Effect of plant growth regulators and explants on organogenesis of soybean (Glycine max L.)

Document Type : Research Paper

Authors

Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Tabriz, Tabriz, Iran.

10.22034/jppb.2026.69640.1391

Abstract

Objective: The soybean plant is one of the most important crops, and it is known as a rich source of vegetable oil and protein in the world. Despite its advantages, soybean production faces different challenges and requires genetic improvement. Soybean transformation provides an attractive advancement for soybean breeding programs, allowing the production of novel and genetically diverse plant materials. Therefore, developing an efficient plant regeneration protocol is necessary for the transformation programs. In this study, a simple, efficient, and repeatable protocol was developed for in vitro organogenesis of the Saman soybean cultivar.
Methods: The soybean seeds were cultured on different strengths of MS medium (full and half-strength MS) in combination with different concentrations of BAP/Kin (0, 0.5, 1, 1.5, 2, 2.5, and 3 mg/L). After germination, cotyledon, hypocotyl, and primary leaf explants were cultured on the shoot induction media with different concentrations of BAP/Kin (0, 1, 1.5, and 2 mg/L) in combination with IBA/NAA (0, 0.1, and 0.5 mg/L) to evaluate their organogenesis potential. In the next step, for shoot elongation, all regenerated buds were transferred to two media (MS and ½ MS) in combination with BAP (0, 0.1, 0.2, and 0.4 mg/L) and GA3 (0, 0.1, 0.5, and 1 mg/L). At the final stage, for root induction, the elongated shoots were cut and cultured in different strengths of the MS medium (full and half-strength MS) in combination with different concentrations of IAA, IBA, and NAA (0, 0.1, 0.5, 1, 1.5, and 2 mg/L). All experiments were conducted as a factorial arrangement based on a completely randomized design.
Results: The results showed that the highest percentage of seed germination (97.18%) was obtained in full-strength MS medium supplemented with 2 mg/L BAP. The maximum number of shoot induction per explant (7.1 shoots) was observed in the medium containing 1.5 mg/L BAP and 0.1 mg/L IAA from the cotyledon explants. The maximum rate of shoot elongation (8.5 cm) was achieved in the half-strength MS medium containing 0.5 mg/L GA3 and 0.2 mg/L BAP. Also, the maximum number of roots was produced in the half-strength of MS medium supplemented with 1.5 mg/L IBA. Finally, the rooted plantlets after acclimatization were transferred into the greenhouse for flowering and pod maturation.
Conclusion: The results of this study can be used for gene transfer and genetic engineering research for the Saman cultivar and other soybean cultivars.

Keywords

Main Subjects

References

Arun M, Chinnathambi A, Subramanyam K, Karthik S, Sivanandhan G, Theboral J, Alharbi SA, Kim CK, Ganapathi A. 2016. Involvement of exogenous polyamines enhances regeneration and Agrobacterium-mediated genetic transformation in half-seeds of soybean. 3 Biotech. 6(2): 148. https://doi.org/10.1007/s13205-016-0448-0
Begum N, Zenat EA, Sarkar MKI, Roy CK, Munshi JL, Jahan MAA. 2019. In vitro micro propagation of soybean (Glycine max) BARI-5 variety. Open Microbiol J. 13(1): 177-187. https://doi.org/10.2174/1874285801913010177
Gamborg OL, Miller RA, Ojima K. 1968. Nutrient requirements of suspension cultures of
soybean root cells. Exp Cell Res. 50(1): 151-158. https://doi.org/10.1016/0014-4827(68)90403-5
Joyner EY, Boykin LS, Lodhi MA. 2010. Callus induction and organogenesis in soybean (Glycine max L.) cv. Pyramid from mature cotyledons and embryos. Open Plant Sci J. 4: 18-21. https://doi.org/10.2174/1874294701004010018
Khan MW, Shaheen A, Zhang X, Zhang J, Dewir YH, Magyar-Tábori, K. 2024. Generation and assessment of soybean (Glycine max (L.) Merr.) hybrids for high-efficiency Agrobacterium- mediated transformation. Life. 14(12): 1649. https://doi.org/10.3390/life14121649
Lee MH, Lee J, Jie EY, Choi SH, Jiang L, Ahn WS, Kim CY, Kim SW. 2020. Temporal and spatial expression analysis of shoot-regeneration regulatory genes during the adventitious shoot formation in hypocotyl and cotyledon explants of tomato (CV. Micro-Tom). Int J Mol Sci. 21(15): 5309. https://doi.org/10.3390/ijms21155309
Li S, Cong Y, Liu Y, Wang T, Shuai Q, Chen N, Gai J, Li Y. 2017. Optimization of Agrobacterium-mediated transformation in soybean. Front Plant Sci. 8(1): 246. https://doi.org/10.3389/fpls.2017.00246
Liu HK, Yang C, Wie ZM. 2004. Efficient Agrobacterium tumefaciens-mediated transformation of soybeans using an embryonic tip regeneration system. Planta. 219(6): 1042-1049. https://doi.org/10.1007/s00425-004-1310-x
Long Y, Yang Y, Pan G, Shen Y. 2022. New insights into tissue culture plant-regeneration mechanisms. Front Plant Sci. 13: 926752. https://doi.org/10.3389/fpls.2022.926752
Ma XH, Wu TL. 2008. Rapid and efficient regeneration in soybean [Glycine max (L.) Merrill] from whole cotyledonary node explants. Acta Physiol Plant. 30(2): 209-216. https://doi.org/10.1007/s11738-007-0109-3
Mangena P. 2021. Analysis of correlation between seed vigour, germination and multiple shoot induction in soybean (Glycine max L. Merr.). Heliyon. 7(9): e07913. https://doi.org/10.1016/j.heliyon.2021.e07913
Mangena P, Mokwala PW, Nikolova RV. 2015. In vitro multiple shoot induction in soybean. Int J Agric Biol. 17(4): 838-842. https://doi.org/10.17957/IJAB/14.0006
Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco cultures. Physiol Plant. 15(3): 473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Muzika NS, Kamai T, Williams LE, Kleiman M. 2024. Characterization of gelling agents in callus inducing media: physical properties and their effect on callus growth. Physiol Plant. 176(2): e14312. https://doi.org/10.1111/ppl.14312
Paes de Melo B, Lourenço-Tessutti IT, Morgante CV, Santos NC, Pinheiro LB, de Jesus Lins CB, Silva MCM, Macedo LLP, Fontes EPB, Grossi-de-Sa MF. 2020. Soybean embryonic axis transformation: combining biolistic and Agrobacterium-mediated protocols to overcome typical complications of in vitro plant regeneration. Front Plant Sci. 11: 1228. https://doi.org/10.3389/fpls.2020.01228
Pasternak TP, Steinmacher D. 2024. Plant growth regulation in cell and tissue culture in vitro. Plants. 13(2): 327. https://doi.org/10.3390/plants13020327
Pratap A, Prajapati U, Singh CM, Gupta S, Rathore M, Malviya N, Tomar R, Gupta AK, Tripathi S, Singh NP. 2018. Potential, constraints and applications of in vitro methods in improving grain legumes. Plant Breed. 00: 1-15. https://doi.org/10.1111/pbr.12590
Raza G, Singh MB, Bhalla PL. 2017. In vitro plant regeneration from commercial cultivars of soybean. BioMed Res Int. 2017: 7379693. https://doi.org/10.1155/2017/7379693
Raza G, Singh MB, Bhalla PL. 2019. Somatic embryogenesis and plant regeneration from commercial soybean cultivars. Plants. 9(1): 38. https://doi.org/10.3390/plants9010038
Rehman A, Rafique MA, Ahmed Z. 2025. Comparative evaluation of soybean genotypes for in vitro regeneration via direct organogenesis. Crop Breed Genet Genom. 7(3): 1-13. https://doi.org/10.20900/cbgg20250011
Sehaole EMK, Mangena P. 2024. N6-benzyladenine (BAP)-based seed preconditioning enhances the shoot regeneration of seedling-derived explants for subsequent indirect gene transfer in soybeans (Glycine max [L.] Merrill.). Int J Plant Biol. 15(2): 254-266. https://doi.org/10.3390/ijpb15020022
Shan Z, Raemakers K, Tzitzikas EN, Ma Z, Visser RGF. 2005. Development of a highly efficient, repetitive system of organogenesis in soybean (Glycine max (L.) Merr). Plant Cell Rep. 24(9): 507-512. https://doi.org/10.1007/s00299-005-0971-7
Sojková J, Žur I, Gregorová Z, Zimová M, Matušíková I, Mihálik D, Kraic J, Moravčíková J. 2016. In vitro regeneration potential of seven commercial soybean cultivars (Glycine max L.) for use in biotechnology. Nova Biotechnol Chim. 15(1): 1-11. https://doi.org/10.1515/nbec-2016-0001
Tiwari R, Singh AK, Rajam MV. 2023. Improved and reliable plant regeneration and Agrobacterium-mediated genetic transformation in soybean (Glycine max L.). J Crop Sci Biotechnol. 26(3): 1-10. https://doi.org/10.1007/s12892-022-00179-9
Vargas-Almendra A, Ruiz-Medrano R, Núñez-Muñoz LA, Ramírez-Pool JA, Calderón-Pérez B, Xoconostle-Cázares B. 2024. Advances in soybean genetic improvement. Plants. 13(21): 3073. https://doi.org/10.3390/plants13213073
Wang Y, Li Z, Chen X, Gu Y, Zhang L, Qiu L. 2022. An efficient soybean transformation protocol for use with elite lines. Plant Cell Tiss Organ Cult. 151: 457-466. https://doi.org/10.1007/s11240-022-02312-6
Xu H, Guo Y, Qiu L, Ran Y. 2022. Progress in soybean genetic transformation over the last decade. Front Plant Sci. 13: 900318. https://doi.org/10.3389/fpls.2022.900318
Zhong H, Li C, Yu W, Zhou HP, Lieber T, Su X, Wang W, Bumann E, Lunny Castro RM, Jiang Y, et al.  2024. A fast and genotype-independent in planta Agrobacterium-mediated transformation method for soybean. Plant Commun. 5(12): 101063. https://doi.org/10.1016/j.xplc.2024.101063
 
Journal of Plant Physiology and Breeding
Volume 16, Issue 1
2026
Pages 93-108

Files

Share

How to cite

Statistics