شناسایی miRNA های دخیل در مسیر گل دهی گیاه چغندرقند (.Beta vulgaris L)

نوع مقاله: کامل علمی - پژوهشی

نویسندگان

1 دکتری بیوتکنولوژی کشاورزی، دانشکده کشاورزی، دانشگاه بوعلی سینا. همدان، ایران.

2 دانشیار گروه بیوتکنولوژی کشاورزی، دانشکده کشاورزی، دانشگاه بوعلی سینا. همدان، ایران.

3 دانشیار گروه اصلاح نباتات، دانشکده کشاورزی، دانشگاه بوعلی سینا. همدان، ایران.

4 استادیار گروه کشاورزی، دانشگاه پیام نور همدان. همدان، ایران.

چکیده

یکی از فرآیندهای کنترلی در بسیاری از گیاهان از جمله چغندرقند انتقال از مرحله رشد رویشی به زایشی می­باشد که توسط شبکه پیچیده­ای از پروتئین­ها تنظیم می­شود. تاکنون چندین مسیر تنظیمی دخیل در مسیر گل­دهی و بولتینگ و ژن­های موثر در آن­ها در گیاه مدل آرابیدوپسیس شناسایی شده است. یک مسیر موثر در انتقال از مرحله رویشی به زایشی مسیر وابسته به سن است که تحت­تاثیر دو miRNA172 و miRNA156 می‌باشد. ژن­های کدکننده miRNA ها در بین گیاهان بسیار گسترده و حفاظت­شده هستند. در این تحقیق، برای شناسایی توالی این دو ژن در ژنوم چغندرقند، توالی آن­ها از چندین گیاه جمع­آوری و با توالی­های Refseq-genomic چغندرقند مورد هم­ردیفی قرار گرفتند. نواحی با توالی مشترک شناسایی و اطلاعات آن­ها به طور دقیق مورد بررسی قرارگرفت. نتایج نشان داد که در ژنوم چغندرقند برای ژن کدکننده miR156 فقط 127 جفت باز از کروموزوم شماره 2 و ژن کدکننده miR172 فقط 110 جفت ­باز از کروموزوم شماره 9 دارای توالی­های کامل پیش­ساز miRNA بودند. جداسازی ژن کدکننده miR156 از ژنوم گیاه چغندرقند، با طراحی آغازگرهای اختصاصی از نواحی ابتدا و انتهای ژن انجام گرفت. ژن کدکننده miR156 توسط تکنیک واکنش زنجیره­ای پلیمراز و با استفاده از آغازگرهای اختصاصی MIRF و MIRR از روی DNA ژنومی چغندرقند تکثیر شد. همانگونه که انتظار می­رفت قطعه DNA 127 جفت باز تکثیر شد و در نهایت از طریق توالی­یابی مشابهت 99 درصدی توالی تکثیر شده با توالی شناسایی شده ژن کدکننده miR156 به دست آمد و صحت توالی شناسایی شده به عنوان ژن کدکننده miR156 در چغندرقند تأیید گشت. شناسایی و  مطالعه این ژن­های مؤثر در فرآیند گل­دهی راه را برای قابلیت بررسی عملکرد دقیق آن­ها در فرآیندهای گل­دهی و بولتینگ چغندرقند خواهد گشود.

کلیدواژه‌ها


عنوان مقاله [English]

Identification of miRNAs involved in flowering pathway in sugar beet (Beta vulgaris L.)

نویسندگان [English]

  • M. Asgari 1
  • A. Mirzaie-asl 2
  • M.R. Abdollahi 3
  • L. Khodaie 4
1 Ph.D. in Biotechnology, Agriculture Faculty, Bu-Ali Sina University, Hamedan, Iran.
2 Associate Professor of Agriculture Faculty, Bu-Ali Sina University, Hamedan, Iran.
3 Associate Professor of Agriculture Faculty, Bu-Ali Sina University, Hamedan, Iran.
4 Assistant Professor of Payamenoor University, Hamadan, Iran.
چکیده [English]

Transition from vegetative to reproductive stage is one of the important procedures in most crops such as sugar beet which is regulated by the complex network of proteins. So far, several regulatory pathways involved in flowering and bolting and related genes have been identified in the Arabidopsis as a model plant. Age-related pathway is one of the important pathways involved in transition from vegetative to reproductive stage that is influenced by miR156 and miR172. In order to identify the sequence of the above mentioned genes in sugar beet genome, their sequences were collected from several plants and were aligned with sugar beet Refseq-genomic sequence. Regions with a common sequence were identified and evaluated in detail. For miR156 coding gene, only 127 bp in chromosome 2, and for miR172 coding gene only 110 bp in chromosome 9 had the full-length of miR precursor sequence in the sugar beet genome. In order to isolate the miR156 coding gene from the sugar beet genome, specific primers were designed. The miR156 coding gene was amplified from genomic DNA of sugar beet by polymerase chain reaction technique using specific primers of MIRF and MIRR. As expected, the 127 bp gene was amplified and showed 99% similarity with the sequence of the miR156. Identifying and studying these effective genes in the flowering process will open the way for the ability to verify their exact performance in sugar beet flowering and bolting processes.

کلیدواژه‌ها [English]

  • sugar beet
  • Coding genes
  • flowering pathway
  • miR156
  • miR172
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