year 21, Issue 81 (3-2022)                   J. Med. Plants 2022, 21(81): 79-91 | Back to browse issues page


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Mirjalili M H, Esmaeili H. Callus induction and withanolides production through cell suspension culture of Withania coagulans (Stocks) Dunal. J. Med. Plants 2022; 21 (81) :79-91
URL: http://jmp.ir/article-1-3233-en.html
1- Department of Agriculture, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, 1983969411, Tehran, Iran , m-mirjalili@sbu.ac.ir
2- Department of Agriculture, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, 1983969411, Tehran, Iran
Abstract:   (2919 Views)
Background: Withania coagulans (Stocks) Dunal is a well-known medicinal plant due to its many healing properties. Objective: The aim of the present study was to induce friable callus and subsequently establish the plant cell suspension cultures for the production of two important withanolides i.e. withaferin A (WFA) and withanolide A (WNA). Methods: In vitro callus induction was carried out from young leaf and internodal explants cultured on MS medium fortified with various concentrations (0, 1.0, 1.5, 2.0 and 2.5 mg/L) of auxins (2,4-D, NAA, and IAA) solely or in combination with BAP (0., 0.5 and 1.0 mg/L) in a factorial experiment based on a completely randomized design with five replications. The plant cell culture was then established for the production of both withanolides. Results: The percentage of callogenesis from the leaf (25.0-96.0 %) was higher than internodal explants (23.2-85.4 %). The high percentage of friable calli was achieved from leaf explants cultured on MS medium fortified with 2.5 mg/L 2,4-D + 0.5 mg/L BAP. Cell suspension culture was established from derived friable callus cultured on MS medium supplemented with 1.5 mg/L IAA + 0.5 mg/L BAP. The highest accumulation of biomass (172 g/L fresh weight and 15 g/L dry weight) and the production of both withanolides were observed in the fourth week of the culture period. The plant cells produced 0.08 and 21 µg/L WFA and WNA at this time, respectively. Conclusion: These results can be used for future research on biosynthesis pathways of withanolides as well as their production in bioreactors.
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Type of Study: Research | Subject: Medicinal Plants
Received: 2021/12/22 | Accepted: 2022/02/19 | Published: 2022/03/1

References
1. Schonbeck-Temesy, E Solanaceae In Rechinger, KH. (Ed.). Flora Iranica, No. 100, Akademische Druck-u. Verlagsanstalt, Graz, Austria. 1972, pp: 29-26.
2. Hepper F.N Old World Withania (Solanaceae): a taxonomic review and key to the species. loc. cit. 1991, pp: 211-27.
3. Williamson E Major herbs of Ayurveda, Churchill Livingstone, London. 2002: 323.
4. Gupta GL and Rana AC. Withania somnifera (Ashwagandha): a review. Pharmacognosy Reviews. 2007; 1(1): 129-136.
5. Glotter E. Withanolides and related ergostane-type steroids. Natural Product Reports 1991; 8(4): 415-40. [DOI:10.1039/np9910800415]
6. Yang H, Shi G and Dou QP. The tumor proteasome is a primary target for the natural anticancer compound Withaferin A isolated from "Indian winter cherry". Molecular Pharmacology 2007; 71(2): 426-37. [DOI:10.1124/mol.106.030015]
7. Kuboyama T, Tohda C and Komatsu K. Neuritic regeneration and synaptic reconstruction induced by withanolide A. British Journal of Pharmacol. 2005; 144(7): 961-71. [DOI:10.1038/sj.bjp.0706122]
8. Tohda C, Kuboyama T and Komatsu K. Search for natural products related to regeneration of the neuronal network. Neurosignals 2005; 14(1-2): 34-45. [DOI:10.1159/000085384]
9. Mirjalili HM, Fakhr‐Tabatabaei SM, Bonfill M, Alizadeh H, Cusido RM, Ghassempour A and Palazon J. Morphology and withanolide production of Withania coagulans hairy root cultures. Engineering in Life Sciences 2009; 9(3): 197-204. [DOI:10.1002/elsc.200800081]
10. Mirjalili MH, Fakhr-Tabatabaei SM, Alizadeh H, Ghassempour A and Mirzajani F. Genetic and withaferin A analysis of Iranian natural populations of Withania somnifera and W. coagulans by RAPD and HPTLC. Natural Product Communications 2009; 4(3): 1934578X0900400307. [DOI:10.1177/1934578X0900400307]
11. Mirjalili MH, Moyano E, Bonfill M, Cusido RM and Palazon J. Overexpression of the Arabidopsis thaliana squalene synthase gene in Withania coagulans hairy root cultures. Biologia Plantarum. 2011; 55(2): 357-60. [DOI:10.1007/s10535-011-0054-2]
12. Mirjalili MH, Moyano E, Bonfill M, Cusido RM and Palazón J. Steroidal lactones from Withania somnifera, an ancient plant for novel medicine. Molecules. 2009; 14 (7): 2373-2393. [DOI:10.3390/molecules14072373]
13. Verpoorte R, Contin A and Memelink J. Biotechnology for the production of plant secondary metabolites. Phytochemistry Reviews. 2002; 1(1): 13-25. [DOI:10.1023/A:1015871916833]
14. Rao SR and Ravishankar GA. Plant cell cultures: chemical factories of secondary metabolites. Biotechnology Advances 2002; 20(2): 101-53. [DOI:10.1016/S0734-9750(02)00007-1]
15. Namdeo AG and Ingawale DK. Ashwagandha: Advances in plant biotechnological approaches for propagation and production of bioactive compounds. Journal of Ethnopharmacology 2021; 271: 113709. [DOI:10.1016/j.jep.2020.113709]
16. Sabir F, Sangwan NS, Chaurasiya ND, Misra LN and Sangwan RS. In vitro withanolide production by Withania somnifera L. cultures. Zeitschrift für Naturforschung C. 2008; 63 (5-6): 409-412. [DOI:10.1515/znc-2008-5-616]
17. Nagella P and Murthy HN. Establishment of cell suspension cultures of Withania somnifera for the production of withanolide A. Bioresource Technology 2010; 101 (17): 6735-6739. [DOI:10.1016/j.biortech.2010.03.078]
18. Sivanandhan G, Selvaraj N, Ganapathi A and Manickavasagam M. Elicitation approaches for withanolide production in hairy root culture of Withania somnifera (L.) Dunal. In Biotechnology of Plant Secondary Metabolism. 2016; 1-18. [DOI:10.1007/978-1-4939-3393-8_1]
19. Tripathi D, Rai KK, Rai SK and Rai SP. An improved thin cell layer culture system for efficient clonal propagation and in vitro withanolide production in a medicinal plant Withania coagulans Dunal. Industrial Crops and Products. 2018; 119: 172-82. [DOI:10.1016/j.indcrop.2018.04.012]
20. Sivanandhan G, Selvaraj N, Ganapathi A and Lim YP. Up-regulation of Squalene synthase in hairy root culture of Withania somnifera (L.) Dunal yields higher quantities of withanolides. Industrial Crops and Products 2020; 154: 112706. [DOI:10.1016/j.indcrop.2020.112706]
21. Murashige T and Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 1962; 15(3): 473-497. [DOI:10.1111/j.1399-3054.1962.tb08052.x]
22. Yousefian Z, Hosseini B, Rezadoost H, Palazón J and Mirjalili MH. Production of the anticancer compound withaferin a from genetically transformed hairy root cultures of Withania somnifera. Natural Product Communications 2018; 13(8): 1934578X1801300806. [DOI:10.1177/1934578X1801300806]
23. Rani G and Grover IS. In vitro callus induction and regeneration studies in Withania somnifera. Plant Cell, Tissue and Organ Culture. 1999; 57(1): 23-7. [DOI:10.1023/A:1006329532561]
24. George EF, Hall MA and De Klerk G-J. Plant propagation by tissue culture. Volume I. The background. Plant Propagation by Tissue Culture. 2008; 1: 205-226. [DOI:10.1007/978-1-4020-5005-3_1]
25. Rani G, Virk GS and Nagpal A. Callus induction and plantlet regeneration in Withania somnifera (L.) Dunal. In vitro Cellular & Developmental Biology-Plant. 2003; 39(5): 468-74. [DOI:10.1079/IVP2003449]
26. Singh S, Tanwer BS and Khan M. Callus induction and in vivo and in vitro comparative study of primary metabolites of Withania somnifera. Adv. Appl. Sci. Res. 2011; 2(3): 47-52.
27. Nagella P and Murthy HN. Establishment of cell suspension cultures of Withania somnifera for the production of withanolide A. Bioresource Technology 2010; 101(17): 6735-9. [DOI:10.1016/j.biortech.2010.03.078]
28. Chakraborty N, Banerjee D, Ghosh M, Pradhan P, Gupta NS, Acharya K and Banerjee M. Influence of plant growth regulators on callus mediated regeneration and secondary metabolites synthesis in Withania somnifera (L.) Dunal. Physiology and Molecular Biology of Plants. 2013; 19(1): 117-25. [DOI:10.1007/s12298-012-0146-2]
29. Skoog F and Miller C. Chemical regulation of growth and organ formation in plant tissues cultured. In: Vitro Symp Soc Exp Biol. 1957; 11: 118-131.
30. Ikeuchi M, Sugimoto K and Iwase A. Plant callus: mechanisms of induction and repression. The Plant Cell 2013; 25 (9): 3159-3173. [DOI:10.1105/tpc.113.116053]
31. Zhong JJ. Biochemical engineering of the production of plant-specific secondary metabolites by cell suspension cultures. Plant Cells 2001:1-26. [DOI:10.1007/3-540-45302-4_1]
32. Yue W, Ming QL, Lin B, Rahman K, Zheng CJ, Han T and Qin LP. Medicinal plant cell suspension cultures: pharmaceutical applications and high-yielding strategies for the desired secondary metabolites. Critical Reviews in Biotechnology 2016; 36(2): 215-32. [DOI:10.3109/07388551.2014.923986]
33. Nagella P and Murthy HN. Effects of macroelements and nitrogen source on biomass accumulation and withanolide-A production from cell suspension cultures of Withania somnifera (L.) Dunal. Plant Cell, Tissue and Organ Culture (PCTOC). 2011; 104(1): 119-24. [DOI:10.1007/s11240-010-9799-0]
34. Ahlawat S, Saxena P, Ali A, Khan S and Abdin MZ. Comparative study of withanolide production and the related transcriptional responses of biosynthetic genes in fungi elicited cell suspension culture of Withania somnifera in shake flask and bioreactor. Plant Physiology and Biochemistry 2017; 114: 19-28. [DOI:10.1016/j.plaphy.2017.02.013]
35. Sivanandhan G, Mariashibu TS, Arun M, Rajesh M, Kasthurirengan S, Selvaraj N and Ganapathi A. The effect of polyamines on the efficiency of multiplication and rooting of Withania somnifera (L.) Dunal and content of some withanolides in obtained plants. Acta Physiologiae Plantarum. 2011; 33(6): 2279-88. [DOI:10.1007/s11738-011-0768-y]

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