In collaboration with Scientific Association of Iranian Medicinal Plants

Document Type : Research Paper

Authors

1 MSc., Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

2 Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

Abstract

In this research, Echinacea purpurea L. hairy roots were cultured in 1000 ml bubble column bioreactor containing 500 ml WPM liquid medium and the effects of inoculum densities (3, 6 and 9g l-1) and aeration rates (0.1, 0.2 and 0.4vvm) on biomass and cichoric acid production were investigated. Fresh and dry weights and also the cichoric acid content in hairy roots were measured after 30 days. The results showed that the highest amount of biomass (16.4g l-1 fresh weight and 2.084g l-1 dry weight) and cichoric acid (16.74mg g-1 DW) were produced in the inoculum density of 6 gl-1. The investigation of aeration rates on biomass and cichoric acid production showed that the maximum fresh weight (15.4g l-1) and dry weight (2.467g l-1) were obtained in aeration rate of 0.4vvm and the maximum cichoric acid content (12.74mg g-1 DW) was obtained in aeration rate of 0.2vvm. Overall, the inoculum density and aeration rate had considerable effects on the hairy root growth and development in bubble column bioreactor and they should be optimized for obtaining the highest hairy root biomass and secondary metabolites.

Keywords

- Abbasi, B.H., Liu, R., Saxena, P. and Liu, C.Z., 2009. Cichoric acid production from hairy root cultures of Echinacea purpurea grown in a modified airlift bioreactor. Journal of Chemical Technology and Biotechnology, 84(11): 1697-1701.
- Abbasi, B.H., Stiles, A.R., Saxena, P.K. and Liu, C.Z., 2012. Gibberellic acid increases secondary metabolite production in Echinacea purpurea hairy roots. Applied Biochemistry and Biotechnology, 168(7): 2057-2066.
- Abbasi, B.H., Tian, C.L., Murch, S.J., Saxena, P.K. and Liu, C.Z., 2007. Light-enhanced caffeic acid derivatives biosynthesis in hairy root cultures of Echinacea purpurea. Plant Cell Reports, 26(8): 1367-1372.
- Abdoli, M., 2013. Study on Cichoric acid production via optimization of hairy roots culture and polyploidy induction in purple coneflower (Echinacea purpurea L.).Ph.D thesis, Department of Plant Breeding and Biotechnology, Facultuy of Agriculture, Tarbiat Modares University, Iran.
- Abdoli, M., Moieni A. and Naghdi Badi, H., 2013. Influence of KNO3, CaCl2 and MgSO4 concentrations on growth and cichoric acid accumulation in hairy root culture of purple coneflower (Echinacea purpurea L.). Journal of Medicinal Plants, 45: 75-84.
- Bakkali, A.T., Jaziri, M., Foriers, A., Vander Heyden, Y., Vanhaelen, M. and Homes, J. 1997. Lawsone accumulation in normal and transformed cultures of henna, Lawsonia inermis. Plant Cell, Tissue and Organ Culture, 51(2): 83-87.
- Banerjee, S., Rahman, L., Uniyal, G.C. and Ahuja, P.S., 1998. Enhanced production of valepotriates by Agrobacterium rhizogenes induced hairy root cultures of Valeriana wallichii DC. Plant Science, 131(2): 203-208.
- Baque, M.A., Moh, S.H., Lee, E.J., Zhong, J.J. and Paek, K.Y. 2012. Production of biomass and useful compounds from adventitious roots of high-value added medicinal plants using bioreactor. Biotechnology Advances, 30(6): 1255-1267.
- Bourgaud, F., Gravot, A., Milesi, S. and Gontier, E., 2001. Production of plant secondary metabolites: a historical perspective. Plant Science, 161(5): 839-851.
- Chattopadhyay, S., Farkya, S., Srivastava, A.K. and Bisaria, V.S., 2002. Bioprocess considerations for production of secondary metabolites by plant cell suspension cultures. Biotechnology and Bioprocess Engineering, 7: 138-149.
- Choi, Y.E., Kim, Y.S. and Paek, K.Y., 2006. Types and designs of bioreactors for hairy root culture. Plant Tissue Culture Engineering, 6: 161-172.
- Cui, X.H., Chakrabarty, D., Lee, E.J. and Paek, K.Y., 2010. Production of adventitious roots and secondary metabolites by Hypericum perforatum L. in a bioreactor. Bioresource Technology, 101(12): 4708-4716.
- Curtis, W.R., 2000. Hairy roots, bioreactor growth: 827-841. Spier, R.E., (Ed.). Encyclopedia of Cell Technology. Wiley-Interscience, 1249p.
- Eibl, R. and Eibl, D., 2008. Design and use of the wave bioreactor for plant cell culture. Plant Tissue Culture Engineering, 6: 203-227.
- Huang, S.Y., Hung, C.H. and Chou, S.N., 2004. Innovative strategies for operation of mist trickling reactors for enhanced hairy root proliferation and secondary metabolite productivity. Enzyme and Microbial Technology, 35: 22-32.
- Huang, T.K. and McDonald, K.A., 2012. Bioreactor systems for in vitro production of foreign proteins using plant cell cultures. Biotechnology Advances, 30(2): 398-409.
- Jeong, J.A., Wu, C.H., Murthy, H.N., Hahn, E.J. and Paek, K.Y., 2009. Application of an airlift bioreactor system for the production of adventitious root biomass and caffeic acid derivatives of Echinacea purpurea. Biotechnology and Bioprocess Engineering, 14: 91-98.
- Kayser, O. and Quax, W.J., 2007. Medicinal Plant Biotechnology: From Basic Research to Industrial Applications. Weinheim, Wiley-VCH Press, 576p.
- Kino-Oka, M., Hitaka, Y., Taya, M. and Tone, S., 1999. High-density culture of red beet hairy roots by considering medium flow condition in a bioreactor. Chemical Engineering Science, 54(15-16): 3179-3186.
- Kittipongpatana, N., Hock, R.S. and Porter, J.R., 1998. Production of solasodine by hairy root, callus, and cell suspension cultures of Solanum aviculare Forst. Plant Cell, Tissue and Organ Culture, 52: 133-143.
- Kwok, K.H. and Doran, P.M., 1995. Kinetic and stoichiometric analysis of hairy roots in a segmented bubble column reactor. Biotechnology Progress, 11(4): 429-435.
- Lee, C.W. and Shuler, M.L., 2000. The effect of inoculum density and conditioned medium on the production of ajmalicine and catharanthine from immobilized Catharanthus roseus cells. Biotechnology and Bioengineering, 67: 61-71.
- Lee, J. and Scagel, C.F., 2013. Chicoric acid: chemistry, distribution, and production. Frontiers in Chemistry, 1: 1-17.
- Liu, C.Z., Wang, Y., Zhao, B., Guo, C., Ouyang, F., Ye, H. and Li, G., 1999. Development of a nutrient mist bioreactor for growth of hairy roots. In Vitro Cellular & Developmental Biology Plant, 35: 271-274.
- Liu, R., Li, W., Sun, L. and Liu, C., 2012. Improving root growth and cichoric acid derivatives production in hairy root culture of Echinacea purpurea by ultrasound treatment. Biochemical Engineering Journal, 60: 62-66.
- Liu. C.Z., Abbasi, B.H., Gao, M., Murch, S.J. and Saxena, P.K., 2006. Caffeic acid derivatives production by hairy root cultures of Echinacea purpurea. Journal of Agricultural and Food Chemistry, 54(22): 8456-8460.
- Min, J.Y., Jung, H.Y., Kang, S.M., Kim, Y.D., Kang, Y.M., Park, D.J., Prasad, D.T. and Choi, M.S., 2007. Production of tropane alkaloids by small-scale bubble column bioreactor cultures of Scopolia parviflora adventitious roots. Bioresource Technology, 98: 1748-1753.
- Murthy, H.N., Lee, E.J. and Paek, K.Y., 2014. Production of secondary metabolites from cell and organ cultures: strategies and approaches for biomass improvement and metabolite accumulation. Plant Cell, Tissue and Organ Culture (PCTOC): 1-16.
- Paek, K., Chakrabarty, D. and Hahn, E., 2005. Application of bioreactor systems for large scale production of horticultural and medicinal plants. Plant Cell, Tissue and Organ Culture, 81(3): 287-300.
- Ramakrishnan, D., Salim, J. and Curtis, W.R., 1994. Inoculation and tissue distribution in pilot-scale plant root culture bioreactors. Biotechnology Techniques, 8(9): 639-644.
- Romero, F.R., Delate, K., Kraus, G.A., Solco, A.K., Murphy, P.A. and Hannapel, D.J., 2009. Alkamide production from hairy root cultures of Echinacea. In Vitro Cellular & Developmental Biology-Plant, 45: 599-609.
- Schlernitzauer, A., Oiry, C., Hamad, R., Galas, S., Cortade, F., Chabi, B., Casas, F., Pessemesse, L., Fouret, G. and Feillet-Coudraym, C., 2013. Chicoric acid is an antioxidant molecule that stimulates AMP Kinase pathway in L6 Myotubes and extends lifespan in Caenorhabditis elegans. Plos One, 8: 78-88.
- Sivakumar, G., 2006. Bioreactor technology: A novel industrial tool for high-tech production of bioactive molecules and biopharmaceuticals from plant roots. Biotechnology Journal, 1: 1419-1427.
- Wang, B., Zhang, G., Zhu, L., Chen, L. and Zhang, Y., 2006. Genetic transformation of Echinacea purpurea with Agrobacterium rhizogenes and bioactive ingredient analysis in transformed cultures. Colloids and Surfaces Biointerfaces, 53: 101-104.
- Weathers, P., Wyslouzil, B., Wobbe, K., Kim, Y. and Yigit, E., 1999. The biological response of hairy roots to O2 levels in bioreactors. In Vitro Cellular & Developmental Biology-Plant, 35: 286-289.
- Yesil Celiktas, O., Gurel, A. and Vardar-Sukan, F., 2010. Large scale cultivation of plant cell and tissue culture in bioreactors. Transworld Research Network, 1-54.