Volume: 43 Issue: 1
Year: 2023, Page: 291-298, Doi: https://doi.org/10.51248/.v43i01.2298
Introduction and Aim: Gnidia glauca (Fresen.) Gilg of thymelaeaceae is enlisted as an endangered plant. Hemolytic triterpenoid saponins of oleanane and ursane classes are present in G. glauca. The purpose of the present study is to increase the production of hemolytic triterpenoids in vitro using salicylic acid as an elicitor in G. glauca cell suspensions.
Materials and Methods: Explants were collected and surface sterilized, then inoculated onto Murashige and Skoog medium for callus culture, further establishing suspension cultures that were treated with salicylic acid. Saponins produced in the suspension culture were extracted and analyzed by HPLC and FTIR. The hemolytic activity of saponins was confirmed by spectrophotometric assay in erythrocyte suspensions.
Results: Product enhancement of hemolytic triterpenoid saponins was observed in cell suspensions treated with salicylic acid that also induced concomitant change in medium pH compared to control set of culture. Results of HPLC and FTIR analyses confirm the presence of triterpenoid saponins in suspension culture extracts. The optimal concentration of salicylic acid was found to be 200 μM for product enhancement, pH alteration. Hemolytic saponins concentration was observed to be increased with increasing concentrations of elicitor supplemented to the medium.
Conclusion: Cell suspension is a viable alternative technique to produce the phytochemicals in vitro using elicitors that avoids destructive harvesting of endangered plants like G. glauca. Results of the present study demonstrate the large scale production of hemolytic saponins through cost-effective approaches.
Keywords: Saponin; FTIR; HPLC; salicylic acid; hemolytic assay.
1. Sannabommaji, T., Kumar, V., Poornima, D. V., Basappa, G., Gajula, H., Rajashekar, J. Antibacterial Activity of Gnidia glauca (Fresen.) Gilg. phytochemical extract against rice bacterial blight pathogen. Ind. J. Pure. App. Biosci. 2019; 7(5): 287-296.
2. Sannabommaji, T., Kumar, V., Poornima, D.V., Gajula, H., Rajashekar, J., Manjunatha, T., et al., Phytochemical Analysis with Special Reference to Leaf Saponins in Gnidia glauca (Fresen.) Gilg. In: Nitish Kumar (Ed), Biotechnological Approaches for Medicinal and Aromatic Plants Conservation, Genetic Improvement and Utilization; Springer Nature Singapore Pte Ltd. 2018; pp 978-981.
3. Vo, N.N.Q., Fukushima, E.O., Muranaka, T. Structure and hemolytic activity relationships of triterpenoid saponins and sapogenins. 2017; J. Nat. Med. (207) 71: 50-58.
4. Sannabommaji, T., Kumar, V., Poornima, D. V., Basappa, G., Gajula, H., Rajashekar, J. Exploration of Antibacterial Properties of Gnidia glauca (Fresen.) Gilg. Leaf Saponin Fraction. Int. J. Pharm. Phyto. Res. 2020; 12(1): 66-71.
5. Ghosh, S., Parihar, S. P., Dhavale, D. D., Chopade, B. A. Commentary on the therapeutic potential of Gnidia glauca: A novel medicinal plant. Med Chem. 2015; 5: 8.
6. Gowrish, A., Vagdevi, H. M., Rajashekar, H., Kumar, V. M. L., Shobha, B. S. In-vitro cytotoxic and antioxidant activity of Gnidia glauca (Fresen.) Gilg Root extract. J. APP. Chem. 2013; 2(5): 1362-1369.
7. Vijendra, P.D., Jayanna, S.G., Kumar, V., Sannabommaji, T., Rajashekar, J., Gajula, H. Product enhancement of triterpenoid saponins in cell suspension culture of Leucas aspera Spreng. Ind. Crop. Prod. 2020; 156: 112857.
8. Rajashekar, J., Kumar, V., Poornima, D.V., Hari, G., Sannabommaji, T., Raghuramulu, D. Dose-dependent effect of silver nitrate on enhanced production of gymnemic acid in cell suspension cultures of Gymnema Sylvestre R.Br. Biomed. 2021; 41(1): 16-22.
9. Narayani, M., Srivastava, S. Elicitation: A stimulation of stress in in-vitro plant cell/tissue cultures for enhancement of secondary metabolite production. Spri. Sci+. Buss. Med B. V. 2017; 16(6): 1227-1252.
10. Murashige, T., Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant. 1962; 15(3): 473-497.
11. Veerashree, V., Anuradha, C.M., Kumar, V. Elicitor-enhanced production of gymnemic acid in cell suspension culture of Gymnema Sylvestre R.Br. Plant cell. Tiss. Organ. Cult. 2012; 108(1): 27-35.
12. Estrada, K.R., Limon, H.V., Hidalgo, D., Moyano, E., Golenioswki, M., Cusido, R. M., et al., Elicitation an effective strategy for the biotechnology production of bioactive high-added value compounds in plant cell factories. Mole. 2016; 21(2): 182.
13. Bajad, P. N., Pardeshi, A. B., Pagore, V. P. Extraction, isolation and quantification of saponin from Dodonaea viscosa JACQ. The Pharma. Inn. J. 2019; 8(5): 41-44.
14. Hari, G., Kumar, V., Vijendra, P.D., Rajashekar, J., Sannabommaji, T., Basappa, G. A combination of elicitor and precursor enhances psoralen production in Psoralea corylifolia Linn. suspension cultures. Ind. Crop. Prod. 2018; 15(124): 685-691.
15. Noreen, S., Ashraf, M., Hussain, M., Jamil, A. Exogenous application of salicylic acid enhances antioxidative capacity in salt-stressed sunflower (Helianthus annuus L.) plants. Pak. J. Bot. 2009; 41(1): 473-479.
16. Arfan, M., Athar, H. R., Ashraf, M. Does the exogenous application of salicylic acid through the rooting medium modulate growth and photosynthetic capacity in two differently adapted spring wheat cultivars under salt stress? Jour. Plant. Phys. 2007; 164: 685-694.
17. Isah, T. Stress, and defence’s response in plant secondary metabolites production. Isah. Biol. Res. 2019; 52: 39.
18. Ashtiani, S.R., Hasanloo, T., Bihamata, M.R. Enhanced production of silymarin by Ag+ elicitor in cell suspension cultures of Silybum marianum. Pharm. Bio. 2010; 48(6): 708-715.
19. Kareru, P.G., Keriko, J.M., Gachanja, A.N., Kenji, G.M. Direct detection of triterpenoid saponins in medicinal plants. Afr. J. Trad. Comp. Alt. Med. 2008; 5(1): 56-60.
20. Almutairia, M.S., Ali, M. Direct detection of saponins in crude extracts of soapnuts by FTIR. Nat. Prod. Res. 2015; 29(13): 1271-1275.
21. Sarikahya, N.B., Nalbantsoy, T, H., Gokturk, R.S., Sumbul, H., Kirmizigul. Immunomodulatory, hemolytic, and cytotoxic activity potentials of triterpenoid saponins from eight Cephalaria species. Phytmed. 2018; 38(1): 135-144.
22. Voutquenne, L., Lavaud, C., Massiot, G., Men-Olivier, L. L. Structure-activity relationship of hemolytic saponins. Pharm. Bio. 2002; 40(4): 253-262.
23. Duc, M.H., Phuong, T.P., Trung, Q. B., Anh, T .L. N., Quyen, T.N., Trang, T.K., et al., Stem cell-based therapy for human diseases. Signal. Trans. Targ. Ther. 2022; 7: 272.
24. Zivile, G., Gabrielis, K., Regina, L., Jelena, R., Marcin, J. Quick and effective method of bone marrow mesenchymal stem cell extraction. Open Med. 2015; 10: 44-49.
Vishala E., Vadlapudi Kumar, Ruksana F., Poornima D.V., Anuradha C.M., Prathap H.M., Pratap G.K., Savitharani M., Manjunatha T.. Hemolytic saponins product enhancement in Gnidia glauca (Fresen.) Gilg
cell suspension cultures. Biomedicine: 2023; 43(1): 291-298