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

نوع مقاله : علمی پژوهشی

نویسندگان

1 دانشجوی کارشناسی‌ارشد اصلاح‌نباتات، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران.

2 استادیار، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران.

3 دانشیار، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران.

چکیده

microRNA (میکروارنا)، دسته‌ای از مولکول‌های کوچک و غیر کد کننده بوده که بیان ژن را در حیوانات و گیاهان تنظیم می‌کند. زنیان (Trachyspermum ammi)، گیاه شناخته شده‌ای از نظر خواص دارویی می‌باشد. تاکنون گزارشی از شناسایی میکروارنا برای گیاه دارویی زنیان در پایگاه داده mirbase ثبت نشده است. از این رو در مطالعه حاضر به‌منظور پیش‌بینی میکروارنا و ژن‌های هدف، از رویکرد محاسباتی مبتنی بر جستجوی همسانی از طریق الگوریتم Blastx در برابر پایگاه داده mirbase استفاده شد. پارامترهای درصد باز GC، حداقل انرژی آزاد تاخوردگی، شاخص حداقل انرژی آزاد تاخوردگی و ساختار ثانویه صورت گرفت و توالی‌های کاندید پیش‌ساز میکروارنا شناسایی شدند. به‌منظور بررسی بیان ژن‌های منتخب با استفاده از PCR time Real یک آزمایش در قالب طرح کاملاً تصادفی در گلدان روی اکوتیپ اراک زنیان در سه سطح صفر، 12 و 24 ساعت پس از اعمال متیل جاسمونات صورت گرفت. در مجموع تعداد نه میکروارنای mi156، miR160، miR166، miR168، miR171، miR172، miR396، miR477 وmiR827 شناسایی شد. نتایج تخمینی نشان داد که آنها 931 ژن متعلق به چندین خانواده ژنی با عملکردهای بیولوژیکی متفاوت در زنیان را تنظیم می‌کنند. جاسمونات و مشتقات آن مولکول‌های سیگنال دهنده گیاهی هستند. از این رو، بیان miR160 و miR166 با تکنیک PCR time Real مورد ارزیابی قرار گرفت. نتایج نشان داد که pri-miR160 و pri-miR166 در پاسخ به تیمار متیل جاسمونات افزایش پیدا می‌کنند. این موضوع نشان می‌دهد که pri-miR160 و pri-miR166 با انتقال هورمون مرتبط است.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Identification of miRNAs and their target genes in Trachyspermum ammi

نویسندگان [English]

  • Alireza Payriz 1
  • Leila Nejadsadeghi 2
  • Daryoosh Nabati Ahmadi 3

1 M. Sc. Student of Plant Breeding, Production Engineering and Plant Genetic Department, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran

2 Assistant Professor, Plant Production and Genetics Department, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran

3 Associate Professor, Plant Production and Genetics Department, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran

چکیده [English]

MicroRNAs are main groups of small, non-coding molecules that regulate gene expression in animals and plants. Ajwain (Trachyspermum ammi) is plants known for their medicinal properties. To date, no miRNAs have beenidentified in T. ammi. Therefore, in the present study, a computational approach based on homology search through Blastx algorithm against mirbase database was used to predict miRNA and their target genes. The parameters of GC percentage, minimum folding free energy, minimum folding free energy index and secondary structure were determined and the sequences of miRNA precursor candidate were identified. In order to investigate the expression of selected genes using Real time PCR, an experiment was performed in a completely randomized design on the ajwain Arak ecotype at three levels of 0, 12 and 24 hours after methyl jasmonate application. A total of nine miRNAs including miR156, miR160, miR166, miR168, miR171, miR172, miR396, miR477 and miR827 were identified. It was estimated that they regulate 931 of T. ammi genes, which belong to several gene families with different biological functions. Jasmonate and its derivatives are plant signaling molecules. Therefore, miR160 and miR166 expression was evaluated by Real time PCR technique. The results showed that pri-miR160 and pri-miR166 was up-regulated in response to methyl jasmonate treatment, that indicated pri-miR160 and pri-miR166 were associated with hormone transfer.

کلیدواژه‌ها [English]

  • Transcriptome Trachyspermum ammi
  • methyl jasmonate
  • microRNA
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