A well-known herbal remedy for a variety of conditions is withania sominifera. Using a combination of indole butyric acid (IBA) at 1.0 mg/l and indole acetic acid (IAA) at 0.5 mg/L, adventitious roots were successfully initiated from leaf explants of in vitro propagated plantlets in MS medium in the current study. The best conditions for inducing hairy roots were found to be a co-culture period of 24 hours with Agrobacterium rhizogenes. Within 7 days of treatment, hairy roots started to appear. The addition of 50 M acetosyringone and a 1-hour infection time was found to increase the transformation frequency from 72.11 0.5% to 81.22 0.24%. Following a 30-day period of culture, the induced roots were moved to a shake flask system, and the growth index was discovered to be 33.35 for adventitious roots and 37.24 for hairy roots. The induced roots were moved to a shake flask culture system for a 30-day period of culture, and the growth indices for adventitious roots and hairy roots were found to be 33.35 and 37.24, respectively. The roots were extracted after being collected, and they were then screened for phytochemicals using GC-MS, TLC, and HPLC. Alkaloids, flavanoids, tannins, saponins, phenols, glycosides, terpenoids, reducing sugar, and anthraquinones were found in these analyses. The presence of Withaferin A and Withanolide A was confirmed by HPLC of the methanolic extracts of the roots. Withaferin A was present in transformed roots at a concentration of 69.21 g/g dry weight, which was two times greater than that of adventitious root cultures at 32.45 g/g dry weight. This work emphasises the importance of Withania sominifera poshita variety adventitious and hairy root culture, which may be utilised for systematic and reliable metabolite production that can be used for scientific and commercial applications.
| Published in | American Journal of Plant Biology (Volume 8, Issue 2) |
| DOI | 10.11648/j.ajpb.20230802.13 |
| Page(s) | 36-42 |
| Creative Commons |
This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited. |
| Copyright |
Copyright © The Author(s), 2023. Published by Science Publishing Group |
Withania sominifera Poshita Variety, Adventitious Roots, Hairy Roots
| [1] | Ansari, A. Q., et al., Extraction and determination of antioxidant activity of Withania somnifera Dunal. Eur J Exp Biol, 2013. 3 (5): p. 502-507. |
| [2] | Kaur, K., et al., Evaluation of the anti-proliferative and anti-oxidative activities of leaf extract from in vivo and in vitro raised Ashwagandha. Food and chemical toxicology, 2004. 42 (12): p. 2015-2020. |
| [3] | Ahmad, M., et al., Neuroprotective effects of Withania somnifera on 6-hydroxydopamine induced Parkinsonism in rats. Human & experimental toxicology, 2005. 24 (3): p. 137-147. |
| [4] | Adaikkappan, P., M. Kannapiran, and A. Anthonisamy, Antimycobacterial activity of Withania somnifera and Pueraria tuberosa against Mycobacterium tuberculosis H37Rv. J. Acad. Indus. Res, 2012. 1 (4): p. 153-156. |
| [5] | Singariya, P., P. Kumar, and K. Mourya, Comparative primary phyto-profile and microcidal activity of Cenchrus ciliaris (Anjan grass) and Withania somnifera (winter cherry). International Journal of Research in Ayurveda and Pharmacy (IJRAP), 2012. 3 (2): p. 303-308. |
| [6] | Wankhede, S., et al., Examining the effect of Withania somnifera supplementation on muscle strength and recovery: a randomized controlled trial. Journal of the International Society of Sports Nutrition, 2015. 12 (1): p. 43. |
| [7] | Ilayperuma, I., W. Ratnasooriya, and T. Weerasooriya, Effect of Withania somnifera root extract on the sexual behaviour of male rats. Asian Journal of Andrology, 2002. 4 (4): p. 295-298. |
| [8] | Chandrasekhar, K., J. Kapoor, and S. Anishetty, A prospective, randomized double-blind, placebo-controlled study of safety and efficacy of a high-concentration full-spectrum extract of ashwagandha root in reducing stress and anxiety in adults. Indian journal of psychological medicine, 2012. 34 (3): p. 255. |
| [9] | Imtiyaz, S., et al., Withania somnifera: a potent unani aphrodisiac drug. International Research Journal of Pharmaceutical And Applied Sciences, 2013. 3 (4): p. 59-63. |
| [10] | Saiyed, A., et al., Medicinal properties, phytochemistry and pharmacology of Withania somnifera: an important drug of Unani Medicine. Journal of Scientific & Innovative Research, 2016. 5 (4): p. 156-60. |
| [11] | Yadav, B., et al., In vitro anticancer activity of the root, stem and leaves of Withania somnifera against various human cancer cell lines. Indian Journal of Pharmaceutical Sciences, 2010. 72 (5): p. 659. |
| [12] | Sharma, V., et al., A validated and densitometric HPTLC method for the quantification of withaferin-A and withanolide-A in different plant parts of two morphotypes of Withania somnifera. Chromatographia, 2007. 66 (9-10): p. 801-804. |
| [13] | Misra, H., Registration of a new variety Poshita of Withania somnifera. J. Medi. Aro. Plant Sci., 2001. 23: p. 97-98. |
| [14] | Mirjalili, M. H., et al., Steroidal lactones from Withania somnifera, an ancient plant for novel medicine. Molecules, 2009. 14 (7): p. 2373-2393. |
| [15] | Misra, L., et al., Unusually sulfated and oxygenated steroids from Withania somnifera. Phytochemistry, 2005. 66 (23): p. 2702-2707. |
| [16] | Sangwan, R. S., et al., Withanolide A is inherently de novo biosynthesized in roots of the medicinal plant Ashwagandha (Withania somnifera). Physiologia plantarum, 2008. 133 (2): p. 278-287. |
| [17] | Nagella, P. and H. N. Murthy, Establishment of cell suspension cultures of Withania somnifera for the production of withanolide A. Bioresource Technology, 2010. 101 (17): p. 6735-6739. |
| [18] | Viji, M. O. and N. Wilson, Effect of Piriformospora Indica on Secondary Metabolite Production in Hairy Root Cultures of Withania Sominifera Poshita Variety. International Journal of Emerging Technology and Advanced Engineering, 2017. 7 (9). |
| [19] | Nilsson, O. and O. J. P. P. Olsson, Getting to the root: the role of the Agrobacterium rhizogenes rol genes in the formation of hairy roots. 1997. 100 (3): p. 463-473. |
| [20] | Pavlova, O., T. Matveyeva, and L. J. R. J. o. G. A. R. Lutova, rol-Genes of Agrobacterium rhizogenes. 2014. 4 (2): p. 137-145. |
| [21] | Schmülling, T., J. Schell, and A. J. T. E. j. Spena, Single genes from Agrobacterium rhizogenes influence plant development. 1988. 7 (9): p. 2621-2629. |
| [22] | Murashige, T. and F. Skoog, A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia plantarum, 1962. 15 (3): p. 473-497. |
| [23] | De Klerk, G.-J., et al., Effectiveness of indoleacetic acid, indolebutyric acid and naphthaleneacetic acid during adventitious root formation in vitro in Malus ‘Jork 9’. 1997. 49 (1): p. 39-44. |
| [24] | Chabaud, M., et al., Agrobacterium rhizogenes-mediated root transformation. 2006. |
| [25] | Tzfira, T., et al., Agrobacterium rhizogenes-mediated DNA transfer in Pinus halepensis Mill. 1996. 16 (1): p. 26-31. |
| [26] | Sivanesan, I. and B. R. J. A. J. o. B. Jeong, Induction and establishment of adventitious and hairy root cultures of Plumbago zeylanica L. 2009. 8 (20). |
| [27] | Halket, J. M., et al., Chemical derivatization and mass spectral libraries in metabolic profiling by GC/MS and LC/MS/MS. Journal of Experimental Botany, 2004. 56 (410): p. 219-243. |
| [28] | Wei, K., et al., Auxin-Induced Adventitious Root Formation in Nodal Cuttings of Camellia sinensis. International Journal of Molecular Sciences, 2019. 20 (19): p. 4817. |
| [29] | Anoopkumar, A., et al., A novel intervention on the inhibiting effects of Catunaregam spinosa induced free radical formation and DNA damage in Aedes aegypti (Diptera: Culicidae): a verdict for new perspectives on microorganism targeted vector control approach. INTERNATIONAL JOURNAL OF TROPICAL INSECT SCIENCE, 2020. |
| [30] | Anoopkumar, A., E. M. Aneesh, and A. V. Sudhikumar, Exploring the mode of action of isolated bioactive compounds by induced reactive oxygen species generation in Aedes aegypti: a microbes based double-edged weapon to fight against Arboviral diseases. International Journal of Tropical Insect Science, 2020: p. 1-13. |
| [31] | Anoopkumar, A., et al., Screening of a few traditionally used medicinal plants for their larvicidal efficacy against Aedes aegypti Linn (Diptera: Culicidae), a dengue fever vector. SOJ Microbiol Infect Dis, 2017. 5 (4): p. 1-5. |
| [32] | Yang, H., G. Shi, and Q. P. Dou, The tumor proteasome is a primary target for the natural anticancer compound Withaferin A isolated from “Indian winter cherry”. Molecular pharmacology, 2007. 71 (2): p. 426-437. |
| [33] | Shukla, D. D., N. Bhattarai, and B. Pant, In-vitro mass propagation of Withania somnifera (L.) Dunal. Nepal journal of Science and Technology, 2010. 11: p. 101-106. |
| [34] | Thomas, T. D. and B. Philip, Thidiazuron-induced high-frequency shoot organogenesis from leaf-derived callus of ia medicinal climber, Tylophora Indica (Burm. F.) merrill. In Vitro Cellular & Developmental Biology-Plant, 2005. 41 (2): p. 124-128. |
| [35] | Fatima, N., N. Ahmad, and M. Anis, In Vitro Propagation and Conservation of Withania somnifera (Dunal) L, in Protocols for In Vitro Cultures and Secondary Metabolite Analysis of Aromatic and Medicinal Plants, Second Edition. 2016, Springer. p. 303-315. |
APA Style
Viji Mo, Neeba Wilson. (2023). Enhanced Production of Secondary Metabolites Through Adventitious and Hairy Root Cultures of Withania sominifera cv. Poshita. American Journal of Plant Biology, 8(2), 36-42. https://doi.org/10.11648/j.ajpb.20230802.13
ACS Style
Viji Mo; Neeba Wilson. Enhanced Production of Secondary Metabolites Through Adventitious and Hairy Root Cultures of Withania sominifera cv. Poshita. Am. J. Plant Biol. 2023, 8(2), 36-42. doi: 10.11648/j.ajpb.20230802.13
AMA Style
Viji Mo, Neeba Wilson. Enhanced Production of Secondary Metabolites Through Adventitious and Hairy Root Cultures of Withania sominifera cv. Poshita. Am J Plant Biol. 2023;8(2):36-42. doi: 10.11648/j.ajpb.20230802.13
Share This Article
Twitter
Linked In
Facebook
Copy
Download@article{10.11648/j.ajpb.20230802.13,
author = {Viji Mo and Neeba Wilson},
title = {Enhanced Production of Secondary Metabolites Through Adventitious and Hairy Root Cultures of Withania sominifera cv. Poshita},
journal = {American Journal of Plant Biology},
volume = {8},
number = {2},
pages = {36-42},
doi = {10.11648/j.ajpb.20230802.13},
url = {https://doi.org/10.11648/j.ajpb.20230802.13},
eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajpb.20230802.13},
abstract = {A well-known herbal remedy for a variety of conditions is withania sominifera. Using a combination of indole butyric acid (IBA) at 1.0 mg/l and indole acetic acid (IAA) at 0.5 mg/L, adventitious roots were successfully initiated from leaf explants of in vitro propagated plantlets in MS medium in the current study. The best conditions for inducing hairy roots were found to be a co-culture period of 24 hours with Agrobacterium rhizogenes. Within 7 days of treatment, hairy roots started to appear. The addition of 50 M acetosyringone and a 1-hour infection time was found to increase the transformation frequency from 72.11 0.5% to 81.22 0.24%. Following a 30-day period of culture, the induced roots were moved to a shake flask system, and the growth index was discovered to be 33.35 for adventitious roots and 37.24 for hairy roots. The induced roots were moved to a shake flask culture system for a 30-day period of culture, and the growth indices for adventitious roots and hairy roots were found to be 33.35 and 37.24, respectively. The roots were extracted after being collected, and they were then screened for phytochemicals using GC-MS, TLC, and HPLC. Alkaloids, flavanoids, tannins, saponins, phenols, glycosides, terpenoids, reducing sugar, and anthraquinones were found in these analyses. The presence of Withaferin A and Withanolide A was confirmed by HPLC of the methanolic extracts of the roots. Withaferin A was present in transformed roots at a concentration of 69.21 g/g dry weight, which was two times greater than that of adventitious root cultures at 32.45 g/g dry weight. This work emphasises the importance of Withania sominifera poshita variety adventitious and hairy root culture, which may be utilised for systematic and reliable metabolite production that can be used for scientific and commercial applications.},
year = {2023}
}
TY - JOUR T1 - Enhanced Production of Secondary Metabolites Through Adventitious and Hairy Root Cultures of Withania sominifera cv. Poshita AU - Viji Mo AU - Neeba Wilson Y1 - 2023/07/06 PY - 2023 N1 - https://doi.org/10.11648/j.ajpb.20230802.13 DO - 10.11648/j.ajpb.20230802.13 T2 - American Journal of Plant Biology JF - American Journal of Plant Biology JO - American Journal of Plant Biology SP - 36 EP - 42 PB - Science Publishing Group SN - 2578-8337 UR - https://doi.org/10.11648/j.ajpb.20230802.13 AB - A well-known herbal remedy for a variety of conditions is withania sominifera. Using a combination of indole butyric acid (IBA) at 1.0 mg/l and indole acetic acid (IAA) at 0.5 mg/L, adventitious roots were successfully initiated from leaf explants of in vitro propagated plantlets in MS medium in the current study. The best conditions for inducing hairy roots were found to be a co-culture period of 24 hours with Agrobacterium rhizogenes. Within 7 days of treatment, hairy roots started to appear. The addition of 50 M acetosyringone and a 1-hour infection time was found to increase the transformation frequency from 72.11 0.5% to 81.22 0.24%. Following a 30-day period of culture, the induced roots were moved to a shake flask system, and the growth index was discovered to be 33.35 for adventitious roots and 37.24 for hairy roots. The induced roots were moved to a shake flask culture system for a 30-day period of culture, and the growth indices for adventitious roots and hairy roots were found to be 33.35 and 37.24, respectively. The roots were extracted after being collected, and they were then screened for phytochemicals using GC-MS, TLC, and HPLC. Alkaloids, flavanoids, tannins, saponins, phenols, glycosides, terpenoids, reducing sugar, and anthraquinones were found in these analyses. The presence of Withaferin A and Withanolide A was confirmed by HPLC of the methanolic extracts of the roots. Withaferin A was present in transformed roots at a concentration of 69.21 g/g dry weight, which was two times greater than that of adventitious root cultures at 32.45 g/g dry weight. This work emphasises the importance of Withania sominifera poshita variety adventitious and hairy root culture, which may be utilised for systematic and reliable metabolite production that can be used for scientific and commercial applications. VL - 8 IS - 2 ER -
Email: