Preprint / Version 1

In vitro production of steroidal saponin, total phenols and antioxidant activity in callus suspension culture of Paris polyphylla Smith: an important Himalayan medicinal plant

Authors

  • Janhvi Rawat Department of Biotechnology, Graphic Era Deemed to be University, Dehradun, Uttarakhand, India
  • Ahmed El-Gazzar Department of Veterinary Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, Alexandria University, Alexandria, Egypt
  • Mohamed Mahmoud Department of Biochemistry, College of Science, King Saud University, Riyadh, Saudi Arabia
  • Ajay Thakur Genetics and Tree Propagation Division, Forest Research Institute, Dehradun, Uttarakhand, India
  • Arvind Negi School of Agriculture, Graphic Era Hill University, Dehradun, Uttarakhand, India
  • Jigisha Anand Department of Biotechnology, Graphic Era Deemed to be University, Dehradun, Uttarakhand, India
  • Sumit Purohit Department of Biotechnology, Uttarakhand Biotechnology Council, Pantnagar, Uttarakhand, India
  • Balwant Rawat School of Agriculture, Graphic Era Hill University, Dehradun, Uttarakhand, India
  • Shweta Pandey School of Agriculture, Graphic Era Hill University, Dehradun, Uttarakhand, India
  • Gaber Batiha Department of Pharmacology and Therapeutics, Damanhour University, Damanhour, Egypt

Keywords:

Paris polyphylla, dioscin, diosgenin, steroidal saponin, antioxidant activity

Abstract

Paris polyphylla Smith (Melanthiaceae) family, which is native to the Himalayan region, has received a lot of attention recently due to its extensive history of usage in traditional medicine. The production of steroidal saponin from callus suspension cultures of P. polyphylla was observed in the current study. The current study attempted to develop a P. polyphylla plant callus suspension culture through optimization of cultivation technique for callus suspension, quantification of total phenolic components and estimation of the extract’s antioxidant activity. A light-yellow callus was formed within six weeks of cultivating rhizomes on Murashige and Skoog (MS) media supplemented with Thidiazuron (TDZ). Furthermore, the effect of TDZ, Methyl Jasmonate (MeJA), and Yeast Extract (YE) on callus growth, steroidal saponin (dioscin and diosgenin), total phenolic content, total flavonoids, total tannin, and total antioxidant activity was also measured. The medium containing 0.5 μM TDZ depicted the maximum callus biomass (2.98 g fresh weight). Significantly high phenolic and tannin content was observed in the MS medium containing 50 μM MeJA, whereas, no significant increase was observed in total tannin production in any treatment. Three in vitro assays, DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2′-azino-bis (3-ethylbenzothiazoline- 6-sulfonic acid)) and FRAP (ferric ion reducing antioxidant potential) and FC (Folin-Ciocalteu), were used to assess antioxidant potential of callus. Maximum antioxidant analysis reported in 1.0 μM TDZ (6.89 mM AAE/100 g) containing medium followed by 50 μM MeJA (6.44 mM AAE/100 g). The HPLC analysis showed a high presence of dioscin and diosgenin (5.43% and 21.09%, respectively) compared to the wild sample (2.56% and 15.05%, respectively). According to the results, callus produced on media supplemented with 50 μM MeJA have significant phenolic contents and elevated antioxidant activity; nevertheless, callus growth was greater in the presence of 0.5 μM TDZ. The findings of the current study have commercial implications since greater biomass production will result in active phytochemicals that the pharmaceutical and nutraceutical sectors are in need desperately. Keywords: Paris polyphylla, dioscin, diosgenin, steroidal saponin, antioxidant activity

Author Biography

Ahmed El-Gazzar, Department of Veterinary Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, Alexandria University, Alexandria, Egypt

Department of Experimental Pathology and Tumor Biology, Graduate School of Medical Sciences, Nagoya City University, Nagoya, Japan