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In Vitro Plant Regeneration from Hypocotyl and Leaf Explants of Soybean (glycine Max L.)

S.M.H. Kabir
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

M.A. Baten
Department of Environmental Science, Bangladesh Agricultural University, Mymensingh, Bangladesh

M.S. Haque
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

In order to establish an efficient plant regeneration system for soybean in Bangladesh condition, a research work was conducted to find out better explant (hypocotyl and leaf) and hormone combination. Callus initiation, shoot and root development were observed at various frequencies. Hypocotyl explants were superior to the leaves considering all the parameters for callus induction. The best result for callus induction was observed in MS medium supplemented with 1.5 mg/l Kinetin and 2.0 mg/l NAA. Plantlet regeneration from leaf and hypocotyl derived calli of soybean was also investigated. The best shoot induction occurred in MS medium supplemented with 3.0 mg/l SAP and 0.5 mg/l NAA. Leaf derived calli performed better than those derived from hypocotyls did. The elongated shoots developed roots on MS medium where 1.5 mg/l IBA performed the best for rooting. Shoots developed from leaf calli performed better. The rooted plants were transferred to plastic container for hardening. Plantlet survival rate was 70%.

Keywords: Regeneration, Soybean, Leaf and Hypocotyl explant, NAA, SAP, Kinetin

Location: Biotechnology Laboratory, Department of Biotechnology, Bangladesh Agricultural University, Mymensingh

Start Date: 01-10-2004

End Date: 30-05-2005

Program Area: Variety and Species

Commodity: Soybean

Objectives:

To standardize the regeneration protocol of soybean in Bangladesh condition and find out the suitable explant for regeneration.

Methodology:

The research work was carried out in Biotechnology Laboratory, Department of Biotechnology, Bangladesh Agricultural University, Mymensingh, Bangladesh, from October 2004 to May 2005. Soybean seeds cv. G 2 (Bangladesh Soybean-4) were collected from Soybean and Oilseed Laboratory, Bangladesh Agricultural University, Mymensingh. Seeds were washed in tap water and then surface sterilized with 70% (v/v) ethanol for 1 min. and then rinsed with sterile distilled water. Afterwards seeds were soaked in 0.1% HgCI2 plus a drop of Tween-20 solution and shaken gently for 12 minutes; followed by three rinses in sterilized distilled water. The surface sterilized seeds were placed in test tubes containing 12 ml MS medium (Murashige and Skoog, 1962). Seed germination was occurred on hormone free MS medium. For callus induction and maintenance, MS medium was used with different combinations of NAA (0.0, 1.0, 1.5 and 2.0 mg/1) and Kinetin (0.0, 1.0, 1.5 and 2.0 mg/1) supplementation. MS medium supplemented with NAA (0.5 mg/1) and BAP (0.0, 1.0, 2.0 and 3.0 mg/1) was used for shoot induction. For root formation from shoots different concentrations of IBA (0.0, 1.0, 1.5, 2.0 and 3.0 mg/1) were added to MS medium. The pH of media was adjusted at 5.8 in all the cases. All cultures were incubated under complete darkness at 25±2°C for a week after which incubates were kept under 12h photoperiod at 25±2°C. Cultures were transferred to fresh callusing medium periodically and were routinely examined for different morphogenic changes. Six to seven weeks after inculation the calli attained a size of about 20-25 mm in diameter and were transferred to shoot induction medium. Well developed shoots were rescued aseptically and again cultured on freshly prepared root induction medium to induce root. Healthy plantlets with leaves and roots were successfully planted in soil through gradual acclimation. The experiment was arranged in Completely Randomized Design (CRD) with four replications. The analysis of variances for different parameters was performed and means were compared by the Duncan's Multiple Range Test (DMRT).

Source of Information: Bangladesh J. Agril. Sci. 34(1): 23-30, January, 2007

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

From the parameters discussed above it was clear that leaf derived shoots showed higher performance in root formation and 1.5 mg/l IBA is the best hormone concentration both in leaf and hypocotyl. Liu eta!. (1998) observed root formation with 2.0 mg/l IBA. The survival rate of the transplanted plantlets was 70%. The protocol developed from the present research might be useful for improving the soybean cultivars of Bangladesh using different biotechnological approaches.

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