In vitro studies on micropropagation based on axillary shoot proliferation in palash (Butea monosperma)
Article Main
Abstract
Butea monosperma, commonly called the Flame of the Forest, is a leguminous plant of significant ecological, medicinal, and socio-economic value, found across tropical and subtropical Asia. Despite its importance in traditional medicine and rural economies, large-scale propagation is limited by poor seed viability, dormancy, and low success rates with conventional vegetative methods. The present investigation was undertaken to develop a reliable and efficient in vitro micropropagation protocol for B. monosperma based on axillary shoot proliferation, by analyzing the effects of various cytokinins and their combinations with auxins on nodal explants. After eight weeks of culture, 2.5 µM 6-benzyladenine (BA) yielded the highest shoot induction rate (83.33%) with an average of 12.44 ± 0.43 shoots and a shoot length of 4.00 ± 0.17 cm. Meta topolin was the second most effective. An enhanced shoot multiplication response was observed when BA (2.5 µM) was combined with NAA (0.5 µM), resulting in up to 20.89 ± 0.58 shoots per explant and 94.44% shoot proliferation response. Rooting frequency was highest in IBA-treated cultures (94.44%), with improved root number and quality observed in half-strength MS medium supplemented with 0.5-1.0 µM IBA. Acclimatization studies showed a maximum survival of 80.83 ± 2.40% in Soilrite™, indicating its suitability for ex vitro establishment. This optimized micropropagation system demonstrates high proliferation potential and provides a reliable method for conservation, large-scale propagation, and sustainable use of B. monosperma. The results hold significant implications for medicinal applications and species restoration.
Article Details
Article Details
Keywords
Acclimatization , Axillary shoot proliferation, Butea monosperma, Cytokinins, Micropropagation, Nodal explants
References
Ahmad, A., Ahmad, N. & Anis, M. (2018). Preconditioning of nodal explants in thidiazuron-supplemented liquid media improves shoot multiplication in Pterocarpus marsupium (Roxb.). https://doi.org/10.1007/978-981-10-8004-3_8
Ahmed, M. R., Anis, M., Alatar, A. A. & Faisal, M. (2017). In vitro clonal propagation and evaluation of genetic fidelity using RAPD and ISSR marker in micropropagated plants of Cassia alata L.: a potential medicinal plant. Agroforestry Systems, 91(4), 637-647. https://doi.org/10.1007/s10457-016-9992-1
Ahmed, S., Anis, M. & Shahzad, A. (2014). Micropropagation studies of Senna alata (L.) Roxb. an anti allergenic plant. Indian Journal of Applied Research, 4(10), 54–57.
Aileni, M., Damodar, M. & Narashimha, M. E. (2014). In vitro seed germination and development of Butea monosperma (Lam.) Taub. var. lutea (Willt.): A step for rehabilitation. International Journal of Multidis- Ciplinary and Current Research, 2, 297–301.
Anis, M., Husain, M. K. & Shahzad, A. (2005). In vitro plant regeneration from nodal segments of Pterocarpus marsupium Roxb. Journal of Plant Biology, 32(1), 7–12.
Badhepuri, M. K., Beeravelli, P. R., Arolla, R. G., Jogam, P., Rohela, G. K. & Singisala, N. R. (2024). Micropropagation and genetic fidelity analysis using SCoT and ISSR markers in Muehlenbeckia platyclada (F. Muell.) meisn. Plant Cell, Tissue and Organ Culture, 157(3), 51. https://doi.org/10.1007/s11240-024-02763-z
Badhepuri, M. K., Manokari, M., Raj, M. C., Jogam, P., Dey, A., Faisal, M. & Shekhawat, M. S. (2023). Meta-Topolin enhanced direct shoot organogenesis and regeneration from leaf explants of coleus forskohlii (Willd.) Briq. Industrial Crops and Products, 197, 116584. https://doi.org/10.1016/j.indcrop.2023.116584
Bhargava, S. K. (1986). Estrogenic and postcoital anticonceptive activity in rats of butin isolated from Butea monosperma seed. Journal of Ethnopharmacology, 18(1), 95-101. https://doi.org/10.1016/0378-8741(86)90046-2
Bhat, M.H., Fayaz, M., Kumar, A., Dar, A.A. & Jain, A.K. (2022). Development of an efficient micropropagation system for Dioscorea bulbifera L. and phytochemical profile of regenerated plants. Journal of Genetic Engineering and Biotechnology, 20(1), 107. https://doi.org/10.1186/s43141-022-00382-9
Burkill, I. H. (1935). A dictionary of the economic products of the Malay Peninsula.
Chaturvedi, R., Razdan, M. K. & Bhojwani, S. S. (2004). In vitro clonal propagation of an adult tree of neem (Azadirachta indica A. Juss.) by forced axillary branching. Plant Science, 166(2), 501-506.
Dharavath, R., Narra, M., Ellendula, R., Velivela, Y. & Abbagani, S. (2017). Rapid in vitro plant regeneration from nodal explants and assessment of genetic fidelity using inter simple sequence repeats markers in Butea monosperma (Lam.) Taub. var. lutea (Witt.). Asian Journal of Biotechnology, 9, 80-87.
Farooq, I., Qadri, Z. A., Rather, Z. A., Nazki, I. T., Banday, N., Rafiq, S. & Mansoor, S. (2021). Optimization of an improved, efficient and rapid in vitro micropropagation protocol for Petunia hybrida Vilm. Cv.“Bravo”. Saudi Journal of Biological Sciences, 28(7), 3701-3709. https://doi.org/10.1016/j.sjbs.2021.05.018
Fatima, N. & Anis, M. (2021). Regulation of in vitro morphogenesis by modulation of culture conditions in withania somnifera L. using cotyledonary node explants. In: Siddique, I. Propagation and genetic manipulation of plants. Springer, Singapore. https://doi.org/10.1007/978-981-15-7736-9_9
Fayaz, M., Singh, S., Bhat, I.A., Bhat, M.H., Khanday, F.A. & Dar, A.A. (2025). Efficient micropropagation of bistorta amplexicaulis (D. Don) Greene: An important medicinal plant of western himalaya. International Journal of Analytical Chemistry, 69(1), 4457383. https://doi.org/10.1155/ianc/4457383
Gupta, P.G., Chauhan, N.S., Milind Pande, M.P. & Anupam Pathak, A.P. (2012). Phytochemical and pharmacological review on Butea monosperma (Palash). International journal of Agronomy and Plant Production, 3 (7), 255-258.
Husain, M. K., Anis, M. & Shahzad, A. (2008). In vitro propagation of a multipurpose leguminous tree Pterocarpus marsupium Roxb. using nodal explants. Acta Physiologiae Plantarum, 30(3), 353–359.
Hussain, S.A., Ahmad, M., Anis, M. & Alatar, A. A. (2018). Influence of Meta topolin on in vitro organogenesis in Tecoma stans L., assessment of genetic fidelity and phytochemical profiling of wild and regenerated plants. Plant Cell, Tissue and Organ Culture, 138, 339–351. https://doi.org/10.1007/s11240-019-01631-5
Hussain, S.A., Ahmad, N., Anis, M. & Hakeem, K. (2019). Development of an efficient micropropagation system for Tecoma stans [(L.) Juss. ex Kunth], using thidiazuron and effects on phytochemical constitution. In Vitro Cellular & Developmental Biology Plant, 55, 442–453. https://doi.org/10.1007/s11627-019-10001-x
Jadhav, S.N., Ved, D.K., Ghate, U., Reddy, K.N. & Reddy, C.S. (2001). Proceedings of the workshop and conservation assessment and management planning for medicinal plants of Andhra Pradesh. Bangalore: FRLHT.
Jaiswal, S., Singh, R. & Arya, I. D. (2015). Micropropagation of adult tree of Pterocarpus marsupium Roxb. using nodal explants. Journal of Biological Sciences, 15(1), 39–44.
Khanam, M.N., Javed, S.B., Ahmad, N. & Singh, N. (2021). Substitution of benzyladenine with meta-Topolin improved shoot regeneration and rooting in Wedelia chinensis (Osbeck) Merr. and quantitative estimation of flavonoid. In I. Siddique (Ed.), Propagation and genetic manipulation of plants. Springer Nature Singapore, https://doi.org/10.1007/978-981-15-7736-910
Kumar, N. (2018). Biotechnological approaches for medicinal and aromatic plants: Conservation, genetic improvement and utilization. Springer.
Mahender, A., Mahesh, D. M. & Murthy, E. N. (2014). In vitro seed germination and development of Butea monosperma (Lam.) Taub. Var. lutea (Willt.): a step for rehabilitation. International Journal of Multidisciplinary and Current Research, 2, 297-301.
Mishra, A., Verma, S. & Mishra, A.P. (2012). A plant review: Butea monosperma (Lam.) Kuntze. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 700-714.
Murashige, T. & Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant, 15, 473-497.
Nithya, V. & Kamalam, M. (2021). Standardization of a protocol for micropropagation of Eupatorium glandulosum L. an important medicinal plant. Plant Cell, Tissue and Organ Culture, 146(2), 339-344.https://doi.org/10.1007/s11240-021-02073-8
Pandey, A., Tamta, S. & Giri, D. (2010). In vitro propagation of Clitoria ternatea L.: A rare medicinal plant. Journal of Medicinal Plants Research, 4(10), 934–939.
Pandey, R., Gautam, A.K., Bhargavan, B., Trivedi, R., Swarnkar, G., Nagar, G.K. & Singh, D. (2010). Total extract and standardized fraction from the stem bark of Butea monosperma have osteoprotective action: evidence for the nonestrogenic osteogenic effect of the standardized fraction. Menopause, 17(3), 602-610.https://doi.org/10.1097/gme.0b013e3181d0f7f0
Polhill, R. M. (1981). Papilionoideae. Advances in legume systematics part 1, 191-208.
Prashanth, D., Asha, M. K., Amit, A. & Padmaja, R. (2001). Anthelmintic activity of Butea monosperma.
Raghuvanshi, S. S. & Pande, K. (1985). B Chromosome in Butea monosperma, a tree. Caryologia, 38(2), 161-168.
Ramesh, M., Umate, P., Venugopal Rao, K. & Sadanandam, A. (2005). Micropropagation of Terminalia bellirica Roxb. a sericulture and medicinal plant. In Vitro Cellular & Developmental Biology Plant, 41(3), 320-323.
Reddy, C.R.K., Jha, B., Fujita, Y. & Ohno, M. (2008). Seaweed micropropagation techniques and their potentials: an overview. Journal of Applied Phycology, 20(5), 609-617.
Rohela, G.K., Jogam, P. & Mir, M.Y. (2020). Indirect regeneration and genetic fidelity analysis of acclimated plantlets through SCoT and ISSR markers in Morus alba L. cv. Chinese white. Biotechnology Reports, 27, e00417. https://doi.org/10.1016/j.btre.2020.e00417
Rout, G. R. (2005). Micropropagation of Clitoria ternatea Linn. (Fabaceae)—An important medicinal plant. In Vitro Cellular & Developmental Biology - Plant, 41(4), 516–519.
Rout, G. R., Senapati, S. K. & Aparajeta, S. (2008). Micropropagation of Acacia chundra (Roxb.) DC. Hort Sci, 35(1), 22-6.
Sahare, K.N., Anandharaman, V., Meshram, V.G., Meshram, S.U., Gajalakshmi, D., Goswami, K. & Reddy, M.V.R. (2008). In vitro effect of four herbal plants on the motility of Brugia malayi microfilariae. Indian Journal of Medical Research, 127(5), 467-471.
Sanjappa, M. (1987). Revision of the genera butea Roxb. Ex Willd. and Melzotropls Volgt (Fabaceae). Nelumbo, 199-225.
Siddique, I., & Yadav, V. (2021). A potent, quick and refined plant regeneration protocol for Dianthus caryophyllus via nodal segment explants. In I. Siddique (Ed.), Propagation and genetic manipulation of plants (pp. 1–16). Springer Nature Singapore Pte Ltd. https://doi.org/10.1007/978-981-15-7736-9
Srivastava, S., Ray, D. P. & Giri, S. K. (2013). Extraction of palas dye for its use in textile coloration. International Journal of Agriculture, Environment and Biotechnology, 6(3), 497-502.
Tandon, R., Shivanna, K.R. & Mohan Ram, H.Y. (2003). Reproductive biology of Butea monosperma (Fabaceae). Annals of Botany, 92(5), 715-723.
Yarra, R., Bulle, M., Mushke, R. & Murthy, E.N. (2016). In vitro conservation and genetic homogeneity assessment of Butea monosperma (Lam.) Taub. Var. lutea (Witt.) Maheshwari A potential pharmaceutical legume tree. Journal of Applied Research on Medicinal and Aromatic Plants, 3(4), 195-199.
Ahmed, M. R., Anis, M., Alatar, A. A. & Faisal, M. (2017). In vitro clonal propagation and evaluation of genetic fidelity using RAPD and ISSR marker in micropropagated plants of Cassia alata L.: a potential medicinal plant. Agroforestry Systems, 91(4), 637-647. https://doi.org/10.1007/s10457-016-9992-1
Ahmed, S., Anis, M. & Shahzad, A. (2014). Micropropagation studies of Senna alata (L.) Roxb. an anti allergenic plant. Indian Journal of Applied Research, 4(10), 54–57.
Aileni, M., Damodar, M. & Narashimha, M. E. (2014). In vitro seed germination and development of Butea monosperma (Lam.) Taub. var. lutea (Willt.): A step for rehabilitation. International Journal of Multidis- Ciplinary and Current Research, 2, 297–301.
Anis, M., Husain, M. K. & Shahzad, A. (2005). In vitro plant regeneration from nodal segments of Pterocarpus marsupium Roxb. Journal of Plant Biology, 32(1), 7–12.
Badhepuri, M. K., Beeravelli, P. R., Arolla, R. G., Jogam, P., Rohela, G. K. & Singisala, N. R. (2024). Micropropagation and genetic fidelity analysis using SCoT and ISSR markers in Muehlenbeckia platyclada (F. Muell.) meisn. Plant Cell, Tissue and Organ Culture, 157(3), 51. https://doi.org/10.1007/s11240-024-02763-z
Badhepuri, M. K., Manokari, M., Raj, M. C., Jogam, P., Dey, A., Faisal, M. & Shekhawat, M. S. (2023). Meta-Topolin enhanced direct shoot organogenesis and regeneration from leaf explants of coleus forskohlii (Willd.) Briq. Industrial Crops and Products, 197, 116584. https://doi.org/10.1016/j.indcrop.2023.116584
Bhargava, S. K. (1986). Estrogenic and postcoital anticonceptive activity in rats of butin isolated from Butea monosperma seed. Journal of Ethnopharmacology, 18(1), 95-101. https://doi.org/10.1016/0378-8741(86)90046-2
Bhat, M.H., Fayaz, M., Kumar, A., Dar, A.A. & Jain, A.K. (2022). Development of an efficient micropropagation system for Dioscorea bulbifera L. and phytochemical profile of regenerated plants. Journal of Genetic Engineering and Biotechnology, 20(1), 107. https://doi.org/10.1186/s43141-022-00382-9
Burkill, I. H. (1935). A dictionary of the economic products of the Malay Peninsula.
Chaturvedi, R., Razdan, M. K. & Bhojwani, S. S. (2004). In vitro clonal propagation of an adult tree of neem (Azadirachta indica A. Juss.) by forced axillary branching. Plant Science, 166(2), 501-506.
Dharavath, R., Narra, M., Ellendula, R., Velivela, Y. & Abbagani, S. (2017). Rapid in vitro plant regeneration from nodal explants and assessment of genetic fidelity using inter simple sequence repeats markers in Butea monosperma (Lam.) Taub. var. lutea (Witt.). Asian Journal of Biotechnology, 9, 80-87.
Farooq, I., Qadri, Z. A., Rather, Z. A., Nazki, I. T., Banday, N., Rafiq, S. & Mansoor, S. (2021). Optimization of an improved, efficient and rapid in vitro micropropagation protocol for Petunia hybrida Vilm. Cv.“Bravo”. Saudi Journal of Biological Sciences, 28(7), 3701-3709. https://doi.org/10.1016/j.sjbs.2021.05.018
Fatima, N. & Anis, M. (2021). Regulation of in vitro morphogenesis by modulation of culture conditions in withania somnifera L. using cotyledonary node explants. In: Siddique, I. Propagation and genetic manipulation of plants. Springer, Singapore. https://doi.org/10.1007/978-981-15-7736-9_9
Fayaz, M., Singh, S., Bhat, I.A., Bhat, M.H., Khanday, F.A. & Dar, A.A. (2025). Efficient micropropagation of bistorta amplexicaulis (D. Don) Greene: An important medicinal plant of western himalaya. International Journal of Analytical Chemistry, 69(1), 4457383. https://doi.org/10.1155/ianc/4457383
Gupta, P.G., Chauhan, N.S., Milind Pande, M.P. & Anupam Pathak, A.P. (2012). Phytochemical and pharmacological review on Butea monosperma (Palash). International journal of Agronomy and Plant Production, 3 (7), 255-258.
Husain, M. K., Anis, M. & Shahzad, A. (2008). In vitro propagation of a multipurpose leguminous tree Pterocarpus marsupium Roxb. using nodal explants. Acta Physiologiae Plantarum, 30(3), 353–359.
Hussain, S.A., Ahmad, M., Anis, M. & Alatar, A. A. (2018). Influence of Meta topolin on in vitro organogenesis in Tecoma stans L., assessment of genetic fidelity and phytochemical profiling of wild and regenerated plants. Plant Cell, Tissue and Organ Culture, 138, 339–351. https://doi.org/10.1007/s11240-019-01631-5
Hussain, S.A., Ahmad, N., Anis, M. & Hakeem, K. (2019). Development of an efficient micropropagation system for Tecoma stans [(L.) Juss. ex Kunth], using thidiazuron and effects on phytochemical constitution. In Vitro Cellular & Developmental Biology Plant, 55, 442–453. https://doi.org/10.1007/s11627-019-10001-x
Jadhav, S.N., Ved, D.K., Ghate, U., Reddy, K.N. & Reddy, C.S. (2001). Proceedings of the workshop and conservation assessment and management planning for medicinal plants of Andhra Pradesh. Bangalore: FRLHT.
Jaiswal, S., Singh, R. & Arya, I. D. (2015). Micropropagation of adult tree of Pterocarpus marsupium Roxb. using nodal explants. Journal of Biological Sciences, 15(1), 39–44.
Khanam, M.N., Javed, S.B., Ahmad, N. & Singh, N. (2021). Substitution of benzyladenine with meta-Topolin improved shoot regeneration and rooting in Wedelia chinensis (Osbeck) Merr. and quantitative estimation of flavonoid. In I. Siddique (Ed.), Propagation and genetic manipulation of plants. Springer Nature Singapore, https://doi.org/10.1007/978-981-15-7736-910
Kumar, N. (2018). Biotechnological approaches for medicinal and aromatic plants: Conservation, genetic improvement and utilization. Springer.
Mahender, A., Mahesh, D. M. & Murthy, E. N. (2014). In vitro seed germination and development of Butea monosperma (Lam.) Taub. Var. lutea (Willt.): a step for rehabilitation. International Journal of Multidisciplinary and Current Research, 2, 297-301.
Mishra, A., Verma, S. & Mishra, A.P. (2012). A plant review: Butea monosperma (Lam.) Kuntze. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 700-714.
Murashige, T. & Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant, 15, 473-497.
Nithya, V. & Kamalam, M. (2021). Standardization of a protocol for micropropagation of Eupatorium glandulosum L. an important medicinal plant. Plant Cell, Tissue and Organ Culture, 146(2), 339-344.https://doi.org/10.1007/s11240-021-02073-8
Pandey, A., Tamta, S. & Giri, D. (2010). In vitro propagation of Clitoria ternatea L.: A rare medicinal plant. Journal of Medicinal Plants Research, 4(10), 934–939.
Pandey, R., Gautam, A.K., Bhargavan, B., Trivedi, R., Swarnkar, G., Nagar, G.K. & Singh, D. (2010). Total extract and standardized fraction from the stem bark of Butea monosperma have osteoprotective action: evidence for the nonestrogenic osteogenic effect of the standardized fraction. Menopause, 17(3), 602-610.https://doi.org/10.1097/gme.0b013e3181d0f7f0
Polhill, R. M. (1981). Papilionoideae. Advances in legume systematics part 1, 191-208.
Prashanth, D., Asha, M. K., Amit, A. & Padmaja, R. (2001). Anthelmintic activity of Butea monosperma.
Raghuvanshi, S. S. & Pande, K. (1985). B Chromosome in Butea monosperma, a tree. Caryologia, 38(2), 161-168.
Ramesh, M., Umate, P., Venugopal Rao, K. & Sadanandam, A. (2005). Micropropagation of Terminalia bellirica Roxb. a sericulture and medicinal plant. In Vitro Cellular & Developmental Biology Plant, 41(3), 320-323.
Reddy, C.R.K., Jha, B., Fujita, Y. & Ohno, M. (2008). Seaweed micropropagation techniques and their potentials: an overview. Journal of Applied Phycology, 20(5), 609-617.
Rohela, G.K., Jogam, P. & Mir, M.Y. (2020). Indirect regeneration and genetic fidelity analysis of acclimated plantlets through SCoT and ISSR markers in Morus alba L. cv. Chinese white. Biotechnology Reports, 27, e00417. https://doi.org/10.1016/j.btre.2020.e00417
Rout, G. R. (2005). Micropropagation of Clitoria ternatea Linn. (Fabaceae)—An important medicinal plant. In Vitro Cellular & Developmental Biology - Plant, 41(4), 516–519.
Rout, G. R., Senapati, S. K. & Aparajeta, S. (2008). Micropropagation of Acacia chundra (Roxb.) DC. Hort Sci, 35(1), 22-6.
Sahare, K.N., Anandharaman, V., Meshram, V.G., Meshram, S.U., Gajalakshmi, D., Goswami, K. & Reddy, M.V.R. (2008). In vitro effect of four herbal plants on the motility of Brugia malayi microfilariae. Indian Journal of Medical Research, 127(5), 467-471.
Sanjappa, M. (1987). Revision of the genera butea Roxb. Ex Willd. and Melzotropls Volgt (Fabaceae). Nelumbo, 199-225.
Siddique, I., & Yadav, V. (2021). A potent, quick and refined plant regeneration protocol for Dianthus caryophyllus via nodal segment explants. In I. Siddique (Ed.), Propagation and genetic manipulation of plants (pp. 1–16). Springer Nature Singapore Pte Ltd. https://doi.org/10.1007/978-981-15-7736-9
Srivastava, S., Ray, D. P. & Giri, S. K. (2013). Extraction of palas dye for its use in textile coloration. International Journal of Agriculture, Environment and Biotechnology, 6(3), 497-502.
Tandon, R., Shivanna, K.R. & Mohan Ram, H.Y. (2003). Reproductive biology of Butea monosperma (Fabaceae). Annals of Botany, 92(5), 715-723.
Yarra, R., Bulle, M., Mushke, R. & Murthy, E.N. (2016). In vitro conservation and genetic homogeneity assessment of Butea monosperma (Lam.) Taub. Var. lutea (Witt.) Maheshwari A potential pharmaceutical legume tree. Journal of Applied Research on Medicinal and Aromatic Plants, 3(4), 195-199.
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How to Cite
In vitro studies on micropropagation based on axillary shoot proliferation in palash (Butea monosperma). (2026). Journal of Applied and Natural Science, 18(2), 916-928. https://doi.org/10.31018/jans.v18i2.7474
