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In Vitro Regeneration of Mungbean (vigna Radiata L.) from Different Explants

M. K. Khatun
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

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

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

Abstract/ Executive Summary:

To regenerate plantlets both directly and indirectly, different explants (cotyledon, hypocotyls, root tip and shoot tip) of mungbean were cultured on medium supplemented with different concentrations and combinations of BAP (0, 1.0 and 5.0 mgL^-1) and NAA (0, 0.5 and 2.5 mgL^-1). Cotyledon explants performed best in callus induction (90.0%) at the combination of 1 mgL^-1 BAP and 2.5 mgL^-1 NAA for both the varieties (BINA mung 5 and BINA mung 7). The calli derived from cotyledon, hypocotyls and root tip were cultured in MS medium supplemented with different concentrations of Kn, BAP and/or NAA for shoot induction. Regeneration was achieved only from cotyledon calli at a frequency of 62.50% on 5 mgL-1 BAP and 0.05 mgL^-1 NAA. Shoot tip cultured for direct regeneration in the same media containing 5 mgL-1 BAP and 0.05 mgL^-1 NAA resulted in 90.0% shoot differentiation in BINA mung 7. The regenerated shoots rooted on MS medium with 0.2 mgL^-1 NAA (90.0).

Keywords: Callus, Plantlet, Regeneration, Mungbean, Seedling explants

Location: Bangladesh Institute of Nuclear Agriculture, Mymensingh, Bangladesh

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Mungbean

Objectives:

a. To find out suitable explant and medium for regeneration, and

b. To develop a stable, reproducible and efficient protocol for the in vitro regeneration of mungbean.

Methodology:

Seeds of the variety BINA mung 5 and BINA mung 7 were collected from Bangladesh Institute of Nuclear Agriculture. Healthy seeds were washed in tap water. The seeds were then sterilized with 70% ethanol for one minute and then rinsed with sterile distilled water. Afterwards, the seeds were soaked in 0.1% HgCl₂plus two drops of a liquid detergent (Tween-20) solution and then agitated gently for 15-20 minutes, followed by 4 rinses in sterilized distilled water. The seeds were aseptically germinated on half MS (Murashige and Skoog, 1962) medium. Cotyledons, hypocotyls and root tips were excised aseptically from 5 to 7 days old seedlings and cultured on MS medium containing different concentrations and combinations of BAP (0, 1.0 and 5.0 mgL^-l) and NAA (0, 0.5 and 2.5 mgL^-1). Subcultures were carried out at 21 days interval and finally the calli were transferred to MS medium with different concentrations of cytokinins (BAP and Kn) and auxin (NAA) singly or in combinations. Shoot tips were also cultured on the same media. The regenerated shoots were rooted in MS media with different concentrations of NAA (0, 0.1, 0.2 and 0.5 mgL^-1) or IAA (0, 0.1, 0.2 and 0.5 mgL^-1). The pH of the medium was adjusted to 5.8 prior to the addition of agar @ 8.0 g/L and the medium was autoclaved at 121^oC for 30 minutes. Twenty explants were cultured per treatment and each treatment was replicated 4 times. All cultures were maintained at 26±1⁰C under white fluorescent tubes with a 16h photoperiod. The data collected were analyzed and the means were using Duncan’s Multiple Range Test (DMRT).

Source of Information: Progress. Agric. 19(2) : 13-19, 2008 ISSN 1017-8139

Article Link (Online Journal): http://www.banglajol.info/index.php/PA/article/view/16908/11891

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In conclusion, for the creation of genetic variability in crop plants it is very important to regenerate plants via callus. With this obvious reasons it is suggested that in future tissue culture programme of mungbean some more cytokinin`s additives viz; 2-ip, zeatin, coconut water, ascorbic acid, glutamine may be used for successful plantlet regeneration from calli. Here a direct in vitro regeneration protocol was developed. However, further study is needed with different explants to standardize the protocol of regeneration from calli. The protocol developed here could be used for future improvement of mungbean through in vitro culture and genetic transformation.

Knowledge Management: Journal