Skip to main navigation menu Skip to main content Skip to site footer
Plant Science Today (PST)

Research Articles

Vol. 12 No. sp3 (2025): Advances in Plant Health Improvement for Sustainable Agriculture

Direct and indirect organogenesis in soybean for efficient shoot induction through balanced levels of auxins and cytokinins

DOI
https://doi.org/10.14719/pst.10257
Submitted
25 June 2025
Published
06-10-2025

Abstract

Soybean (Glycine max L.) is an essential oilseed crop widely recognised for its high protein and oil content. Soybean (Co1) is a high-yielding variety and well-suited for cultivation in Tamil Nadu and nearby regions. Direct and indirect organogenesis of soybean using different growth regulators is helpful for regeneration, micropropagation and genetic transformation. We evaluated the impact of various plant growth regulators (BAP, KIN, ZT and TDZ) for direct organogenesis using split seeds and cotyledonary nodes of soybean (Co1). Among the different combinations, B5 medium with BAP 1.0 mg/L (for multiple shoots) and KIN 1.0 mg/L (for shoot length) revealed the better outcomes for the direct shoot rejuvenation in the split-seeds. The highest frequency of multiple shoots from the cotyledonary node was induced in MS medium containing TDZ at 0.5 mg/L. Different levels of 2,4-D have been assessed for callus induction from various explants and a higher percentage of callus was developed in hypocotyls. MS media containing 2,4-D (2.0 mg/L) and KIN (0.2 mg/L) leads to better callus development and the friable callus was further used for shoot induction using BAP, TDZ and ZT. After 2 - 3 weeks, initiation of shoot formation from embryonic callus was observed in MS medium supplemented with 1.0 mg/L ZT and 0.5 mg/L IAA. Shoot induction from split seeds showed better response than the cotyledonary node and callus. These findings are expected to be useful for the regeneration of soybean (Co1) to develop transgenic lines for enrichment of nutritional value.

References

  1. 1. USDA. Oil seeds: World markets and trade. 2020. https://apps.fas.usda.gov/psdonline/circulars/oilseeds.pdf.
  2. 2. Hill K, Schaller GE. Enhancing plant regeneration in tissue culture: a molecular approach through manipulation of cytokinin sensitivity. Plant Signal Behav. 2013;8(10):212–24. https://doi.org/10.4161/psb.25709
  3. 3. Sugimoto K, Meyerowitz EM. Regeneration in Arabidopsis tissue culture. In: De Smet I, editor. Plant organogenesis. Totowa, NJ: Humana Press. 2013. p. 265–75. https://doi.org/10.1007/978-1-62703-221-6_18
  4. 4. Abbasi Z, Hooshyar S, Bagherieh-Najjar MB. Improvement of callus production and shoot regeneration using various organs of soybean (Glycine max L. Merr) by response surface methodology. In Vitro Cell Dev Biol Plant. 2016;52:537–45. https://doi.org/10.1007/s11627-016-9778-1
  5. 5. Pareddy D, Chennareddy S, Anthony G, Sardesai N, Mall T, Minnicks T, et al. Improved soybean transformation for efficient and high-throughput transgenic production. Transgenic Res. 2020;29:267–81. https://doi.org/10.1007/s11248-020-00198-8
  6. 6. Dan Y, Reichert NA. Organogenic regeneration of soybean from hypocotyl explants. I Cell Dev Biol Plant. 1998;34:14–21. https://doi.org/10.1007/BF02823117
  7. 7. Arun M, Subramanyam K, Theboral J, Ganapathi A, Manickavasagam M. Optimized shoot regeneration for Indian soybean: The influence of exogenous polyamines. Plant Cell Tiss Org Cult. 2014;117:305–9. https://doi.org/10.1007/s11240-014-0431-6
  8. 8. Patel E, Kalia A, Gill BS. Direct organogenesis from cortical cells of hypocotyl segments in soybean. In Vitro Cell Dev Biol Plant. 2023;59(1):140–6. https://doi.org/10.1007/s11627-023-10329-5
  9. 9. Radhakrishnan R, Ranjithakumari BD. Callus induction and plant regeneration of Indian soybean (Glycine max (L.) Merr. cv. CO3) via half seed explant culture. J Agric Technol. 2007;3(2):287–97.
  10. 10. Sairam RV, Franklin G, Hassel R, Smith B, Meeker K, Kashikar N, et al. A study on the effect of genotypes, plant growth regulators and sugars in promoting plant regeneration via organogenesis from soybean cotyledonary nodal callus. Plant Cell Tiss Org Cult. 2003;75:79–85. https://doi.org/10.1023/A:1024649122748
  11. 11. Bidabadi SS, Jain SM. Cellular, molecular and physiological aspects of in vitro plant regeneration. Plants. 2020;9(6):702. https://doi.org/10.3390/plants9060702
  12. 12. Gamborg OL, Murashige T, Thorpe TA. Plant tissue culture media. In Vitro Cell Dev Biol-Plant. 1976;12:473–8. https://doi.org/10.1007/BF02796489
  13. 13. Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant. 1962;15:473–97. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  14. 14. Arun M, Chinnathambi A, Subramanyam K, Karthik S, Sivanandhan G, Theboral J, et al. Involvement of exogenous polyamines enhances regeneration and Agrobacterium-mediated genetic transformation in half-seeds of soybean. Biotech. 2016;6:1–12. https://doi.org/10.1007/s13205-016-0448-0
  15. 15. Mederos Molina S. In vitro growth and multiplication of Hypericum canariense L. Horticulturae. Acta Hortic. 289, 133–34. https://doi.org/10.17660/ActaHortic.1991.289.30
  16. 16. Moura M. Conservation of Hypericum foliosum aiton, an endemic Azorean species, by micropropagation. In Vitro Cell Dev Biol-Plant. 1998;34:244–8. https://doi.org/10.1007/BF02822715
  17. 17. Hesar AA, Kaviani B, Tarang A, Zanjani SB. Effect of different concentrations of kinetin on regeneration of (Matthiola incana). Plant Omics. 2011;4(5):236–8.
  18. 18. Lu CY. The use of thidiazuron in tissue culture. In Vitro Cell Dev Biol-Plant. 1993;29:92–6. https://doi.org/10.1007/BF02632259
  19. 19. Huetteman CA, Preece JE. Thidiazuron: A potent cytokinin for woody plant tissue culture. Plant Cell Tiss Org Cult. 1993;33:105–19. https://doi.org/10.1007/BF01983223
  20. 20. Shan Z, Raemakers K, Tzitzikas EN, Ma Z, Visser RG. Development of a highly efficient, repetitive system of organogenesis in soybean (Glycine max (L.) Merr). Plant Cell Rep. 2005;24:507-12. https://doi.org/10.1007/s00299-005-0971-7
  21. 21. Kim DG, Kantayos V, Kim DK, Park HG, Kim HH, Rha ES, et al. Plant regeneration by in vitro tissue culture in Korean soybean (Glycine max L.). Korean J Plant Res. 2016;29(1):143–53. https://doi.org/10.7732/kjpr.2016.29.1.143
  22. 22. Ma XH, Wu TL. Rapid and efficient regeneration in soybean (Glycine max (L.) Merrill) from whole cotyledonary node explants. Acta Physiol Plant. 2008;30:209–16. https://doi.org/10.1007/s11738-007-0109-3
  23. 23. Kiran G, Kaviraj CP, Jogeswar G, Kishor PK, Rao S. Direct and high frequency somatic embryogenesis and plant regeneration from hypocotyls of chickpea (Cicer arietinum L.), a grain legume. Curr Sci. 2005;89(6):1012–8.
  24. 24. Kumari BD, Settu A, Sujatha G. Somatic embryogenesis and plant regeneration in soybean (Glycine max (L.) Merr.). Indian J Biotechnol. 2006;5:243–5.
  25. 25. Zulkarnain Z. The effect of different levels and sources of auxin and cytokinin to callus formation on soybean anther culture. J Agrotek Trop. 2014;3(2):58–67.
  26. 26. Wen SS, Ge XL, Wang R, Yang HF, Bai YE, Guo YH, et al. An efficient Agrobacterium-mediated transformation method for hybrid Poplar 84K (Populus alba X P. glandulosa) using calli as explants. Int J Mol Sci. 2022;23(4):2216. https://doi.org/10.3390/ijms23042216
  27. 27. Gordon SP, Heisler MG, Reddy GV, Ohno C, Das P, Meyerowitz EM. Pattern formation during de novo assembly of the Arabidopsis shoot meristem. Development. 2007;134:3539–48. https://doi.org/10.1242/dev.010298
  28. 28. Kaur P, Bansal KC. Efficient production of transgenic tomatoes via Agrobacterium-mediated transformation. Biol Plant. 2010;54:344–8. https://doi.org/10.1007/s10535-010-0060-9
  29. 29. Yaroshko O, Pasternak T, Larriba E, Pérez-Pérez JM. Optimization of callus induction and shoot regeneration from tomato cotyledon explants. Plants. 2023;12(16):2942. https://doi.org/10.3390/plants12162942
  30. 30. Sandhya D, Jogam P, Venkatapuram AK, Savitikadi P, Peddaboina V, Allini VR, et al. Highly efficient Agrobacterium-mediated transformation and plant regeneration system for genome engineering in tomato. Saudi J Biol Sci. 2022;29(6):103292. https://doi.org/10.1016/j.sjbs.2022.103292
  31. 31. Sugimoto K, Gordon SP, Meyerowitz EM. Regeneration in plants and animals: Dedifferentiation, transdifferentiation, or just differentiation?. Trends Cell Biol. 2011;21(4):212–8. https://doi.org/10.1016/j.tcb.2010.12.004
  32. 32. Liu J, Sheng L, Xu Y, Li J, Yang Z, Huang H, et al. WOX11 and 12 are involved in the first-step cell fate transition during de novo root organogenesis in Arabidopsis. Plant Cell. 2014;26(3):1081-93. https://doi.org/10.1105/tpc.114.122887
  33. 33. Duclercq J, Sangwan-Norreel B, Catterou M, Sangwan RS. De novo shoots organogenesis: from art to science. Trends Plant Sci. 2011;16(11):597–606. https://doi.org/10.1016/j.tplants.2011.08.004

Downloads

Download data is not yet available.