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شناسایی ژن‌های مسیر بیوسنتزی اسکلت ترپنوئید در میوه گیاه دارویی هندوانه ابوجهل (Citrullus colocynthis (L.) Schrad.) با استفاده از تکنیک توالی‌یابی RNA | ||
| تحقیقات گیاهان دارویی و معطر ایران | ||
| مقاله 13، دوره 35، شماره 4 - شماره پیاپی 96، مهر و آبان 1398، صفحه 691-702 اصل مقاله (954.75 K) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22092/ijmapr.2019.125294.2510 | ||
| 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): 98.1000/1735-0905.1398.35.691.96.4.1588.65 توالی‌یابی RNA در حال حاضر یک انتخاب مؤثر و پرسرعت برای مطالعه ترانسکریپتوم گونه‌های گیاهی غیر مدل است که برای شناسایی شبکه‌های ژنی و الگوهای بیان ژن‌های تولیدکننده متابولیت‌های ثانویه در اندام‌های مختلف گیاهان استفاده می‌شود. اصلی‌ترین ترکیب‌ها در بافت‌های میوه و برگ گیاه دارویی هندوانه ابوجهل (Citrullus colocynthis (L.) Schrad.) را ترپنوئیدها، فلاونوئیدها و آلکالوئیدها تشکیل می‌دهند. در این مطالعه، ترانسکریپتوم میوه گیاه هندوانه ابوجهل با استفاده از تکنیک توالی‌یابی RNA، بن‌سازه Iluumina HiSeq2500 اجرا گردید. بعد از کنترل کیفیت با استفاده از نرم‌افزارهای FastQC و Trimmomatic تعداد 21، 952 و 885 خوانش دارای کیفیت بالا تولید شد و با استفاده از برنامه Evidential-gene به‌صورت نوپدید یکپارچه‌سازی شد که منتهی به تولید 55 و 311 تک‌ژن دارای N50 برابر 927 جفت باز گردید. توالی تک‌ژن‌های یکپارچه شده در پایگاه KAAS بارگذاری شد. در مجموع 13، 657 تک‌ژن تفسیر شد که این تعداد در 134 مسیر زیستی قرار گرفتند. مسیر "اسکلت ترپنوئید" با تعداد 93 تک‌ژن از پرتعدادترین مسیرهای شناسایی شده از میان 1، 552 تک‌ژن مسیر بیوسنتز متابولیت ثانویه بود. با بررسی تک‌ژن‌های مرتبط با مسیر بیوسنتزی اسکلت ترپنوئید، 29 شناسه ژنی (K) شناسایی شد که تمام ژن‌های دو مسیر اصلی بیوسنتزی اسکلت ترپنوئیدی: مسیر سیتوزولی موالونیک اسید (MVA) و پلاستیدی متیل-ارتریتول-فسفات (MEP) از ابتدای مسیر تا تولید ایزوپنیل دی‌فسفات (IPP) را شامل می‌شود. شناسایی ژن‌های مسیر بیوسنتزی اسکلت ترپنوئید امکان تحقق توسعه تجاری محصول گیاه دارویی که پایه‌ای برای پژوهش‌های آینده مربوط به شناسایی مسیرهای بیوسنتزی سایر متابولیت‌های اختصاصی، مهندسی متابولیت، اصلاح مولکولی و به‌نژادی گیاهان دارویی است را فراهم می‌کند. | ||
| کلیدواژه‌ها | ||
| گیاه دارویی؛ توالی‌یابی نسل آینده؛ ترانسکریپتوم؛ یکپارچه‌سازی نوپدید؛ پایگاه اطلاعاتی | ||
| عنوان مقاله [English] | ||
| Identification of terpenoid backbone biosynthetic pathway genes in fruit of Citrullus colocynthis (L.) Schrad. medical plant by RNA sequencing | ||
| نویسندگان [English] | ||
| M. Dorafshan1؛ M. Soltani Howyzeh2؛ V. Shariati3 | ||
| 1M.Sc. student, Genetic and Plant Breeding Departemant, Ahvaz Branch, Islamic Azad University, Ahvaz, Iran | ||
| 2Genetic and Plant Breeding Departemant, Ahvaz Branch, Islamic Azad University, Ahvaz, Iran | ||
| 3National Institute of Genetic Engineering and Biotechnology, Tehran, Iran | ||
| چکیده [English] | ||
| Nowadays, RNA sequencing is an effective and fast choice to study the transcripts of non-model plant species used to identify gene networks and patterns of gene expression producing secondary metabolites in different plant organs. Terpenoids, flavonoids, and alkaloids are the main compounds in fruits and leaves of Citrullus colocynthis (L.) Schrad. In this study, the transcriptome of C. colocynthis fruit was performed using RNA-Seq technique, Iluumina HiSeq2500 platform. After quality control using FastQC and Trimmomatic software, 21,952,885 high-quality reads were produced and the de novo assembly with the Evidential-gene program resulted in the production of 55,311unigenes with an N50 equal to 927 base pairs. The sequence of assembled unigenes was loaded on the KAAS database. A total of 13,657 unigenes were annotated, matched into 134 biosynthetic pathways. The "terpenoid backbone" biosynthetic pathway with 93 unigenes was one of the most numerous identified pathways among 1,552 unigenes of secondary metabolites biosynthetic pathway. By examining the unigenes associated with the biosynthetic pathway of the terpenoid backbone, 29 gene identifiers (K number) of the pathway were detected, which contained all identified genes of the two main biosynthetic pathways, Mevalonic Acid (MVA) and Methylerythritol Phosphate (MEP) pathways, from the beginning of the path to the production of isoprenyl diphosphate (IPP). Identification of the genes in the biosynthetic pathway of the terpenoid backbone makes it possible to realize the commercial development of the medicinal plant products, providing the basis for further research on the identification of biosynthetic pathways of other specific metabolites, metabolite engineering, molecular breeding, and medical plants breeding. | ||
| کلیدواژه‌ها [English] | ||
| Medicinal plant, next generation sequencing, transcriptome, de novo assembly, database | ||
| مراجع | ||
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