با همکاری مشترک دانشگاه پیام نور و انجمن بیوتکنولوژی جمهوری اسلامی ایران

نوع مقاله : علمی پژوهشی

نویسندگان

1 فارغ تحصیل کارشناسی ارشد مهندسی کشاورزی، گروه بیوتکنولوژی کشاورزی، دانشکده کشاورزی، دانشگاه تربیت مدرس، تهران، ایران

2 عضو هیات علمی گروه زیست فراورده‌های گیاهی، پژوهشکده بیوتکنولوژی کشاورزی، پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری، تهران، ایران.

3 دانشیار گروه بیوتکنولوژی کشاورزی، دانشکده کشاورزی، دانشگاه تربیت مدرس، تهران، ایران

چکیده

در این مطالعه تاثیر سه غلظت از محرک متیل‌جاسمونات بر تولید متابولیتهای ثانویه و فعالیت آنتی‌اکسیدانی در ریشه‌مویین Origanum vulgare بررسی شد. به این منظور، سویه A13 باکتری Agrobacterium rhizogenes برای تهیه لاین‌های پایدار ریشه‌مویین از ریزنمونه‌ها مورد استفاده قرار گرفت. در این راستا، دو محیط هم‌کشتی ( MS و MS تغییر یافته)، دو زمان تلقیح (5 و 10 دقیقه) و دوغلظت استوسیرینگون ( صفر و 100 میکرومولار) آزمایش شدند. پس از تکثیر، ریشه‌های‌مویین با غلظت های مختلف متیل‌جاسمونات (1/0، 2/0 و 5/0 میکرو مولار) تیمار شدند. بعد از 24 و 96 ساعت فعالیت آنزیم‌های آنتی‌اکسیدانی اندازه‌گیری شد. علاوه بر آن، بیان ژن‌هایOvTPS2 وOvDXR ، دو ژن کلیدی مسیر MEP، با تکینک Real time PCR بررسی شد. بیشترین نرخ تراریختی در ریزنمونه‌های کشت شده در MS تغییر یافته، بعد از 10 دقیقه غوطه‌وری در سوسپانسیون باکتری همراه با استوسیرینگون به غلضت 100 میکرومولار بود. ریشه‌های‌مویین تیمار شده با متیل‌جاسمونات در مقایسه با شاهد بطور معنی‎‌داری بیوماس کمتری تولید کردند. همچنین نتایج نشان داد که غلظت‌های مختلف متیل‌جاسمونات، زمان برداشت و همچنین اثر متقابل آن‌ها بطور معنی‌داری بر میزان فعالیت آنزیم‌های کاتالاز، پراکسیداز و سوپراکسیددیسموتاز تاثیر دارند. مقدار بیان ژن OvDXR پس از 24 ساعت از تیمار با 2/0 میکرومولار متیل‌جاسمونات، افزایش معنی‌داری داشت، در حالیکه در همین نمونه بیان OvTPS2 بطور معنی‌داری کاهش یافت. بیان هر دو ژن OvDXR و OvTPS2 پس از 96 ساعت از تیمار 5/0 میکرومولار متیل‌جاسمونات افزایش معنی‌داری نشان داد. کاربرد متیل‌جاسمونات بعنوان یک محرک کارآمد در سیستم کشت ریشه‌مویین O. vulgare می‌تواند برای تولید متابولیت‌های ثانویه ارزشمند باشد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Expression analysis of key genes involved in terpenes biosynthesis and antioxidant enzymes in hairy root of Origanum vulgare elicited with methyl jasmonate

نویسندگان [English]

  • Faeze Aibabaei 1
  • Forough Sanjarian 2
  • Nima Ahmadi 3

1 1. M.Sc. Student. Department of Agricultural Biotechnology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

2 2. Assistant Prof. Plant Bioproducts Group, Department of Agricultural Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

3 3. Associate Prof. Department of Agricultural Biotechnology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

چکیده [English]

In the current study, the effects of three levels of methyl jasmonate concentration were studied on the hairy root cultures of Origanum vulgare. For this purpose, A. rhizogenes strain A13 was used to create stable hairy root lines from shoot explants. In this regard, two co-cultivation media (MS and modified MS), two inoculation times (5 and 10 min) as well two concentrations of acetosyringone (0 and 100 µM) were attentively studied. Propagated hairy roots were treated with different concentrations of methyl jasmonate (0.1, 0.2, and 0.5 µM). Sampling after 24 and 96 h of methyl jasmonate application, the activities of antioxidant enzymes were evaluated. Moreover, the gene expression level of OvDXR and OvTPS2, two important genes involved in the MEP pathway, were studied using real-time PCR technique. The highest transformation rate was observed in the explants cultured modified MS which were incubated in the bacterial suspension with acetosyringone for 10 minutes. The biomass of hairy root cultures was significantly decreased by methyl jasmonate compared to control samples. In addition, the results indicated that the concentration of methyl jasmonate, harvesting time, and their interactions significantly affected the activity of catalase, peroxidase, and SOD enzymes. The expression of OvDXR significantly increased after 24 h of treatment by 0.2 µM methyl jasmonate, while in the same sample, the expression of OvTPS2 decreased noticeably. The expression of OvDXR and OvTPS2 genes was significantly increased by 0.5 µM methyl jasmonate after 96 h. The application of methyl jasmonate as an efficient elicitor in hairy root culture system of O. vulgare is suggested for the production of valuable metabolites.

کلیدواژه‌ها [English]

  • Catalase
  • Gene expression analysis
  • Peroxidase
  • Transformation rate
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