شناسایی ژن‌های کلیدی مؤثر در استقرار و تداوم حافظه تنش گرمایی در گیاهچه‌‌‌های آرابیدوپسیس با استفاده از داده‌‌‌های ریزآرایه

نوع مقاله : علمی پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد بیوتکنولوژی کشاورزی، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه علوم کشاورزی و منابع طبیعی

2 استادیار، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه علوم کشاورزی و منابع طبیعی خوزستان. ملاثانی. ایران.

3 استادیار، پژوهشکده آبزی پروری آب‌های جنوب کشور، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، اهواز، ایران

چکیده

گیاهان قادرند از طریق به خاطر سپردن تنش‌‌ گرمای قبلی (پرایمینگ)، نسبت به تنش‌‌های کشنده بعدی (برگشتی) تحمل گرمایی بدست آورند. اثر پرایمینگ که برای ساعت‌‌ها، روزها یا حتی نسل‌‌ها پس از تنش گرمایی برگشتی حفظ می‌‌شود، حافظه تنش گرمایی نامیده می‌‌شود. هدف از این مطالعه شناسایی ژن‌‌های کلیدی موثر در استقرار و تداوم حافظه تنش گرمایی است. در این مطالعه، داده‌‌های ریزآرایه پروفایل بیانی نمونه‌‌های آرابیدوپسیس از بانک داده‌‌های (Gene expression omnibus) GEO جمع‌‌آوری و ژن‌‌هایی با بیان افتراقی بر مبنای فعالیت بیشتر رونویسی‌‌شان تحت تنش برگشتی نسبت به تنش اول (مقایسه تیماری P+T/P) و همچنین القاء بیان پایدار تا 52 ساعت پس از فراغت از تنش (فاز حافظه) شناسایی شدند. سپس ژن‌‌های شناسایی شده به وسیله ابزارهای بیوانفورماتیک جهت دسته‌‌بندی هستی‌‌شناسی (Gene Ontology) و شبکه‌‌های برهم‌‌کنش پروتئینی (Protein-protein interaction networks) مورد بررسی قرار گرفتند. بررسی هستی‌‌شناسی عبارت‌‌ها نشان داد که ژن‌‌هایی با بیان افزایشی عمدتا با پاسخ سلولی و خوگیری به گرما و تاخوردگی پروتئین مرتبط بودند. از طریق خوشه‌‌بندی شبکه برهم‌‌کنش پروتئینی در عبارت‌‌ مربوط به "پاسخ به گرما "در مقایسه تیماری P+T/P، تعدادی از ژن‌‌های کلیدی موثر در استقرار حافظه تنش گرما نظیر HSP70T-2،HSP90 ، HSP60، AR192، HSP70، BIP2، J2، CLPB4، HOP3، HSP101، HSFA3، ROF1، HSFA2، HSP70B، CLPB3، MBF1C، FES1A شناسایی شدند. همچنین بر اساس تداوم بیان افتراقی ژن‌‌ها تا 52 ساعت پس از فراغت از تنش اول (فاز پرایمینگ) مشخص شد ژن‌‌هایی که در پایداری حافظه تنش گرما دخیل هستند عمدتا متعلق به اعضای خانواده پروتئین‌‌های شوک گرمایی کوچک (sHSPs) نظیر HSP17.6، HSP21، HSP17.6II، HAS32، HSP17.4، HSP18.2 و HSP22 بودند. علاوه بر این، در بررسی مسیرهای زیستی از طریق پایگاهKEGG (دانشنامه ژن‌‌ها و ژنوم‌‌های کیوتو) مشخص شد که ژن‌‌های حافظه تنش گرما عمدتا در مسیرهای پردازش پروتئین در شبکه اندوپلاسمی و فسفریلاسیون اکسیداتیو نقش داشتند. همچنین بررسی عناصر تنظیمی سیس در ناحیه پروموتری ژن‌‌های حافظه تنش نشان داد که خانواده فاکتورهای رونویسی bZIP، AP2;B3;RAV، MYB/SANT، HD-ZIP و GATA; tify دارای بیشترین جایگاه اتصال در ناحیه بالادست ژن‌‌های مذکور بودند. در مجموع این یافته‌‌ها اطلاعات مفیدی در خصوص آنالیز عملکردی و تنظیمی ژن‌‌های موثر در استقرار و تداوم حافظه تنش گرمایی و برهم کنش شبکه‌‌های پروتئینی آن‌‌ها ارائه داد که می‌‌توان از آنها در راستای بهبود ظرفیت تحمل گیاه تحت تنش شدید گرما استفاده کرد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Identification of key genes involved in the establishment and maintenance of heat stress memory in Arabidopsis seedlings using microarray data

نویسندگان [English]

  • Atena AlKian Abadi 1
  • Hengameh Taheri 2
  • Ayeh Sadat Sadr 3
1 M.Sc. in Agricultural biotechnology, Department of Plant Production and Genetics, Faculty of Agriculture, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Iran
2 Assistant Professor, Department of Plant Production and Genetics, Faculty of Agriculture, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Iran
3 Assistant Professor, South of Iran Aquaculture Research Institute (SIARI), Iranian Fisheries Science Research Institute, Agricultural Research Education and Extension Organization (AREEO), Ahvaz, Iran
چکیده [English]

Plants are able to acquire thermotolerance to the subsequent lethal stress through memorizing previous heat stress (HS) (Priming). A priming effect that can be sustained for several hours, days, or even generations after reverse heat stress, is called heat stress memory. The aim of this study was to identify effective key genes in establishing and maintaining heat stess memory. To achieve this, microarray data of the expression profile of Arabidopsis samples were retrieved from the GEO (Gene expression omnibus) database and differentially expressed genes (DEGs) were identified based on their higher transcriptional activation following recurring stress (in P+T/P treatment comparison) and their sustained induction even 52 hours after stress relief (during memory phase).The identified genes were further analyzed by bioinformatics tools for gene ontology (GO) classification and protein-protein interaction (PPI) networks. GO terms analysis disclosed that the up-regulated DEGs were mainly associated with cellular response to heat, heat acclimation and protein folding. By clustering of PPI networks in the term related to response to heat (in P+T/P treatment comparison), several candidate genes involved in thermomemory were identified including HSP70T-2, HSP91, AR192, HSP60, HSP70, BIP2, J2, CLPB4, HOP3, HSP101, ROF1, HSFA3, HSFA2, HSP70B, CLPB3, FES1A, MBF1C. Also, based on the sustained differential expression of genes even 52 hours after the priming phase, it was determined that genes responsible for maintaining heat stress memory were mainly members of the small heat shock protein family (sHSPs) such as HSP17.6, HSP21, HSP17.6II, HAS32, HSP17.4, HSP18.2 and HSP22. KEGG (Kyoto Encyclopedia of Genes and Genome) pathway analysis revealed that the HS memory genes were mainly involved in protein processing in the endoplasmic reticulum (ER) and oxidative phosphorylation. Furthermore, the analysis of cis-regulatory elements in the promoter regions of the thermomemory genes revealed that the transcription factors families of bZIP, AP2;B3;RAV, MYB/SANT, HD-ZIP and GATA; tify had the highest binding sites in their upstream regions. In summary, these findings provide useful information about functional and regulatory analysis of genes involved in the establishment and maintenance of heat stress memory, as well as their protein network interactions. This information can be used to improve the heat tolerance capacity of plants under extreme heat stress.

کلیدواژه‌ها [English]

  • Arabidopsis
  • Heat stress memory
  • Microarray
  • Protein interaction network
  • Thermopriming

مراجع

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