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بررسی پایداری برخی ژن‌های مرجع در ارقام سبز و قرمز گیاه ریحان شیرین (Ocimum basilicum L.) تحت تنش‌های غیر زیستی | ||
| فصلنامه علمی زیست فناوری گیاهان زراعی | ||
| دوره 10، شماره 4 - شماره پیاپی 33، شهریور 1400، صفحه 67-78 اصل مقاله (463.3 K) | ||
| نوع مقاله: علمی پژوهشی | ||
| شناسه دیجیتال (DOI): 10.30473/cb.2021.58408.1850 | ||
| نویسندگان | ||
| مژگان شاهیوند1؛ رضا میر دریکوند* 2؛ مسعود گماریان1؛ کامران سمیعی3 | ||
| 1گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه آزاد اسلامی واحد اراک، اراک | ||
| 2گروه زراعت و اصلاح نباتات، واحد خرم آباد، دانشگاه آزاد اسلامی. خرم آباد.ایران. | ||
| 3استادیار، گروه مهندسی کشاورزی، واحد کنگاور، دانشگاه آزاد اسلامی. کنگاور. ایران | ||
| چکیده | ||
| روش PCR در زمان واقعی یک تکنیک بسیار قدرتمند برای تجزیه و تحلیل بیان ژن در موجودات گوناگون است. با این حال، نرمال‌سازی داده‌های بیان ژن حاصل از این تکنیک و همچنین به‌دست آوردن نتایج قابل اعتماد تا حد زیادی به انتخاب ژن‌های مرجع مناسب و پایدار بستگی دارد. در این مطالعه، پایداری بیان شش ژن مرجع پرکاربرد (EF-1α ، 18S rRNA ، ACTIN ، β-Tubulin ، HSP و GAPDH) در ارقام سبز و قرمز ریحان شیرین تحت پنج تنش غیرزیستی (سرما، خشکی، گرما، شوری و تاریکی) مورد بررسی قرار گرفت. پایداری ژن‌های مرجع مذکور با استفاده از نرم‌افزارهای BestKeeper و NormFinder مورد تجزیه و تحلیل قرار داده شد. نتایج نشان داد که تمامی ژن‌های مرجع مورد بررسی دارای سطوح بیان متفاوتی در تنش‌های غیرزیستی در گیاه ریحان شیرین هستند. ژن‌های مرجع مورد بررسی از نظر میزان بیان تفاوت معنی‌داری بین ارقام سبز و قرمز نشان ندادند. بالاترین و پایین‌ترین مقادیر بیان ژن در ارقام سبز و قرمز ریحان شیرین تحت تنش‌های غیرزیستی به ترتیب برای ژن‌های مرجع β-Tubulin و18S rRNA محاسبه شد. نتایج بررسی با نرم‌افزار BestKeeper به ترتیب ژن‌های HSP و ACTIN را به‌عنوان ژن مرجع پایدار مشخص کرد. نرم افزار NormFinder ژن β-Tubulin را به‌عنوان ژن مرجع پایدار شناسایی کرد. رتبه‌بندی نهایی نتایج نرم افزار‌های BestKeeper و NormFinder ژن ACTIN را به عنوان پایدارترین ژن مرجع برای مطالعات بیان ژن در ارقام سبز و قرمز ریحان شیرین با استفاده از تکنیک PCR در زمان واقعی مشخص کرد. | ||
| کلیدواژه‌ها | ||
| بررسی بیان؛ تنش‌های محیطی؛ رتبه‌بندی پایداری؛ ژنومیکس؛ PCR در زمان واقعی | ||
| عنوان مقاله [English] | ||
| Evaluation of expression stability of some reference genes in green and red cultivars of sweet basil (Ocimum basilicum L.) under abiotic stresses | ||
| نویسندگان [English] | ||
| Mojgan shahivand1؛ Reza Mir drikvand2؛ masod gomarian1؛ Kamran Samiei3 | ||
| 1Department of Agronomy and Plant Breeding, Arak Branch, Islamic Azad University, Arak, Iran. | ||
| 2Department of Agronomy and Plant Breeding, Khorramabad Branch, Islamic Azad University, Khorramabad, Iran | ||
| 3Department of Agriculture, Kangavar Branch, Islamic Azad University, Kangavar, Iran. | ||
| چکیده [English] | ||
| The Real-time PCR is a very powerful technique for the analysis of gene expression in various organisms. However, normalizing gene expression data and obtaining reliable results largely depends on the selection of stable reference genes. In this study, the expression stability of six general reference genes (EF-1α, 18S rRNA, ACTIN, β-Tubulin, HSP and GAPDH) was investigated in green and red cultivars of sweet basil under five abiotic stresses (cold, drought, heat, salt and light). The stability of these reference genes was analyzed using BestKeeper and NormFinder softwares. Results showed that all reference genes had different expression levels in both green and red cultivars of sweet basil. There was no significant difference in expression between green and red cultivars. The highest and lowest expression levels were calculated for β-Tubulin and 18S rRNA reference genes, respectively. The results of BestKeeper software identified HSP and ACTIN genes as stable reference genes, respectively. The NormFinder software identified β-Tubulin genes as stable reference gene. Final ranking of the results of BestKeeper and NormFinder softwares identified ACTIN gene as the most stable reference gene for the gene expression studies in green and red cultivars of sweet basil using real-time PCR. | ||
| کلیدواژه‌ها [English] | ||
| Environmental stresses, Expression analysis, Expression ranking, Gemomics, Real-Time PCR | ||
| مراجع | ||
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