بررسی‌ شناسایی مسیرها و lncهای دخیل در گل‌دهی نخل خرما با استفاده از تکنیک RNA-Seq

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه اصلاح نباتات و بیوتکنولوژی،دانشکده کشاورزی، دانشگاه زابل، زابل، ایران

2 گروه اصلاح نباتات و بیوتکنولوژی، دانشکده کشاورزی ،دانشگاه زابل، زابل، ایران

3 گروه اصلاح نباتات و بیوتکنولوژی، دانشکده تولید گیاهی، دانشگاه علوم کشاورزی و منایع طبیعی گرگان، گرگان، ایران

4 کارشناس بخش تحقیقات اصلاح و تهیه نهال و بذر، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی هرمزگان، سازمان تحقیقات، آموزش و ترویج

5 محقق پست دکتری، گروه بیوتکنولوژی، موسسه تحقیقاتی آیپک آلمان، آلمان

10.22103/jab.2024.23423.1568

چکیده

چکیده:
هدف: نخل خرما با نام علمی (Phoenix dactylifera L) گیاهی تک‌لپه‌ و سازگار به نواحی گرمسیری و نیمه‌گرمسیری می‌باشد. رشد گل یکی از مراحل اصلی در ظهور ارگان متمرکز به تولیدمثل جنسی در گیاهان گل‌دار بوده و فرایندهای گلدهی توسط مجموعه‌ای بسیار پیچیده از مسیرهای کنترل شده توسط ژن‌های هویت مریستم گل، چرخه گل، هویت اندام گل و برخی lncRNAها حفاظت شده تکاملی تنظیم می‌شود. مطالعات مختلف روی موجودات به طور مداوم بیان بالایی از lncRNAها را در اندام‌های تولیدمثلی نشان می‌دهد.
مواد و روش‌ها‌: به‌منظور دستیابی به شناسایی مسیرها و lncهای دخیل در گل‌دهی نخل خرما با استفاده از تکنیک RNA-Seq، نمونه‌های‌ جوانه گل از نخل‌های خرمای ایستگاه‌ تحقیقاتی میناب (استان هرمزگان) جمع‌آوری و به پژوهشکده زیست‌فناوری کشاورزی طبرستان منتقل شدند. RNAی کل از نمونه‌های جمع‌آوری‌شده از جوانه‌ گل‌ ارقام مختلف نر و ماده استخراج و به نسبت مساوی با یکدیگر مخلوط شده و در انتها، دو تکرار از هر نمونه‌ ادغام شده به دست آمد و برای توالی یابی ارسال شد. پس از دریافت نتایج توالی‌یابی آنالیزهای موردنیاز با استفاده از نرم‌افزارهای مناسب انجام شد.
نتایج: نتایج تحقیق حاضر نشان داد که lncRNAهای زیادی در مسیرهای گل‌دهی نخل خرما در سه مسیر تناوب نوری (سیزده مورد)، خودانگیزی (یک مورد) و مسیر جیبرلین (چهار مورد) نقش داشتند درحالی‌که در مسیر بهاره‌سازی هیچ lncRNA شناسایی نشد. در ادامه lncRNAهای شناسایی شده بر اساس توالی با lncRNAهای گزارش شده در سایر گیاهان مورد مقایسه قرار گرفتند که نتایج نشان داد که lncRNA شناسایی شده مرتبط با مسیر تناوب نوری، با موارد گزارش شده در گیاهانی چون نخل روغنی، شبدر قرمز، زیتون و گندم مشابه بودند و افزایش بیان آن منجر به گل‌دهی سریع و مستقل از تناوب نوری در گیاهان می‌شود. یک lncRNA شناسایی شده مرتبط با مسیر جیبرلین، با lncRNA گزارش شده در پنبه که مرتبط با ژن LFY بود، مشابهت داشت. چهار lncRNA شناسایی شده مرتبط با مسیر خودانگیزی، با lncRNAهای موجود در سیب، ارزن و نخودفرنگی مشابه بود و با ژن FCA دخیل در مسیر خودانگیزی مرتبط بود.
نتیجه‎گیری: فرایند گل‌دهی در گیاهان تنها در فصل‌های معینی از سال، به‌واسطه شبکه‌های تنظیمی حاصل از سیگنال‌های محیطی و منتج از فرایند ژن‌های دخیل در چهار مسیر تنظیمی گل‌دهی، شامل تناوب نوری، بهاره‌سازی، خودانگیزی و جیبرلین صورت می‌گیرد. شناسایی و مقایسهlncRNAهای دخیل در مسیرهای مختلف گل‌دهی و شناسایی ژن‌های مرتبط با آنها زمینه را برای انجام تحقیقات آتی کاربردی در این گیاه ارزشمند فراهم می‌نماید.

کلیدواژه‌ها

عنوان مقاله [English]

Identification of pathways and lncRNAs involved in date palm flowering using RNA-Seq technique

نویسندگان [English]

  • Shahin Madadi 1
  • Salehe Ganjali 2
  • Leila Fahmideh 3
  • Hamed Hasanzadeh Khankahdani 4
  • Hadis Kord 5
  • Reza Haghi Dareh Deh 5
1 Ph. D. student of Biotechnology, Department of Plant Breeding and Biotechnology, University of Zabol, Zabol, Iran
2 Assistant Professor Department of Plant Breeding and Biotechnology, University of Zabol, Zabol, Iran
3 Associate Professor of the Department of Plant Breeding and Biotechnology, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran
4 Horticultural Crops Research Department, Hormozgan Agricultural and Natural Resources Research and Education Center, Agricultural Research, Education and Extension Organization (AREEO), Bandar Abbas, Iran.
5 Post-Doctoral Research, Department IPK Gatersleben Gemany
چکیده [English]

Objective
The date palm, known scientifically as Phoenix dactylifera L, grows well in tropical and subtropical regions. The process of flower development initiates sexual reproduction in plants. Specific genes regulate this process in the floral meristem, cycle, organ identity, and certain lncRNAs. Studies on animals have shown that lncRNAs are notably active in reproductive organs.
Materials and methods
We aimed to explore the pathways and lncRNAs involved in date palm flowering. Minab date palm flower bud samples were collected from the Tropical Fruit Research Station in Hormozgan Province, Iran. The samples were then transferred to the Tabaristan Agricultural Biotechnology Research Institute for analysis. RNA was extracted from various cultivars' male and female flower buds and combined equally. Two replicates were produced for each mixed sample. Subsequently, it was sent for sequencing.
Results
We scrutinized the sequencing outcomes to delve into the mechanisms of plant flowering which is dependent on seasonal cues and is influenced by environmental factors and specific genes. Our research focused on four key pathways: photoperiod, vernalization, self-stimulation, and gibberellin. Our investigation uncovered long non-coding RNAs (lncRNAs) associated with date palm flowering. We identified 13 lncRNAs in the photoperiod pathway, one in the self-stimulation, and four in the gibberellin pathway. Interestingly, no lncRNAs were detected in the vernalization pathway. We proceeded to compare these lncRNAs with those found in other plant species. The lncRNAs in the photoperiod pathway resembled those in oil palm, red clover, olive, and wheat. Their increased expression leads to accelerated flowering, irrespective of photoperiod. Moreover, we pinpointed a lncRNA in the gibberellin pathway that is similar to the one found in cotton which is linked to the LFY gene. Furthermore, our study revealed four lncRNAs in the self-stimulation pathway, resembling lncRNAs in apple, millet, and pea, all of which are associated with the FCA gene.
Conclusions
The flowering process in plants occurs only in certain seasons of the year, through regulatory networks resulting from environmental signals and involving the genes associated with the four regulatory pathways of flowering: photoperiod, vernalization, self-stimulation, and gibberellin. The identification and comparison of lncRNAs involved in different flowering pathways, along with the identification of related genes, provide the basis for future applied research in this valuable plant.

کلیدواژه‌ها [English]

  • long non-coding RNAs
  • next generation sequencing
  • gibberellin pathway

مراجع

جاودان اصل مریم، رجبی معماری حمید، نباتی احمدی داریوش، رهنما افراسیاب (1395) مقایسه روش‌های مختلف استخراج RNA ژنومی از گیاه دارویی بومادران (Achillea millefolium). مجله تولیدات گیاهی 39 (2) ، 114-105.
جعفری احمدآبادی سید علی اصغر، عسکریهمت حشمتاله، محمدآبادی محمدرضا (1402) تاثیر شاهدانه بر بیان ژن DLK1 در بافت‌ قلب بره‌های کرمانی. مجله بیوتکنولوژی کشاورزی 15(1) ، 234-217.
روح الامین سپیده، زاهدی بهمن، نظریان-فیروزآبادی فرهاد، ساعی علی (1394) تجزیه و تحلیل بیان ژن‌های تنظیم کننده کلیدی بیوسنتز آنتوسیانین در انار. مجله علوم باغبانی 186 ،86-84
شکری سمیرا، خضری امین، محمدآبادی محمدرضا، خیرالدین حمید (1402) بررسی بیان ژنMYH7  در بافت‌های ران، دست و راسته بره‌های پرواری نژاد کرمانی. مجله بیوتکنولوژی کشاورزی 15(2) ، 236-217.
شجاعی سیدحبیب، مصطفوی خداداد، بی‌همتا محمدرضا و همکاران (1400) پایداری در هیبریدهای ذرت بر اساس تکنیک گرافیکی بای پلات .GGE مجله زراعت 12(2) ، 394.
محمدآبادی محمدرضا، گلکار افروز، عسکری حصنی مجید (1402) اثر رازیانه (Foeniculum vulgare) بر بیان ژن فاکتور 1 رشد شبه انسولین در بافت شکمبه گوسفند کرمانی. مجله بیوتکنولوژی کشاورزی 15(4) ،256-239.
 
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مجله بیوتکنولوژی کشاورزی
دوره 16، شماره 4
دی 1403
صفحه 139-166

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