Iranian Journal of Horticultural Science

Iranian Journal of Horticultural Science

Optimization of hairy roots induction in chicory (Cichorium intybus L.) and effects of auxin and carbon source on their growth

Document Type : Full Paper

Authors
Roghayeh Fathi 1
Mehdi Mohebodini 2
Esmaeil Chamani 3
1 Former M.Sc. Student, Faculty of Agriculture and Natural resources, University of Mohaghegh Ardabili, Ardabil, Iran
2 Associate Professr, Faculty of Agriculture and Natural resources, University of Mohaghegh Ardabili, Ardabil, Iran
3 Professor, Faculty of Agriculture and Natural resources, University of Mohaghegh Ardabili, Ardabil, Iran
10.22059/ijhs.2017.225587.1171
Abstract
Hairy root cultures are an effective method for production of secondary metabolites, because hairy roots are genetically and biologically stable and they are able to produce metabolite within a short time without need to use hormones. Chicory (Cichorium intybus L.) is one of the important medicinal plants that contains a number of important medicinal compounds.In this research, hairy root induction was established through the mediation of the A4 strain of Agrobacterium rhizogenes. In first experiment, the effects of type and age of explant and co-culture times on the efficiency of hairy root induction were investigated. In the second experiment, the effect of three different media (Murashige and Skoog solid, liquid and liquid half strength) were investigated on growth rate and biomass accumulation hairy roots. In third experiment, effects of type and age of explant were investigated on the total phenolic content of hairy roots. In fourth experiment, the effect of various concentrations of NAA (0, 0.5, 1 and 1.5 mg/l) and sucrose (3, 4, 5 and 6%) were investigated on biomass accumulation. The obtained results showed maximum hairy root induction (60 percent) and number of roots (11.61 roots per explant) induced from 5-days-old cotyledons. The half strength medium of liquid Murashige and Skoog was the best medium for growth of hairy roots. Also, maximum total phenolic content in hairy roots was obtained by 28-days-old leaves. Medium supplemented with combination of 0.5 mg/l NAA and 3% sucrose showed the best effect on biomass accumulation.
Keywords
Co-culture
NAA
RolB gene
secondary metabolites
Subjects
  1. Azarmehr, B., Karimi, F., Taghizade, M. & Mousavi Gargari, S. L. (2012). Comparative study of growth and secondary metabolite production ability in transformed hairy roots from Cichorium intybus L. Journal of Plant Research, 26, 476-485. (in Farsi)
  2. Brijwal, A. & Tamta, S. (2015). Agrobacterium rhizogenes mediated hairy root induction in endangered Berberis aristata DC. SpringerPlus, 2015(4), 443-453.
  3. Cho, H. J., Widholm, J. M., Tanaka, N., Nakanishi, Y. & Murooka, Y. (1998). Agrobacterium rhizogenes mediated transformation and regeneration of the legume Astragalus Sinicus (Chinese milk). Plant Science, 138, 53-65.
  4. Crozier, A., Ashihara, H. & Clifford, M. N. (Eds). (2006) Plant secondary metabolites occurrence, structure and role in the human diet. Blackwell Publishing.
  5. Dhakulkar, S., Ganapathi, T., Bhargava, S. & Bapat, T. (2005). Induction of hairy roots in Gmelina arborea Roxb. and production of verbascoside in hairy roots. Plant Science, 169, 812-818.
  6. Emad, M., Garibi, F., Rasuli, S. & Khanjanzadeh, S. (2013). Medicinal-industrial plant chicory. Noandish.
  7. Ewa, C., Aneta, K. & Werner, P. (2002). Functional properties of fructans. In: Proceedings of Ninth seminar on inulin, 18-19 April., Budapest, Hungary, p. 122-131.
  8. Gamborg, O. L., Miller, R. A. & Ojima, K. (1968). Nutrient requirements of suspension cultures of soybean root cells. Experimental Cell Research, 50(2), 151-158.
  9. Gangopadhyaya, M., Dewanjeeb, S., Chakrabortyc, D. & Bhattacharyaa, S. (2011). Role of exogenous phytohormones on growth and plumbagin accumulation in Plumbago indica hairy roots and conservation of elite root clones via synthetic seeds. Industrial Crops and Products, 33, 445-450.
  10. Hasanlu, T., Rezazadeh, S. & Rahnama, H. (2008). Hairy roots sources for the production of valuable pharmaceutical compounds. Journal of Medicinal Plants, 29, 1-17. (in Farsi)
  11. Kabirnetaj, S., Zolala, J., Nematzadeh, G. A. & Shokri, E. (2012). Optimization of hairy root cultue establishment in chicory plants (Cichorium intybus L.) through inoculation by Agrobacterium rhizogenes. Iranian journal of agricultural biotechnology, 4, 61-75. (in Farsi)
  12. Khan, S., Irfan, Q. M., Kamaluddin, A. T. & Abdin, M. Z. (2007). Protocol for isolation of genomic DNA from dry and fresh roots of medicinal plants suitable for RAPD and restriction digestion. African Journal of Biotechnology, 6, 175-178.
  13. Kodjo, D., Atsou, V. A., Melin, C., Bland, N., Oudin, A., Courdavault, V., Creche, J. & Lanoue, A. (2013). Optimized genetic transformation of Zanthoxylum zanthoxyloides by Agrobacterium rhizogenes and the production of chelerythrine and skimmiamine in hairy root cultures. Engineering in life sciences, 14, 95-99.
  14. Lee, J. & Scagel, C. F. (2010). Chicoric acid levels in commercial basil (Ocimum basilicum) and Echinacea purpurea products. Journal of Functional Foods, 2, 77-84.
  15. Linsmaier, E. M. & Skoog, F. (1965) Organic growth factor requirements of tobacco tissue cultures. Physiology Plant, 18,100-127
  16. Lourenco, P. M. L., Castro, S. D., Martins, T. M. & Domingos, A. C. (2002). Growth and proteolytic activity of hairy roots from Centaurea calcitrapa: effect of nitrogen and sucrose. Enzyme and Microbial Technology, 31, 242-249.
  17. Malamy, J. E. & Ryan, K. S. (2001). Environmental regulation of lateral root initiation in Arabidopsis. Plant Physiology, 127, 899-909.
  18. Murashige, T. & Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiolgia Plantarum 15, 473-476.
  19. Nguyen, C., Bourgaud, F., Forlot, P. & Giri, A. (1992). Establishment of hairy root cultures of Psoralea species. Plant Cell Reports, 11, 424-427.
  20. Pakdin Parizi, A., Farsi, M., Nematzadeh, G. A. & Mirshamsi, A. (2014). Impact of different culture media on hairy roots growth of Valeriana officinalis L. Acta agriculturae Slovenica, 103, 299-305.
  21. Park, S. U., Li, X., Eom, S. H., Lee, C. Y. & Lee, S. Y. (2010). E-P-Methoxycinnamic acid production in hairy root cultures of Scrophularia buergeriana miquel. Archives of Biological Sciences, Belgrade, 62, 649-652.
  22. Samadi, A., Carapetian, J., Heidary, R., Jafari, M. & Hssanzadeh, A. (2012). Hairy root induction in Linum mucronatum ssp. an anti-tumor lignans production plant. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 40(1), 125-131.
  23. Shinde, A., Malpathak, N. & Fulzele, D. (2010). Impact of nutrient components onproduction of the phytoestrogens daidzein and genistein by hairy roots of Psoralea corylifolia. Journal of Natural Medicines, 64, 346-353.
  24. Sivakumar, G., Yu, K.W. & Paek, K. Y. (2005). Production of biomass and ginsenoides from adventitious roots of Panax ginseng in bioreactor cultures. Engineering in Life Science, 5, 333-342.
  25. Sonald, S. F. & Laima, S. K (2001). Phenolics and cold tolerance of Brassica napus. Plant Agric, 1, 1-5.
  26. Sujatha, G., Zdravkovic-Korac, S., Calic, D., Flamini, G. & Ranjitha Kumari, B. D. (2013). High-efficiency Agrobacterium rhizogenes-mediated genetic transformation in Artemisia vulgaris: Hairy root production and essential oil analysis. Industrial Crops and Products, 44, 643-652.
  27. Taper, H. S. & Roberfroid, M. B. (2002). Inulin/oligofructose and anticancer therapy. British Journal of Nutrition, 87(2), 283-286.
  28. Wu, C. H., Dewir, Y. H., Hahn, E. J. & Paek, K. Y. (2006). Optimization of culturing conditions for the production of biomass and phenolics from adventitious roots of Echinacea angustifolia. Journal of Plant Biology, 49, 193-199.
  29. Yang, Y. (2009). Process Optimization of Extracting Phenols from Cichorium intybus cv. Puna with Response Surface Methodology. Journal of Northwest Forestry University, 24, 118-120.
  30. Yu, S., Kwok, K. H. & Doran, P. M. (1996). Effect of sucrose, exogenous product concentration, and other culture conditions on growth and steroidal alkaloid production by Solanum aviculare hairy roots. Enzyme and Microbial Technology, 18, 238-243.
  31. Zhi-Bi, H. & Min, D. (2006). Hairy Root and Its Application in Plant Genetic Engineering. Journal of Integrative Plant Biology, 48(2), 121-127.
Iranian Journal of Horticultural Science
Volume 49, Issue 3
Autumn 2018
Pages 657-667

  • Receive Date 20 January 2017
  • Revise Date 06 September 2017
  • Accept Date 06 September 2017