شناسایی و آنالیز پروموتر ژن‌های کلیدی دخیل در تحمل به تنش خشکی در مرحله زایشی جو با استفاه از آنالیز داده های ریزآرایه

نوع مقاله : علمی پژوهشی

نویسندگان

1 گروه بیوتکنولوژی و اصلاح نباتات، واحد علوم و تحقیقات تهران، دانشگاه آزاد اسلامی، تهران، ایران

2 گروه پژوهشی زیست شناسی سیستم‌ها، پژوهشگاه بیوتکنولوژی کشاورزی، سازمان تحقیقات، آموزش و ترویج کشاورزی

3 گروه بیوتکنولوژی و اصلاح نباتات، علوم تحقیقات تهران، دانشگاه آزاد اسلامی، تهران، ایران

4 دانشیار فیزیولوژی گیاهی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان، ایران

چکیده

خشکی مهم‌ترین تنش محیطی است که باعث کاهش عملکرد محصولات گیاهی می‌شود. تحقیقات در راستای ایجاد ارقام متحمل به تنش خشکی از اهمیت فوق العاده‌ای برخوردار است. در این تحقیق تلاش شده است تا با آنالیز داده‌های تولید شده از طریق فناوری ریزآرایه، ژن‌های مهم پاسخگو به تنش خشکی و ژن‌های هاب در مرحله زایشی جو شناسایی‌شده و آنالیز پروموتور انجام گیرد. به همین منظور با استفاده از نرم‌افزار FlexArray تمامی ژن‌های دارای بیان افتراقی 5/2 ≥ و 5/2- ≥ در بین دو سری از آزمایشات ریزآرایه انجام شده در جو شناسایی شدند. نتیجه این تجزیه و تحلیل، شناسایی 559 ژن پاسخ‌دهنده به تنش خشکی در مرحله زایشی بود. ژن‌های هاب با استفاده از سه الگوریتم محاسباتی Cyto-Hubba در نرم-افزار Cytoscape مشخص شدند. این امر منجر به شناسایی 10 ژن غیر تکراری شد که به‌عنوان مؤثرترین ژن‌ها در پاسخ به تنش خشکی در نظر گرفته شدند. براساس بررسی هستی‌شناسی ژن‌های دارای بیان افتراقی و ژن‌های هاب، تنظیم رونویسی از گروه‌های اصلی بود که نشان دهنده اهمیت عوامل رونویسی در مکانیسم تحمل به خشکی می‌باشد. در میان عوامل رونویسی می‌توان به HvCBF6، HvDRF1.3، LFL1، VP1، WRKY71 و ABI5 (متعلق به خانواده‌های AP2، WRKY و bZIP) اشاره کرد. آنالیز پرموتر نشان داد که برخی از خانواده‌های عوامل رونویسی از جمله AP2، AT-hook family، bHLH، NAC، bZIP و MYB قابلیت اتصال به 85 درصد از پرموترهای شناسایی‌شده را دارند. مطالعه این عوامل رونویسی، می‌تواند به شناخت هر چه بیشتر سازوکار تحمل به تنش خشکی در جو کمک کند.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Identification and promoter analysis of the key drought tolerance involved genes in reproductive stage in barley using microarray data analysis.

نویسندگان [English]

  • Seyede mehri Javadi 1
  • Zahra-Sadat Shobbar 2
  • Asa Ebrahimi 3
  • Maryam Shahbazi 4
1 Department of Biotechnology and Plant Breeding, Tehran Science and Research Branch, Islamic Azad University, Tehran, Iran
2 Department of Systems Biology, Agricultural Biotechnology Research Institute of Iran (ABRII), Education and Extension Organization (AREEO)
3 Department of Biotechnology and Plant Breeding, Tehran Science and Research Branch, Islamic Azad University, Tehran, Iran
4 Associate Professor, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran
چکیده [English]

Drought is the most important environmental stress that reduces crop yield. Therefore, research toward developing tolerant varieties is of great importance. In this study, microarray data analysis was used for identification of drought stress responsive genes and relevant hub genes in the reproductive stage of barley, and then their promoter analysis was performed. To achieve the goal, all the differentially expressed genes (DEGs) at drought conditions with fold changes ≥+2.5 and ≤-2.5 were identified between two microarray data-series in barley using FlexArray software. Bioinformatics analysis indicated that 559 genes were drought responsive at reproductive stage. The hub genes were distinguished using three Cyto-Hubba computational algorithms by Cytoscape software. Based on the hub analysis results, 10 unique (non-redundant) genes were identified as the most effective genes in response to drought stress. According to the gene ontology analysis of DEGs and hub genes, regulation of transcription were among the major groups indicating the importance of transcription factors (TFs) at drought tolerance mechanism. Amongst the hubs, several TFs such as HvCBF6, HvDRF1.3, LFL1, VP1, ABI5 and WRKY71 genes (belonged to AP2, WRKY and bZIP families) were observed. Promoter analysis was also revealed that some TF families including AP2, AT-hook family, bHLH, NAC, bZIP and MYB had binding site in 85% of promoters of the drought responsive genes and the hub genes in barley. Studying these transcription factors can help in better identification of drought tolerance mechanism in barley.

کلیدواژه‌ها [English]

  • Barley (Hordeum vulgare)
  • drought stress
  • hub gene
  • gene ontology
  • promoter analysis

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