تأثیر محیط‌های کشت و تنظیم‌کننده‌های مختلف بر استقرار درون‌شیشه‌ای و تولید متابولیت‌های ثانویه گیاه Vaccinium arctostaphylos L.

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد، دانشکده کشاورزی و منابع طبیعی، دانشگاه محقق اردبیلی، اردبیل

2 دانشیار، دانشکده کشاورزی و منابع طبیعی، دانشگاه محقق اردبیلی، اردبیل

3 استاد، دانشکده کشاورزی و منابع طبیعی، دانشگاه محقق اردبیلی، اردبیل

4 استادیار، دانشکده کشاورزی و منابع طبیعی، دانشگاه محقق اردبیلی، اردبیل

چکیده

این تحقیق به­منظور بررسی تأثیر نوع محیط کشت پایه و تنظیم­کننده­های رشد گیاهی مختلف بر استقرار و رشد ریزنمونه­های قره­قاط و میزان متابولیت‌های ثانویه در شرایط کشت درون­شیشه­ای انجام گردید. برای این منظور، ریزنمونه‌های تک­‌گره­ای و جوانه­های انتهایی قره­قاط از مناطق جنگلی استان اردبیل تهیه و از طریق تیمار با محلول g/l3 بنومیل، محلول H2O2 (5%)­، اتانول 70% و در نهایت محلول هیپوکلریت­سدیم 5/2% با 10pH=  به­مدت 12دقیقه به ترتیب ضدعفونی شدند. ریزنمونه­ها روی محیط کشت­های MS، AN و WPM حاوی  mg/l­1/0 NAA یا IBA و سطوح مختلف BAP، زآتین و TDZ کشت شدند. نتایج نشان داد که درصد ریزنمونه‌های رشد کرده و تعداد برگ در هر ریزنمونه به‌طور معنی‌داری تحت تأثیر نوع محیط کشت پایه، ترکیب هورمونی محیط کشت و اثر متقابل بین آنها قرار گرفت. تعداد برگ در هر ریزنمونه و رشد ریزنمونه‌ها در  محیط کشت MS به‌طور معنی‌داری بیشتر از محیط کشت­های پایه AN و WPM بود. بیشترین درصد ریزنمونه‌های رشد کرده و تعداد برگ در هر ریزنمونه در محیط کشت پایه MS حاوی (mg/l 1/0) IBA به­همراه 5/0 و 1 میلی‌گرم بر لیتر زآتین یا 5/0 میلی‌گرم بر لیتر BAP به­دست آمد.­همچنین از نظر میزان متابولیت‌های ثانویه بین تیمار­های مختلف اختلاف معنی­داری وجود داشت. بیشترین میزان ترکیبات آنتوسیانینی و فلاونوئید به­ترتیب در محیط کشت MS حاوی 2 میلی­گرم بر­لیتر BAP و 1/0 میلی­گرم بر­لیتر IBA و MS حاوی 2 میلی­گرم بر لیتر TDZ و 1/0 میلی­گرم بر لیتر NAA به­دست آمد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Effect of culture media and plant growth regulators on in vitro growth and production of secondary metabolites in Vaccinium arctostaphylos L.

نویسندگان [English]

  • Mehran Noruzpour 1
  • Nasser Zare 2
  • Rasool Asghari 3
  • Parisa Sheikhzadeh 4
1 M.Sc. Student, Department of Agronomy and Plant Breeding, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
2 Associate Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
3 Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
4 Assistant Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
چکیده [English]

The current study was conducted to evaluate the effect of basal culture media and plant growth regulators on establishment and growth of Whortleberry explants and production of secondary metabolites in in vitro conditions. Accordingly, single node explants and apical buds of Whortleberry were collected from forestry areas (Soha, Ardabil, Iran), surface sterilized by benomyl (3 g/l), H2O2 (5%), Ethanol (70%) and sodium hypochlorite (2.5%, pH=10) for 12 min, respectively. The explants were cultured on MS, AN, and WPM media supplemented with 0.1 mg/l NAA or IBA, and different levels of BAP, Zeatin and TDZ. Results showed that growth of explants (%) and number of leaves per explant were affected significantly by type of basal culture medium, plant growth regulators combination and their interaction. Number of leaves per explant and explant’s growth in MS medium was significantly higher than in AN, and WPM media. The highest shoots (%) and number of leaves were obtained in MS basal medium supplemented with 0.1 mg/l IBA plus 0.5 and 1 mg/l Zeatin or 0.5 mg/l BAP. Moreover, there were significant differences among the treatments for secondary metabolite contents. The highest amount of anthocyanin and flavonoid were obtained in MS medium supplemented with 2 mg/l BAP+ 0.1 mg/l IBA and MS medium plus 2 mg/l TDZ and 0.1 mg/l NAA.

کلیدواژه‌ها [English]

  • Basal MS medium
  • Caucasian whortleberry
  • In vitro micro-propagation
  • flavonoids
  • Medicinal plant

مراجع

  1. Akhondzadeh, Sh. (2000). Encyclopedia of Medicinal Plants of Iran. Arjmandi Publishing, ­144 p.
    (in Farsi)
  2. Al-Farsi, M., Alsalvar, C., Morris, A., Baron, M. & Shadih, F. (2005). Comparison of antioxidant activity,­ anthocyanins, carotenoids and phenolics of three native fresh and sun-dried date (Phoenix dactylifera L.) varieties grown in Oman. Journal of Agricultural and Food Chemistry, 53(19) 7592-7599.
  3. Alfermann, A.W.­ & Petersen, M. (1995). Natural products formation by plant cell biotechnology. Plant Cell, Tissue and Orgaince Culture, 43, 199-205.
  4. Anderson, W.C. (1980). Tissue culture propagation of red and black raspberries, Rubus idaeus and R. occidentalis. Acta Horticulturae, 112, 13-20.
  5. Bohn, H. & Rink, E.­ (1988). Betalains. In: F. Constabel & I.K. Vasil (Editors). Cell Culture and Somatic Cell Genetics of Plants. New York, Academic Press, 5, 449-463.
  6. Castilho,­ P., Liu, K., Rodrigues, A., Feio, S., Tomi, F. & Casanova, J. (2006). Composition and antimicrobial activity of the essential oil of Clinopodium ascendens (Jordan) Sampaio from Madeira. Flavour and Fragrance Journal, 22, 139-144.
  7. Chang, C., Yang, M., Wen, H. & Chern, J. (2002). Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of Food and Drug Analysis, 10, 178-182.
  8. Cüce, M., Bektaş, E. & Sökmen, A. (2013). Micropropagation of Vaccinium arctostaphylos L.­            via lateral-bud culture. Turkish Journal of Agriculture and Forestry,37(1), 40-44
  9. Daryani, P., Zare, N., Chamani. E., Sheikhzadeh Mossadeg, P. & Javadi mojaddad, D. (2016). Eevaluation of the effects of different basal medium and plant growth regulators on in vitro growth of hazelnut. Journal of Horticultural Science, 30, 417-422. (in Farsi)
  10. Debnath, S. C. & McRae, K. B. (2002). An efficient adventitious shoot regeneration system on excised leaves of micropropagated lingonberry (Vaccinium vitis-idaea L.). The Journal of Horticultural Science and Biotechnology, 77, 744-752.
  11. Debnath, S. C. (2002). In vitro culture of lingonberry (Vaccinium vitis-idaea L.):The influence of cytokinins and media types on propagation. Small Fruits Review, 1, 3-19.
  12. Debnath, S. C. (2005). Micropropagation of lingonberry: Influence of genotype, explant orientation and overcoming TDZ-induced inhibition of shoot elongation using zeatin. Horticultural Science, 40, 185-188.
  13. Farsi, M. (2006). Principles of Plant Breeding. Mashhad University Press. (pp. 234-239). (in Farsi)
  14. George, E. F., Micheal, A. H. & Greet-Jan, D. K. (2008). The components of plant tissue culture media I, macro and micro nutrients. In: George, E.F., Micheal, A.H. & Greet-Jan, D.K. (Eds), Plant Propagation by Tissue Culture. V.1, The Background. (pp. 65-113) Springer.
  15. Hazrati Jahan, R., Zare, N., Dezhsetan, S. & Sheikhzadeh Mosaddegh, P. (2017). Enhanced Taxol production in cell suspension cultures of hazelnut (Corylus avellana L.) by combination of elicitor and precursor. Iranian Journal of Medicinal and Aromatic Plants, 33, 73-89. (in Farsi)
  16. Ivanova, M. & Van Staden, J. (2009). Nitrogen source, concentration, and NH4+: NO3− ratio influence shoot regeneration and hyperhydricity in tissue cultured Aloe polyphylla. Plant Cell, Tissue and Organ Culture, 99(2), 167-174.
  17. Kazeroonian, R., Kalatejari, S., Mousavi, A. & Tohidfar, M. (2017). Reaction of various explants of a Chrysanthemum morifolium cultivar to plant growth regulators in vitro. Iranian Journal of Horticultural Science, 48(3), 527-534.
  18. Kumar, J. & Gupta, P.K. (2008). Molecular approaches for improvement of medicinal and aromatic plants. Plant Biotechnology Reports, 2(2), 93-112.
  19. Lenee, P. & Chupeau, Y. (1989). Development of nitrogen assimilating enzymes during growth of cells derived from protoplasts of sunflower and tobacco. Plant Science, 59, 109-117.
  20. Lloyd, G. & McCown, B. (1980). Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. In: Combined Proceedings of. International Plant Propagators. Society USA, 30, 421-427.
  21. Machakova, I., Zazimalova, E. & George, E. F. (2008). Plant growth regulators. I: Introduction; auxins, their analogues and inhibitors. In: George, E.F., Micheal, A.H. & Greet-Jan, D.K. (Eds) Plant Propagation by Tissue Culture. V.1, the Background, (pp. 175-204). Springer.
  22. May, P. & Cellier, K. M. (1973). The fruitfulness of grape buds. II. The variability in bud fruitfulness in ten cultivars over four seasons. Annals Amelior Plant, 23, 13-26.
  23. Meiners, J., Schwab, M. & Szankowski, I. (2007). Efficient in vitro regeneration systems for Vaccinium species. Plant Cell, Tissue and Organ Culture, 89, 169-176.
  24. Meyer H. J. & Van Staden J. (1995). The in vitro production of an anthocyanin from callus culture of Oxalis linearis. Plant Cell, Tissue and Organ Culture, 40, 55-58.
  25. Mok, M. C., Gabelman, W. H. & Skoog, F. (1976). Carotenoid synthesis in tissue cultures of Daucus carota. Journal of the American Society for Horticultural Science­, 10, 9-42.
  26. Mulabagal, V. & Tsay, H. S. (2004). Plant cell cultures an alternative and efficient source for the production of biologically important secondary metabolites. International Journal of Applied Science and Engineering Research, 2(1), 29-48.
  27. Murashige, T. & Skoog, F. (1977). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15, 473-497.
  28. Mustafa, C., Ersan, B. & Atalay, S. (2013). Micropropagation of Vaccinium arctostaphylos L. via lateral- bud culture. Turkish Journal of Agriculture and Forestry, 37, 40-44.
  29. Nourafcan, H. & Ansari, F. (2017). The effect of MS and B5 media on growth indices of lemon ‘Verbena’ in in vitro condition. Iranian Journal of Horticultural Science, 48(1), 249-252.
  30. Okamoto, G., Tada, H., Suyama, A., Hayashi, Y. & Hirano, K. (2001). Effect of shoot vigor on the development of transmitting tissue and pollen tube growth in pistils of tetraploid grape, cv. Pione. Vitis, 40(3), 105-110.
  31. Ostrolucka, M. G., Libiakova, G., Ondruskova, E. & Gajdosova, A. (2004). In vitro propagation of Vaccinium species. Acta Universitias Latviensis Biology, 676, 207-212.
  32. Paprstein, F. & Sedlak, J. (2015). In vitro multiplication of lingonberry. Horticultural Science, 42 (2), 102-106.
  33. Pezzuto, J. M. (1995). Natural product cancer chemoprotective agents In: Arnason, J.T., Mata, R. & Romeo, J. T. (Eds). Recent Advances in Phytochemistry. Phytochemistry of Medicinal Plants. New York. Plenum Press, pp. 19-45.
  34. Radman, R., Saez, T., Bucke, C. & Keshavarz, T. (2003). Elicitation of plant and microbial systems. Biotechnology Applied Biochemistry, 37, 91-102.
  35. Rajendran, L., Ravishankar, G. A., Venkataraman, L. V. & Prathiba, K. R. (1992). Anthocyanin production in callus cultures of Daucus carota L. as influenced by nutrient stress and osmoticum. Biotechnology Letters, 14, 707-712.
  36. Ramachandra, S. & Ravishankar, G. A. (2002).­ Chemical factories of secondary metabolites. Biotechnology Advances, 20,101-153.
  37. Ramage, C. M. & Williams, R. R. (2001). Mineral nutrition and plant morphogenesis. In vitro. Cellular & Developmental Biology Plant, 38, 116-124.
  38. Ramirez-Estrada, K., Vidal-Limon, H., Hidalgo, D., Moyano, E., Golenioswki, M., Cusido, R. M. & Palazon, J. (2016) Elicitation, an effective strategy for the biotechnological production of bioactive high-added value compounds in plant cell factories. Molecules, 21(2),182
  39. Sabeti, H. (1994). Forests, Trees and Shrubs of Iran. Yazd University, Second Edition. (in Farsi)
  40. Sae-Lee, N., Kerdchoechuen, O. & Laohakunjit, N. (2014). Enhancement of phenolics, resveratrol and antioxidant activity by nitrogen enrichment in cell suspension culture of Vitis vinifera. Molecules, 19, 7901-7912.
  41. Sato, F., Hashimoto, T. & Hachiya, A. (2001). Metabolic engineering of plant alkaloid biosynthesis. Proceedings of the National Academy of sciences of the United States of America, 98(1), 367-372.
  42. Seitz, H. U. & Hinderer, W. (1988). Anthocyanins. In: Constabel, F. & Vasil, I. (Eds). Cell Culture and Somatic Cell Genetics of Plants. Vol 5. Phytochemicals in Plant Cell Cultures. Academic Press, San Diego, (pp. 49-76) Springer­.
  43. Siddiqui, Z. H., Mujib, A., Aslam, J. & Hakeem, K. R. (2013). In vitro production of secondary metabolites using elicitor in Catharanthus roseus: A case study. In: Hakeem, K.R., Ahmad, P. & Ozturk, M. (Eds). Crop Improvement, New Approaches and Modern Techniques. (pp. 401–419). Springer Science, Berlin, Germany.
  44. Stals, H. & Inze, D. (2001). When plant cells decide to divide. Trends in Plant Science, 8, 359-364.
  45. Taiz, E. & Zeiger, L. (2003). Plant Physiology. (3rd ed.). Sinauer Associates Publisher, 690p.
  46. Tripathi, L. & Tripathi, J. N. (2003). Role of biotechnology in medicinal plants. Tropical Journal of Pharmaceutical Research, 2(2), 243-253.
  47. Wada, S., Niedz, R. P. & Reed, B. M. (2015). Determining nitrate and ammonium requirements for optimal in vitro response of diverse pear species. In Vitro Cellular & Developmental Biology-Plant, 51, 19-27.
  48. Wagner, G­. J. (1979). Content and vacuole extra vacuole distribution of neutral sugars free amino acids and anthocyanins in protoplast. Plant Physiology­, 64, 88-93.
  49. Wang, G. Y., Yuan, M. F. & Hong, Y. (2002). In vitro flower induction in roses. In Vitro Cellular and Developmental Biology-Plant, 38, 513-518.

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  • تاریخ دریافت: 12 اسفند 1396
  • تاریخ بازنگری: 30 تیر 1397
  • تاریخ پذیرش: 30 تیر 1397

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