی miRNAهای حفاظت‌شده و ژن­ های هدف آنها با استفاده از داده­ های توالی‌یابی نسل جدید در گل محمدی (Rosa damascena Mill.) انجام شد. اگرچه روش‌های بیوانفورماتیک به‌عنوان کارآمدترین راهبرد برای شناسایی miRNA هدف توسعه‌یافته‌اند، راهبرد‌های آزمایشی با کارایی بالا هنوز بسیار مورد تقاضا هستند. برای شناسایی miRNAهای جدید، پیش‌بینی و تعیین هستی­ شناسی ژن‌های هدف درگیر در مسیر تولید رایحه و رنگ در گل محمدی به ترتیب از ابزارهای بیوانفورماتیکی Cmii، psRNATarget، WEGOو Blast2GO به ­علاوه روش‌ آزمایشگاهی (Real-time PCR) استفاده شد. با استفاده از داده‌های EST و RNA-seq مبتنی بر همولوژی گیاه Rosa lucieae در نهایت چهار خانواده miRNA، شامل: miR5021، miR2673، miR156 و miR838 به‌عنوان miRNAهای نامزد انتخاب شدند. در مرحله بعد برای ارزیابی کمی Real-time PCR برای اعتبارسنجی سطح بیان miR5021 منتخب در دو نمونه (سفید و صورتی) و مرحله بیولوژیکی (جوان و غنچه) گل محمدی، محلول‌پاشی با غلظت­ های صفر (شاهد) و 300 میکرومولار متیل ­جاسمونات به مدت 48 ساعت بیشترین بیان نسبی   miR5021 متعلق به نمونه صورتی‌رنگ را در مرحله نموی جوان نشان داد. این یافته‌ها مطالعات چشم‌انداز آینده را در مورد سازوکارهای تنظیمی miRNAها درRosa damascena سرعت می‌بخشد.
 

کلیدواژه‌ها

موضوعات

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