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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 (941.17 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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- Akasaka, Y., Mii, M. and Daimon, H., 1998. Morphological alterations and root nodule formation in Agrobacterium rhizogenes-mediated transgenic hairy roots of peanut (Arachis hypogaea L.). Annals of Botany, 81: 355-362. - Akhondzadeh, S. and Daliri, A., 2004. Herbal medicine in sleep disorders. Journal of Medicinal Plants, 3(9): 79-86. - Barik, D.P., Mohapatra, U. and Chand, P.K., 2005. Transgenic grass pea (Lathyrus sativus L.): factors influencing Agrobacterium-mediated transformation and regeneration. Plant Cell Reports, 24: 523-531. - Baron, C., Domke, N., Beinhofer, M. and Hapfelmeier, S., 2001. Elevated temperature differentially affects virulence, virB protein accumulation, and t-pilus formation in different Agrobacterium tumefaciens and Agrobacterium vitis strains. Journal of Bacteriology, 183: 6852-6861. - Baskaran, P. and Jayabalan, N., 2009. Psoralen production in hairy roots and adventitious roots cultures of Psoralea coryfolia, Biotechnology Letters, 31: 1073-1077. - Bauer, A., Brönstrup, M., 2014. Industrial natural product chemistry for drug discovery and development. Natural Product Reports, 31: 35-60. - Bensaddek, L., Gillet, F., Nava-Saucedo, J.E. and Fliniaux, M.A., 2001. The effect of nitrate and ammonium concentrations on growth and alkaloid accumulation of Atropa belladonna hairy roots. Journal of Biotechnology, 85: 35-40. - Bent, S., Padula, A., Moore, D., Patterson, M. and Mehling, W., 2006. Valerian for Sleep: A Systematic Review and Meta-Analysis. The American Journal of Medicine, 119(12): 1005-1012. - Bhatt, I.D., Dauthal, P., Rawat, S., Gaira, K.S., Jugran, A., Rawal, R.S. and Dhar, U., 2012. Characterization of essential oil composition, phenolic content, and antioxidant properties in wild and planted individuals of Valeriana jatamansi Jones. Science Horticulture, 136: 61-68. - Chabaud, M., Carvalho-Niebel. F. and Barker, D.G., 2003. Efficient transformation of Medicago truncatula cv. Jemalong using the hyper virulent Agrobacterium tumefaciens strain AGL1. Plant Cell Reports, 22(1): 46-51. - Damnjanović, I., Kitić, D., Zlatković-Guberinić, S., Milosavljević, J. and Conic, I., 2010. 'Contemporary aspects of using Valeriana Officinalis'. Acta Medica Medianae, 49: 65-70. - Dini Torkamani, MR., Abaspour, N., Jafari, M. and samadi, A., 2014. Induction and Optimization of Hairy Root Growth Condition for Valeriana officinalis L. Through Inoculation by Agrobacterium rhizogenes. Journal of Cell & Tissue, 5(1): 23-30 - Fernandez, S., Wasowski, C., Paladini, A. and Mariel, M., 2007. Sedative and Sleep-enhancing Properties of Linarin, a Flavonoid-isolated from Valeriana officinalis L. Pharmacology Biochemistry and Behavior, 77(2): 399-404. - Filizadeh, Y. and Goodarzi, G., 2010. Essential Oils from Hairy Root Cultures and Field Cultivated Roots of Valerian (Valeriana sisymbriifolium). Journal of Medicinal Plants, 35: 120-128. - Fitzgerald, J.A., Könemann, S., Krümpelmann, L., Županič, A. and Vom Berg, C., 2020. Approaches to Test the Neurotoxicity of Environmental Contaminants in the Zebrafish Model - From Behavior to Molecular Mechanisms. Environmental toxicology and chemistry, 40(4): 989-1006. - Geier, T. and Sangwan, R.S., 1996. Histology and chimera segregation reveal cell-specific differences in the competence for shoot regeneration and Agrobacterium-mediated transformation in Kohleria internode explants. Plant Cell Reports, 15: 386-390. - Giri, A., Ravindra, S.T., Dhingra, V., and Narasu, M. L., 2001. Influence of different strains of Agrobacterium rhizogenes on induction of hairy roots and artemisinin production in Artemisia annua. Current Science, 81: 378-382. - Granicher, F., Christen, P. and Kapetandis, I., 1992. High yield production of valepotriates by hairy root cultures of Valeriana officinalis L. var. sambucifolia Miken. Plant Cell Reports, 11(7): 339-342. - Gitz, D.C., Gitz, L.L., Mcclure, J.W. and Huerta, A.J., 2004. Effects of a PAL inhibitor on phenolic accumolation and UV-B tolerance in Spirodela intermedia. Journal of Experimental Botany, 55: 919-927. - Hagel, J.M. and Facchini, P.J., 2013. Benzylisoquinoline alkaloid metabolism–a century of discovery and a brave new world. Plant and Cell Physiology, 54(5): 647-72. - Hobbs, C., 1989. Valerian: a literature review. Herbalgram, 21(3):19-34. - Hu, Z.B. and Alferman, A.W., 1993. Diterpenoid production in hairy root culture of Salvia miltiorrhiza. Phytochemistry, 32: 699-703. - Houghto, P.J., 1999. The scientific basis for the reputed activity of Valerian. Journal of Pharmacy and Pharmacology, 5: 505-12. - Jenifer, U., Francina Cecilia, K., and Ravindhran, R., 2012. In vitro adventitious root and hairy root cultures in Boerhaavia diffusa L., International Journal of oral care Research, 4(1): 65 –7. - Jugran, A.K., Bahukhandi, A., Dhyani, P., Bhatt, I.D., Rawal, R.S., Nandi, S.K. and Palni, L.M.S., 2015. The effect of inoculation with mycorrhiza: AM on growth, phenolics, tannins, phenolic composition and antioxidant activity in Valeriana jatamansi Jones. Journal of Soil Science and Plant Nutrition, 15(4):1036-1049. - Kamada, H., Okamura, N., Sataka, M., Harada, H. and Shimomura, K., 1986. Alkaloid production by hairy root cultures in Atropa belladonna. Plant Cell Reports, 5(4): 523-531. - Kamboj, VP., 2000. Herbal medicine. Current Science, 78: 35–7. - Khalili S., Moieni, A. and Abdoli, M., 2014. Influence of different strains of Agrobacterium rhizogenes, culture medium, age and type of explant on hairy root induction in Echinacea angustifolia. Iranian Journal of Genetics and Plant Breeding, 3(1): 49-56. - Kim, K.H., Lee, Y.H., Kim, D. and Park, YH., 2004. Agrobacterium-mediated genetic transformation of Perilla frutescens. Plant Cell Reports, 23: 386-390. - Kwiatkowski, C., 2010. 'Evaluation of yield quality and weed infestation of common valerian (Valeriana officinalis L.) in dependence on weed control method and forecrop'. Acta Agribotanica. 63:179-188. - Królicka, A., Staniszewska, I., Bielawski, K., Maliński, E., Szafranek, J. and Łojkowska, E., 2001. Establishment of hairy root cultures of Ammi majus. Plant Science, 160: 259-264. - Lee, SK., Cui, B., Mehta, R. and Kinghorn, AD., 1998. Cytostatic mechanism and antitumor potential of novel 1H-cyclopenta[b]benzofuran lignans isolated from Aglaia elliptica. Chemico-Biological Interactions. 115: 215–228. - Lee, S.Y., Kim, S.G., Song, W.S., Kim, Y.K., Park, N.I. and Park, S.U., 2010. Influence of different strains of Agrobacterium rhizogenes on hairy root induction and production of alizarin and purpurin in Rubia akane Nakai, Romanian Biotechnology, 15: 5405-5409. - Lei, C., Zhen-Yu W. and Xiu-Hua, Z., 2012. Optimization of elicitors and precursors to enhance valerate production in adventitious roots of Valeriana amurensis. Plant Cell, Tissue and Organ Culture, 108: 411–420. - Mehrota, T., Kukreja, AK., Khanuja, S. and Mishra, B.N. 2008. Genetic transformation studies and scale up of hairy root culture of Glycyyhiza glabra in bioreactor. Electronic Journal of Biotechnology, 11:2: 1-7. - Murashige, T. and Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15: 473-497. - Namdeo, A.G., 2007. Plant cell elicitation for production of secondary metabolites. Pharmacognosy Reviews, 11: 69-79. - Ooi, J., Li, Y., Rogers, C. and Cantorna, M., 2016. Vitamin D regulates the gut microbiome and protects mice from dextran sodium sulfate-induced colitis. The Journal of Nutrition, 143(10):1679-86. - Ohara, A., Akasaka, Y., Paimon, H. and Mii, M., 2000. Plant regeneration from hairy roots induced by infection with Agrobacterium rhizogenes in Crotalaria Juncea L. Plant Cell Reports, 56: 563-568. - Ozyigit, I.I., Dogan, I. and Tarhan, E.A., 2013. Agrobacterium rhizogenes-mediated transformation and its biotechnological applications in crops. In Crop Improvement. pp: 1-48. - Palazon, J., Pinol, M.T., Cusido, R.M., Morales, C. and Bonfill, M., 1997. Application of transformed root technology to the production of bioactive metabolites. Recent development in Plant Physiology, 1: 125-143. - Panda, M., Mehta, U., and Hazra, S., 2017. Optimizing culture conditions for establishment of hairy root culture of Semecarpus anacardium L. Biotechnology, 7(1):21 - Park, S.U. and Facchini, P.J., 2000. Agrobacterium rhizogenes-mediated transformation of opium poppy, Papaver somniferum L., and California poppy, Eschscholzia californica Cham., root cultures. Journal of Experimental Botany, 51: 1005-1016. - Penzkofer, M., Ziegler, E. and Heuberger, H., 2014. 'Contents of essential oil, valerenic acids and extractives indifferent parts of the rootstock of medicinal valerian (Valeriana officinalis L)'. Journal of Applied Research on Medicinal and Aromatic Plants, 1: 98–106. - Prasad, A., Patel, P., Pandey, S., Niranjan, A. and Misra, P., 2020. Growth and alkaloid production along with expression profiles of biosynthetic pathway genes in two contrasting morphotypes of prickly and prickleless Solanum viarum Dunal. Protoplasma, 257(2): 561-572. - Rahnama, H., Hasanloo, T., Shams, M.K. and Sepehrifar, R., 2008. Silymarin production in hairy root culture of Silybum marianum Gearth. Iranian Journal of Biotechnology, 6: 113-118. - Rao, R.S. and Ravishankar, G.A., 2002. Plant tissue cultures; chemical factories of secondary metabolites. Biotechnology Advances, 20: 101-153. - Rezaie, A., Pashazadeh, M., Ahmadizadeh, Ch., Jafari, B. and Jalilzadeh, H.M., 2010. Study of Sedative and Anxiolytic Effect of Herbal Extract of Nardostachys jatamansi in Comparison with Dizepam in Rats. Journal of Medicinal Plants, 9(36):169 -74. - Saravankumar, A., Aslam, A. and Shajahan, A., 2012. Development and optimization of hairy root culture systems in Withania somnifera (L.) Dunal for withaferin-A production. African Journal of Biotechnology, 11(89):16412-16420. - SAS Institute, 2002. JMP statistics and graphics guide. SAS Institute Inc., Cary, NC. - Sen, C., Gordillo, G., Roy, S., Kirsner, R., Lambert, L., Hunt, K, Gottrup, F., Gurtner, G. and Longaker, M., 2009. Human skin wounds: a major and snowballing threat to public health and the economy. Wound Repair Regeneration, 17(6): 763-71. - Shirazi, Z., Piri, K., Mirzaie, A. and Hasanloo, T., 2012. Glycyrrhizin and isoliquiritigenin production by hairy root culture of Glycyrrhiza glabra. Journal of Medicinal Plants Research, 6(31): 4640-4646. - Solemani, T., Keyanfar, M., Piri, H., and Hasanloo, T., 2012. Morphological Evaluation of hairy roots induced in Artemisia annua L and investigating elicitation effects on the hairy roots biomass production. International Journal of Agriculture: Research and Review, 2: 1005-1013. - Tariverdizadeh, N., Mohebodini, M., Chamani E. and Ebadi, A., 2018. Effects of explant age and strain of Agrobacterium rhizogenes on hairy root induction in Fenugreek (Trigonella foenumgraecum L.). Iranian Journal of Genetics and Plant Breeding, 7(1): 50-58. - Thwe, A., Valan Arasu, M., Li, X., Park, C.H., Kim, S.J., Al-Dhabi, N.A. and Park, SU., 2016. Effect of different Agrobacterium rhizogenes strains on hairy root induction and phenylpropanoid biosynthesis in tartary buckwheat (Fagopyrum tataricum gaertn). Frontiers in Microbiology, 7: 1-10 - Vamenani, R., Pakdin-Parizi, A., Mortazavi, M. and Gholami, Z., 2020. Establishment of hairy root cultures by Agrobacterium rhizogenes mediated transformation of Trachyspermum ammi L. for the efficient production of thymol. Biotechnology and Applied Biochemistry, 67(3): 389-395. - Verpoorte, R., Contin, A. and Memelink, J., 2002. Biotechnology for the production of plant secondary metabolites, Phytochemistry Reviews, 1: 13-25. - Wiśniewski, J., Szczepanik, M., Kołodziej, B. and Król, B., 2016. Plantation methods effects on common valerian (Valeriana officinalis) yield and quality. The Journal of Animal & Plant Sciences, 26(1): 177-184. - World Health Organization (WHO), 2008. "Traditional medicine" Fact sheet number: 134 (December) ".http://www.who.int/mediacentre/factsheets/fs13 4/en/. - Yoshimatsu, K. and Shimomura, K., 1992. Transformation of opium poppy (Papaver somniferum L.) with Agrobacterium rhizogenes MAFF 03-01724. Plant Cell Reports, 11: 132-136. - Zhou, M.L., Zhu, X.M., Shao, J.R., Tang, Y.X. and Wu, Y.M., 2011. Production and metabolic engineering of bioactive substances in plant hairy root culture. Applied Microbiol Biotechnology, 90: 1229-1239. | ||
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آمار تعداد مشاهده مقاله: 103 تعداد دریافت فایل اصل مقاله: 53 |
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