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تأثیر نوع محیط کشت پایه و سویه‌های مختلف باکتریایی بر رشد ریشه‌های موئین گیاه دارویی سنبل‌الطیب (Valeriana officinalis L.) | ||
| تحقیقات گیاهان دارویی و معطر ایران | ||
| مقاله 8، دوره 41، شماره 1 - شماره پیاپی 123، فروردین 1404، صفحه 96-111 اصل مقاله (862.88 K) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22092/ijmapr.2025.352320.2886 | ||
| نویسندگان | ||
| مهدی حیدری رهنی1؛ یوسف فیلی زاده* 2؛ محمد نصری3 | ||
| 1دانشکده تحصیلات تکمیلی کشاورزی دانشگاه آزاد اسلامی واحد ورامین-پیشوا، ورامین، ایران | ||
| 2گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه شاهد، تهران، ایران | ||
| 3گروه زراعت و آگرواکولوژی دانشکده تحصیلات تکمیلی دانشگاه آزاد اسلامی واحد ورامین-پیشوا، ورامین، ایران | ||
| چکیده | ||
| سابقه و هدف: ریشه گیاه دارویی سنبل‌الطیب (Valeriana officinalis L.) به دلیل داشتن ترکیبات والپوتریات، والرنون و اسید والرنیک، به عنوان مسکن قوی، تسکین‌دهنده اعصاب، آرام‌بخش، ضد اضطراب و بی‌خوابی استفاده می‌شود. کشت ریشه‌های موئین گیاهان چندساله به دلیل کوتاه کردن زمان تشکیل متابولیت‌های ثانویه همراه با حفظ خصوصیات زیستی و نژادی، روشی کارآمد در تولید ترکیبات مؤثر گیاهی محسوب می‌شود. بر همین اساس، بهینه‌سازی شرایط رشد ریشه‌های موئین گیاه دارویی سنبل‌الطیب با استفاده از 3 سویه مختلف باکتری آگروباکتریوم رایزوژنز (A13، R1601، LBA9402)، روش‌های مختلف تلقیح و چهار نوع محیط کشت (MS Full Strength; MS-KH2PO4, NH4NO3, KNO3; MS- KH2PO4, NH4NO3, KNO3, CaCl2; MgSO4, microelements, Na2EDTA, FeSO4 and Vitamins) در آزمایشی بررسی شد. مواد و روش: این تحقیق به صورت فاکتوریل در قالب طرح کاملاً تصافی با 8 تکرار در شرایط آزمایشگاهی انجام شد. بذر سنبل‌الطیب بعد از کشت در ظروف آزمایشگاهی محتوی محیط کشت پایه MS، برای جوانه‌زنی به اتاقک رشد با درجه حرارت 25 درجه سانتی‌گراد، رطوبت 20 درصد و 45 میکرومول متر مربع در ثانیه در 16 ساعت روشنایی و 8 ساعت تاریکی منتقل گردید. ریزنمونه‌های هیپوکوتیل و برگ‌های لپه‌ای بهمنظور تلقیح با باکتری از گیاهچههای استریل 4 هفتهای تهیه شدند. القای ریشه‌های موئین از سویه‌های آگروباکتریوم به دو روش اسپری و دیسک برگی و تأیید تراریختی و عدم آلودگی با واکنش PCR برای تکثیر اختصاصی ژن rol-B و VirD انجام شد. ریشه‌های موئین در ریزنمونه‌های برگ لپه‌ای و هیپوکوتیل بعد از 10 روز شروع به ظاهر شدن کردند و تعداد انشعاب فرعی ریشه موئین در طول یک سانتی‌متر و تعداد ریشه‌های موئین ریزنمونه‌ها به صورت روزانه و وزن خشک آنها بعد از 60 روز اندازه‌گیری گردید. نتایج: نتایج این تحقیق نشان داد که سویه‌های آزمایشی باکتری A. rhizogenes قادر به القای ریشه موئین در گیاه سنبل‌الطیب هستند. نتایج تجزیه واریانس آزمایش ریزنمونه هیپوکوتیل و برگ‌های لپه‌ای نشان داد که سویه‌های مختلف، محیط کشت و انتخاب روش تلقیح تأثیر معنی‌داری بر تولید ریشه موئین ایجاد کردند. نتایج این تحقیق نشان داد که ریزنمونه برگ در سویه باکتریایی A13 و محیط کشت MS-KH2PO4, NH4NO3, KNO3, CaCl2 بهترین ترکیب تحریک رشد ریشه موئین در گیاه دارویی سنبل‌الطیب بود. بیشترین درصد تراریختی ریزنمونه‌ها (92 درصد)، وزن خشک ریشه‌های موئین (272 میلی‌گرم) و تعداد ریشه‌های موئین ریزنمونه‌ها (19.20 عدد) نیز مربوط به همین ترکیب تیماری بود. نتایج نشان داد که اختلاف معنی‌داری در خصوصیات ظاهری ریشه‌های موئین تولید شده بوسیله سویه‌های مختلف باکتریایی مشاهده نگردید. نتیجه‌گیری کلی: این آزمایش نشان داد که تولید ریشه‌های موئین در گیاه دارویی سنبل‌الطیب به‌طور چشمگیری بستگی به انتخاب سویه مناسب باکتریایی، محیط کشت مطلوب و شرایط رشد بهینه دارد. از آنجایی که سیستم ریشه‌های موئین سنبل‌الطیب در فرایندهای پیچیده فیزیولوژیکی قادر به حفظ اسید والرنیک موجود در بافت‌های رویشی است، بنابراین، یافتن محیط رشدی منطبق با بیشترین رشد رویشی و تولید مطلوب متابولیت‌های ثانویه در گیاه در تحقیقات آینده ضروریست. از سوی دیگر، بهینه‌سازی شرایط کشت برای تولید ریشه‌های مویین گیاه دارویی سنبل‌الطیب و بالا بردن متابولیت‌های دارویی با ارزش از اهمیت ویژه‌ای برخوردار است. | ||
| کلیدواژه‌ها | ||
| سنبل‌الطیب؛ اگروباکتریوم رایزوژنز؛ محیط کشت؛ وزن خشک ریشه‌های مویین | ||
| عنوان مقاله [English] | ||
| Determination of different growth media and Agrobacterium rhizogenes strains on the hairy root growth of Valerian (Valeriana officinalis L.) | ||
| نویسندگان [English] | ||
| Mehdi Heydari-Rahni1؛ Yousef Filizadeh2؛ Mohammad Nasri3 | ||
| 11Department of Agronomy, Varamin-Pishva Branch, Islamic Azad University, Varamin, Iran. | ||
| 2Department of Agronomy and Plant Breeding, Agricultural College, Shahed University, Tehran, Iran | ||
| 3Department of Agronomy, Varamin-Pishva Branch, Islamic Azad University, Varamin, Iran. | ||
| چکیده [English] | ||
| Background and Objectives: Valeriana, a genus in the Caprifoliaceae family, includes several species, notably Valeriana officinalis L. (valerian). Valerian root extract contains various bioactive compounds, including valerenic acid, valeranone, valepotriates, and gamma-aminobutyric acid, which have been widely used as sedatives to treat sleep disorders such as insomnia. This study aimed to evaluate the induction of valerian hairy roots using three different strains of Agrobacterium rhizogenes (A13, R1601, and LBA9402) under four different growth media compositions: (1) MS full strength, (2) MS supplemented with KH2PO4, NH4NO3, and KNO3, (3) MS supplemented with KH2PO4, NH4NO3, KNO3, and CaCl2, and (4) a medium containing MgSO4, microelements, Na2EDTA, FeSO4, and vitamins. Methodology: This study was a factorial experiment based on a completely randomized design with eight replications under laboratory conditions. Seeds of V. officinalis were obtained from a commercial supplier (Pakan Seed Company, Isfahan, Iran) and germinated under greenhouse conditions. Before sowing, seeds were washed with tap water and distilled water. Seeds that settled at the bottom of the washing vessel were selected as they were expected to be the most viable. They were soaked overnight in water before being washed with a 10% detergent solution for 10 minutes, followed by surface disinfection with 70% ethanol for 30 seconds and 6% NaOCl for 5 minutes. After rinsing four times with sterile water, the seeds were germinated on solid Schulz medium, without growth regulators, at 25°C under a 16-hour light/8-hour dark photoperiod (45 µE m⁻² S⁻¹). Hairy root induction was performed using spray and leaf disc methods, and transformation was confirmed by polymerase chain reaction (PCR) analysis for the presence of rolB and virD genes. Hairy root induction rate, lateral branches per centimeter, number of hairy roots per explant, and dry weight of hairy roots were measured after 60 days. Essential oil was extracted by hydro-distillation and analyzed using gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). Results: The study confirmed that the tested A. rhizogenes strains could induce hairy roots in valerian. The fastest hairy root emergence was observed in explants inoculated with strain A13, occurring within 10 days. The highest induction rate (92%), hairy root dry weight (272 mg), and number of hairy roots per explant (19.2) were obtained in leaf explants inoculated with the A13 strain and cultured in MS medium supplemented with KH2PO4, NH4NO3, KNO3, and CaCl2. No significant differences were observed in the morphological traits of hairy roots induced by different A. rhizogenes strains. Conclusion: The findings indicate that growth media composition, appropriate A. rhizogenes strain, and optimized growth conditions enhanced hairy root biomass production. The A13 strain was the most effective in promoting mass hairy root production, particularly in MS medium supplemented with KH2PO4, NH4NO3, KNO3, and CaCl2. Given the complex physiological processes involved, valerian hairy root cultures can preserve valeric acid found in vegetative tissues. Future research should focus on identifying the optimal growth media conditions that maximize vegetative growth and secondary metabolite production in valerian. | ||
| کلیدواژه‌ها [English] | ||
| Valeriana officinalis, Agrobacterium rhizogense, culture media, hairy root dry weight | ||
| مراجع | ||
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