Development Of Optimized Liquid Culture System For Hybanthus Enneaspermus (L.) F. Muell Using Response Surface Methodology
Keywords:
Hybanthus enneaspermus, Media optimization, Response surface methodology, Central composite design
Abstract
Hybanthus enneaspermus (L.) F. Muell. is a pharmacologically important ethnobotanical herb. Over exploitation from its natural habitats has led to its scarcity. To fulfil the growing pharmaceutical demands, development of novel in vitro culture methods for H. enneaspermusis necessary. In the present study, the central composite design (CCD) of response surface methodology (RSM) has been applied for liquid media component optimization for shoot regeneration from leaf explants. Research Method: An experimental model comprising of four independent variables of growth regulators (Kinetin (KN), 6-benzylamonipurine (BAP), indole acetic acid (IAA), and Salicylic acid (SA) and their two dependents responses (shoot regeneration percentage (Y1) and the average number of shoots per explant (Y2) were calculated. Findings: The maximum responses for Y1 and Y2, was 88.33 ± 0.75 and 72.5 ± 0.84 respectively were obtained with the growth regulator concentrations - KN2.31µM, BAP 4.44 µM, IAA 2.84 µM, and SA 100mg.MS salt at full strength and salicylic acid at concentration 100 mg/l were found to be most suitable for shoot regeneration in liquid culture media. H. enneaspermus plants with high aurantiamide acetate and coumarin levels were obtained using SA containing liquid culture media. Limitation: Large scale application of this procedure needs to be cost-effective owing to the cost involved in designing the same. Value: total flavonoids and phenols content along with chlorophyll accumulation were found to increase in plants cultured on liquid MS media.References
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2. Behera, P. R., Jena, R. C., Das, A., Thirunavoukkarasu, M., & Chand, P. K. (2016). Genetic stability and coumarin content of transformed rhizoclones and regenerated plants of a multi-medicinal herb, Hybanthus enneaspermus (L.) F. Muell. Plant Growth Regulation, 80, 103-114.
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5. Karthi, C., & Velayutham, P. (2017). Direct organogenesis and mass propagation from the nodal explants of Hybanthus enneaspermus (L.) F. Muell. Plant Cell Biotechnology and Molecular Biology, 18(5&6), 290-296.
6. Murugan, M., & Kamaraj, M. (2018). In vitro propagation and conservation of useful ethnomedicinal plant of Hybanthus enneaspermus (Linn.) F. Muell. belonging to the violaceae family. International Journal of Current Research in Life Sciences, 7(7), 2493-2499.
7. Parthasarathy, S. P., Anusuya, S., Rajalakshmi, S., Megha, D., Appunu, C., Alagumanian, S., & Manickavasagam, M. (2024). Elucidating the efficacy of functionalized multi-walled carbon nanotube in the biogenesis of L-Dopa and antioxidant metabolites in cell cultures of Hybanthus enneaspermus. Plant Physiology and Biochemistry, 206, 108310.
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10. Premkumar, G., Arumugam, N., Muthuramkumar, S., Varatharaju, G., & Rajarathinam, K. (2013). Improved micropropagation in Hybanthus enneaspermus L. Muell.
11. Premkumar, G., Karuppanapandian, T., Sureshpandian, C., Arumugam, N., Selvam, A., & Rajarathinam, K. (2020). Optimization of a liquid culture system for shoot regeneration and achieving an enriched level of scopadulcic acid b in the leaf organ cultures of Scoparia dulcis L. by response surface methodology. In Vitro Cellular & Developmental Biology-Plant, 56, 60-71.
12. Rajsekhar, P., Bharani, R., Angel, K. J., Ramachandran, M., & Rajsekhar, S. P. V. (2016). Hybanthus enneaspermus (L) F. Muell: A phytopharmacological review on herbal medicine. J. Chem. Pharm. Res, 8(1), 351-355.
13. Ramya, A. K. N., & Devika, R. (2022). A Current Pharmacological Impacts and Perspective of Hybanthus enneaspermus (Linn.) F. Muell. Pharmacognosy Research, 14(4).
14. Shekhawat, M. S., & Manokari, M. (2016). Somatic embryogenesis and in vitro flowering in Hybanthus enneaspermus (L.) F. Muell.–A rare multipotent herb. Asian Pacific Journal of Reproduction, 5(3), 256-262.
15. Shekhawat, M. S., & Manokari, M. (2018). In vitro multiplication, micromorphological studies and ex vitro rooting of Hybanthus enneaspermus (L.) F. Muell.–a rare medicinal plant. Acta Botanica Croatica, 77(1), 80-87.
16. Singleton, V. L., Orthofer, R., & Lamuela-Raventós, R. M. (1999). [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. In Methods in enzymology (Vol. 299, pp. 152-178): Elsevier.
17. Velayutham, P., & Karthi, C. (2020). Rapid and Mass Propagation of Hybanthus enneaspermus (L.) F. Muell. from Shoot Tip and Nodal Explants.
18. Velayutham, P., Karthi, C., Nalini, P., & Jahirhussain, G. (2012). In vitro Regeneration and Mass Propagation of Hybanthus enneaspermus (L.) F. Muell. from the stem explants through callus culture. Journal of Agricultural Technology, 8(3), 1119-1128.
19. Vyas, S., Rao, M. S., Suthar, R. K., & Purohit, S. D. (2008). Liquid culture system stimulates in vitro growth and shoot multiplication in four medicinally important plants. Medicinal and Aromatic Plant Science and Biotechnology, 2(2), 96-100.
20. Nirmal, D., Teraiya , S., & Joshi, P.(2023). Liquid culture system: An efficient approach for sustainable micropropagation. Current Agriculture Research Journal, 11 , 28-42.
21. Sathish, S., Vasudevan, V., Karthik, S., Pavan, G., & Manikavasagan, M.(2019). Enhanced L-Dopa production from elicited cell suspension culture of Hybanthus enneaspermus(L.)F.Muell. Plant Biotechnology Reports, 13, 613-621.
22. Sathish, S., Vasudevan, V., Karthik,S., Elayaraja, D., Pavan, G., Ajithan, C., &Manikavasagan, M.(2020). Elicitors induced L-Dopa accumulation in adventitious root cultures of Hybanthus enneaspermus (L.) F . Muell.Vegetos, 33, 304-312.
23. Kaur, P., Gupta, R. C., Dey, A., Malik, T., & Pandey, D. K. (2020). Optimization of salicylic acid and chitosan treatment for bitter secoiridoid and xanthone glycosides production in shoot cultures of Swertia paniculata using response surface methodology and artificial neural network. BMC Plant Biology, 20(1).
2. Behera, P. R., Jena, R. C., Das, A., Thirunavoukkarasu, M., & Chand, P. K. (2016). Genetic stability and coumarin content of transformed rhizoclones and regenerated plants of a multi-medicinal herb, Hybanthus enneaspermus (L.) F. Muell. Plant Growth Regulation, 80, 103-114.
3. Chang, C.-C., Yang, M.-H., Wen, H.-M., & Chern, J.-C. (2002). Estimation of total flavonoid content in propolis by two complementary colometric methods. Journal of food and drug analysis, 10(3), 3.
4. Jeyasri, R., Muthuramalingam, P., Karthick, K., Shin, H., Choi, S. H., & Ramesh, M. (2023). Methyl jasmonate and salicylic acid as powerful elicitors for enhancing the production of secondary metabolites in medicinal plants: an updated review. Plant Cell, Tissue and Organ Culture (PCTOC), 153(3), 447-458.
5. Karthi, C., & Velayutham, P. (2017). Direct organogenesis and mass propagation from the nodal explants of Hybanthus enneaspermus (L.) F. Muell. Plant Cell Biotechnology and Molecular Biology, 18(5&6), 290-296.
6. Murugan, M., & Kamaraj, M. (2018). In vitro propagation and conservation of useful ethnomedicinal plant of Hybanthus enneaspermus (Linn.) F. Muell. belonging to the violaceae family. International Journal of Current Research in Life Sciences, 7(7), 2493-2499.
7. Parthasarathy, S. P., Anusuya, S., Rajalakshmi, S., Megha, D., Appunu, C., Alagumanian, S., & Manickavasagam, M. (2024). Elucidating the efficacy of functionalized multi-walled carbon nanotube in the biogenesis of L-Dopa and antioxidant metabolites in cell cultures of Hybanthus enneaspermus. Plant Physiology and Biochemistry, 206, 108310.
8. Patankar, P. K. Invitro Induction of Shoot in Medicinally Important Plant Hybanthus Enneaspermus (Linn.).
9. Patel, D., Kumar, R., Sairam, K., & Hemalatha, S. (2013). Hybanthus enneaspermus (L.) F. Muell: a concise report on its phytopharmacological aspects. Chinese journal of natural medicines, 11(3), 199-206.
10. Premkumar, G., Arumugam, N., Muthuramkumar, S., Varatharaju, G., & Rajarathinam, K. (2013). Improved micropropagation in Hybanthus enneaspermus L. Muell.
11. Premkumar, G., Karuppanapandian, T., Sureshpandian, C., Arumugam, N., Selvam, A., & Rajarathinam, K. (2020). Optimization of a liquid culture system for shoot regeneration and achieving an enriched level of scopadulcic acid b in the leaf organ cultures of Scoparia dulcis L. by response surface methodology. In Vitro Cellular & Developmental Biology-Plant, 56, 60-71.
12. Rajsekhar, P., Bharani, R., Angel, K. J., Ramachandran, M., & Rajsekhar, S. P. V. (2016). Hybanthus enneaspermus (L) F. Muell: A phytopharmacological review on herbal medicine. J. Chem. Pharm. Res, 8(1), 351-355.
13. Ramya, A. K. N., & Devika, R. (2022). A Current Pharmacological Impacts and Perspective of Hybanthus enneaspermus (Linn.) F. Muell. Pharmacognosy Research, 14(4).
14. Shekhawat, M. S., & Manokari, M. (2016). Somatic embryogenesis and in vitro flowering in Hybanthus enneaspermus (L.) F. Muell.–A rare multipotent herb. Asian Pacific Journal of Reproduction, 5(3), 256-262.
15. Shekhawat, M. S., & Manokari, M. (2018). In vitro multiplication, micromorphological studies and ex vitro rooting of Hybanthus enneaspermus (L.) F. Muell.–a rare medicinal plant. Acta Botanica Croatica, 77(1), 80-87.
16. Singleton, V. L., Orthofer, R., & Lamuela-Raventós, R. M. (1999). [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. In Methods in enzymology (Vol. 299, pp. 152-178): Elsevier.
17. Velayutham, P., & Karthi, C. (2020). Rapid and Mass Propagation of Hybanthus enneaspermus (L.) F. Muell. from Shoot Tip and Nodal Explants.
18. Velayutham, P., Karthi, C., Nalini, P., & Jahirhussain, G. (2012). In vitro Regeneration and Mass Propagation of Hybanthus enneaspermus (L.) F. Muell. from the stem explants through callus culture. Journal of Agricultural Technology, 8(3), 1119-1128.
19. Vyas, S., Rao, M. S., Suthar, R. K., & Purohit, S. D. (2008). Liquid culture system stimulates in vitro growth and shoot multiplication in four medicinally important plants. Medicinal and Aromatic Plant Science and Biotechnology, 2(2), 96-100.
20. Nirmal, D., Teraiya , S., & Joshi, P.(2023). Liquid culture system: An efficient approach for sustainable micropropagation. Current Agriculture Research Journal, 11 , 28-42.
21. Sathish, S., Vasudevan, V., Karthik, S., Pavan, G., & Manikavasagan, M.(2019). Enhanced L-Dopa production from elicited cell suspension culture of Hybanthus enneaspermus(L.)F.Muell. Plant Biotechnology Reports, 13, 613-621.
22. Sathish, S., Vasudevan, V., Karthik,S., Elayaraja, D., Pavan, G., Ajithan, C., &Manikavasagan, M.(2020). Elicitors induced L-Dopa accumulation in adventitious root cultures of Hybanthus enneaspermus (L.) F . Muell.Vegetos, 33, 304-312.
23. Kaur, P., Gupta, R. C., Dey, A., Malik, T., & Pandey, D. K. (2020). Optimization of salicylic acid and chitosan treatment for bitter secoiridoid and xanthone glycosides production in shoot cultures of Swertia paniculata using response surface methodology and artificial neural network. BMC Plant Biology, 20(1).
Published
2024-06-06
How to Cite
Chandran Sureshpandian, & Gandhi Premkumar. (2024). Development Of Optimized Liquid Culture System For Hybanthus Enneaspermus (L.) F. Muell Using Response Surface Methodology. Revista Electronica De Veterinaria, 25(1), 3592 - 3603. https://doi.org/10.69980/redvet.v25i1.1661
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