Effect of Inoculum Density and Different Media on the Growth of Hairy Roots and Production of Withanolide-A from Withaniasomnifera
DOI:
https://doi.org/10.12723/mjs.43.2Keywords:
Withaniasomnifera, Withanolide-A, inoculum density, Media strength, MS media, Gamborg’s media, Nitsch and Nitsch Media, Chu’s mediaAbstract
Withaniasomnifera (L.) Dunal. (Indian ginseng) is one of the most important medicinal plants used as a crude drug for its preventive and therapeutic purposes. Among the diverse constituents of Withania, withanolides are found to be the major components responsible for their biological and pharmacological actions. The present study deals with the effect of inoculum density and different media on the growth of hairy roots and withanolide-A production from Withaniasomnifera. An inoculum size of 10 g/L FW favoured the biomass accumulation(120.42 g/L of FW and 11.98 g/L DW) and withanolide-A production(11.96 mg/g DW) inthe tested range of 2.5, 5.0, 10.0 and 20.0 g/L FW. Among different media tested [Murashige and Skoog (MS), Gamborg’s (B5), Nitsch and Nitsch (NN) and Chu’s (N6)], MS medium favoured both biomass accumulation (121.15 g/L FW and 11.96 g/L DW) and withanolide-A production (11.50 mg/g DW).
References
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[24] Z. W. Pan, H. Q. Wang, J. J. Zhong,“Scale-up study on suspension cultures of Taxus chinensis cells for production of taxanediterpene,” Enz.Microb. Technol. Vol. 27, pp. 714-723, 2000.
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[26] J. Y. Min, H. Y. Jung, S. M. Kang, Y. D. Kim, Y. M. Kang, D. J. Park, D. T. Prasad, M. S. Choi, “Production of tropane alkaloids by small scale bubble column bioreactor cultures of Scopoliaparviflora adventitious roots, ”Bioresource Technol., vol. 98, pp. 1748–1753, 2007.
[2] J. Zhao, N. Nakamura, M. Hattori, T. Kuboyama, C. Tohda, K. Komatsu, “Withanolidederivatives from roots of Withaniasomnifera and their neurite outgrowth activities,” Chem. Pharm. Bull., vol. 50, pp. 760–765,2002.
[3] T. Kuboyama, C. Tohda, K. Komatsu,“Neuritic regeneration and synaptic reconstruction induced by withanolide-A,” Br. J.Pharmacol.,vol.144, pp. 961–971, 2005.
[4] C. Tohda, K. Komatsu, T. Kuboyama, “Scientific basis of anti-dementia drugs of constituents from ashwaghanda (Withaniasomnifera),” J. Tad. Med., vol. 22, pp. 176–1822005.
[5] R. Verpoorte, A. Contin, and J.Memelink,“Biotechnology for the production of plant secondary metabolites,” Phytochem. Rev.,vol. 1, pp. 13–25, 2002.
[6] S. Ramachandra Rao, G. A. Ravishankar, “Plant cell cultures: chemical factories of secondary metabolites,” Biotech. Adv., vol.20, pp. 10–153, 2002.
[7] S. Banerjee, A. A. Naqvi, S. Mandal, and P. S. Ahuja, “Transformation of Withaniasomnifera (L.) Dunal by Agrobacterium rhizogenes: Infectivity and phytochemical studies,” Phytother. Res., vol. 8, pp. 452-455, 1994.
[8] M. Furmanowa, D.Gajdzis-Kuls, J.Ruszkowska, Z. Czarnocki, G. Obidoska, A. Sadowska, R. Rani, S. N. Upadhyay, “Invitro propagation of Withaniasomnifera and isolation of withanolides with immunosuppressive activity,” Planta Med., vol.67, pp. 146–149, 2001.
[9] H.N. Murthy, C. Dijkstra, P. Anthony, D. A. White, M. R. Davey, J. B. Power, E. J. Hahn,K. Y. Paek, “Establishment of Withaniasomnifera hairy root cultures for the production of withanolide A,” J. Int. Plant Biol., vol. 50, pp. 975–981, 2008.
[10] S. Ray,S.Jha, “Withanolide synthesis in cultures of Withania somnifera transformed with Agrobacterium tumefaciens,” Plant Science, vol. 166, pp. 1-7, 1999.
[11] S. Ray, S. Jha, “Production of withaferin A from shoot cultures of Withaniasomnifera,”Planta Med., vol. 67, pp. 432-436, 2001.
[12] S. Ray, B. Ghosh, S.Sen, and S. Jha, “Withanolide production by root cultures of Withania somnifera transformed with Agrobacterium rhizogenes,” Planta Med., vol. 62, pp. 571-573, 1996.
[13] G. Roja, M. R. Heble, “Tissue cultures of Withania somnifera: morphogenesis and withanolide synthesis,” Phytother. Res.,vol. 5, pp. 185–187, 1991.
[14] R. S. Sangwan, N. S. Chaurasiya, P. Lal, L. Misra, G. C. Uniyal, R. Tuli, and N. S. Sangwan,“Withanolide A biogeneration in vitro shootcultures of ashwagandha (Withania somniferaDunal), a main medicinal plant in Ayurveda,” Chem. Pharm. Bull., vol. 55, pp. 1371–1375, 2007.
[15] G. Vitali, L. Conte, M. Nicoletti, “Withanolide composition and invitro culture of Italian Withaniasomnifera,”Planta Med., vol. 62, pp. 287–288, 1996.
[16] T. Murashige, and F.Skoog, “A revised medium for rapid growth andbioassays with tobacco tissue cultures,” Physiol. Plant. Vol. 15, pp. 473–497, 1962.
[17] O. L. Gamborg, R. A. Miller, K. Ojima,“Nutrient requirements ofsuspension cultures of soybean root cells,” Exp. Cell Res.,vol. 50, pp. 151–158, 1968.
[18] J. P. Nitsch, and C. Nitsch, “Haploid plants from pollen grains,” Science, vol.163, pp. 85–87, 1969.
[19] C. C. Chu, “The N6 medium and its applications to anther culture of cereal crops. In: Proceedings of symposium plant tissueculture,” Science Press, Beijing, pp. 43–50, 1978.
[20] M. Ganzera, M. I. Choudhary, I. A. Khan, “Quantitative HPLC analysis of withanolides in Withaniasomnifera,”Fitoterapia, vol. 74, pp. 68–76,2003.
[21] C. W. T. Lee, and M. L. Shuler, “The effect of inoculum density and conditioned medium on the production of ajmalicine and catharanthine from immobilized Catharanthusroseus cells,” Biotechnol. Bioeng., vol. 67, pp. 61–67,2000.
[22] [22]C. H. Wu, Y. H. Dewir, E. J. Hahn, K. Y.Paek,“Optimization ofculturing conditions for the production of biomass and phenolicsfrom adventitious roots of Echinacea angustifolia,” J. Plant Biol.Vol. 49, pp. 193–199, 2006.
[23] C. S. Jeong, H. N. Murthy, E. J. Hahn, H. L. Lee, K. Y. Paek, “Inoculumsize and auxin concentration influence the growth of adventitiousroots and accumulation of ginsenosides in suspension cultures of ginseng (Panax ginseng C.A. Meyer),”Acta. Physiol. Plant.Vol. 31, pp. 219–222, 2009.
[24] Z. W. Pan, H. Q. Wang, J. J. Zhong,“Scale-up study on suspension cultures of Taxus chinensis cells for production of taxanediterpene,” Enz.Microb. Technol. Vol. 27, pp. 714-723, 2000.
[25] F. Betsui, N. Tanaka-Nishikawa, K. Shimmomura, “Anthocyaninproduction in adventitious root cultures of RaphanussativusL.cv. Peking Koushin,” Plant Biotechnol., vol. 21, pp. 387–391, 2004.
[26] J. Y. Min, H. Y. Jung, S. M. Kang, Y. D. Kim, Y. M. Kang, D. J. Park, D. T. Prasad, M. S. Choi, “Production of tropane alkaloids by small scale bubble column bioreactor cultures of Scopoliaparviflora adventitious roots, ”Bioresource Technol., vol. 98, pp. 1748–1753, 2007.
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