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شناسایی ژنهای مسیر بیوسنتزی تریترپن و سزکوییترپن در میوه هندوانه ابوجهل (Citrullus colocynthis L.) با استفاده از فناوری توالییابی نسل جدید (NGS) | ||
| تحقیقات گیاهان دارویی و معطر ایران | ||
| مقاله 4، دوره 36، شماره 3 - شماره پیاپی 101، مرداد و شهریور 1399، صفحه 390-403 اصل مقاله (434.01 K) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22092/ijmapr.2020.128273.2662 | ||
| نویسندگان | ||
| معصومه درافشان1؛ مهدی سلطانی حویزه* 2؛ وحید شریعتی3 | ||
| 1دانشجوی کارشناسی ارشد، گروه ژنتیک و بهنژادی گیاهی، واحد اهواز، دانشگاه آزاد اسلامی، اهواز، ایران | ||
| 2مربی، گروه ژنتیک و بهنژادی گیاهی، واحد اهواز، دانشگاه آزاد اسلامی، اهواز، ایران | ||
| 3استادیار، پژوهشگاه ملی مهندسی ژنتیک و زیستفناوری، تهران، ایران | ||
| چکیده | ||
| شناسه دیجیتال (DOR): 98.1000/1735-0905.1398.36.390.101.3.1588.41 شناسایی ژنهای بیوسنتز کننده متابولیتهای اختصاصی گیاهان دارویی امروزه با سرعت و دقت فراوان با استفاده از فناوریهای جدید مطالعه ترنسکریپتوم مانند توالییابی RNA انجام میشود. این مطالعه بهمنظور شناسایی ژنهای اختصاصی مسیر بیوسنتز تریترپنها و سزکوییترپنهای موجود در بافت میوه هندوانه ابوجهل (Citrullus colocynthis L.) انجام شد. با استخراج RNA از بافت میوههای هندوانه ابوجهل برداشت شده از منطقه اندیمشک واقع در استان خوزستان در سال 1396، تکنیک توالییابی RNA با استفاده از بنسازه Iluumina HiSeq 2500 اجرا شد. مراحل بیوانفورماتیکی شامل یکپارچهسازی نوپدید با استفاده از نرمافزار Evidential-gene و تفسیر کارکردی با استفاده از پایگاه اطلاعاتی KAAS انجام گردید. از تعداد 21952885 توالی دارای کیفیت بالا، تعداد 55311 تکژن یکپارچه تولید شد و این تکژنها بهصورت موازی در پایگاههای اطلاعاتی مختلف بارگذاری شدند. در پایگاه KAAS تعداد 17359 تکژن در 134 مسیر گیاهی آنوتیت (Annotate) شدند. از میان مسیرهای متابولیتهای ثانویه مختلف و مهم در بافت میوه گیاه هندوانه ابوجهل، به مسیر ژنی "تریترپنها" و" سزکوییترپنها" تعداد 39 تکژن و 8 ژن اورتولوگ اختصاص داشت. آنالیز ترنسکریپتوم این گیاه دارویی با هدف شناسایی ژنهای مسیرهای بیوسنتزی متابولیتهای ثانویه جنبههای پژوهشی و اجرایی مختلفی ازجمله مهندسی مسیر بیوسنتز متابولیتهای دارویی گیاهی را زمینهسازی میکنند. | ||
| کلیدواژهها | ||
| کوکوربیتاسین؛ توالییابی RNA؛ متابولیتهای ثانویه؛ یکپارچهسازی نوپدید؛ ترپنها | ||
| عنوان مقاله [English] | ||
| Identification of triterpene and sesquiterpene biosynthetic pathway genes in Citrullus colocynthis L. fruit using Next Generation Sequencing (NGS) technology | ||
| نویسندگان [English] | ||
| M. Dorafshan1؛ M. Soltani Howyzeh2؛ V. Shariati3 | ||
| 1M.Sc. student, Department of Genetics and Plant Breeding, Ahvaz Branch, Islamic Azad University, Ahvaz, Iran | ||
| 2Faculty member, Department of Genetics and Plant Breeding, Ahvaz Branch, Islamic Azad University, Ahvaz, Iran | ||
| 3Faculty Member, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran | ||
| چکیده [English] | ||
| Identification of the genes biosynthesizing medicinal plants-specific metabolites is now performed with great speed and accuracy using new transcriptome study technologies such as RNA sequencing. The present study was carried out to find the specific genes in the biosynthetic pathway of triterpenes and sesquiterpenes in the fruit tissue of colocynth (Citrullus colocynthis L.). After RNA extraction from the tissue of colocynth fruits harvested from Andimeshk region in Khuzestan province in 2017, RNA sequencing technique was performed using the Illumina HiSeq2500 platform. The bioinformatics steps including de novo assembly, using the Evidential-gene software, and functional annotation, using the KAAS database, were performed. In the KAAS database, 17359 unigenes were annotated in 134 plant pathways. Among the different and important secondary metabolites pathways in the fruit tissue of colocynth, 39 unigenes and 8 orthologous genes were assigned to the triterpenes and sesquiterpenes gene pathways. Transcriptome analysis of this medicinal plant with the aim of identifying the genes of secondary metabolites biosynthetic pathways underlies various research and practical aspects such as biosynthetic pathway engineering of herbal medicines. | ||
| کلیدواژهها [English] | ||
| Cucurbitacine, RNA sequencing, secondary metabolites, De novo assembly, terpenes | ||
| مراجع | ||
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