القای کالوس و ایجاد کشت سوسپانسیون سلولی مریم‌گلی سهندی (Salvia sahendica) برای تولید تری‌ترپنوئیدهای دارویی بتولینیک، اولئانولیک و اورسولیک اسید

نوع مقاله : علمی - پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد، گروه کشاورزی، پژوهشکده گیاهان و مواد اولیه دارویی، دانشگاه شهید بهشتی، تهران

2 دانشیار، گروه کشاورزی، پژوهشکده گیاهان و مواد اولیه دارویی، دانشگاه شهید بهشتی، تهران

چکیده

بتولینیک، اولئانولیک و اورسولیک اسید، تری‌ترپنوئیدهای باارزشی هستند که اثرات دارویی متعددی ازجمله ضدتومور، ضدمیکروب، ضدالتهاب و ضدویروس ایدز برای آن‌ها گزارش شده است. گیاه مریم‌گلی سهندی یک گیاه انحصاری و چندساله جنس مریم‌گلی است که در مناطق محدودی از شمال‌غرب ایران رویش دارد. در این تحقیق القای کالوس و ایجاد کشت سوسپانسیون سلولی این گیاه برای تولید این تری‌ترپنوئیدها مطالعه شده است. از ریزنمونه‌های برگِ دانهال‌های درون شیشه‌ای برای القای کالوس بر روی محیط‌کشت موراشیگ و اسکوگ (MS) به‌همراه غلظت‌های مختلف توفوردی (5/0، 1، 5/1 و 2 میلی‌گرم در لیتر) در ترکیب با بنزیل آدنین (5/0، 1 و 5/1 میلی‌گرم در لیتر) در قالب طرح آزمایشی فاکتوریل کامل تصادفی استفاده شد. بیش‌ترین درصد القای کالوس (100%) در محیط‌کشت حاوی 1 میلی‌گرم در لیتر توفوردی و 5/0 میلی‌گرم در لیتر بنزیل آدنین به‌دست آمد. مطالعه الگوی رشد سلولی نشان داد که بیش‌ترین میزان وزن تر (2/13 گرم) و خشک (2/2 گرم) در هفته چهارم به‌دست آمده و بعد از آن رشد سلول‌ها کاهش یافت. ارزیابی الگوی تولید تری ترپنوئیدها (میلی‌گرم در 100 گرم ماده خشک) نیز حاکی از بیش‌ترین میزان تولید اورسولیک اسید (81/173)، بتولینیک اسید (2/162) و اولئانولیک اسید (32/174) در هفته سوم بود. نتایج نشان داد که میزان تولید بتولینیک و اورسولیک اسید در هفته سوم کشت به‌ترتیب 4/10 و 5/1 برابر بیشتر از مقدار آن‌ها در گیاه وحشی است. این نتایج می‌تواند برای تولید انبوه این ترکیبات دارویی مهم در شرایط کنترل شده به‌ویژه بیوراکتور مورد توجه قرار گیرد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Callus Induction and Cell Suspension Culture Establishment of Salvia sahendica for The Production of Medicinal Triterpenoids Betulinic, Oleanolic and Ursolic Acid

نویسندگان [English]

  • Ahad Hedayati 1
  • Mohammad Hossein Mirjalili 2
  • Ziba Bakhtiar 1
1 MSc Student, Department of Agriculture, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, Tehran
2 Associate Professor, Department of Agriculture, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, Tehran
چکیده [English]

Betulinic, oleanolic and ursolic acid are highly valuable pentacyclic triterpenoids because of their wide spectrum of biological activities such as anti-inflammatory, hepatoprotective, antitumor, anti-HIV, antimicrobial, antifungal, anti-ulcer, gastroprotective, hypoglycemic, and antihyperlipidemic. Salvia sahendica Boiss. & Buhse is an aromatic, endemic perennial herb which grows in the restricted regions of Northwest of Iran. In the present study, callus induction and cell suspension culture establishment of the plant for the production of triterpenoids were studied. Callus induction was conducted from in vitro-grown leaf explant cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of 2,4-D (0.5, 1.0, 1.5 and 2.0 mg l−1) and BA (0.5, 1.0 and 1.5 mg l−1) base on factorial complete random design experiment. Maximum callus induction (100%) was obtained in MS medium supplemented with 1 mg l−1 2,4-D and 0.5 mg l−1 BA. The study of cell growth curve showed that the highest fresh weight (13.2g) and dry weight (2.2g) obtained after 4 week of culture. High content of ursolic acid (173.81 mg/100g DW), betulinic acid (162.2 mg/100g DW) and oleanolic acid (174.32 81 mg/100g DW) were obtained at third week of culture. Results showed that the content of betulinic acid and ursolic acid at third week of culture was 10.4 and 1.5-fold than the aerial part of wild growing plant. These results can be considered for scaling-up of triterpenoids production in bioreactors

کلیدواژه‌ها [English]

  • Lamiaceae
  • Tissue culture
  • Cell culture
  • Secondry metabolite
  • Triterpenoids

مراجع

Ali, MS., Ibrahim, SA., Jalil, S. and Choudhary, MI. 2007. Ursolic acid: a potent inhibitor of superoxides produced in the celular system. Phytotherapy Research, 21: 558-561.
Azarmehr, B., Karimi, F., Taghizade, M. and Mousavi Gargari, SL. 2014. Comparative study of growth and secondary metabolite production ability in transformed hairy roots from Cichorium intybus. Journal of Plant Researches, 26(4): 476-485.
Bagheri, A. and Saffari, M. 2008. Principle of Plant Tissue Culture (Translation). Ferdowsi University Press. Mashad, 406 pp.
Bakhtiar, Z., Mirjalili, MH., Sonboli, A. and Moridi Farimani, M. 2014. In vitro propagation, genetic and phytochemical assessment of Thymus persicus-a medicinally important source of pentacyclic triterpenoids. Biologia, 69: 594-603.
Bakhtiar, Z. 2012. Optimizing of micropropagation and cell suspension culture of Thymus persicus for the production of pentacyclic triterpenoids betulinic acid, oleanolic acid and ursolic acid. MSc thesis. Department of Agriculture, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University. 160 pp.
Bakhtiar, Z., Mirjalili, M. H. and Sonboli, A. 2016. In vitro callus induction and micropropagation of Thymus persicus (Lamiaceae), an endangered medicinal plant. Crop Breeding and Applied Biotechnology, 16: 48-54.
Bolta, Z., Baricevic, D., Bohance, B. and Andrensek, S. 2000. Apreliminary inverstigation of ursolic acid in cell suspension culture of Salvia officinalis. Plant Cell, Tissue and Organ Culture, 62: 57-63.
Chen Qi-he, C., Jing, L., Zhang -feng, H., Guo-qing, H. and Ming-liang, F. 2009. The betulinic acid production from betulin through biotransformation by fungi. Enzyme and Microbial Technology, 45: 175-180.
Croteau, RM., Felton, F., Krap, A. and Kjonaas, H. 1981. Relationship of camphor biosynthesis to leaf development in sage (Salvia officinalis). Plant Physiology, 13: 59-64.
Ghahraman, A. 2008. Flora of Iran. Research Institute of Forests and Rangelands Press, Tehran, 16 pp.
Hasanloo, T., Rezazadeh, S. and Rahnama, H. 2008. Hairy root source for the production of valuable pharmaceutical compounds. Journal of Medicinal Plants Research, 8: 1-17.
Hedge, A. 1982. Labiateae (Salvia). In: Flora Iranica, Rechinger, K. H. (Ed.). Akademische drucku Verlangsanstalt, Graz Austria, 150: 403-476.
Hey Wood, VH. 1978. Flowing plants of the world. Oxford University Press. 335 pages.
Jie, L. 1995. Pharmacology of oleanolic acid and ursolic acid. Journal of Ethnopharmacology, 49: 57-68.
Kazakova, OB., Medvedeva, NI., Baikova, IP., Tolstikov, GA., Lopatina, TV., Yunusov, MS. and Zaprutko, L. 2010. Synthesis of triterpenoid acylates: effective reproduction inhibitors of influenza A (H1N1) and papilloma viruses. Russian Journal of Bioorganic Chemistry, 36: 841-848.
Krystyna, I., Wolska, AM., Grudniak, B., Fiecek, A., Kraczkiewicz, D. and Kurek, K. 2010. Antibacterial activity of oleanolic and ursolic acids and their derivatives. Central European Journal of Biology, 5: 553-543.
Lari Yazdi, H., Goudarzi, M., Yazdani, D. and Chehregai, A. 2005. Essential oils composition of leaves and flowers of Salvia syriaca L. and S. reuterana Boiss. from Borujerd-Iran. Journal of Medicinal Plants, 4(16): 15-21.
Lokhande, VH., Nikam, TD. and Penna, S. 2010. Biochemical, physiological and growth changes in response to salinity in callus cultures of Sesuvium portulacastrum L., Plant Cell Tissue and Organ Culture, 102: 17-25.
Lotfipour, F., Samiee, M. and Nazemiyeh, H. 2007. Evaluation of the antibacterial activity of Salvia sahendica and Phlomis caucasica. Journal of Pharmaceutical Science, 1: 29-34.
Norrizah, J. S., Yaseer Suhaimi, M., Rohaya, A. and Nik Roslan, N. A. R. 2012. Ursolic acid and oleanolic acid productions in elicited cell suspension cultures of Hedyotis corymbosa. Biotechnology, 11: 238-242.
Mevis, I., Smetanska, IM., Muller, CT. and Ulrichs, C. 2011. Specific polyphenolic compounds in cell culture of Vitis vinifera L. cv. Gamay Freaux. Applied Biochemistry and Biotechnology, 2: 148-161.
Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15: 473-497.
Ramachandra Rao, S. and Ravishankar, GA. 2002. Plant cell cultures: chemical factories of secondary metabolites. Biotechnology Advances, 20: 101-153.
Razborsek, MI., Voncina, BD., Dolecek, V. and Voncina, E. 2008. Determination of oleanolic, betulinic and ursolic acid in Lamiaceae and mass spectral fragmentation of their trimethylsilylated derivatives. Chromatographia, 67: 433-440
Rezanejad, F. and Tarrahi, R. 2013. The effect of light and plant growth regulators on callogenesis andanthocyanin accumulation in calli of different explants in Rosa gallica. Journal of Plant Researches, 26(2): 184-195.
Salehi, P., Sonboli, A., Neghad Ebrahimi, S. and Yousefzadi, M. 2007. Antibacterial and antioxidant activities of the essential oils and various extracts of Salvia sahendica in different phenological stages. Chemistry of Natural Compound, 43: 328-330.
Santos-Gomes, PC., Seabra, RM., Andrade, PB. and Fernandes-Ferreira, M. 2002. Phenolic antioxidant compounds produced by in vitro shoots of sage (Salvia officinalis L.). Plant Science, 162: 981-987.
Shirazi, Z., Piri, Kh., Mirzaie Asl,A., Hasanloo, T. and Ghiasvand, T. 2014. The effect of methyl jasmonate and salicylic acid elicitors on production amount of Glycyrrhizin and Isoliquiritigenin in hairyroots of Licorice (Glycyrrhiza glabra L.). Journal of Plant Researches, 27(3): 440-449.
Srivastava, P., Sisodia, V. and Chaturvedi, R. 2010. Effect of culture conditions on synthesis of triterpenoids in suspension cultures of Lantana camara L. Bioprocess and Biosystem Engineering, 34: 75-80.
Srivastava, P. and Chaturvedi, R. 2010. Simultaneous determination and quantification of three pentacyclic triterpenoids betulinic acid, oleanolic acid, and ursolic acid in cell cultures of Lantana camara L., In Vitro Cellular and Developmental Biology-Plant, 46: 549-557.
Srivastava, P., Kasoju, N., Bora, U. and Chaturvedi, R. 2010. Accumulation of betulini oleanolic, and ursolic acids in in vitro cell cultures of Lantana camara L. and their significant cytotoxic effects on HeLa cell lines. Biotechnology and Bioprocess Engineering, 15: 1038-1046.
Stward, N., Martin, R., Engasser, JM. and Goergen, JL. 1999. A new methosdology for plant cell viability assessment using intracellular esterase activity. Plant Cell Report, 19: 171-176.
Tan, N., Kaloga, M., Radtke, OA., Kiderlen, AF., Oksuz, S., Ulubelen, A. and Kolodziej, H. 2002. Abietane diterpenoids and triterpenoic acids from Salvia cilicica and their antileishmanial activities. Phytochemistry, 61: 881-884.
Xiao CM, L. and JW De, AG. 2005. A new sesquiterpene from transformation of curdione by cell suspension culture of Platycodon grandiflorum. Chinese Chemical Letters, 16: 1487-1488.
دوفصلنامه فن آوری زیستی در کشاورزی(علمی-پژوهشی)
دوره 8، شماره 2
آذر 1396
صفحه 39-47

فایل ها

هم رسانی

ارجاع به این مقاله

آمار