Investigation of some effective factors in the Invitro soybean regeneration (Glycine max L.)

Document Type : Research Paper


1 Department of agricultural biotechnology, faculty of agriculture, islamic azad university, maku, iran

2 Assistant Professor, Department of Biotechnology and Plant Breeding, Faculty of Agriculture, Islamic Azad University, Maku, Iran

3 دانشیار گروه به نژادی و بیوتکنولوژی گیاهی، دانشکده کشاورزی، دانشگاه تبریز، تبریز، ایران


Soybean (Glycine max L.) is one of the most important agricultural products that is of special importance not only because of its capacity in oil production but also because of its role in human and animal nutrition. One of the effective methods in overcoming the breeding problems of this crop is the use of in vitro culture techniques and molecular methods.  The use of plant tissue culture techniques is one of the effective methods in overcoming the breeding problems of this crop. Tissue culture and plant regeneration in the handling process depend on several factors. The aim of this study was to optimize the effective factors such as genotype, type of micro sample, type of gelling agent and combination of growth regulators in soybean regeneration.
Materials and methods
After germination and seedling growth of Saman and DPX cultivars in culture medium, hypocotyl and cotyledon micro samples in culture medium containing BAP hormone treatments (0, 1 and 2 mg/l) And IBA (0, 0.1 and 0.5 mg/l) were cultured. Subsequently, the samples were recovered in culture media containing IBA (0, 0.5, 1 and 1.5 mg/l) with agar (5, 6 and 7 g) or gellate (2 and 3 g) were cultured. Separately, the effect of growth regulators on soybean cultivars was performed as a CRD-based factorial experiment, the effect of gelling agents on organogenesis and rooting experiment of cultivars was performed in a completely randomized design with three replications and each replicate containing six samples.
The results of this study showed that in the cotyledon sample of Saman cultivar under hormonal treatment of 2 μg/l BAP and 0.1 μg/l IBA, the highest rate of organogenesis and number of regenerated branches were observed. Also, the presence of 7 g of agar in the culture medium showed the highest rate of organogenesis and the number of regenerated branches. In addition, a concentration of 1.5 micrograms per liter of IBA induced the highest rooting rate.
Saman cultivar showed better results in regeneration and rooting than DPX cultivar. Findings of this study can be used for gene transfer and genetic engineering studies for Saman cultivar and other soybean cultivars.


Abbasi Z, Hooshyar S, Bagherieh-Najjar MB (2016) Improvement of callus production and shoot regeneration using various organs of soybean (Glycine max L. Merr) by response surface methodology. In Vitro Cel Dev Biol Plant 52, 537-545.
Abdolinasab M (2018) Optimization of Callogenesis and Regeneration of Cucumber (Cucumis Sativus L.) UtilizingCotyledon, Hypocotyl and Leaf Explants. Agric Biotechnol J 10, 75-92 (In Persian).
Arun M, Chinnathambi A, Subramanyam K et al. (2016) Involvement of exogenous polyamines enhances regeneration and agrobacterium-mediated genetic transformation in half-seeds of soybean. Biotechnol J 6, 148- 160.
Asghari M (2017) New (non-classical) plant growth hormones and regulators: the key to plant growth management and sustainable production of a healthy crop in agriculture. Urmia: Urmia University Press, first edition, 346 pages (In Persian).
Delcheh KS, Kashefi B, Mohammadhassan R (2014) A review optimization of tissue culture medium medicinal plant: Thyme Int J Farm Alli Sci 3, 1015-1019.
Farza L, Siddique MF, Galani S and Mehdi F (2019) Assessing the effect of phytohormone on growth and germination of soybean (Glycin max L.) from cotyledonary node. Pak J Bot 51, 103-107.
Godfray HCJ, Beddington RJ, Crute IR, Haddad Let al. (2010) Food security: the challenge of feeding 9 billion people. Science 327, 812-818.
Hazell P. and Wood S (2008).Drivers of change in global agriculture. Philosophical transactions of the Royal Soc Biol Sci 363, 495-515.
Li S, Cong Y, Liu Y et al. (2017) Optimization of agrobacterium-mediated transformation in soybean. Plant Sci 8, 246-261.
Mangena P, Mokwala PW and Nikolova RV (2015) In vitro multiple shoot induction in soybean. Int J Agric Biol 17, 838-842.
Mello-Farias PC and Chaves ALS (2008) Advances in agrobacterium-mediated plant
transformation with enphasys on soybean. Sci Agric 65, 95-106.
Mohammadhassan R, Kashefi B, Delcheh KS (2014) Agrobacterium-based vectors: a review. Intl J Farm Alli Sci 3, 1002-1008.
Nejati Sarvi N, Babaian Jelodar N, Bagheri NA, Pakdin Parizi A (2019) Investigation of the effect of explants and different levels of NAA and 2-4-D hormones for callus induction in soybean Glycine Max L. 6th National Conference on Medicinal Plants Traditional medicine and organic agriculture, Hamedan (In Persian).
Ozel, CA, Khawar KM and Arslan O (2008) A comparison of the gelling of isubgol, agar and gelrite on in vitro shoot regeneration and rooting of variety Samsun of tobacco (Nicotiana tabacum L.). Sci Hortic 117, 174-181.
Phat P, Rehman SU, Jung HI and Ju J (2015) Optimization of soybean (Glycine max L.) regeneration for korean cultivars. Pak J Bot 47, 2379-2385.
Raza G, Singh MB, Bhalla PL (2017) In Vitro plant regeneration from commercial cultivars of soybean. BioMed Res Int 1, 1-9.
Sharifi A, Bagheri A and Vesal S (2004) effect of explants and varoous levels of auxin and cytokinin on soybean regeneration. Agri Sci Indus J 18, 117-125 (In Persian).
 Soto N, Ferreira A, Abad CD and Enriquez, GA (2013) Regeneration in vitro de plantas de soya de la variedad cubana Incasoy-36. Biotecnol Apl 30, 29-33.
Sukweenadhi J, Choi JY, Kim YJ et al. (2019) June. Callus induction and in vitro mass culture of adventitious roots from leaf segment explants of Dendropanax morbifera Lev. In IOP Conf Ser Earth Environ Sci 293, 012-24).
VargasVML, Avalos RMG, De la Pena C and Figueroa FRQ (2008) Plant tissue culture. Biol Plant 19, 875-904.
Verma K, Saini R, Rani A (2014) Recent advances in the regeneration and genetic transformation of soybean. Innov Biol 1, 15-26.
Zhang F, Chen C, Ge H (2014) Efficient soybean regeneration and Agrobacterium-mediated transformation using a whole cotyledonary node as an explant.  Biotechnol Appl Biochem 61, 620-625.